NUCISnetworks.iamz.ciheam.org/nuts/pdfs/NUCIS 12_2004.pdf · 2015. 11. 25. · FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004 1 NUCIS NEWSLETTER Information Bulletin of the

1FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

N U C I SN E W S L E T T E R

Number 12 September 2004Information Bulletin of the Research Network on Nuts (FAO-CIHEAM)

IRTA - Mas Bové ● Coordination Centre of the Research Network on Nuts

FAO CIHEAMNut Network

CIHEAM

INIAINSTITUTO NACIONAL DE INVESTIGACIÓN Y TECNOLOGÍA AGRARIA Y ALIMENTARIA

Young almond orchard growing on the slopes of Sierra de María, Almería, Andalusia, Spain

EDITORIAL

Activities 2003-2004The activities of the Nut Network havebeen affected by the assessment andevaluation phase of the European Sys-tem of Cooperative Research Networks inAgriculture (ESCORENA). This systemwas established in 1974 by FAO and Eu-ropean research institutions, and current-ly consists of eleven active networks. Dueto budgetary reductions, FAO is no longerin a position to provide the ESCORENA

secretariat for servicing the Networks norof supporting activities. Nonetheless,most of the planned activities under theFAO-CIHEAM Interregional CooperativeResearch Network on Nuts could be im-plemented through the support providedby the FAO’s Regional Office for Europeand some FAO technical services(AGPS) and financial support providedby INIA of Spain, INRA of France andthe co-sponsor and partner CIHEAM per-mits the most planned activities to be fi-nally developed for exemple the publica-tion of this Newsletter and the Inventoryof Walnut Germplasm, Research and Re-

ferences. In addition, some researchersfrom developing or transition countrieshave been supported to participate in dif-ferent meetings and congresses.

During 2003 and part of 2004 a number ofother activities was implemented withinthe framework of the FAO-CIHEAM Inter-regional Cooperative Research Networkon Nuts. In June 2003, the XIII GREMPAAlmond and Pistachio Meeting was heldin Mirandela, Tras os Montes, Portugaland the proceedings will be published inthe Options Méditerranéennes Cahiers byNovember 2004. In June 2004, the VI

2 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

ISHS International Congress on Hazelnutwas held in Tarragona, Spain and theproceedings will be published in ActaHorticulturae. During 2004 two moreSymposia are planned: (i) the III ISHS In-ternational Symposium on Chestnut to beheld in October in Portugal, Chaves, VilaReal; and (ii) the V ISHS InternationalWalnut Symposium to be held in Sorren-to, Italy in November.

The Network’s Coordinator participated intwo Scientific Committee Meetings andthe Convention of the International TreeNut Council (INC) held in Berlin (Ger-many) and Las Vegas (USA) respectively.The importance of promoting the healthycomponent of nut consumption in the hu-man diet through advertisement wasstressed. This is considered essential forsuccessful marketing and to sustain de-mand. Worldwide nut production is in-creasing over the years and its supply istaken by the also increasing world con-sumption. It seems that nut markets canbe further developed. The INC expressedan interest to collaborate and work close-ly with the Nut Network. The issues ofpreventing and reducing kernel’s aflatoxincontamination through better crop prehar-vest and postharvest management is ofparamount importance for the INC.

ESCORENA’s reviewThe ESCORENA networks can be classi-fied into three groups according to thefield of agricultural activity: crop produc-tion, animal production and environment.The 23rd Regional Conference for Europe(Nicosia, Cyprus, May 2002) recognizedthe importance of ESCORENA as the re-search networks brought added value notonly to European scientists and their re-search institutions, but also to partnersand beneficiaries beyon European bor-ders. Furthermore, the Conference en-dorsed the recommendation of the 32nd

Session of the European Commission onAgriculture (ECA) that an assessmentand evaluation be undertaken for eachnetwork. During 2003, this assessmentwas carried out by M. Larbier, a visitingscientist seconded to FAO by INRA, andhis report and proposals for the futurewere submitted to the 33rd Session ofECA held in Rome in March 2004. OurReseach Network on Nuts was conside-red among those ten to be retained andfor having good potential for continuity inthe future. This conclusion was reacheddue to dynamic coordination, adaptationof research towards the aim of sustaina-ble development, importance of the pro-gramme and quality of the scientific andtechnical communication means used.The ECA was also informed of CIHEAM’sintention to continue to cooperate and co-sponsor three of these ten networks, in-cluding our Nut Network. The report sub-

mitted to the ECA included an estimate ofthe necessary funds needed to ensure fu-ture functioning of ESCORENA. Conside-ring the budgetary constraints faced byFAO, the proposal to establish a multi-do-nor Trust Fund was made. The ECA re-port was submitted to the 24th FAO Regio-nal Conference for Europe (Montpellier,May 2004) but no decission on this issuewas taken. It seems that the financialsupport towards our Network by themember countries under circumstancesof financial difficulties and uncertainty isessential to foster activities.

The Nut Network’s futureNut tree production, trade and industry isan important economical activity relatedto sustainable agriculture, often in margi-nal lands, in both European and NearEastern Regions. Nuts are of major im-portance and are typical components ofthe traditional and healthy Mediterraneandiet. To accomplish sustainable develop-ment and food security, a future combi-ned effort in R&D, environmental mana-gement and communication is needed.

Due to overall budgetary constraints, theNetwork on Nuts has in recent years, un-dergone a restructuring exercise which isnow fully implemented. Thus, the Networkhas reorganized and simplified its structu-re and is trying to focus more on develop-ment oriented issues rather than its pre-vious strong genetic resources collection,description and improvement. In addition,as a result of the assessment carried outin 2003 mentioned above, two workinggroups: Genetic Resources and Econo-mics were suggested for elimination.

Considering the current and future rele-vance of the Network, it is generally seenby its members as a useful tool for bettercollaboration, and the opportunities to

come together are viewed as very impor-tant for project development. Accordingly,recent Network developments are focu-sed on research issues which are of prio-rity importance for developing countriesand for countries in transition, particularlyin the Balkans, Caucasus and North Afri-ca, but also beyond, wherever researchresults are relevant to stimulate growthand alleviate poverty and hunger. The re-levance and potential usefulness of ourNut Network is based on the availabilityof considerable genetic resources andtechnical experience for the developmentof nut tree crops in Europe and the NearEast regions. In addition, useful contribu-tions to rural development and the globalgenetic resources understanding can bemade. Major research areas of broad inter-est, which should be addressed are: foodsafety, biodiversity, biotechnology and sus-tainable management of natural resources,including low-input agriculture. To streng-then FAO’s support to our Network it is ne-cessary to adopt a “project approach” ofactivities. This should be carried out incollaboration with relevant FAO Units.

Genetic resources inventoriesThree Inventories on Germplasm, Re-search and References have been alrea-dy published: Almond (1997, RTS 51),Hazelnut (2000, RTS 56) and Chestnut(2001, RTS 65). These inventories pu-blished in the REU Technical Series(http://www.fao.org/regional/europe/PUB),are important compilations of the current-ly available species genetic resourcesand information on ongoing research pro-jects and bibliography. In addition, twomore inventories are being compiled andare at different stages of completion. TheInventory on Walnut being compiled by E.Germain is to be published shortly andthe Inventory on Pistachio is being colla-ted by N. Kaska and B.E. Ak. All thesecatalogues are being supported by FAO’sRegional Office for Europe and the Seedand Plant Genetic Resources Service(AGPS) together with CIHEAM-IAMZ.

In addition, a draft Descriptors List forHazelnut has been developed by mem-bers of the Network and has now beensubmitted to IPGRI for assessment andeventual editing and publication.

Changes in the FAO Regional Officefor EuropeAs mentioned above, Mr. Michel Larbier,a visiting scientist seconded from INRA,France, took the task to evaluate andsuggest a proposal to re-organize ESCO-RENA. The efforts made by M. Larbier inreviewing the whole ESCORENA systemare acknowledged. Ms. Jutta Krause, Re-gional Representative for Europe (FAORegional Office for Europe in Rome) isthe FAO contact person for ESCORENA.

Almond trees starting bloom at IRTA,E.E. Lleida, Spain


EDITORIAL .......................................... 1

ARTICLES AND REPORTS

• Consumers’ acceptance of industrial productsmade with hazelnuts and almonds .............................. 4

• Differential flower and fruit damagesby spring frosts in almond .......................................... 5

• Prunus webbii as a way to look intoself-compatibility in almond ....................................... 8

• Application of molecular markers in almondbreeding programmes ................................................ 9

• Fusicoccum canker in almond orchards –biotechnology tools for early selectionof tolerant genotypes ................................................ 12

• Virus screening for Australian almonds .................... 15

• The pistachio industry in Italy: current situationand prospects ........................................................... 16

• Pistachio production in Tunisia ................................ 19

• Breeding monoecious pistachio cultivars ................. 21

• Chilling stress and injury in pistachio female flowers .......................................................... 23

• Underground drip system: the new irrigation method for pistachio and almond orchard in Turkey ............. 24

• Iranian agricultural system to develop higher quality and healthier pistachio production ........................... 26

• Pistacia vera L. 1753 in Iran ..................................... 26

• Pistacia atlantica desf. 1800 in Iran .......................... 27

• Pistacia khinjuk stocks 1852 in Iran ......................... 30

• Germplasm collection of pistacia at the Jacob Blaustein Institute for desert research. Phenotypic traits and molecular markers .................................... 31

• Where in a pistachio tree is Xanthomonas and how did it get there? .......................................... 33

• Chestnut production in Andalusia ............................. 33

• The world macadamia industry and research activities ................................................................... 36

• Carob tree development in China .............................. 39

NOTES AND NEWS

• In memorian: my professor Dale Kester ................... 41

CONGRESSES AND MEETINGS

• XIII GREMPA meeting on pistachios and almonds ... 41

• VI International Congress on Hazelnut ...................... 42

TO BE HELD ................................................................. 45

BIBLIOGRAPHY ........................................................... 46

BACKPAGE ................................................................... 52

CONTENTS PageThe Nut Network on the webIn addition to the already existing informa-tion on the Nut Network at http://www.iamz.ciheam.org/ingles/nuts.htm,and on the previous FAO web page crea-ted in 2000, the FAO European System ofCooperative Research Networks in Agri-culture (ESCORENA) and the Nut Net-work web page has now been closed dueto lack of commitment from coordinatorsto make use of the interactive system fordata entry and lack of resources fromFAO to take over this role. However, es-sential information on network coordina-tors, major network publications (e.g.REU Technical Series), ESCORENA re-ports and links to existing network websi-tes such as the nut network is included inthe REU website (see http://www.fao.org/world/regional/REU/content/escorena/index_en.htm ).

NUCIS on the webA short version of the Newsletter (edito-rial, contents and back page) from issuenumber 6 and onwards, is available onthe Internet web pages of both FAO (http://www.fao.org/regional/europe/public-e/nucis.htm) and CIHEAM (http://www.iamz.ciheam.org/ingles/nuts.htm# publi-cations). A full electronic version of NU-CIS is available from issue number 9 andonwards (http://www.iamz.ciheam.org/in-gles/nucis9.pdf). The contents of thisNewsletter can be browsed through andalso copied and printed.

Contributions to NUCISAs in past NUCIS editorials, it is againstressed that this Newsletter must be aneffective vehicle of communication for allthe Network members. The pages of thisbulletin are open to all readers who wouldlike to suggest ideas or to express theiropinion about the work developed by theNetwork (activities carried out and plan-ned) or to publish short articles and re-ports on relevant horticultural subjects ofgeneral interest. A sufficient number ofcontributions are received from the Medi-terranean Basin and overseas for the arti-cles and reports section. However, thesections on news and notes and also oncongresses and meetings are usually dif-ficult to cover due to the scarce informa-tion received and thus, contributions aremost welcomed. Otherwise, the Editorhas to report on the issues he is aware of,but certainly there must be many more is-sues on-going throughout the year whichmerit reporting. Also, the place for ‘grey’bibliography (references and documentswhich are difficult to search like Mastersor Ph. Theses) is scarcely filled.

The NUCIS Newsletter is distributedworldwide free of charge to 1.400 readersfrom over 60 countries. The disseminationof information originated by the Network

is of paramount importance and throughthis bulletin has been largely successful.The first NUCIS was published in 1993,this issue of the NUCIS Newsletter isnumber 12 and during these eleven yearsgreat editing effort has been made. I ack-nowledge and gratefully thank all contri-butors for their efforts and interest to pro-duce and send me valuable information.The exchange of information betweenNetwork members through the pages ofthis Newsletter is the basis for developingcollaboration. The editing task in the ele-ven NUCIS issues already published hasbeen huge (NUCIS 1, 9 pages; 2, 20 pa-ges; 3, 24 pages; 4, 28 pages; 5, 36 pa-ges; 6, 52 pages, 7, 44 pages, 8, 46 pa-ges 9, 68 pages, 10, 48 pages, 11, 48 pa-ges and 12, 52 pages). In order to reducethe time consuming formal editing contri-butors who send articles, news, notes, bi-bliographic references, etc. to the diffe-rent sections are asked to provide themwell organized and elaborated. Informa-tion should be sent in satisfactory En-glish. Contributions could be sent throughInternet using the Editor’s email. The al-ternative is to provide them on disketteand also in printed format. This bulletin isreproduced in black and white only, inclu-ding pictures. Please send your contribu-tions for the next issue, number 13 by theend of October 2005; however it cannotbe guarantied that the article will be pu-blished due to the uncertainty of funding.We apologize to readers for the delay inprinting this issue due to late funding. Wethank all who have contributed to this is-sue.

The Editor

The designations employed and thepresentation of material in this publica-tion do not imply the expression of anyopinion whatsoever on the part of theFood and Agriculture Organization ofthe United Nations concerning the legalstatus of any country, territory, city orarea or of its authorities, or concerningthe delimitation of its frontiers or boun-daries.

This publication contains the collectiveviews of an international group of ex-perts and does not necessarily repre-sent the decisions or the stated policyof the Food and Agriculture Organiza-tion of the United Nations, the Interna-tional Centre for Advanced Mediterra-nean Agronomic Studies nor of the Or-ganization for the Economic Coopera-tion and Development.

Contributions should be written conci-sely in English. Please send contribu-tions on paper and diskette (Microsoft®

Word or Word Perfect®). Authors areresponsible for the content of their pa-pers. Reproduction of the articles is au-thorized, provided that the original sour-ce is clearly stated.


ARTICLESAND REPORTS

CONSUMERS’ ACCEPTANCEOF INDUSTRIAL PRODUCTS

MADE WITH HAZELNUTSAND ALMONDS

INTRODUCTIONHazelnuts and almonds are used to makeindustrial products like snacks, turrons,marzipan or chocolates. Although there iswide information about their chemicalcomposition and the influence of manyprocesses on the products´ final charac-teristics, there is a lack of informationabout consumers’ acceptance of these ty-pes of products when hazelnut and al-mond varieties change.

For these reasons IRTA started in 1999 aresearch programme in order to analyzethe importance of selecting different nutvarieties to make different industrial pro-ducts, as a consequence of consumers’acceptance. This information could be re-levant not only to producers but also forthe confectionery industry. Results for dif-ferent nut products have been publishedin several scientific reports (see referen-ces). An overall analysis of hazelnut andalmond commodities is made in this arti-cle. Variety is used as synonym of culti-var.

METHODOLOGYFrom 1999 to 2002 four hazelnut varieties(‘Negret’, ‘Pauetet’, ‘Tomboul’ and ‘Tondadi Giffoni’) and eight almond varieties(‘Desmayo Largueta’, ‘Francolí’, ‘Glorie-ta’, ‘Marcona’, ‘Masbovera’, ‘Nonpareil’and ‘Planeta’) were used to make severalproducts: dark chocolate with nuts, whitechocolate with nuts, brown chocolate withnuts, bonbons made with roasted nuts en-robed with burnt sugar and coated withdark chocolate, marzipan and hard andsoft turrons.

Chocolates and bonbons were made incollaboration with the industry “L’Art de laXocolata” (El Vendrell, Tarragona), whilemarzipan and turrons were made in theindustry “Turrons i Mel Alemany SL” (Osde Balaguer, Lleida). Hazelnut and al-mond samples came from IRTA´s collec-tions, while ‘Tomboul’ hazelnuts and‘Nonpareil’ almonds were purchased onthe market.

Each commodity was assessed for indus-trial facility, final quality and consumers’acceptance. Commodity acceptance wasevaluated using the consumers panel

from IRTA (table 1), including over 670people (almost 300 families), and distribu-ted all around Catalonia (Spanish NE).

Consumer’s answers were weighted byage and education level in order to extra-polate conclusions to Catalonian people.

MAIN RESULTSResults show that there are statistical dif-ferences in variety acceptance, depen-ding on the industrial product studied. So-metimes these differences can be solvedchanging the technological process, al-though this has been observed for roas-ted hazelnuts and not for other products.

For example, dark chocolate (Figure 1)was the most accepted, mainly for ‘Paue-tet’ hazelnut variety and for ‘Masbovera’almond variety. ‘Negret’ and ‘Tomboul’hazelnuts and ‘Desmayo Largueta’ and‘Francolí’ almonds showed a very goodtechnological behaviour to make bonbons(figure 2). ‘Marcona’ almonds were thebest accepted to make marzipan (Figure3) and hard turron (Figure 4), while ‘Ne-gret’ variety was the most accepted forhard turron made with hazelnuts. Finally,soft turrons (Figure 5) were most accep-

Table 1.- Consumers distribution by sex, age, education level and city size in boththe IRTA panel and the Catalonian people.

IRTA’s panel Catalonia

Sex: % %Men 49.7 48.8Women 50.3 51.2

Age:< 30 years old 23.5 36.730 to 65 years old 68.6 46.1> 65 years old 7.9 17.2

City habitants:< 30.000 40.4 43.6> 30.000 59.6 56.4

Education level:Basic 34.9 69.3Middle 22.9 20.7High 42.2 10.0

Figure 2.- Chocolate coating to make bonbons.“L’Art de la Xocolata” industry

Figure 3.- Cutting marzipan.“Turrons i mels Alemany” industry

ted when they were made with ‘Marcona’or ‘Desmayo Largueta’ almond varieties.

The overall analysis of the results alsoshows that sensorial characteristics fromhazelnuts and almonds must be conside-red to make an industrial product. Inmost of them, sensorial profile from roas-ted nuts should be known while in a fewof them the most important is the profilefrom raw nuts. Even though, when a pro-duct includes sugar in its recipe the diffe-rences in taste are not relevant enough,but it is still important that nut taste canbe identified.

Figure 1.- Chocolate with nuts.“L’Art de la Xocolata” industry


Figure 4.- Hard turron ‘Alicante’.Labelled and vacuum packaged tablets

Figure 5.- Soft turron ‘Jijona’.Labelled and vacuum packaged tablets

Raw hazelnuts and almonds homogeneityis very important. Even though, this pointshould be studied in more detail for suchkind of products where nuts are groun-ded, not roasted or sugar enriched. Maindefects to be considered in raw nut bulksare rancidness, bitterness or burnt ker-nels. Some hazelnut and almond varietiesseem to show serious difficulties to beadapted to some industrial processes,due to a very high fragility or strength, anexcess of splits generation, an inadequa-te surface to volume ratio, bad roastingaptitude, etc.

Finally, both hazelnuts and almondsseem to show the same acceptation atconsumers level for those particular pro-ducts studied. This could confirm thatthey can substitute each other dependingon the marked price, as it is usually assu-med. However, it should be further confir-med for other products.

ACKNOWLEDGMENTSResearch supported by Instituto Nacionalde Investigación y Tecnología Agraria yAlimentaria (INIA) of Spain, project num-ber SC99-002.

REFERENCES

Romero, A., Tous, J.; Guerrero, L.;Gou, P.; Guardia, Mª D., 1999. Aplica-ciones del análisis sensorial en el tosta-do de avellana en grano. FruticulturaProfesional (Especial Frutos Secos II),104: 71-77.

Gou, P.; Díaz, I.; Guerrero, L.; Valero,A.; Arnau, J.; Romero, A. 2000. Physi-co-chemical and sensory property chan-ges in almonds of ‘Desmayo Largueta’variety during toasting. Food Sci. Tech.Int., 6 (1): 1-7.

Romero, A.; Tous, J., 2000. Nutricionalvalue of hazelnut (Corylus avellana, L.).Nucis-Newsletter, 9 (12): 25-27.

Tous, J.; Romero, A., 2002. Almondand hazelnuts products. Consumer´spreferences. Nucis-Newsletter, 10 (12):12-13.

Romero, A.; Tous, J.; Plana, J.; Guar-diola, MD.; Díaz, I., 2002. How varietychoice affects Spanish consumer´s ac-ceptance of marzipan and chocolatesmade with almonds. Ed. ISHS. ActaHorticulturae, 591: 117-123.

Romero, A.; Tous, J.; Plana, J., 2003.Importancia de la variedad de almendraen la aceptación de turrones tipo ‘Ali-cante’ y ‘Jijona’. II Congreso Nacionalde Ciencia y Tecnología de los Alimen-tos. Orihuela (Alicante). Ed. Universi-dad Miguel Hernández (UMH). VolumenII: 611-614.

Romero, A.; Tous, J.; Plana, J. 2004.How cultivar choice affects spanishconsumers’ acceptance of chocolates,bonbons and hard turron made with ha-zelnuts. 6th Int. Congress on Hazelnut.Tarragona-Reus, 14-18 June.

A.Romero and J.TousIRTA, Centre Mas Bové. Apartat 415, 43280-

Reus (Tarragona, SpainE-mail: [email protected]

DIFFERENTIAL FLOWER ANDFRUIT DAMAGES BY SPRING

FROSTS IN ALMOND

INTRODUCTIONFlowers and young fruits of almost all de-ciduous fruit trees are vulnerable to frostdamage (Andrews et al., 1983). Conse-quently spring frost injury is a major limi-ting factor in the production and the distri-bution of horticultural crops (Ashworth,1992). The anatomical, physiological andbiological aspects of this damage havebeen reviewed in fruit trees (Rodrigo,2000). In Prunus, fruit damage is detecta-ble by observing ovule browning, ice for-mation (Saunier, 1960), and browning ofthe petals and pistils.

Almond (Prunus amygdalus, Batsch) isan early blooming species. As a result, al-mond cultivation was restricted to regionswith low risk of spring frosts. However,the expansion of the culture into inlandMediterranean areas, where the occur-

rence of spring frost is common and over-lapping with bloom of most almond culti-vars, increased the risk of reducing oreven nullifying yield (Socias i Company etal., 1999). Thus, almond breeding aimedto create and select late blooming culti-vars (Kester and Assay, 1975). Neverthe-less, this solution is not enough to over-come spring frost damages, and frosthardness is also considered a selectionobjective in our breeding programme (So-cias i Company et al, 1998).

In almond, little is known about the mini-mum threshold temperature causing da-mages in flowers and young fruits. Howe-ver, differences among several commer-cial almond cultivars were described byFelipe (1988), concluding that at –2.5ºCsome cultivars suffered heavy productionlosses. Our aim was to assess springfrost damage in the flower and youngfruits of several almond selections andtheir consequences on final fruit set.

MATERIALS AND METHODSThe study was carried out during the win-ter-spring of 2003 on 21 self-compatibleselections from CITA´s breeding pro-gramme at Zaragoza. They were graftedonto the almond x peach hybrid rootstock‘Garnem’ (Gómez Aparisi et al., 2001).The situation of this orchard, in relation tospring frost, was considered of high risk,because of the frequent incidence of frost(Felipe, 1988).

To evaluate the damages caused in flo-wers and young fruits, samples were col-lected two days after frost occurrence,placed in polyethylene bags, and taken tothe laboratory. Damage evaluation wasbased on the morphological symptoms,such as brown discolouration of the stylebase in flowers or browning of the ovulein young fruits. To assess the effect offrost on fruit set, all bud flowers werecounted on two branches of each selec-tion before bloom, thus much before frostincidence. Fruit set was recorded in June.Bud density was also measured accor-ding to Socias i Company (1988).

RESULTS AND DISCUSSIONA frost of –2.5°C during five hours tookplace in the early morning of March 18,2003. On that day, the almond selectionswere at different phenological stages andcan be separated in two groups (Fig. 1).The first group comprised 12 selectionswhose bloom was already over, thus thefrost affected young fruits. The secondgroup comprised 9 selections that on thatday were opening early flowers; conse-quently, the frost only affected buds andearly flowers. The behaviour of bothgroups was different and needs to beexamined differently for each group of se-lections.


Early blooming selections: Frost dama-ge on small fruits ranged from 20% (G-6-39) to 77% (H-3-39) and produced a hea-vy reduction of fruit set, which was practi-cally nil in some (G-1-38 and H-3-37) andeconomically insufficient in others (G-2-23, G-2-7, G-5-18, G-6-24, and H-3-39).These could be due to the fact that theseselections had a young developing fruit,which is considered the most vulnerablestage to frost (Proebsting and Mills,1978b).

In addition, some selections showed anintermediate fruit set, although not com-mercially acceptable (G-6-39, I-1-95, andI-2-12), and selection G-1-58 showedvery high fruit set (33.5%), of a commer-cial level (Kester and Griggs, 1959). The-se differences, observed in selectionsthat were at the same phenological stage,could be due to a differential resistance ofthese selections to frost damage. Thesame observation was reported for otheralmond cultivars previously studied (Feli-pe, 1988), as spring frost damage on re-productive organs depends mainly on thecultivar (Proebsting and Mills, 1978a) andon the intensity and duration of the lowtemperature (Mazur, 1969). In fact, diffe-rences in yield losses have been obser-ved among almond cultivars at differentcritical low temperatures (Felipe, 1988).This suggests that selection G-1-58 hassuffered lighter losses than the other se-lections of this group with a temperatureof –2.5°C.

A high negative correlation was observedbetween the percentage of fruit damageand fruit set (-0.65), showing how theamount of damaged fruits significantly re-duces fruit set. There was also a high ne-gative correlation between the percenta-ge of damaged fruits and bud density (-0.63), showing that the effect of springfrosts is higher in selections with a lowerbud density. Moreover, a significant posi-tive correlation was found between buddensity and fruit set (0.42), thus, fruit setof selections with a medium to high buddensity is higher than that of selectionswith a low bud density (Fig 2). This couldbe due to the fact that a high potential toproduce bud flowers may compensate thespring frost damages, and, together witha good frost resistance, as in the case ofselection G-1-58, could maintain the fruitset at a commercially acceptable level.

Late blooming selections: flower andbud frost damage ranged from 2% (G-2-2) to 47% (G-3-5), generally lower thanthe fruit damage on selections of the ear-ly flowering group, confirming that frostdamage is highly dependent on the stageof bud development, as selections of thelate flowering group (Fig 1) were in theearly blooming stages, C, D and a few E

(Felipe, 1977) when the frost happened.Also, the flower hardness of each selec-tion (genotype) reflects the differencesamong selections at the same phenologi-cal stage (Westwood, 1993).

In general, fruit set decreased when theamount of damaged flowers increased(Figure 3), which explains the significantnegative correlation between fruit set andpercentage of damaged flowers (-0.37).In addition, in spite of the high percenta-ge of damaged flowers, selection G-3-5shows a medium fruit set (Figure 3). Thiscould be due to the high bud density ofthis selection, which has compensatedthe loss of flowers and maintained a me-dium fruit set.

CONCLUSIONIn general, the amount of damages cau-sed by frost in selections at the beginningof bloom (late flowering group) wereslight in comparison to the damages cau-sed in early blooming selections (earlyflowering group), confirming that frost da-mage is highly dependent on the pheno-logical stage of the bud/flower/fruit, theearly developing fruit being the most sus-ceptible stage (Proebsting and Mills,1978b). Furthermore, differences obser-ved among selections for the amount offlower and fruit losses show different frost

hardness for each selection. Consequent-ly, only tolerant or resistant genotypes(cultivars or selections) can produce acrop when frost takes place at or laterthan blooming time.

In some cases frost produced the com-plete loss of the crop, and in others animportant reduction of fruit set and, con-sequently, of yield. Moreover, selectionshaving a high bud density gave a fruit setfrom medium to acceptable. Hence, ahigh bud density seems to compensatespring frost damages.

These data were not enough to explainthe effect of spring frost damage on fruitset, because fruit set in almond dependson several factors, including other wea-ther conditions (such as rain), insect acti-vity among inter-compatible and overlap-ping blooming cultivars or the ability fornatural autogamy in self-compatible culti-vars (Socias i Company and Felipe,1992). However, a wide variability of res-ponses to frost were observed in theseselections, showing that even with thisfrost some genotypes can reach a 25-40% fruit set, considered as the economi-cally acceptable threshold for a commer-cial crop (Kester and Griggs, 1959). Con-sequently, selection for frost resistancecan be efficiently undertaken to obtain

Figure 1: Blooming time and fruit set of the studied selections


REFERENCES

Andrews, P. K., Proebsting, E.L.,Gross, D.C., 1983. Differential thermalanalysis and freezing injury of deaccli-mating peach and sweet cherry tissues.J. Amer. Soc. Hort. Sci. 108, 755-759.

Ashworth, E.N., 1992. Formation andspread of ice in plant tissues. Hort. Rev.13, 215-255.

Felipe, A. J., 1988. Observaciones so-bre comportamiento frente a heladastardías en almendro. Rap. EUR, 11557.145-148.

Felipe, A.J., 1977. Almendro. Estadosfenológicos. Inf. Técn. Econ. Agrar., 27:8-9.

Gómez Aparisi, J., Carrera, M., Felipe,A.J., Socias i Company, R., 2001. ‘Gar-nem’, ‘Monegro’ y ‘Felinem’: nuevos pa-trones híbridos almendro x melocotone-ro resistentes a nematodos y de hojapara frutales de hueso. Inf. Técn. Econ.Agrar., 97V(3): 282-288.

Kester, D.E., Asay, R.D., 1975. Al-monds. In: Advances in Fruit Breeding.Janick, J. and Moor, J. N (eds). PurdueUniv. Press, West Lafayette (IN), pp.387-419.

Kester, D.E., Griggs, W.H., 1959. Fruitsetting in almond: the effect of cross-pollinating various percentages of flo-wers. Proc. Amer. Soc. Hort. Sci. 74:214-219.

Mazur, P., 1969. Freezing injury inplants. Annu. Rev. Plant Physiol. 20,419-448.

Proebsting, E.L., Mills, H.H., 1978a. Asynoptic analysis of peach and cherryflower bud hardiness. J. Amer. Soc.Hort. Sci. 103, 842-845.

Proebsting, E.L., Mills, H.H., 1978b.Low temperature resistance of develo-ping flower buds of six deciduous fruitspecies. J. Amer. Soc. Hort. Sci. 103,192-198.

Rodrigo. J., 2000. Spring frosts in deci-duous fruit trees- morphological dama-ge and flower hardiness. Scientia Hort.85, 155-173.

Saunier, R., 1960. La lutte contre lesgelées printanières chez les arbres frui-tiers (I). Pomol. Fr. 2: 5-12.

Socias i Company, R., 1988. La densitéflorale comme critère variétale chezl’amandier. Rap. EUR, 11557. 145-148.

Socias i Company, R., Felipe, A.J.,1992. Self-compatibility and autogamyin ‘Guara’ almond. J. Hort. Sci. 67: 313-317.

Socias i Company, R., Felipe, A.J. andGomez-Aparisi, J., 1999. A major genefor flowering time in almond. PlantBreed., 118: 443-448.

Socias i Company, R., Felipe, A.J., Gó-mez Aparisi, J., García, J.E., Dicenta,F., 1998. The ideotype concept in al-mond. Acta Hort., 470: 51-56.

Westwood, M.N., 1993. Temperate-zone Pomology: Physiology and Cultu-re. Timber Press, Portland.

O. Kodad and R. Socias i CompanyUnidad de Fruticultura CITA, DGA

Apartado 727, 50080 Zaragoza, SpainE-mail: [email protected]

cultivars adapted to the harsh weatherconditions of many almond growing re-gions in continental climates.

ACKNOWLEDGMENTSThis work was supported by the projectAGL2001-1054-C03-02 of the SpanishCICYT. OK gratefully acknowledges agrant from Instituto Nacional de Investi-gación y Tecnología Alimentaría (INIA).Technical assistance by J. Búbal, O.Frontera and meteorological data from A.Cabezas are highly appreciated.

Figure 2. Percentage of damaged young fruits, bud density and fruit setof selections of the early flowering group.

Figure 3. Percentage of damaged flowers, bud density and fruit set of selectionsof the late flowering group.


Prunus webbiiAS A WAY TO LOOK INTO

SELF-COMPATIBILITYIN ALMOND

INTRODUCTIONAlmond [Prunus amygdalus Batsch or P.dulcis Miller (D.A. Webb)], derives fromone or more wild almond species (suchas P. fenziliana, P. bucharica, P. kurami-ca, P. triloba) that evolved in the desertsof Central Asia. By 450 BC, almond star-ted spreading in the Mediterranean basinand countries like Italy, Spain, France,Portugal, Morocco, Tunisia became im-portant centres of production (Kester andRoss, 1996).

The fact that almond is mainly a self-in-compatible species accounts for the greatdiversity found and also for the differen-ces among populations of different origins(with distinct histories of hybridization).Furthermore, as pointed out by Socias iCompany (2002) it is expected that hybri-dizations with wild Mediterranean species(such as P. webbii) may have taken placeduring almond expansion, resulting in thealmond populations that can be currentlyfound in this region.

SELF-INCOMPATIBILITYMany flowering plants display self-incom-patibility (SI), a genetically determinedcharacteristic which prevents inbreedingby promoting outcrossing. Self-incompati-bility is determined by a single multialleliclocus, the S locus. Whenever there is amatch between the S alleles expressed inthe pistils and those of the pollen, fertili-zation is hampered. In the families Sola-naceae, Rosaceae and Schrophularia-ceae, the products of the S locus in thepistil are S-RNases that are thought todestroy the RNA of incompatible pollen(Wang et al., 2003).

Even though SI is believed to have pla-yed an important role in the success ofAngiosperms, it represents a limitation toplant breeders when fruit production isimportant. In almond, SI is almost 100%effective, which means that unless com-patible cultivars are present in an or-chard, nut production will never be achie-ved. It has long been the desire of al-mond breeders to obtain self-compatible(SC) cultivars that are, at the same time,good almond producers.

SELF-COMPATIBILITY IN ALMONDSome self-compatible almond varietieshave been identified and the self-compa-tibility trait is usually attributed to the pre-sence of a self-fertility allele SF (Raynaudand Grasselly, 1985). The Italian cultivar‘Tuono’ is the best characterized SC al-

mond cultivar and analysis of stylar RNa-se activity indicate that SF is inactive in‘Tuono’ styles and in its SC progeny(Boskovíc et al., 1999). The SF gene se-quence segregating with SC was isolatedfrom ‘Tuono’ (Ma and Oliveira, 2000,2001; Channuntapipat et al., 2001)though so far it is not known why there isno corresponding RNase activity. Bree-ding tests confirmed segregation of SF se-quence and lack of RNase activity withthe SC phenotype (Ortega and Dicenta,2003). Apparently, other SC almond culti-vars display only one S-RNase in theirstyles (i.e. they lack one S-RNase) butthe corresponding genes have still notbeen isolated.

THE ORIGIN OF SELF-COMPATIBILITYIN ALMOND‘Tuono’ is originally from Italy, from theregion of Apuglia (Herrero et al., 1977)where other Prunus such as P. webbii co-exist. Because P. webbii is reported as aSC species, it has been assumed that SCtrait was introgressed into almond fromthis wild relative (Godini, 1979, 2000),probably through Sf (Socias i Company,2002). A consequence of this assumptionis that P. webbii would have to carry theSf allele. Furthermore, if Sf was the onlycause of self-compatibility, all SC P. web-bii would contain this (or similarly non-functional) allele(s). If, on the other hand,P. webbii would have functional S allelesstill being SC, then other loci should alsobe implicated in SI/SC in this species.

THE Sf ALLELE IN P. webbiiWhen testing for the presence of Sf (usingSf specific primers) in six ecotypes of P.webbii (one from Spain and five fromApuglia) two were shown to carry the Sf

allele (Sanchez and Oliveira, unpublishedresults), which could account for theirself-compatibility. Since no RNase activi-ty in the pistil can be attributed to Sf, atleast half of the pollen grains would beable to grow in the pistil and set seed.Such Prunus webbii plants may have, atsome point, crossed with cultivated al-mond originating cultivars as ‘Tuono’ thatcarry Sf. It must be noted that the hybridi-zation must have occurred several gene-rations ago since, as assessed by mole-cular markers, ‘Tuono’ is more closely re-lated with almond than with P. webbii(Martins et al., 2003), that is to say, ‘Tuo-no’ is not an F1 hybrid and further crosseswith cultivated almond must have beentaken place in the origin of this cultivar.The fact that other ecotypes of P. webbiido not contain the Sf allele but are still SCdismisses the notion that the presence ofSf is a sine qua non condition for self-compatibility.

OTHER S ALLELES IN P. webbiiBy using primers designed for the conser-

ved regions of Rosaceae S alleles (Signalpeptide, C1 and C5) on P. webbii geno-mic DNA, it was possible to isolate otherS alleles besides Sf in this species (San-chez and Oliveira, 2003). Thus, for four ofthe six ecotypes tested, two S alleleswere isolated. As expected one of thesealleles was Sf in the two ecotypes alreadyshown to carry Sf. For the two remainingecotypes, only one S allele was found.The fact that P. webbii genome containsS-alleles does not constitute in itself asurprise. If SI was the original condition inPrunus and SC plants are derived from SIplants, it could be expected that all plantsof this genus may carry S-RNase genesthough some may be in a relic non-func-tional form. Nevertheless, testing S-geneexpression on pistil RNA from four varie-ties by RT-PCR, amplification was suc-cessful. This means that at least some P.webbii S-alleles are pistil-expressed sug-gesting that these genes may be functio-nal, a feature that could be confirmed bytesting the RNase activity of P. webbiipistils.

S-ALLELESAND SELF-COMPATIBILITYHow plants carrying the Sf allele could beself-compatible was already explained.Similarly plants carrying a single S allelemay be accounted for if we consider thatnot being able to isolate a second S-alleleis due to either its absence from the ge-nome or (more likely) an allele mutated tosuch an extreme that it is no longer re-cognisable by the primers and therefore itis most likely non-functional. The possibi-lity that these individuals are homozygousfor the S-locus (since P. webbii is SC thiswould be possible) and therefore to iden-tical S-alleles are present which wouldnot be distinguishable by PCR. As for theecotypes that carry two distinct S-allelesthat as judged by their sequence seem tocode putatively functional S-RNases, thecause of SC may be due to a lack of (orreduced) expression of the S-genes (oneor both) or SC may be a result of the ac-tion of other genes outside of the S-locus.It has long been known that the geneticbackground also affects the self-incompa-tibility response.

Self-fertile varieties of SI species areknown in other plants and in many casesmutations in the S-RNase coding regionor low expression levels of this proteincan explain self-fertility (Kowyama et al.,1994; Royo et al., 1994; Sassa et al.,1997). However other factors such asmutations on the putative S-pollen coun-terpart or at modifier loci can account forSC in the presence of fully functional S-RNases (Ai et al., 1991; Golz et al., 1998;Tao et al., 2002 and Tsukamoto et al.,2003). Similar situations are not to be dis-carded in Prunus webbii.


ALMOND GENES IN P. webbiiAnalysis of P. webbii S-allele sequencesshowed that three ecotypes carry allelesidentical to almond functional S-alleles.The observation that self-compatible P.webbii carries alleles known to confer SIis perhaps unexpected but supports thehypothesis put forward by Socias i Com-pany (2002) that at least to some extent,gene flow probably occurs (or has occur-red in the past) between the cultivated al-mond (P.amygdalus or P. dulcis) andneighbouring P. webbii trees. Alternative-ly, but less likely, these S-alleles werepresent in a common ancestor of thesetwo species. It is interesting to note thatthe Spanish ecotype shares alleles withthe ecotypes collected in Italy. This couldreflect the ancient nature of the S-allelesthat would be present in the original po-pulation of P. webbii or shows exchangeof plant material between these two Euro-pean regions.

CONCLUSIONIn conclusion, S-RNase coding genescould be isolated from a self-compatiblespecies, Prunus webbii, suggesting thatthis species may have been self-incom-patible in the past. The fact that the Sf

allele known to confer self-compatibility inalmond was found in some ecotypes of P.webbii provides evidence to the sugges-tion of this species being the origin of theSf allele of ‘Tuono’. This gene introgres-sion must be a result of ancient hybridiza-tion events. Furthermore, probably S-gene flow also occurred in the oppositedirection since almond S-alleles are pre-sent in P. webbii. Finally, self-compatibleplants do not necessarily carry the Sf alle-le previously identified, so other causesfor SC may exist that could be exploitedin breeding SC almonds.

REFERENCES

Ai, Y., Kron, E. and Kao, T.-H. (1991).S-alleles are retained and expressed ina self-compatible cultivar of Petunia hy-brida. Mol. General Genet. 230, 353-358.

Boskovíc, R., Tobutt, K.R., Duval, H.and Batlle, I. (1999). A stylar ribonu-clease assay to detect self-compatibleseedlings in almond progenies. TheorAppl Genet 99: 800–810.

Channuntapipat, C., Sedgley, M. andCollins G. (2001). Sequences of the cD-NAs and genomic DNAs encoding theS1, S7, S8, and SF alleles from almond,Prunus dulcis. Theor Appl Genet103:1115–1122.

Godini A. (1979). Ipotesi sulla compar-sa dell’autocompatibilità nel mandorla.Sci Tec Agr 19: 1–10.

Godini, A. (2000). About the possiblerelationships between Amygdalus web-bii Spach and Amygdalus communis L.Nucis Newsletter 9.

Golz, J.F., Clarke, A.E., Newbigin, E.and Anderson, M. (1998). A relic S-RNase is expressed in the styles of self-compatible Nicotiana sylvestris. Plant J.16, 591-599.

Herrero, M., Cambra, M. and Felipe, A.J. (1977) lnterpolinización de varieda-des de almendro. An. Inst. Nac. Invest.Agrar.,Ser. Prod. Veg. 7: 99-103.

Kester, D.E.; Ross, N.W. (1996). His-tory In Almond: Production Manual.W.C. Micke (ed) Publication 3364, Uni-versity of California, Oakland.

Kowyama, Y., Kunz, C., Lewis, I., New-bigin, E., Clarke, A.E. and Anderson,M.A. (1994). Self-compatibility in aLycopersicon peruvianum variant(LA2157) is associated with a lack ofstyle S-RNase activity. Theor. Appl. Ge-net. 88, 859-864.

Ortega, E. and Dicenta, F. (2003). Inhe-ritance of self-compatibility in almond:breeding strategies to assure self-com-patibility in the progeny. Theor. Appl.Genet. 106 (5): 904 – 911.

Ma R.C. and Oliveira M.M. (2000). Mo-lecular identification of S-genotypes inalmond (Prunus dulcis). Proceedings ofthe ISHS symposium on molecular mar-kers for characterizing genotypes andidentifying cultivars in horticulture. ActaHorticulturae 546: 575-580.

Ma R.C. and Oliveira M.M. (2001). Mo-lecular cloning of the self-compatibilitygenes S1 and S3 from almond Prunusducis cv.Ferragnes. Sex Plant Reprod14:163-167.

Tao, R., Habu, T., Namba, A., Yamane,H., Fuyuhiro F., Iwamoto, K. and Sugiu-ra, A. (2002). Inheritance of SF-RNasein Japanese apricot (Prunus mume) andits relation to self-compatibility. TheorAppl Genet 105: 222–228.

Crossa Raynaud, P and Grasselly, Ch.(1985). Existence de groupes d’intersté-rilité chez l’amandier. Options Medite-rranean CIMEAM/ IAM2 85/1: 43-45.

Royo, J., Kunz, C., Kowyama, Y., An-derson, M.A., Clarke, A.E. and Newbi-gin, E. (1994) Loss of a histidine resi-due at the active site of S locus ribonu-clease is associated with self-compati-bility in Lycopersicon peruvianum. Proc.Natl Acad. Sci. USA 91, 6511-6514.

Sanchez. A.M. and Oliveira, MM (2003).S-alleles in self-compatible Prunuswebbii. CIHEAM Cahiers Options Médi-terranéenes, Proceedings of the XIIIGREMPA Meeting on Pistachios and Al-monds (in press).

Sassa, H., Hirano, H., Nishio, H. andKoba, T. (1997). Style-specific self-compatible mutation caused by deletionof the S-Rnase gene in Japanese pear(Pyrus serotina). Plant J. 12, 223-227.

Socias i Company. (2002). The relation-ship of Prunus webbii revisited. Nucisnewsletter 11: 17-19.

Tsukamoto, T., Ando, T., Kokubun, H.,Watanabe, H., Sato, T., Masada, M.,Marchesi, E. and and Kao, T.- H.(2003). Breakdown of self-incompatibili-ty in a natural population of Petuniaaxillaris caused by a modifier locus thatsuppresses the expression of an S-RNase gene. Sexual Plant Reproduc-tion 15: 255–263.

Wang, Y., Wang, X., Skirpan, A.L. andKao, T.-H. (2003). S-RNase-mediatedself-incompatibility Journal of Experi-mental Botany, 54 (380): 115-122.

A. Sanchez 1 and M. Oliveira 1, 2

1. Laboratory of Plant Genetic Engineering,IBET/ITQB, Quinta do Marquês, 2784-505

Oeiras, Portugal2. Departamento de Biologia Vegetal,

Faculdade de Ciências da Universidade deLisboa, Campo Grande 1749-016 Lisboa,

PortugalE-mail: [email protected]

APPLICATION OF MOLECULARMARKERS IN ALMOND

BREEDING PROGRAMMES

INTRODUCTIONDeveloping new cultivars is a long and te-dious process in almond [Prunus dulcis(Mill.) D.A. Webb], involving the genera-tion of large populations of seedlings fromwhich the best genotypes are selected.Whereas the capacity of breeders to ge-nerate big populations from crosses is al-most unlimited, its management, scree-ning and selection of seedlings are themain limiting factors in the generation ofnew releases. The development of mole-cular markers to be used in early selec-tion of seedlings in the almond breedingprogrammes, is interesting for the selec-tion of these new cultivars.

Most of the work to develop molecularmarkers in almond has been carried outin setting up the different techniques andmapping the genome of the species, ne-vertheless the application in breedingprogrammes has been more reduced. Dif-ferent types of molecular markers, inclu-ding isoenzymes, RFLPs, RAPDs, andSSRs, have been applied for the geneticcharacterization of germplasm, the esta-blishment of genetic relationships betwe-en cultivars and species, and the cons-truction of genetic maps. In addition, me-thodologies for the analysis of marker-as-sisted selection include the use of map-ping populations segregating for desiredcharacters and bulk segregant analysis.


Currently there is a good knowledgeabout the almond genome, which toge-ther with the quantity and suitability ofplant material raised from almond bree-ding programmes, can allow the develop-ment of useful molecular tools to apply tothe selection of genitors and seedlings.

In this work, a review of the application ofmolecular markers in almond breeding ispresented, with special emphasis on thedevelopment of marker assisted selectionstrategies.

MOLECULAR CHARACTERIZATIONAND GENETIC RELATIONSHIPSTraditionally, the identification and cha-racterization of almond cultivars has beenbased on morphological and agronomicaltraits. However, such traits are not alwaysavailable for analysis and are affected bychanging environmental conditions. Mole-cular marker technology offers severaladvantages over the use of conventionaltraits. Molecular markers also offer apowerful tool to study genome evolution,and for understanding genome structureand determinants of genetic diversity(Wünsch and Hormaza, 2002).

Isoenzymes: Isoenzymes were amongthe first genetic markers to be widely utili-zed. They have been used for cultivaridentification in almond because of theirenvironmental stability, their codominantexpression, and their good reproducibili-ty. Nevertheless, their utilization is limitedby the small number of loci that can beanalysed with conventional enzyme stai-ning methods, as well as a low variationin some loci (Arulsekar et al., 1986;Hauagge et al., 1987; Cerezo et al., 1989;Sathe et al., 2001). More recently, isoen-zymes in combination with DNA-basedmarkers were applied to develop geneticmaps for woody perennials (see “Geneticmapping” below) and for the genetic cha-racterization of multiple embryos in al-mond (Martínez-Gómez and Gradziel,2003).

RFLPs: Restriction fragment length poly-morphism (RFLP) markers are based onthe differential hybridization of clonedDNA to bulk DNA fragments from restric-tion-enzyme digestion. RFLP markers arecodominant. RFLPs can detect a virtuallyunlimited number of markers, thus provi-ding an efficient method for discoveringlinkages among markers and for cons-tructing genetic maps. RFLP have beenassayed in almond for map-based selec-tion (Viruel et al., 1995). However, RFLPanalysis has important limitations: it is la-borious and time-consuming and it ofteninvolves the use of radioisotopes.

RAPDs: Random amplified polymorphicDNA (RAPD) markers are based on the

PCR amplification of random locations inthe genome. RAPDs are characterized byusing arbitrary primers. A single oligonu-cleotide is used for the amplification ofgenomic DNA. In contrast to isoenzymesand RFLPs, RAPDs are dominant mar-kers. This feature, as well as their varia-ble degree of repeatability and problemsin transferring across populations, limitstheir use primarily to map construction.RAPD techniques have been successfullyused in almond for identifying cultivars,estimating genetic diversity and asses-sing possible origins for selected genoty-pes (Bartolozzi et al., 1998; Martins et al.,2003).

SSRs: PCR-based, simple sequence re-peat (SSR) markers (microsatellites) arebecoming the marker of choice for finger-printing and genetic diversity studies for awide range of plants. Because of theirhigh polymorphism, abundance, and co-dominant inheritance, they are well suitedfor the assessment of genetic variabilitywithin crop species, and for the geneticrelationships among species (Powell etal., 1996). In the case of Prunus species,primer pairs flanking SSRs have beencloned and sequenced in different spe-cies including peach, apricot, and cherry(Aranzana et al., 2003). These SSR mar-kers have been used for the molecularcharacterization and identification of al-mond cultivars (Martínez-Gómez et al.,2003a) and related Prunus species (Fig.1) (Martínez-Gómez et al., 2003b); and

elaboration of genetic maps (Joobeur etal., 2000; Bliss et al., 2002) (see “Geneticmapping” below).

Electrophoresis in polyacrilamide with ra-dioactive and silver staining was the firstmethod used in the analysis of the PCRamplified fragment of DNA obtained fromthe SSR markers. Electrophoresis in Me-taphor® agarose was a method used asalternative to the polyacrilamide due to itseasier application (Fig. 2). More recently,new methods for the PCR amplified DNAhave been developed including the use ofautomated sequencers.

Figure 1. Genetic relationship among peach and almond cultivars and related Prunus species obtained bya study with SSR markers (Martínez-Gómez et al., 2003b). Unrooted dendogram obtained by Neighbour

Joining cluster analysis

Figure 2. Allelic segregation of UDP96-013 SSRmarker in almond cultivars using Metaphor®

agarose and Polyarcilamide. First and lastsamples correspond to a 1 kb DNA ladder

Polyacrylamide

Metaphor® Agarose


GENETIC MAPPINGSeveral intraspecific and interspecific al-mond maps have been developed usingdifferent types of molecular markers. Theuse of PCR-based markers made it possi-ble to map and tag a wide range of traits(Arús et al., 1994; 1999). The analysis ofcosegregation among markers eases link-age analysis between markers and majoror quantitative loci controlling horticultu-rally important traits.

Different research groups have releasedlinkage maps using morphological traits,isoenzymes, RFLPs and RAPDs in al-mond (Viruel et al., 1995; Joobeur et al.,1998; 2000; Jáuregui et al., 2001; Bliss etal., 2002). In addition, SSRs have beenused for mapping in almond (Joobeur etal., 2000; Bliss et al., 2002; Aranzana etal., 2003). The similar arrangement ofmarkers observed in different Prunusmaps suggests a high level of syntenywithin the genus (Aranzana et al., 2003).This homology among Prunus speciespartly explains the low level of breedingbarriers to interspecific gene introgres-sion and supports the opportunity for suc-cessful gene transfer between closely re-lated species (Gradziel et al., 2001).

MARKER-ASSISTED SELECTIONSelection by molecular markers is parti-cularly useful in fruit, nut, and other treecrops with a long juvenile period, andwhen the expression of the gene is reces-sive or the assessment of the character isdifficult. Marker-assisted selection (MAS)is emerging as a very promising strategyfor increasing selection gains. If sufficientmapping information is available, MAScan highly shorten the number of genera-tions required to “eliminate” the undesiredgenes of the donor in backcrossing pro-grammes (Arús and Moreno-González,1993). Marker loci linked to major genescan be used for selection, which is moreefficient than direct selection for the tar-get gene (Arús and Moreno-González,1993).

The principal approach for the analysis ofmarker-trait association in almond is theuse of mapping populations segregatingfor the characters of interest. The diffe-rent linkage maps developed in almondinclude markers associated with severaltraits of horticultural value. Mapping

quantitative characters by identifyingquantitative trait loci (QTL) is also beco-ming an important tool in tree breeding.QTLs are generally recognized by com-paring the degree of covariation for poly-morphic molecular markers with phenoty-pic trait measurements. Important charac-ters and QTLs that are presently beingmapped in almond include bloom time,self-incompatibility, shell hardness, orkernel taste (Asins et al., 1994; Viruel etal., 1995; Ballester et al., 1998, 2001;Jobeur et al., 1998; 2002; Bliss et al.,2002) (Table 1). The high degree of ge-nome synteny observed among Prunusspecies should also facilitate the succes-sful transfer of sets of markers and co-ding sequence among species (Aranzanaet al., 2003) (Table 1).

Bulk segregant analysis (BSA), wheretwo pooled DNA samples are formed fromplant sources that have similar geneticbackgrounds but differ in one particulartrait, is another promising approach forthe analysis of molecular marker-horticul-tural trait association. A strategy combi-ning different markers with bulk segre-gant analysis was used to identify mar-kers linked to loci of specific characters inpeach x almond crosses (Warburton etal., 1996), and three RAPD markers as-sociated with a gene conferring delayedbloom in almond (Ballester et al., 2001)(Table 1).

A very promising application of molecularmarker assisted selection is the manipu-lation of self-compatibility in almond. Thisspecies is predominantly self-incompati-ble. Self-incompatibility is of the gameto-phytic type and acts to prevent self-fertili-zation. This character is controlled by asingle locus with multiple codominantalleles (Dicenta and García, 1993), and isexpressed within the styles of flowers asS-RNases glycoproteins (Boskovic et al.,1997; 2003) that are responsible for thesubsequent inactivation of self-pollentube growth. Almond self-incompatibilityalleles (S-alleles) were initially identifiedin the field through controlled crosseswith a series of known S-genotypes (Kes-ter and Gradziel, 1996). More recently,molecular methods have been developedin two areas: identification of stylar S-RNases by electrophoresis in verticalpolyacrilamide gels (Boskovic et al.,

1997, 2003), and the amplification of spe-cific S-alleles using appropriately desig-ned primers for PCR and electrophoresisin horizontal agarose gels (Tamura et al.,2000; Channuntapipat et al., 2003). Thistechnique is being routinely used for theidentification of cross-incompatibility grou-pings for current almond cultivars and forefficiently breeding self-compatibility intonew cultivars (Gradziel et al., 2001b; Or-tega and Dicenta, 2003).

ACKNOWLEDGMENTSThe authors acknowledge the support ofSpanish Ministry of Education and Science.

BIBLIOGRAPHY

Aranzana M.J., Pineda A., Cosson P.,Dirlewanger E., Ascasibar J., CiprianiG., Ryder C.D., Testolin R., Abbott A.,King G.J., Lezzoni A.F. and P. Arús.2003. A set of simple-sequence (SSR)markers covering the Prunus genome.Theor. Appl. Genet. 106: 819-825.

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Arús P. and J. Moreno-González. 1993.Marker-assisted selection. In: Hayward,M.D., Bosemark, N.O., and Romagosa,I. (eds.). Plant Breeding. Principles andProspects. London, United Kingdom:Chapman & Hall. p. 314-331.

Arús P., Messeguer R., Viruel M.A., To-butt K., Dirlewanger E., Santi F., QuartaR. and E. Ritter. 1994. The EuropeanPrunus mapping project. Euphytica 77:97-100.

Arús P., Ballester J., Jauregui B., Jo-obeur T., Truco M.J. and M.C. de Vicen-te. 1999. The European Prunus map-ping project: update of marker develop-ment in almond. Acta Hort. 484: 331-336.

Asins M.J., Mestre, P., García, J.E., Di-centa, F., and E.A. Carbonell. 1994.Genotype ∞ environment interaction inQTL analysis of an intervarietal almondcross by means of genetic markers.Theor. Appl. Genet. 89: 358-364.

Ballester J., Boskovic R., Batlle I., ArúsP., Vargas F. and M.C. de Vicente.1998. Location of the self-incompatibili-ty gene on the almond linkage map.Plant Breed. 117: 69-72.

Ballester J., Socias i Company R., ArúsP. and M.C. de Vicente. 2001. Geneticmapping of a major gene delayingblooming time in almond. Plant Breed.120: 268-270.

Bartolozzi F., Warburton M.L., Arulse-kar S. and T.M. Gradziel. 1998. Geneticcharacterization and relatedness amongCalifornia almond cultivars and bree-ding lines detected by randomly ampli-fied polymorphic DNA (RAPD) analysis.J. Am. Soc. Hort. Sci. 123: 381-387.

Table 1. Markers associated to main agronomic traits in almond.

Trait Symbol Marker Reference

Self-incompatibility SI RFLP Joobeur et al. 1998Self-compatibility Sf RFLP Arús et al. 1999Kernel taste Sw RFLP Bliss et al. 2002Shell hardiness D RFLP Arús et al. 1999Late blooming Lb RAPD Ballester et al. 2001


Bliss F.A., Arulsekar S., Foolad M.R.,Becerra V., Gillen A., Warburton M.L.,Dandekar A.M., Kocsine G.M. and K.K.Mydin. 2002. An expanded geneticlinkage map of Prunus based on an in-terspecific cross between almond andpeach. Genome 45: 520-529.

Boskovic R., Tobutt K.R., Batlle I. andH. Duval. 1997. Correlation of ribonu-clease zymograms and incompatibilitygenotypes in almond. Euphytica 97:167-176.

Boskovic R. , Tobutt K R, Batlle I., Du-val .H, Martínez-Gómez P. and T.M.Gradziel. 2003. Stylar ribonucleases inalmond: correlation with and predictionof self-incompatibility genotypes. PlantBreed. 122: 70-76.

Cerezo M., Socias i Company R. and P.Arús. 1989. Identification of almond cul-tivars by pollen isoenzymes. J. Am.Soc. Hort. Sci. 114: 164-169.

Channuntapipat C., Wirthensohn M.,Ramesh S.A., Batlle I., Arús P., SedgleyM. and G. Collins. 2003. Identification ofincompatibility genotypes in almondusing specific primers based on the in-trons of the S-alleles. Plant Breed. 122:164-168.

Dicenta F. and J.E. García. 1993. Inhe-ritance of self-compatibility in almond.Heredity 70: 313-317.

Gradziel T.M., Martínez-Gómez P., Di-centa F., and D.E. Kester. 2001a. Theutilization of related almond species foralmond variety improvement. J. Am.Pomolog. Soc. 55: 100-109.

Gradziel T.M., Martínez-Gómez P. andA.M. Dandekar. 2001b. The use of S-allele specific PCR analysis to improvebreeding efficiency for self-fertility in al-mond. HortScience 36: 440-440.

Hauagge R., Kester D.E. and R.A.Asay. 1987. Isozyme variation amongCalifornia almond cultivars: I. Inheritan-ce. J. Am. Soc. Hort. Sci. 112: 687-693.

Jáuregui B., de Vicente M.C., Messe-guer R., Felipe A., Bonnet A., SalessesG. and P. Arús. 2001. A reciprocaltranslocation between ‘Garfi’ almondand ‘Nemared’ peach. Theor. Appl. Ge-net. 102: 1169-1176.

Joobeur T., Viruel M.A., de VicenteM.C., Jáuregui B., Ballester J., DettoriM.T., Verde I., Truco M.J., MesseguerR., Battle I., Quarta R., Dirlewanger E.,and P. Arús. 1998. Construction of asaturated linkage map for Prunus usingan almond x peach F2 progeny. Theor.Appl. Genet. 97: 1034-1041.

Joobeur T., Periam N., de Vicente M.C.,King G.J. and P. Arús. 2000. Develop-ment of a second generation linkagemap for almond using RAPD and SSRmarkers. Genome 43: 649-655.

Kester D.E. and T.M. Gradziel. 1996.Almonds (Prunus). In: Moore, J.N. andJanick, J. (eds.). Fruit Breeding. NewYork: Wiley. p. 1-97.

Martínez-Gómez P. and T.M. Gradziel.2003. Sexual polyembryony in almond.Sex. Plant Reprod. 16: 135-139.

Martínez-Gómez P., Arulsekar S., Pot-ter D., T.M. and Gradziel T.M. 2003a.An extended interspecific gene poolavailable to peach and almond breedingas characterized using simple sequencerepeat (SSR) markers. Euphytica 131:313-322.

Martínez-Gómez P., Arulsekar S., Pot-ter D., and T.M. Gradziel. 2003b. Rela-tionships among peach and almond andrelated species as detected by SSRmarkers. J. Amer. Soc. Hort. Sci. 128:667-671.

Martins M., Tenreiro R. and M.M. Olivei-ra. 2003. Genetic relatedness of Portu-guese almond cultivars assessed byRAPD and ISSR markers. Plant CellRep. 22: 71-78.

Ortega E. and F. Dicenta. 2003. Inheri-tance of self-compatibility in almond:breeding strategies to assure self-com-patibility in the progeny. Theor. Appl.Genet. 106: 904-911.

Powell W., Morgante M., Andre C., Ha-nafey M., Vogel J., Tingey S., and A.Rafalski. 1996. Comparison of RFLP,RAPD, AFLP and SSR (microsatellite)markers for germplasm analysis. Mol.Breed. 2: 225-238.

Sathe S.K., Teuber S.S., Gradziel T.M.,and K.H. Roux. 2001. Electrophoreticand immunological analyses of almond(Prunus dulcis L.) genotypes and hy-brids. J. Agr. Food Chem. 49: 2043-2052.

Tamura M., Ushijima K., Sassa H., Hira-no H., Tao R., Gradziel T.M. and A.M.Dandekar. 2000. Identification of self-incompatibility genotypes of almond byallele-specific PCR analysis. Theor.Appl. Genet. 101: 344-349.

Viruel M.A., Messeguer R., de VicenteM.C., Garcia-Mas J., Puigdomènech P.,Vargas F. and P. Arús. 1995. A linkagemap with RFLP and isozyme markersfor almond. Theor. Appl. Genet. 91:964-971.

Wünsch A. and J.I. Hormaza. 2002.Cultivar identification and genetic fin-gerprinting of temperate fruit tree spe-cies using DNA markers. Euphytica125: 56-67.

1R. Sánchez-Pérez, 1F. Dicenta, 2T.M.Gradziel, 3P. Arús, and 1P. Martínez-Gómez.

1CEBAS-CSIC. Departamento de Mejoray Patología Vegetal, Apdo 4195, 30.080,

Murcia, Spain.2University of California-Davis,

Dept. of Pomology, Davis,CA-95616, USA.

3 IRTA. Departament de Genètica Vegetal.Crta. Cabrils s/n, E-08348 Cabrils,

Barcelona, Spain.E-mail: [email protected]

Fusicoccum CANKERIN ALMOND ORCHARDS –BIOTECHNOLOGY TOOLSFOR EARLY SELECTION

OF TOLERANT GENOTYPES

Fusicoccum CANKER ON ALMONDAlmond is an important crop in the Medi-terranean region, reaching about 28% ofthe world production. However, its sus-ceptibility, particularly to certain fungal di-seases such as brown rot blossom blight,constriction canker and almond scab,which can cause severe damage on folia-ge and branches of the tree, can be a ma-jor impediment for almond production.

Regarding Fusicoccum canker or cons-triction canker, this is a damaging andeconomically important fungal disease inalmond orchards in the Mediterranean re-gion, caused by Phomopsis amygdali(Del.) (perfect stage) Tuset and Portilla,formerly classified as Fusicoccum amyg-dali Delacr. (imperfect stage). Besides al-mond, this fungus also infects peachtrees. Symptoms of this disease usuallyappear in early summer and become in-creasingly evident as more infectedshoots appear through late summer. In al-mond mainly two types of symptoms arevisible, on shoots and on May spurs. In-fected twigs and shoots wilt, and even-tually die, because of elongate, brown,sunken cankers, often with a well delimi-ted oval pattern (Grasselly and Duval,1997) (Fig. 1).

The group of leaves that form May spurswilt due to the infection, caused on thatspring or on the previous autumn. It isalso possible to observe cankers on theinternodes of these spurs. Leaves in ge-neral can also be affected, showing ne-crosis that usually appear as big brownspots with circular or irregular shape.Gumming is commonly associated withthe cankers, although it is not a gooddiagnostic characteristic since otheragents may also be responsible for thissign. Constrictions formed at the base ofinfected shoots and leaf symptoms produ-ced well beyond the infection site resultfrom a translocatable toxin, fusicoccin, aphytotoxic terpenoid glucoside. Fusicoc-cin, increases the influx of potassium onguard cells inducing the permanent ope-ning of the stomata and the consequenthigh loss of water can eventually lead toplant death (De Boer, 1997).

The development of Fusicoccum cankeris favoured by high nitrogen levels in thesoil and by wet weather, since this disea-se is transmitted by rain (Rosenberg,1996). Most infections occur during


spring (growing season of the plants) andautumn, when apparently almond treesas well as peach trees are more suscepti-ble to the disease. Pycnidia (darkly pig-mented, flask-shaped, conidia-bearing frui-ting) are produced over the cankered sur-face area. Under wet weather the pycni-dia exude conidia in white tendrils. Theseconidia disseminated by rain, will infectnew plants through scars on leaves, budsor stipules, or directly through youngshoots during their growing stage by latewinter and early spring. By the end ofsummer, the trees can be severely affec-ted exhibiting numerous terminal deadzones spread through the tree canopy(Grasselly and Duval, 1997). These effectscan lead to significant losses on fruit pro-duction in almond cultivars.

The management of infected orchardsshould involve the implementation of fun-gicide application programmes and pru-ning (elimination) of infected limbs to re-duce inoculum. Winter pruning of canke-red shoots, followed by pruning of youngshoots showing symptoms of the diseasein spring, are important practices to helpFusicoccum canker control (Barbé, 1993).Fungicides, applied just before budbreak(February-March) and in autumn, may benecessary in orchards where the diseaseis present (Grasselly & Duval, 1997). Ke-eping nitrogen at low to moderate levelsin the soil contributes to minimize the se-verity of the infection (Rosenberg, 1996).

Wide differences on Fusicoccum toleran-ce between cultivars have been observed(Grasselly and Crossa-Raynaud, 1980;Romero and Vargas, 1981) and shouldbe taken into consideration when new or-chards are planned.

Almond populations are found in mostMediterranean countries, where weatherconditions mainly in coastal orchards areusually favourable to Fusicoccum deve-lopment. Therefore, the use of tolerance

Figure 1. Infected shoots showing brown sunken cankers. (Images kindly provided by I. Batlle)

to Fusicoccum as a selectable trait in al-mond breeding programmes and in theestablishment of new orchards can be ofgreat importance.

MARKER ASSISTED SELECTION (MAS)Genetic markers, as transmissible enti-ties associated to economically importanttraits, can be used by breeders as impor-tant selection tools (Darvasi & Soller,1994). After a decade of marker assistedselection in vegetable crops, increasingavailability of genomics-derived informa-tion is starting to allow the use of molecu-lar markers as tools to support classicalbreeding programmes in fruit tree species.

Identification of molecular markers close-ly linked to genes controlling traits of in-terest are essential in marker assisted se-lection. The closer the linkage, less is theprobability of recombination between themarker and the gene(s) that control thetrait of interest, thus the higher will be theefficiency of selection.

The best perspectives for MAS applica-tion may rely on breeding programmes ai-ming for disease resistance improvement,particularly in systems where several ge-nes are involved, with complex interacti-ons, and where gene pyramiding is desi-rable (Kelly, 1995). Markers for diseaseresistance offer the additional advantageof permitting selection for resistance inthe absence of the pathogen(s) (Me-hlenbacher, 1995). Another possible appli-cation can be on recurrent selection ofthe cultivated genotype when the aim isgene introgression from wild individuals.

The construction of a linkage map of acertain population allows the study of co-segregation of markers with genes res-ponsible for the variability found in thatpopulation, making possible the identifi-cation of major genes or loci of quantitati-ve characters (QTLs) on the map (Arús etal., 1999). Genetic maps developed in al-

mond (or almond x peach) have beenused to identify markers linked to geneswith important effects on the morphologyand physiology of the plant, for exampleself-incompatibility, shell hardness or al-mond bitterness. Integration in thesemaps of new markers closely linked togenes of interest and their routine use oncurrent or future breeding programmes, isof high interest. However, despite its po-tential benefits, MAS applied to almondbreeding will likely be restricted to a limi-ted set of traits and parents where datafor segregating populations are available.Examples of applied MAS reported in theliterature are still scarce, mainly becauseprivate companies are the main users ofMAS, therefore information is more diffi-cult to be available. Moreover, the costsand elaborated technology required formost of the markers available make MASapplication of limited use in breeding pro-grammes.

MAS applied to woody plants such as al-mond would be extremely important,mainly for allowing the selection of cha-racters of interest on an initial stage ofplant development, which would only beexpressed after 2 to 5 years in the field,hence representing a significant reduc-tion on time and area used when compa-red to species with a shorter life cycle.

SEARCH AND IDENTIFICATIONOF MOLECULAR MARKERS –THE CASE OF Fusicoccum LINKEDMARKERS IN ALMONDPlant breeding programmes require relati-vely low-cost methods that can be usedfor hundreds or thousands of individuals,with a reliability of at least 95% (Rafalski& Tingey, 1993), these criteria make PCRbased techniques very attractive. Due totheir low cost, simplicity and easy detec-tion, RAPD (Randomly Amplified Poly-morphic DNA, Williams et al., 1990) mar-kers are frequently chosen for this kind ofstudy. However, a poor specificity andlack of reproducibility can sometimes beassociated to this technique. In order toovercome these deficiencies, RAPD frag-ments can be converted into SCAR mar-kers (Sequence Characterized AmplifiedRegion, Paran & Michelmore, 1993), mo-re reproducible and sometimes codomi-nants, or to CAPS (Cleaved Amplified Po-lymorphic Sequences, Konieczny & Ausu-bel, 1993; Jarvis et al., 1994).

Michelmore et al. (1991) showed thatRAPD bands linked to a trait of interestwould be easily identified through the useof two samples of bulked DNA from a po-pulation segregating for that trait: withineach pool, or bulk, the individuals areidentical for the trait or gene of interestbut are arbitrary for all other genes. Anypolymorphism between the two pools


should be linked to the trait being analy-sed. Mapping on a segregating popula-tion may confirm the markers found. Thistechnique, identified as “bulk segregantanalysis” (BSA), is now commonly usedto map simple traits with RAPD markers.A variation of this technique, known as“bulked DNA analysis”, uses also twosamples of DNA mixtures, one of indivi-duals that express the trait, and anotherof individuals that do not express it, butwithout using a segregating population.

Once a marker is identified as being rela-ted to a trait of interest, the developmentof SCAR or CAPS markers results in ahigher specificity and reproducibility ofthe fragments obtained. RAPD analysisfollowed by SCAR or CAPS developmenthas been used to identify markers asso-ciated to traits of interest in plants, suchas downy mildew resistance in lettuce(Paran & Michelmore, 1993), the locusthat controls flower doubleness in carna-tion (Scovel et al., 1998), a new resistan-ce gene to leaf stripe in barley (Tacconiet al., 2001), leptine content in Solanumphureja (Bouarte-Medina et al., 2002),sex determination in kiwifruit plants (Gillet al., 1998), genotypes of vine rootstocks(Xu et al., 1995), colour of fruit epidermisin apple (Cheng et al., 1996), Ma1 root-knot nematode resistance gene in Myro-balan plum (Lecouls et al., 1999) and re-sistance to root-knot nematodes in peach(Lu et al., 1999).

In our study, the RAPD technique wascombined with bulked DNA analysis toselect markers associated to either tole-rance or sensitivity to Fusicoccum in al-mond. Fourteen almond cultivars classi-fied as tolerant (7) or sensitive (7) to Fu-sicoccum were screened with 120 RAPDprimers. Polymorphic bands were asses-sed for co-segregation with tolerant orsusceptible phenotypes using 30 se-edlings obtained from a cross of ‘Masbo-vera’ (T) x ‘Lauranne’ (S). Three RAPDmarkers were identified, two linked to to-lerance (OPD-19300 and OPA-08900) andone linked to sensitivity (OPD-19500) toFusicoccum (Martins et al., 2001; Martinset al., 2003). SCAR primers were develo-ped and segregation of SCAR markerswas tested, however, polymorphism bet-ween the two groups (tolerant versus sus-ceptible) was lost in this process. The de-velopment of CAPS markers as a possi-ble way to recover the lost polymorphismwill be analysed.

Cosegregation analysis using the ade-quate populations is essential to confirmthe genetic base of the markers linked toa certain trait. In the case of almond, anew segregating population of some 120seedlings derived from a cross ‘Primors-kyi’ (T) x ‘Lauranne’ (S), is currently being

evaluated in the field, at IRTA, CentreMas Bové (Tarragona, Spain), for toleran-ce or sensibility to Fusicoccum. This ma-terial will allow the confirmation of whichfragments correspond to markers asso-ciated to the disease. In this case it wouldbe of high interest to evaluate the possibi-lity of inserting these markers in the refe-rence European Prunus genetic map, al-mond ‘Texas’ x peach ‘Earlygold’ (TxE)(Joobeur et al., 1998).

The genetic nature of tolerance to Fusi-coccum in almond (or related species likepeach) is not yet defined, and further stu-dies are necessary to determine the mo-del that can be applied to almond. Al-though artificial inoculation with Fusicoc-cum is currently being tested in almond(Cabrita et al., 2003), any procedure hasbeen established, so far, to efficientlyevaluate tolerance or sensibility of al-mond plants to Fusicoccum canker. The-refore, the identification of molecular mar-kers linked to tolerance or sensibility toFusicoccum, is of particular importancesince, to our knowledge, this is the onlymethod that will allow the early selectionof tolerant plants from breeding program-mes, as well as the certification of tole-rant cultivars to be implemented in neworchards. Symptoms development of thedisease in plants grown in the field deri-ved from breeding programmes takenabout 4 to 5 years under natural conditi-ons of high atmospheric humidity as it ha-ppens in the Mediterranean coast (I. Bat-lle, personal communication). Finally, it isimportant to highlight that the balanceand coordination among the use of mole-cular technologies and the fieldbasedwork, are essential for the success ofbreeding programmes (Scorza, 1996).

REFERENCES

Arús P, Joobeur T, Jaúregui B, BallesterJ, Viruel MA, de Vicente MC, 1999. Mar-cadores moleculares en especies fruta-les: Prunus. In: Marcide JMO (ed) Iden-tificación Molecular de Germoplasmade Vid. Fundación Premio ARCE, Insti-tuto Madrileño de Investigación AgrariaY Alimentaria (IMIA), Madrid pp 153-165.

Barbé L (1993) Favoriser le renouveaude la culture de l’amandier. Arboricultu-re fruitière. 358: 21-31.

Bouarte-Medina T, Fogelman E, ChaniE, Miller AR, Levin I, Levy D, VeilleuxRE, 2002. Identification of molecularmarkers associated with leptine in reci-procal backcross families of diploid po-tato. Theor Appl Genet 105: 1010-1018.

Cabrita L, Branco V, Neves MA, Leitão J,2003. Barrinho Grado a new source of re-sistance to Phomopsis amygdali in almond.XIII G.R.E.M.P.A. Meeting on Pistachioand Almond, Mirandela p 73 (Abstract).

Cheng FS, Weeden NF, Brown SK,1996. Identification of codominantRAPD markers linked to fruit skin colorin apple. Theor Appl Genet 93: 222-227.

Darvasi A and Soller M, 1994. Optimumspacing of genetic markers for determi-ning linkage between marker loci andquantitative trait loci. Theor Appl Genet89: 351-357.

De Boer B, 1997. Fusicoccum - a key tomultiple 14-3-3 locks? Trends in PlantScience 2: 60-66.

Gill GP, Harvey CF, Gardner RC, Fra-ser LG, 1998. Development of sex-linked PCR markers for gender identifi-cation in Actinidia. Theor Appl Genet97: 439-445.

Grasselly C & Crossa-Raynaud P, 1980.L’amandier. Maisonneuve et Larose,Paris.

Grasselly C & Duval H, 1997. L’Aman-dier. Centre Technique Interprofession-nel des Fruits et Légumes, Paris.

Jarvis P, Lister C, Szabo V, Dean C,1994. Integration of CAPS markers intothe RFLP map, generated using recom-binant inbred lines of Arabidopsis tha-liana. Plant Mol Biol 24: 685-687.

Joobeur T, Viruel MA, de Vicente MC,Jáuregui B, Ballester J, Dettori MT, Ver-de I, Truco MJ, Messeguer R, Batlle I,Quarta R, Dirlewanger E, Arús P, 1998.Construction of a saturated linkage mapin Prunus using an almond x peach F2progeny. Theor Appl Genet 97: 1034-1041.

Kelly JD, 1995. Use of random ampli-fied polymorphic DNA markers in bree-ding from major gene resistance to plantpathogens. HortScience 30: 461-465.

Konieczny A & Ausubel FM, 1993. Aprocedure for mapping Arabidopsis mu-tations using codominant ecotype-spe-cific PCR-based markers. Plant J 4:403-410.

Lecouls AC, Rubio-Cabetas MJ, MinotJC, Voisin R, Bonnet A, Salesses G,Dirlewanger E, Esmenjaud D, 1999.RAPD and SCAR markers linked to theMa1 root-knot nematode resistancegene in Myrobalan plum (Prunus cerasi-fera Ehr.). Theor Appl Genet 99:328-335.

Lu Z-X, Sossey-Alaoui K, Reighard GL,Baird WV, Abbott AG, 1999. Develop-ment and characterization of a codomi-nant marker linked to root-knot nemato-de resistance, and its application topeach rootstock breeding. Theor ApplGenet 99: 115-122.

Martins M, Sarmento D, Batlle I, VargasF, Oliveira MM, 2003. Development ofSCAR/CAPS markers linked to toleran-ce/sensitivity to Fusicoccum in almond.Options Méditerranéennes CIHEAM/IAMZ (in press).

Martins M, Sarmento D, Batlle I, VargasF, Oliveira MM, 2001. Search for mole-cular markers linked to Fusicoccum to-


lerance in almond. Acta Horticulturae577: 87-90.

Mehlenbacher SA, 1995. Classical andmolecular approaches to breeding fruitand nut crops for disease resistance.HortScience 30: 466-477.

Michelmore RW, Paran I, Kesseli RV,1991. Identification of markers linked todisease resistance genes by bulked se-gregant analysis: A rapid method to de-tect markers in specific genomic regi-ons by using segregating populations.Proc Natl Acad Sci 88: 9828-9832.

Paran I & Michelmore RW, 1993. Deve-lopment of reliable PCR-based DNAmarkers linked to downy mildew resis-tance genes in lettuce. Theor Appl Ge-net 85: 985-993

Rafalski JA & Tingey SV, 1993. Geneticdiagnostics in plant breeding: RAPDs,microsatellites and machines. TrendsGenet 9: 275-280.

Romero MA & Vargas FJ, 1981. Contri-bution à la connaissance de la sensibili-té de l’Amandier au Fusicoccum amyg-dali Del. Options Mediterranéennes,IAMZ. 81/1:143-147.

Rosenberg D, 1996. Update on pestmanagement and crop development –Peach constriction disease. ScaffoldsFruit Journal Vol. 5.

Scorza R, 1996. Genome mapping ofhorticultural crops: introduction to thecolloquium. HortScience 31: 1098.

Scovel G, Ben-Meir H, Ovadis M, Itzha-ki H, Vainstein A, 1998. RAPD andRFLP markers tightly linked to the locuscontrolling carnation (Dianthus caryo-phyllus) flower type. Theor Appl Genet96: 117-122.

Socias i Company R & Felipe AJ, 1992.Almond: a diverse germplasm. HortS-cience 27: 717-718, 863.

Tacconi G, Cattivelli L, Faccini N, Pec-chioni N, Stanca AM, Valé G, 2001.Identification and mapping of a new leafstripe resistance gene in barley (Hor-deum vulgare L.). Theor Appl Genet102: 1286-1291.

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M. Martins1,2, M.M. Oliveira1,2

(1) IBET/ITQB, Quinta do Marquês, 2784-505Oeiras, Portugal

(2) Dep. Biologia Vegetal, Fac. Ciênciasde Lisboa, Campo Grande1749-016 Lisboa, Portugal

E-mail: [email protected]

VIRUS SCREENINGFOR AUSTRALIAN ALMONDS

SUMMARYAs part of the Australian almond breedingprogram, work is conducted on breedingparents and progeny to screen for viralpathogens. Transmission of virus of up to23% has been recorded when infectedparents are used. As a service to industrythe University also test the Monash al-mond budwood facility.

INTRODUCTIONAn important aspect of the Australian al-mond breeding programme is the detec-tion of plant viruses in the breeding stockand progenies (Sedgley & Collins, 2002).The two most important viruses present inAustralia in almond (Prunus dulcis Mill.)are Prunus necrotic ringspot virus (PNR-SV) (Calico) and prune dwarf virus (PDV).Infection of nursery stock results in poorbud-take and retarded growth, and in com-mercial orchards growth and crop yieldsare affected. Leaf symptoms of PNRSVinclude chlorotic and necrotic leaf spots,mosaics, ringspots, and line patterns.

There are other viruses, plum pox virus(PPV) (Sharka); tomato black ring virus(TBRV) (Enation); and tomato ringspot vi-rus (ToRSV) (Peach yellow bud mosaic),which are present in Europe and Americabut not in Australia, however imported al-mond material is tested by AQIS for viru-ses on arrival. As a service to the Austra-lian almond industry the University alsoregularly tests the Monash almond bud-wood repository for viruses thus ensuringthat only tested material is used forbudding in nurseries. The Monash facilityconsists of 601 almond trees represen-ting all major cultivars grown in Australiaincluding those newly introduced fromEurope and the USA.

ELISA TESTING AND WOODYINDEXINGWhen screening began for PNRSV andPDV in 1997 the methods used were theantibody-based enzyme-linked immuno-sorbent assay (ELISA) technique and wo-ody indexing, where almond budwood isgrafted onto the indicator plant Prunusserrulata cv. Shirofugen, then monitoredfor diagnostic symptoms. The detection ofPNRSV by ELISA was shown by Bertozzi

et al. (2002) to be most sensitive whenplant material was collected in spring.

All breeding parents and progenies deri-ved from crosses made in the programmefor the first two years, almost 8,000 trees,were tested. A small number have produ-ced a positive response for PNRSV, butnone tested positive for PDV using ELI-SA. Where both parents were positive forPNRSV, transmission rates were high,22.6% & 12.5% for 1997 and 1998 res-pectively (Table 1). Seed transmissionfrom infected mother trees is generallyhigher than from virus-free mother treespollinated by infected pollen. The low le-vels of infected progeny from negativeparents could be due to infections in thefield. It can be seen from these data thatbreeding parents should be virus tested ifpossible, so that potential new cultivarsare clean.

Table 1. The percentage of progeny testing positive for PNRSV.

Progeny Both parents Pollen parent Seed parent Both parents-ve -ve -ve +ve

1997 0.4% 0 6% 22.6%1998 0.1% 1.4% 1.8% 12.5%

Figure 1. Leaf symptoms of PNRSV on almondleaves

Figure 2. Photo showing gumming symptomsof PNRSV on Shirofugen indicator plant after

grafting to infected almond budwood

Figure 3. Photo of ELISA sampling tray showingpositive (yellow) and negative (clear) results for

PNRSV testing


MONASHThe ELISA technique was used to screenall the Monash trees for PNRSV and PDVover 1997, 1998, & 1999 and half thetrees in 2000. PDV was not detected inany of the trees, however PNRSV wasdetected in 34 trees in 1997, 33 trees in1998, and 36 trees in 1999. Where treestest positive one year and negative thenext, this may be due to uneven distribu-tion of the virus in the plant, or seasonalfluctuations of viral concentrations due toextremes of temperature. The increasefrom 1997 to 1999 may be attributed to anincrease in the virus titre of these trees tolevels able to be detected by ELISA, ormay represent new infections.

RT-PCR TECHNIQUEA DNA based technique for virus detec-tion was developed by G. Collins. This te-chnique is based on the reverse transcrip-tion-polymerase chain reaction (RT-PCR)and is more sensitive than ELISA. The te-chnique involves extracting ribonucleicacid (RNA) from fresh leaves and usingspecially designed primers that can de-tect the viral coat protein within the plantcells. This technique was used on trees atMonash in 2000, 2001, 2002 and 2003.Generally RT-PCR confirmed the resultsof ELISA, however some trees were posi-tive for PNRSV where ELISA gave nega-tive results. This is due to the more sensi-tive nature of RT-PCR, and has theadded advantage that plant material canbe tested at any time throughout thegrowing season.

ACKNOWLEDGEMENTSThanks to Wanli Ma for technical assis-tance with the RT-PCR. Thanks to the Al-mond Board of Australia, HorticultureAustralia Limited and Australian Rese-arch Council for funding. Reproducedwith permission from The AustralianNutgrower Journal.

Figure 4. Agarose gel showing some cultivarswith a positive reaction to PNRSV primers.

Lane 1: molecular marker ladder,lane 2: Carmel(-ve), Lane 3: Fritz (+ve),

lane 4: Johnston (-ve), lane 5: Mission (-ve),lane 6: Nonpareil (+ve), lane 7: Nonpareil (-ve),lane 8: Parkinson (+ve), lane 9: Peerless (+ve),

lane 10: Price (+ve), lane 11: Price (-ve),Lane 12: negative control

REFERENCES

Bertozzi, T., Alberts, E., and Sedgley,M. 2002. Detection of Prunus necroticringspot virus in almond: effect of sam-pling time on the efficiency of serologi-cal and biological indexing methodolo-gies. Australian Journal of ExperimentalAgriculture, 42: 207-210.

Sedgley, M., and Collins, G. G. 2002.The Australian almond breeding progra-mme. Acta Horticulturae, 591: 241-244.

M. Wirthensohn1, S. Ramesh1, T. Bertozzi2,E. Alberts3, G. Mekuria4, C. Bennett5,

G. Collins1, and M. Sedgley1

1School of Agriculture & Wine, Universityof Adelaide, Waite Campus, PMB 1 Glen

Osmond, SA 5064 Australia.2Present Address: Evolutionary Biology Unit,

South Australian Museum, North Terrace,Adelaide, SA 5000 Australia.

3South Australian Research and DevelopmentInstitute, GPO Box 397, Adelaide, SA 5001

Australia.4Present address: Department of Natural

Resources and Environment (Victorian Institutefor Dryland Agriculture), Private Bag 260,

Horsham, Vic 3401 Australia.5Almond Board of Australia, Horticulture

House, PO Box 52 Berri, SA 5343 Australia.E-mail: [email protected]

THE PISTACHIO INDUSTRYIN ITALY: CURRENT SITUATION

AND PROSPECTS

INTRODUCTIONPistachio (Pistacia vera L.), is a dioeciousand deciduous species native to westernAsia and Asia Minor. It was introducedinto Italy from Syria in 30 A.D. by the Ro-man Governor of that Province, Lucius Vi-tellio, at approximately the same time itwas introduced in Spain (Minà Palumbo,1882). Its introduction into Sicily, whichnowadays accounts for almost the entirePistachio industry in Italy, probably occur-red some time later, following an initialperiod of cultivation in Campania which atthat time represented the roman “country-side”. Its cultivation in Sicily began tospread during the Arab domination (827-1040 A.D.) together with other major fruitand vegetable species and in parallel withthe improvement of the agronomic techni-ques. As a matter of fact, the Sicilian dia-lectal term for Pistachio tree (Fastuca) isof arab derivation (Fustuq). However,very little is known until recent timesabout its distribution over the region. His-toric references onto Pistachio cultivationare documented by the XVIIth century,even if first official records of the econo-mic importance of Pistachio industry arereferred to the first decades of the XXcentury.

Soon after the second world war the enti-re structure of the Sicilian Pistachio in-dustry underwent significant changes.The relative importance of the traditionalcultivation areas (Agrigento, Caltanis-setta and Palermo) strongly decreasedwith the abandonment of large part of thePistachio cultivation which was partiallycompensated by a parallel increase of theproduction in the Catania Province (Fig. 1).

Currently more than 90% of the total Ita-lian Pistachio area is concentrated in onlyfew territories in the eastern Sicily (wes-tern slopes of Mount Etna volcano), main-ly located in the districts of Bronte andAdrano (Catania Province), onto approxi-mately 4.500 hectares. Few residuals ofthe past cultivation can still be found inCaltanissetta and Agrigento Provinces, insmall and scattered surfaces (as a whole,130 ha in specialized cultivation and 210ha in mixed cultivation).

Despite this ancient origin and the longperiod of cultivation, only few female va-rieties of Pistachio are now grown in Italy(about ten) together with an even morerestricted number of unnamed male se-lections. Among the female cultivars,‘Bianca’ (synonym: Napoletana) is practi-cally the only cultivar grown, other varie-ties, namely ‘Femminella’, ‘Natalora’,‘Agostana’, ‘Silvana’, ‘Insolia’, ‘Cerasola’,‘Cappuccia’, representing not more than3% of the total (Barone et al., 1985) canstill be found, mostly in scattered, aban-doned settlements. This genetic pool isprobably the result of a two step introduc-tion process of genetic material (Baroneet al., 1996) coincident with the earliestintroduction from Syria of the reddish ‘Ce-rasola’ by the Romans and the later intro-duction carried out by the Arabs. The rea-sons of this low number of varieties isthought to be the exclusive use by thegrowers of vegetative propagation sinceancient times due to the long juvenilityperiod of P. vera seedlings, the long life-span of the trees and easy hybridizationswith other Pistacia species. Nevertheless,within the Sicilian Pistachio germplasm,some valuable fruit characteristics suchas colour, flavour and nut quality arehighly appreciated in trade, specially thegreenness of the kernels and the rich oilynut-like flavour (Woodroof, 1967). Most ofthese appreciated characteristics havebeen maintained also by cultivars obtai-ned in the United States from Sicilianseeds imported in the early 1900’s suchas ‘Bronte’ and ‘Trabonella’ (Joley, 1969).

In spite of these valuable characteristicsof the Italian pistachios, the contributionof Italian production represents nowadaysonly less than 0.6% of the world produc-tion. Nevertheless, with respect to pro-duct destination, it is remarcable that Ita-


lian pistachios maintain a dominant mar-ket position for the uses other than directconsumption (snack), since they are al-most entirely used, and highly apprecia-ted, by the confectionery and ice creamindustry.

Due to this fact Italian pistachio exports,between 1000 and 2000 tons per year(Anonymous, 2000), are representedmainly by shelled and peeled pistachios,whereas imports (about 9000 tons) con-sist mainly of in-shell pistachios, consu-med as salted and roasted snacks for do-mestic markets and, partially, for re-ex-portation.

THE BRONTE AREAThis area of the eastern Sicilian provinceof Catania currently represents the mainPistachio growing area in Italy. From theecological point of view this area (Fig. 2)is of great naturalistic importance due tothe proximity to the Etna Volcano and theinclusion in the Regional system of parksand naturalistic reserves (“Parco dell’Etna”,59.000 ha, established in 1987).

Pistachio production in Bronte representsone of the main economic resources ofthe entire territory. A total of 3.300 ha ofspecialized Pistachio orchards are loca-ted in this area and 1.500 ha in the neigh-

bourhood territories (Adrano, Biancavi-lla). The altimetry is comprised between350 (along the Simeto river bank) and900 m a.s.l.. The climate is typically Medi-terranean with fall and winter rainfall (an-nual average: ~550 mm). Average mon-thly temperatures of the area are betweena minimum of 6.0 °C (February) and amaximum of 32.6 °C (July) (Fabbri andValenti, 1998). Most of these Pistachioorchards are defined as “natural Pistachioplantings” because they have long beenobtained by grafting in situ spontaneousTerebinth (Pistacia terebinthus) plantsspread in the particular soils of the area,consisting of rocky, volcanic soils derivedin the centuries from the Mount Etna acti-vity, in steep slopes (Barone et al., 1985).This traditional planting system took ad-vantage of the extraordinary characteris-tics of environmental adaptation of Tere-binth to poor, dry, shallow soils of thearea, where no valid cultural alternativesare available, except for few other fruitspecies such as Prickly Pear and Fig.Thus, “natural Pistachio plantings” arecharacterized by the absence of regularplant spacings and by a wide range ofplant density (50-500 trees per ha). Alsothe age of the trees is consequently high-ly variable. Successive interplanting ofnursery terebinth seedlings is a commonpractice to replace dead plants and to in-crease orchard density. A few local smallnurseries are active in the area of Bronteand supply plant material mainly as se-edling P. terebinthus rootstocks to begrafted directly in the orchard. Smallquantities of grafted P. vera/P.terebinthuspot plants are also produced. In such kindof conditions cultural operations are ne-cessarily carried out by hand, since me-chanization is almost impossible also dueto the training system adopted that canbe defined as an “open vase” with three-four main irregular branches very close tothe soil surface. Pollination is ensured byspontaneous male P. terebinthus plantsor by scattered male P. vera pollinizersand also by natural hybrids between thetwo above mentioned species. Fruitsetproblems are likely to occur, speciallywhen pollination largely relies onto spon-taneous source of pollen, since male/fe-male ratio generally adopted in the Bron-te area is particularly low (1/20). Besidesnatural orchards, new plantations arealso present in the area. In this case ra-tional orchard design (6m x 8m or 8m x8m) and common cultural practices areadopted.

‘Napoletana’ (syn. Bianca) is the only cul-tivar widely utilized both in the “natural” orin the regular plantations and Terebinth isthe only rootstock. Following the descrip-tors’ list for Pistachio (Barone et al.,1997), ‘Bianca’ cultivar can be defined oflow-intermediate vigour. Growth habit is

2500

2000

1500

1000

500

0 1923/28

1953/58

1974/79

1980/85

1986/87

1993/94

2002/2003

(000

)Ton

s

Sicily Bronte area

Figure 1 - Pistachio production in Italy

Figure 2 - The “Parco dell’Etna” area. The Bronte Pistachio area is encircled


spreading; branching habit is intermedia-te. Terminal leaflet length is 10.1 cm, ter-minal leaflet width is 7.7 cm and the ter-minal leaflet/width ratio is 1.3. Peakbloom date occurs in the last week ofApril. Nut size is small to medium; nutlength is 21.6 mm, nut width is 11.6 mm,nut thickness is 9.9 mm, nut shape is ge-nerally elongated, split nuts is generallylow (<25%) and suture opening is narrow.Blank production is about 5%. Averagekernel dry weight is 0.48 g, colour is deepgreen; harvest date occurs between thelast week of August and the first week ofSeptember (Caruso et al., 1986; 1987;1993; Spina, 1982).

Old individual trees of other relic cultivarscan still be found in scattered areas:‘Femminella’, ‘Agostara’ and ‘Natalora’.The first one is very similar to ‘Bianca’and is now accounting for less than 5% ofthe total ‘Bronte’ production; it is charac-terized by a fruit smaller than ‘Bianca’ butit was very appreciated for kernel quality.‘Agostara’ is the earliest flowering and ri-pening cultivar in the area. It is conside-red in danger of extinction. Its fruit issmaller than that of ‘Bianca’. ‘Natalora’,the latest ripening cultivar among the lo-cal germplasm, is a vigorous and uprightcultivar and is considered the worst bythe local growers due to the high blankpercentage. This characteristic is to berelated to the flowering period not coinci-dent with the period of pollen dispersal bythe male trees of the area. Its fruit is big-ger than ‘Bianca’ and presents a moreirregular shape. Splitting percentage, inany case, is generally low.

As a whole, nut size of Sicilian Pistachiocultivars, including others such as ‘Silva-na’, ‘Ghiandalora’, ‘Cappuccia’, ‘Ceraso-la’, ‘Insolia’, is medium to small, nut sha-pe is generally elliptic (elongated) and so-metimes tends to ovoid. All of them havenuts with an attractive green kernel andsimilar sizes and weights. Splitting, al-

though variable from year to year, is ge-nerally low and, therefore, unsatisfactoryfor direct commercialization and con-sumption. Nevertheless, the deep greencolour of kernels and their excellent quali-ty are world-wide appreciated (Woodroof,1967). The overall characteristics of theItalian Pistachio germplasm are, therefo-re, more suitable for industrial transfor-mation (Barone and Caruso, 1996).

The average pistachio farm surface in thearea of Bronte is of approximately 1-2hectares, corresponding to a total ofabout 3000 farms. As a whole, 3000 to3800 tons of in shell pistachio is the pro-ductivity of the Bronte area, correspon-ding to an average yield of about 1-1.2tons per hectare. Exceptionally, maxi-mum yield of 4 tons per hectare has beenrecorded. The average prices paid togrowers for in shell product in the lastcommercial campaigns ranged between 4to 5 €/kg. During “off” years price tendsto be higher (aprox. 7.5 €/kg). Pistachiosare hand harvested with the help of tarps.Fruits are normally hulled with smallequipment (Picture 1), with an operatingcapacity of 400-500 kilos/day, and soldsoon after sun drying process (3-4 days)performed directly in the farm. They arebought by about ten local industries thatproceed to further transformation. Threekinds of end-products are therefore obtai-ned: in shell, shelled (yield 40-45%) andpeeled (Pictures 2-3). The prices of thefirst two categories of products are ofabout 7 and 15 €/kg, respectively. She-lled pistachios are normally commerciali-zed in packages of 25, 12, 1 and 1/2 kg todomestic and foreign markets. The avera-ge operating potential of this kind of pro-cessing plant is of about 500 kilos of she-lled pistachios per day. Some processingplants further process the fruit to obtainpistachio flour, pesto (condiment), creamand paste for pastry and ice-cream. Likein other producing countries, Italian pista-chio industry and market are negativelyaffected by alternate bearing problem. Aparticular cultural practice is generally fo-llowed by the growers to face the alterna-te bearing pistachio habit. It consists,every second year, of the total inflores-cence bud removal carried out during har-vest time, in the “on” year. This traditionalpractice is exerted with the aim of obtai-

ning a complete “off” year in order tomaximize the yield of the “on” year and tominimize the cultural expenses in thebiennium. By this way a biological controlof some main pests (bud borer Acrantus(=Chaetoptelius) vestitus, among theothers) is made by interrupting the naturalcycle of the insects. This practice requi-res about 30 hrs/ha and include also late-ral shoots suppression. Other culturalpractices carried out by the growers con-sist of winter pruning (February andMarch), fertilization (mostly with N and Pfertilizers), chemical weed control, or-chard floor management (with mechanicalhoe) and plant protection. Total culturalrequirements in terms of hand labour (in-cluding harvest) range between a mini-mum of 150 to a maximum of 400 hrs/ha.Among the diseases in the area Cytospo-ra terebinthi, Septoria pistaciae are gene-rally considered the most dangerous(Granata et al., 1993). Megastigmus pis-taciae, together with Acrantus (=Chaetop-telius) vestitus, can be considered themain pests (Greco, 1998). During storagePlodia interpunctella attacks to the fruitshave to be controlled generally by stora-ge room spraying.

PROSPECTSItaly imports annually about 9.000 tons ofroasted and salted (or to be salted inItaly) pistachios for direct consumption(snack) (Anonymous, 2000). About onethird of this import is from USA. For thesnack market, other well-known cultivarsfrom USA (‘Kerman’) and, lastly, alsofrom Australia (‘Sirora’), represent nowa-days the market standard. Italy maintainsa certain importance for Pistachio produc-tion utilized in the confectionery and ice-cream industry as well as for special des-tination such as for “mortadella” (balo-ney). The overall structure of Italian pista-chio industry in the years is relatively sta-ble and no particular changes are expec-ted in the near future. Quality of Sicilianpistachios is undoubtedly of high value,but other Mediterranean countries, withsimilar cultivars and lower productioncosts, are rapidly becoming potentialstrong competitors. Even if, under thepoint of view of environmental suitability,there will be no obstacle to a further diffu-sion of the culture in other areas of Italywithout the natural limits characteristic of

Picture 1. - Electrical hulling machine

Picture 2. - Cultivar ‘Bianca’ pistachio kernels Picture 3. - Cultivar ‘Bianca’ peeled pistachios


Picture 4. - Pistachio shelling machine

the Bronte area, very few new plantationshave been established in the recentyears. The major constraints are, per-haps, the long unproductive period of theculture (full production is normally rea-ched by the 12th or 13th year), togetherwith the lack of sufficient experimentationon new orchard design, alternative root-stocks, mechanization and availability ofless alternate bearing cultivars. The ques-tion that arises is also related to the aimof new plantations: whether for industrialtransformation, following the present spe-cialization, or for direct consumption, asin the rest of the emerging producercountries. In the first case we have gene-tic material, market opportunities andenough know-how to imagine an easy de-velopment. In the second case, other va-rieties should be tested for the Sicilianenvironmental conditions, together withthe best suited cultural techniques. Forthis last option, strong concurrence, es-pecially for the cost of production, has tobe faced. In both cases there is a strongneed for research. At the Dipartimento diColture Arboree of the Università di Pa-lermo researches on alternate bearingphenomenon, rootstocks and germplasmevaluation are underway, in some casesfrom the 80’s. Some results of these re-search activities have been recently pre-sented to the last GREMPA meeting heldin Mirandela, Portugal (Barone et al.,2003; Caruso et al., 2003).

Although currently the pistachio industrysituation, as mentioned before, could beconsidered stable, great opportunitiesare expected from the diffusion of mecha-nization, specially for harvest and othercultural practices. This is more likely tobe implemented in areas of new planta-tions where there are not the environ-mental constraints of the Bronte area.For this last area a strong impulse is ex-pected by obtaining the trade mark DOP(protected origin denomination) for thepistachios of Bronte, acknowledgementthat is underway. Finally, even if coope-ration has recently become stronger thanin the past, a further development of co-operation and, possibly, the establish-ment of an unique Pistachio associationamong the growers are highly advisable topromote the Italian pistachio industry, as al-ready done in other producing countries.

REFERENCES

Anonymous. 2000. Italian PistachioHarvest Nears Zero. Statpub. com.Agric. Commodity Market News.

Barone, E., T. Caruso, L. Di Marco.1985. Il pistacchio in Sicilia: superficicoltivate e aspetti agronomici. L’Infor-matore Agrario 40: 35-42.

Barone, E., L. Di Marco, F.P. Marra, M.Sidari. 1996. Isozymes and canonicaldiscriminant analysis to identify pista-chio (P. vera L.) germplasm. Hortscien-ce, 31 (1): 134-138.

Barone E., Caruso T. 1996. Genetic di-versity within Pistacia vera in Italy. Re-port of the workshop on: Taxonomy,Distribution, Conservation and Uses ofPistacia Genetic Resources. 29-30june, 1995, Palermo. IPGRI, Rome,Italy: 20-28.

Barone E., Padulosi S., Van Mele P.1997. Descriptors for Pistachio (Pista-cia vera L.). International Plant GeneticResources Institute, Rome, Italy.

Barone E., La Mantia M., Marra F.P.,Motisi A., Sottile F., 2003. Manipulationof the vegetative and reproductive cycleof Pistachio (Pistacia vera L.). Procee-dings XIII GREMPA Meeting on Al-monds and Pistachios, 1-5 giugno2003, Mirandela (Portugal). In press.

Caruso, T., L. Di Marco, D. Giovannini,A. Di Pisa. 1986. Caratteristiche carpo-logiche di 4 cultivar di pistacchio colti-vate in Sicilia. Frutticoltura 9-10: 63-66.

Caruso, T., L. Di Marco, A. Raimondo.1987. Caratteristiche carpometriche diquattro cultivar di pistacchio (Pistaciavera L.) individuate in Sicilia. Agricoltu-ra Ricerca 80: 65-68.

Caruso, T., L. Di Marco, R. Lo Bianco,F. Sottile. 1993. The pistachio germ-plasm of Sicily: pomological traits. Pro-ceedings of the IX G.R.E.M.P.A. Mee-ting - Pistachio. 1993. Bronte-Sciacca,20-21 May.

Caruso T., Barone E., Marra F.P., Sotti-le F., La Mantia M., De Pasquale C.2003. Effect of rootstock on growth,yield and fruit characteristics in cv.Bianca Pistachio (Pistacia vera L.)trees. Proceedings XIII GREMPA Mee-ting on Almonds and Pistachios, 1-5giugno 2003, Mirandela (Portugal). Inpress.

Fabbri A., Valenti C. 1997. The SicilianPistachio industry: an overview. ActaHort. 470: 43-49.

Granata G., Sidoti A., Corazza L. 1993.New acquisitions on control of Septoriapistaciae Desm. and Cytospora terebin-thi Bres. Proceedings of the IXG.R.E.M.P.A. Meeting - Pistachio.Bronte-Sciacca, 20-21 May. (Caruso T.,E. Barone and F. Sottile Eds.): 64-67.

Greco F. 1998. Le principali avversitàdel pistacchio nel territorio di Bronte.Amministrazione Comunale di Bronte.

Joley, L.E. 1969. Pistachio. In R.A. Jay-nes (ed.). Handbook of Northern Ameri-can nut trees: 352-355. The NorthernNut Growers Association.

Minà Palumbo, F. 1882. Monografia bo-tanica ed agraria sulla coltivazione deipistacchi in Sicilia. Lorsnaider GiovanniTipografo, Palermo.

Spina P. 1982 - Il pistacchio. Frutticoltu-ra Moderna vol. VI - Edagricole.

Woodroof, J.G. 1967. Pistachio nuts. InTree nuts: Production, Processing, Pro-ducts, 261-287. AVI Publishing, West-port, Conn.

E. Barone and F.P. MarraDipartimento di Colture Arboree -

Università di Palermo - 90128 ItalyE-mail: [email protected]

PISTACHIO PRODUCTIONIN TUNISIA

INTRODUCTIONPistachio (Pistacia vera L.) is a dioeciousspecies, widely present in the Mediterra-nean area. Its introduction in Tunisia mayhave been made by Carthaginians,however other introductions were madeby Romans and Arabs (Jacquy, 1973).Several species are scattered in differentparts of Tunisia (Jacquy, 1973; Nabli,1989): Pistacia terebinthus and Pistacialentiscus grown naturally in the dorsal hillfrom the north-west (Ghardimaou) to thenorth-east (Zaghouan). Pistacia atlanticaspreading from the dorsal hill on thenorth to the Saharian Atlas in the southwith important frequency in the predesertarea of the center and south of the coun-try (Sened, Meknassy, Guettar…). Pista-cia vera, the edible pistachio is foundalso from the north to the south withsome trees four to five hundred years old(Guettar oasis) and others two hundredyears old (Sfax – Mahres regions). Des-pite this long history, development of pis-tachio production in Tunisia starts onlyduring the last 30 years.

CURRENT SITUATIONPistachio is a hardy tree resistant todrought which can valorize poor soils,and therefore a lot of resources were putinto its development. Some developmentprojects were carried out with the supportfrom international organizations like FAOduring the period of the seventies. Fromless than 100 ha of pistachio orchard fiftyyears ago, the growing area is now morethan 43.000 ha localized mainly in the


center and the south of the country (Fig.1). Gafsa, Sidi Bouzid, Kasserine andSfax are the most important producing re-gions with respectively 19.800 ha, 9.188ha, 7.668 ha and 3.290 ha.

Despite this acreage increase the pro-duction is still very weak reaching only1.170 t with important differences betwe-en irrigated and rainfed orchard and alsoregions (table 1). The north region repre-sents less than 1% of the growing surfaceproducing more than 16% of the nationalyield (fig. 2).

To understand this weak yield and cha-racterize the current situation of pistachiocrop in Tunisia, a survey was carried outin the important Tunisian pistachio gro-wing zones (Ghrab et al., 2002b). Themost important observations were:• Inappropriate distribution of male andfemale trees on the orchard. Different ra-tios of male to female varying from 2% to90% were observed reducing the profita-bility of the orchard.• Inadequate planted material that is sup-posed to be ‘Mateur’ as female and malesA40 and A25 cultivars. However the ob-servations made revealed significant phe-notypic differences between trees.• A shift of the flowering periods betweenA25 and A40 male cultivars and ‘Mateur’inducing an inefficient pollination. Thosecultivars were selected on the north ofthe country for their production, qualityand overlapping flowering period howe-ver under the central and southern clima-tic conditions the flowering period of themale cultivars does not overlap with theblooming period of Mateur.• Since pistachio tree needs more thanfive years to bear under the central andsouthern semi-arid conditions, some gro-wers opted for other species and quittheir orchards without any management.• For some pistachio growers, manage-ment operations such as fertilization, pru-ning and pest and disease control were

ignored and considered without any be-nefit for this crop.

CULTIVARS AND ROOTSTOCKSThe most commonly cultivated variety inTunisia is ‘Mateur’ (Fig. 3), selected in theNorth as well as two corresponding polli-nators A40 and A25 (Mlika, 1980). Almostall the development of the pistachio in-dustry in Tunisia is based on this cultivar.Few other local varieties are cultivated insome specific localities from which it tookits names as ‘Meknassy’ cultivars in the re-gion of Meknassy (Sidi Bouzid), ‘El Guettar’from the region of El Guettar (Gafsa) and‘Nouri’ and ‘Thyna’ from Sfax region.

As rootstock, P. vera is almost the onlyused by growers for its good germinationrate. P. atlantica and P. terebinthus, inspite of their good characteristics asrootstock are barely used because oftheir seed germination difficulty.

CULTURAL CONSIDERATIONPropagationT-budding is the most common propaga-tion method. The budding is performed inJuly at the nursery on seedling of P. vera.The following winter trees are planted inthe orchard.

PruningThe training system used for pistachio isusually the open vase, however a lot oforchards were grown without any training.

The annual pruning is also rarely made. Ifit is made it is mainly to remove dried, di-seased, infected, pest damaged, andnon-fruiting old shoots.

PollinationPistachio trees are dioecious, with maleand female flowers borne on separatetrees. The designed orchard should ensu-re an adequate pollination. Male trees areoften planted in the center of a 3m x 3msquare of females, yielding an 8:1 ratio(Jacquy, 1973). Additional male trees aresometimes planted in border rows. Natu-rally, wind transports pollen from maleflowers to female flowers. Efficient polli-nation was assured when the floweringperiods of males is long enough to over-lap with the female flowering. But underTunisian conditions, a high percent ofblack fruit is obtained, and natural windpollination is proved inefficient and artifi-cial pollination was widely adopted(Mlika, 1974). The male panicle is har-vested and kept on a dry, shaded place.The pollen is collected and mixed withtalc (1/9 ratio) and sprayed on the femalepanicle early in the morning. Three to foursprays are made during the female flowe-ring period to insure a good pollinationrate.

Pest and diseasePest and disease decrease quality andquantity of pistachio crop through their di-rect and indirect damages. Few important

Figure 1. Geographical distribution of Tunisianpistachio growing area

Figure 2. Repartition of Pistachio growing area in Tunisia

North Centre

Growing Area

South

70

60

50

40

30

20

10

0

%

% of total production

% of total area

Table 1. Pistachio production in Tunisia (2002).

Surface Number Yield Yield Density(ha) of trees (t) (kg/tree) (trees/ha)

Irrigated crop 1.065 135.500 148,3 1,1 125Rainfed crop 41.983 2.888.618 1.029,5 0,4 70

(Source: DGPA, 2002)


pests and diseases are observed for theTunisian pistachio orchards. Eurytomaplotnikovi and Megastigmus pistaciae:the most pistachio fruit damaging wasps.They damage pistachio by feeding onnuts resulting in fruit blackness and drop.The most important disease is the leafspots, which caused a pre-matured leafing.

Fertilization and irrigationFew growers fertilize and irrigate their or-chards despite that the important growingareas are located on semi-arid zones withannual rainfall ranging from 150 mm to300 mm. This could explain the low yieldobtained. Some growers irrigate their or-chards only during the first years of plan-tation.

HarvestingIn Tunisia all orchards are hand harves-ted from late August to early October de-pending on the climate and the region.

CONCLUSIONIn Tunisia the culture of pistachio existssince antiquity. But, in spite of this, its de-velopment started only thirty years agowith large planting programmes. Howe-ver, its production is still low and rese-arch work should be done for pistachioproduction improvement. Based on thisstatement, different researches on pista-chio are going on in Tunisia such as:• Developing micropropagation techni-ques for Pistacia species for rootstocks

Figure 3. Tree and nuts of ‘Mateur’ cultivar,Mateur, Tunisia

use (Chelli Chaabouni and Gouta, 2002;Chelli Chaabouni and Drira, 2002).• Selection of male trees for Mateur culti-var under the central and southern condi-tions (Ghrab et al., 2002a).• Survey and characterization and con-servation of Tunisian pistachio germ-plasm.• Evaluation of introduced cultivars underTunisian conditions.

AKNOWLEDGMENTSI wish to thank Mrs. Cherif R., Rhouma A.and Triki M. A. for their great help to com-plete the pest and disease part.

REFERENCES

Chelli Chaabouni and Drira N., 2002.Culture Media and Hormone Effects onthe micro-propagation of two Pistachiospecies. III Inter. Symposium on Pista-chios and Almonds. Acta Hort., 591:395-398.

Chelli Chaabouni and Gouta H., 2002.Effect of chemical scarification and gi-berellic acid on in vitro germination ofPistacia atlantica seeds. III Inter. Sym-posium on Pistachios and Almonds.Acta Hort., 591: 73-76.

Ghrab M., Ben Mimoun M., Triki H. andGouta H., 2002a. Evaluation of the per-formances of seventeen male pistachio-tree specimens. III Inter. Symposium onPistachios and Almonds. Acta Hort.,591: 473-478.

Ghrab M., Gouta H., Triki H., Jabeur M.,and Laroussi K., 2002b. L’amandier etle pistachier en Tunisie centrale: étatactuel et perspectives d’amélioration.Document technique, Institut de l’Oli-vier, 1/2002, 16p.

Jacquy R., 1973. La culture de pista-chier en Tunisie. FAO, 97p.

Mlika M., 1974. Contribution à l’étudede la pollinisation chez le pistachier.DEA Faculté des Sciences de Tunisie.

Mlika M., 1980. Contribution à l’étudedu pistachier en Tunisie : choix des va-riétés mâles et femelles à floraison syn-chrone - Anatomie des fleurs. Mémoirede fin d’études de spécialisation INATunisie: 77 p.

Nabli M. A. 1989. Essai de synthèse surla végétation et la phyto-écologie tuni-siennes: I- Eléments de botanique et dephyto-écologie. Faculté des Sciencesde Tunis. 247 p.

Ghrab M.1, Ben Mimoun M.2 and Gouta H.3

1Station d’Arboriculture Fruitière, Institutde l’Olivier, Route Soukra km 1.5, 3003 Sfax,

Tunisie2Institut National Agronomique de Tunisia,43 av. Charles Nicolle, 1082 Tunis, Tunisia

3 Institut de l’Olivier, BP 1087, Sfax 3016,Tunisia


BREEDINGMONOECIOUS PISTACHIO

CULTIVARS

INTRODUCTIONAll Pistacia (Anacardiaceae) species aredioecious and wind is the pollinatingagent (Zohary, 1952). For commercialpistachio orchards, approximately onemale tree is necessary for 8 - 11 females(Maranto and Crane, 1982). As a result,approximately 10% of a typical pistachioorchard is not productive. In the literature,only three cases of exceptional sex typesin Pistacia were reported. Firstly, Özbekand Ayfer (1958) found two hermaphrodi-te trees in Gaziantep province of Turkey.They supposed that these trees were ei-ther seedlings of Pistacia vera L., or hy-brids between P. vera and P. terebinthus.The second report by Crane (1974) des-cribed three trees with exceptional phe-notypes: (I) a branch bearing staminateflowers on a P. atlantica female tree, (II) ahybrid between P. vera and P. atlanticabearing approximately equal numbers ofstaminate and pistillate inflorescencesthat are mostly on separate branches,and (III) a similar hybrid, predominantlystaminate, but several branches bear pis-tillate inflorescences. Recently, Kafkas etal. (2000) reported a few monoecious P.atlantica trees in the Yunt Mountains ofthe Manisa province of Turkey. The distri-bution of staminate and pistillate inflores-cences was found variable betweentrees. They found nine monoecious treeswith three different types: (Type I) allbranches of one of the trees were ‘fullymonoecious’ i.e., they bore a mixture ofmale and female inflorescences. (Type II)three trees had several branches withonly staminate flowers, while the rest ofthe branches bore pistillate inflorescen-ces. (Type III) Five other trees had inflo-rescences of both sexes on several bran-ches, and pistillate inflorescences on theremaining branches. The authors testedpollen germination of monoecious treesand used them for pollination with P.vera. The pollen germination rates weresimilar with normal male trees of P. atlan-tica and the hybridization with P. vera re-sulted normal fruit set and viable seedproduction.

By using the preliminary results on thesemonoecious trees, a project supported byThe Scientific and Technical ResearchCouncil of Turkey (TUBITAK) was startedin the spring of 2001 by planning of a lar-ge scale of crosses with the aim to deve-lop monoecious pistachio cultivars and tounderstand sex mechanism in Pistacia. Inthis paper, current status of the project isreported.


MATERIALS AND METHODSType I (all branches bore a mixture ofmale and female infloresences) and TypeII (a tree has several branches with onlystaminate flowers, while the rest of thebranches bore pistillate inflorescences)monoecious trees found by Kafkas et al(2000) are the main materials of this stu-dy. Figure 1 shows some pictures fromthe fully monoecious tree (Type I). Thesetrees were used both as female and maleparents in this study. Additionally, two fe-male P. vera cultivars (cvs ‘Siirt’ and‘Ohadi’) with a dioecious female P. atlan-tica tree as maternal tree, and dioeciousmale P. vera and P. atlantica trees wereused as pollinators. Totally, twenty diffe-rent crosses were performed betweenfive female and four male parents.

Before the stigma becomes receptive, fe-male inflorescences were closed with apaper bag (Figure 1D). To collect pollenfrom male parents for artificial pollination,staminate branches of four pollinatorswere cut and put in water-containing potsin the laboratory to perform pollen tests andcross-pollination. In the following two days,pollen was collected and stored at -200Cuntil pollination. Wheat flour was used as amixing agent with pollen at a ratio of 1:1.

The harvested nuts were stratified for 60days and then they were germinated inthe greenhouse. Germinated plantletswere transplanted into the five liter plasticbags. The plants were transplanted to theorchard in the autumn of 2003 and dripirrigation is to be set up in 2004.

Besides artificial pollination, the parentsused in this study were described mor-phologically. The progenies between P.atlantica genotypes and four pollinatorswere also characterized. The distributionof male/female branches/inflorescencesin the monoecious trees was observedyear to year by labeling them within thetree as well. When the progenies will re-ach the reproductive stage, they would betested for their sex type, and they will beused as breeding parents to try to obtainmonoecious cultivated pistachios withhigh quality nuts. The segregating popu-lations will also be used to understandsex mechanism in Pistacia and will beused for molecular studies to clonegene(s) that are responsible for sex de-termination.

RESULTS AND DISCUSSIONArtificial pollination was made duringthree years. In 2001, the crosses betwe-en P. atlantica and four pollinators, in2002 the crosses between P. vera andthe pollinators, and in 2003 additionalcrosses were performed to increase plantnumbers. The crosses between P. atlanti-ca and four pollinators were made in Ma-

nisa province, whereas the crossesbetween P. vera and the pollinators wereperformed in Gaziantep Pistachio Re-search Institute placed 1200 km far fromManisa province. The pollens were trans-ported to Gaziantep province in dry ice.Table 1 shows targeted and current plantnumbers in this project. We aimed to ob-tain 500-700 seedlings from the crossesbetween P. vera cvs and monoecious P.atlantica Type I, 100-200 seedlings from

the crosses between P. vera cvs and mo-noecious P. atlantica Type II, 50-100seedlings from rest of the crosses. Totallyit is targetted to have 2000-3400 se-edlings in the project. Currently, we com-pleted all the combinations and we havetotally 3054 seedlings in plastic bags.However, we also did additional crossesin (2003) to increase plant numbers espe-cially between P. vera and monoeciousgenotypes.

Table 1. Targeted and current plant numbers in the crosses.

No Female Male Targeted plant Current plantnumbers numbers

1 P. atlantica x *M-PA-Type I 50-100 1082 P. atlantica x M-PA-Type II 50-100 1083 P. atlantica x P. vera 50-100 1084 P. atlantica x P. atlantica 50-100 1085 M-PA-Type I x M-PA-Type I 50-100 1106 M-PA-Type I x M-PA-Type II 50-100 587 M-PA-Type I x P. vera 50-100 698 M-PA-Type I x P. atlantica 50-100 989 M-PA-Type II x M-PA-Type I 50-100 70

10 M-PA-Type II x M-PA-Type II 50-100 5211 M-PA-Type II x P. vera 50-100 5412 M-PA-Type II x P. atlantica 50-100 9713 P. vera cv. Ohadi x M-PA-Type I 500-700 63014 P. vera cv. Ohadi x M-PA-Type II 100-200 10815 P. vera cv. Ohadi x P. vera 50-100 5816 P. vera cv. Ohadi x P. atlantica 50-100 5617 P. vera cv. Siirt x M-PA-Type I 500-700 65818 P. vera cv. Siirt x M-PA-Type II 100-200 27319 P. vera cv. Siirt x P. vera 50-100 12020 P. vera cv. Siirt x P. atlantica 50-100 111

Total plant numbers 2000-3400 3054

M-PA: Monoecious P. atlantica

Figure 1. (A) The fully monoecious P. atlantica tree, (B) a view from branches of the fully monoecious tree(F: pistillate inflorescences, M: staminate inflorescences), (C) close up view of the staminate and pistillate

inflorescences in a branch, (D) P. vera cv Siirt cultivar tree with bagged branches


During the last three years, we have ob-served also hermaphrodite flowers insome inflorescences of the fully monoe-cious tree. But they were not stable, be-cause the numbers of anthers in a flowerchanged from 1 to 6. Figure 2 shows va-riations in the hermaphrodite flowers ofType I tree. In this tree, only several ofthe inflorescences were fully hermaphro-dite and a few of the others showed notonly hermaphrodite flowers but also hadstaminate and pistillate flowers as well. Insome inflorescences, one side showedstaminate flowers and the other side hadpistillate flowers, and hermaphrodite flo-wers were in the middle of them. Also se-veral stamens having stigma as in thepistils (Figure 2C) were found. In 2001,these inflorescences were baggedwithout pollination and obtained 6 nutsraising five P. atlantica seedlings. In2002, open pollinated nuts of these inflo-rescences were germinated yielding 50seedlings. In 2003, these inflorescenceswere bagged and pollinated with Type Ipollen and now we have 48 seeds.

A pistachio cultivar that has both stamina-te and pistillate or hermaphrodite inflores-cences in a single tree, together with highyield of quality nuts, would eliminate theneed for male trees in the orchard and in-creases the yield per hectare by about10%. Furthermore, protandry is a verycommon problem in pistachio orchards inTurkey and also in some of the other pis-tachio producer countries. Therefore, theimportance of a developing monoeciouspistachio cultivar in pistachio productionand cultivation is clear. In the first cycle,we are not expecting to have a monoecio-us genotype with high yield and qualitynuts because of the hybridization of P.vera with the wild species. But, havingsegregating monoecious populations inthe first cycle and isolating the related ge-nes and transferring them or performingsecond cycle crosses, there may arise acultivated monoecious pistachio cultivarwith high yield and quality.

Sex determination mechanisms in plants

are diverse, and may involve either sexchromosomes such as in Actinidia (Mc-neilage, 1991) or individual sex gene(s)as in Cucumis (Perl-Treves, 1999). InPistacia, the genetic mechanism of sexdetermination is still unknown. But, kno-wing the segregation of sex types in theprogenies of this project may explain sexinheritance and mechanism in Pistacia.

ACKNOWLEDGMENTSThe authors acknowledge the support ofS. Karaca during the cross pollination stu-dy in the Manisa province of Turkey. Thisproject is supported by The Scientific andTechnical Research Council of Turkey(TUBITAK).

REFERENCES

Crane, J.C. 1974. Hermaphroditism inPistacia. California Agriculture, 28: 3-4.

Kafkas, S., Perl-Treves, R. and Kaska,N., 2000. Unusual Pistacia atlanticaDesf. monoecious sex types in the YuntMountains of the Manisa province ofTurkey. Israel Journal of Plant Scien-ces, 48 (4): 277-280.

Maranto, J. and Crane, J.C. 1982. Pis-tachio production. Division of Agricultu-ral Sciences, Univ. of California, Leaflet2279, p.18.

Mcneilage, M.A., 1991. Gender Varia-tion in Actinidia deliciosa, the kiwifruit.Sex. Plant Reprod., 4: 267-273.

Ozbek, S. and Ayfer, M. 1958. A herma-phrodite Pistacia found in the vicinity ofAntep, Turkey. Proc. Am. Soc. Hort.Sci., 72: 240-241.

Perl-Treves, R., 1999. Male and femaleconversion along the cucumber shoot:approaches to studying sex genes andfloral development in Cucumis sativus.(C.C. Ainsworth editor). Sex Determina-tion in Plants. BIOS Scientific Publish-ers, pp. 189-216.

Zohary, M. 1952. A monographical stu-dy of the genus Pistacia. PalestineJournal of Botany, Jerusalem Series, 5:187-228.

S. Kafkas1, I. Acar2, H. Gozel2 and S. Eti11Department of Horticulture, Faculty of

Agriculture, University of Cukurova01330, Adana-Turkey.

2Pistachio Research Institute, GaziantepTurkey.


Figure 2. Variation of hermaphrodite flowers in the fully monoecious tree (St:stamen, P: pistil, Sg: stigma)(A) An hermaphrodite flower with five stamen and one very small pistil (B) A hermaphrodite flower withone stamen and one pistil, stigma with two parts (C) A hermaphrodite flower with one stamen and one

pistil but stamen has stigma as well

CHILLING STRESS AND INJURYIN PISTACHIO FEMALE

FLOWERS

Despite of pistachio resistance to diffe-rent well-known environmental stresses(drought, salt, unfavourable soils), thistree suffers from some serious stresseslike spring chilling. This injury can evennegatively affect crop yield. This stressspecially affects many temperate-zoneorchards of pistachio every year in Iran,causing significant yield losses and gene-ral tree weakness, and severing alternatebearing. Varying regarding on differenttemperatures; these damages occurredthrough decreasing fruit set rate as effec-tive pollination period (through stigmaand style deterioration, decreasing ovaryreception period and flower age, and in-creasing pollen tube growth period), fe-male flower abscission, inflorescence dieback, and flower buds’ death and abscis-sion could occur.

A research project has been directed tostudy the effects caused by this stress,strains, and the level of damages in diffe-rent temperatures. Here, some resultsare briefly presented.

The project involves simulated chillingstress in laboratory, simultaneous tooutdoor temperature decrease, resultingin remarkable effects on pistachio flowe-ring and pollination in early spring 2003,we could compare our lab results withfield observations and records.

In order to study the stress, shoots con-taining flower bud from cultivar “Ohadi” indifferent phenological stages (bud, andblooming and full-bloomed inflorescence)were collected and transferred to a con-trolled low temperature chamber. Tempe-rature was reduced to 2oC in one hour.Every hour, some bud and shoot sampleswere assessed looking for macro- and mi-croscopic visible injuries.

Chilling could adversely affect the repro-ductive structures in different thermal ba-sed and morpho-anatomical levels. Themost severe injuries (from lower to highertemperatures) were complete flower buds,current spring shoot (containing inflores-


cences), inflorescence, pistil, style, andstigma necrosis and deterioration, res-pectively.

Injury level analysisFlower buds: their chilling resistance isthe highest among all reproductive struc-tures and organs. Their resistance, howe-ver, decreases as they finish their dor-mancy period, especially before bloom.Light injury (low to about -4oC for about2h) could lead to their delayed bloom withsome blooming and flowering abnormali-ties. Deeper injury (about -4 to -6oC forabout 6-8h) resulted in the necrosis anddeterioration of very young pistils in clo-sed buds (Picture 1).Inflorescences: in the first visible injury le-vel, the tip vegetative growth (on the inflo-rescence shoot) became darker green incolor. In this stage, the fertilization rateand subsequent fruit set can decrease,possibly because of negative effect of thelow temperature on the pollen tube growthand shortening physiologically effectiveage of the pistil. With more temperaturereduction (low to about -2oC for about 2h),

Picture 1. Healthy control (left) and necrotic bud (right), Immediately before bloom

Y. GholipourIranian Pistachio Research institute (IPRI)

Qazvin Pistachio Research StationE-mail: [email protected]

stigma, and then style showed morpholo-gical necrosis and anatomical deteriora-tion, which resulted in pollen germinationand tube growth inhibition (Picture 2).Complete necrosis of flowers, and then,inflorescence, caused by about -4oC forabout 2h, was observed (Picture 3).

Comparing this simulated pattern of injuryin laboratory, with injuries records fromorchards, showed that we can predict anddetermine injury level and subsequentevents in fruiting and abscissions basedon field temperature data. For example,due to temperature drop, down to 2oC forabout 2 hours, it could be foreseen that adecrease in fruit set would happen as re-sult of delayed pollen tube growth,without visible symptoms. In some areas,lower temperatures caused visible inju-ries of stigma, resulting in problemmes infertilization. We are developing a modelbased on environmental temperature(with other climatic parameters) data andfemales’ injuries, to make our cultivationsready for next years potencial spring chil-ling stress. By the way, since the early

UNDERGROUND DRIP SYSTEM:THE NEW IRRIGATION

METHOD FOR PISTACHIOAND ALMOND ORCHARD

IN TURKEY

It is well known that pistachio trees (Pis-tacia vera L.) are drought resistant. InTurkey the crop was not irrigated until re-cently. During the last decade seminarsand meetings on Pistachio productionand management were given by Prof.Kaska and Ak to farmers. They both triedto convince farmers to irrigate pistachiotrees, when water supply is available.Farmers believe that pistachio treesshould not be irrigated, because theywould die. This belief is only true when awrong irrigation method is applied.

Commercial orchards are budded ontoPistacia vera rootstocks. This rootstock issusceptible to some diseases. We tellgrowers the usefulness of applying asound irrigation system. Some farmershave started to irrigate by tanker system.They have seen that irrigation is usefulfor pistachio trees. They recognized itseffects on reducing alternate bearing.

When irrigating pistachio trees somechanges could occur as below: Treegrowth will be higher, leaf size will be big-ger, the number of current year’s shootlength will be increased, yield will be in-creased, nut size will be higher, splittingpercentage will increase, blank nut ratewill be reduced, and less alternate bea-ring.

season cultural practices may affect phe-nology of female flowers, especially thetime of bud bloom, this model can also beuseful for reliable cultural programming.

Picture 2. Control (left) and injured necrotic stigma and style (right)

Picture 3. Complete flower necrosis (left) and current shoot and inflorescence die back (right)


Irrigation by underground drip systemsare being used on olive plantations inSpain. This system has been started tobe used for pistachio and almond or-chards in Turkey. However farmers arenot confident with this method. The ad-vantages of underground drip irrigationsystem are mentioned as follows: lowerwater consumption, better water distribu-tion, greater uniformity, waste water canbe used, less evaporation, greater trans-piration, location of fertilizer, less calcifi-cation, less diseases, possibility of tillage,

B. E. Ak standing close to a young almond tree

‘Siirt’ and ‘Kirmizi’ cultivars. The depth ofthe watering line is at approximately 20-25 cm deep, under the soil surface.

In Sanliurfa, a private investor, establish-ed a new almond orchard in 2002 byusing this underground drip irrigation sys-tem. There were planted 30 ha of almondtrees budded 6m x 6m onto bitter almondrootstocks. The cultivars were ‘Ferrag-nes’, ‘Ferraduel’, ‘Lauranne’, ‘Bertina’and ‘Felisia’. The new orchard for pista-chio cv ‘Siirt’ and old orchard cv. ‘Kirmizi’and ‘Ketengömlegi’ have been started toirrigate with an underground drip irriga-tion system.

As a result, irrigation is a big revolutionfor pistachio industry in Turkey. In the fu-ture the beneficial effects of irrigation willbe clear and all possible trees will be irri-gated. The researches on this subject arebeing developed at the Pistachio Rese-arch Station in Gaziantep, the Universityof Kahramanmaras and Pistachio Re-search and Application Center, Universityof Harran in Sanliurfa. In addition, almondorchards are expanding at South-easternpart of Turkey. The ecology is very suita-ble and the farmers prefer almond treesbecause of their many advantages.

B.E. AkHarran University, Faculty of Agriculture

Department of Horticulture63200- Sanliurfa (Turkey)

Tel: (414)247 26 97 Fax: (414) 247 44 80E-mail: [email protected]

Young almond orchards growing at Anatolia, Turkey

Tree planting in Turkey

Soil ready for planting

reduction of labour, longer lasting, novandalism.

All above advantages are valid for all kindof fruit trees. In fact, currently this systemis experimentally being used in pistachioorchards at Ceylanpinar State Farm(South Anatolia). The yield seems to be-come more regular, as there is strong al-ternate bearing, especially in the ‘Kirmizi’cultivar. This underground irrigation sys-tem was established in 200 areas wherethe adult trees are 30 years old, using

Underground irrigation system placed


IRANIAN AGRICULTURALSYSTEM TO DEVELOP HIGHER

QUALITY AND HEALTHIERPISTACHIO PRODUCTION

INTRODUCTIONIn recent years, improvement and optimi-zation of different stages of pistachio pro-duction, processing and handling, hasbeen important and has affected most re-search, extension, and governmental pro-grammes and activities. These significantefforts have been directed to increase thequality and health of pistachio product re-garding different aspects: harvest, post-harvest, processing, transportation andmarketing. This is based on internationalstandards, and taken place in order toprotect public health of domestic and fo-reign pistachio consumers. Here, we sum-marize Iranian programmes and activitiesto reach this aim.

GOVERNMENTALAND ADMINISTRATIVE WORKSThe government of Iran, as one of theeffective and active members of WHO, iscommitted to supply healthy and safeproducts to international markets, andtries to use modern technology in variousstages of production:• Organizing the pistachio committee inthe Ministry of Agriculture and by atten-ding the representatives from related mi-nistries and organizations (Ministry ofHealth and Medical Education, Customs,Ports and Shipping, Iranian center for de-velopment of export, Ministry of foreignaffairs, Iranian institute of standards, Pis-tachio research institute etc.)• Organizing the pistachio staff and afla-toxin committees• Preparation and distributing of manualsand technical publications plus 90.000sheets of posters, 85.000 sheets oftracts, more than 500 m wall writings, andseveral extensional films among the pis-tachio growers.• Holding training workshops in order toretraining horticultural exports and pro-moters.• Convening nationwide seminars on afla-toxin in pistachio• Convening executive and applied work-shops and seminars for growers to deve-lop modern and standard cultivation (e.g.pruning, methods of fertilization e.g. mi-croelement application and deep place-ment fertilizer), harvesting (to develop ontime harvest with proper equipment andproper transport), processing (speciallydrying process) etc.• Huge investment to develop establish-ment, mobilization and equipping currentprocessing terminals and 634 new linesof hygienic processing of pistachio.

• Organizing product forewarning net-works with cooperation of research cen-ters, Iranian Ministry of Health and Medi-cal Education and Academic departmentsof plant protection, in pistachio cultivatingprovinces, in order to study and announ-ce the most proper time of harvest. Thisaction has lead to significant decrease ofpistachio early split and aflatoxin con-tents.• Implementing the national project of“pistachio production structural improve-ment in country” within the last few yearsin ten main pistachio cultivation provin-ces. This project covers applying differenttechnical methods to decrease and remo-ve the risk of aflatoxicity of the crop.• Persuasion of pistachio growers tochange their traditional methods of allpistachio cultivation practices, specially adecrease of pesticides (which lead to re-duce to 37.5% per surface unit).• Establishing the nationwide union of pis-tachio production cooperative companiesin order to organizing and activating thecooperative companies in pistachio culti-vation areas.

RESEARCH ACTIVITIESIn recent years many projects have beendeveloped and directed to find and adaptapproaches to improve pistachio quality.A huge effort has been spent to carry outmany research and research-extensionalprojects and activities aiming to developmost proper ways of production and pro-cessing. Their results have been appliedto obtain higher quality product, and nowwe can see the results of all the referredactivities.

H. Hokmabadi 1, B. Gheibi 2, Y. Gholipour 11 Scientific board members, Iranian Pistachio

Research Institute,2 Pistachio Affairs Dept. Ministry of Jahad

Agriculture,E-mail: [email protected]

E-mail: [email protected]: [email protected]

Pistacia vera L. 1753 IN IRAN

INTRODUCTIONThe history of pistachio production andbusiness in Iran is long. The reason isthat the origin of pistachio trees was theIranian plateau and the first pistachiotrees were first grown by Iranians. Thewidest natural pistachio forests havebeen found in Iran. Expanding of pista-chio gardens and improved quality andquantity of Iranian pistachio productshave not acquired all at once. But it is theresult of nonstop efforts and creativity of

Iranian farmers and gardeners during thelast centuries.

The wild species of Pistacia vera is theorigin of all domesticated pistachio culti-vars in Iran. Since thousands of yearsago, the nuts or scions of this wild spe-cies have primarily transferred to Khora-san and then to other parts of Iran. Abris-hami informed that at Safaviehe times thepistachio nuts and scions were introdu-ced to Kerman province for the first time(Abrishami 1994). Today, Kerman andRafsanjan are the main pistachio produ-cing regions in Iran.

The study of the ecological distributionand genetic diversity of Iranian wild pista-chios might be a functional mean to se-lect convenient and resistant rootstocksfor Iranian domestic pistachios (Behboodi2003).

BOTANICAL CHARACTERISTICSOF P. veraThe genus Pistacia belongs to theAnacardiaceae family. There are two sy-nonyms for P. vera, which are P. reticula-ta Willd 1797 and P. trifolia L. 1753. P.vera is a diocious plant and the only dis-tinguishing trait between female and maletrees are their flowers at flowering time.

Trees 3-8 (-10) m high, cortex brownish,becoming gray. Leaves deciduous, coria-ceous, imparipinate. Leaflets 1-2 (3)pairs, sometimes with only one leaflet,glabrous or along the mid rid puberulent,upper surface smooth, lower surface opa-que, ovate or orbiculate, apex abruptlyacuminate, acute, rarely obtuse, 5-10(-12) cm x 3-6 (-8) cm; rachis terete, pu-berulent. Fruiting panicle upright straight.Drupe 10-20 cm x 6-12 mm, oblong-linearor ovate, apiculate (Rechinger 1969).

DISTRIBUTION OF P. VERA FORESTSNative P. vera forests are placed in north-eastern part of Iran, particularly in Sa-rakhs, Zoor-Abad, Jam and Tayebad.

The forest of P.vera in Sarakhs


These forests extend to Bokhara andSamarghand in Turkmenistan from thenorth, to Balkh, Thaleghan, and Harat inAfghanistan from the east. In Sarakhs re-gion, more intense forests are seen atTchahtchahe, Khabeh, and Ameneh Mo-rad-Tappeh. These jungles have sporadicsparse of trees with diffuse spread of pis-tachios. Male and female pistachios aremixed through these forests.

The extend of P. vera forests have beenestimated about 75.000 ha and the mainforest of Sarakhs is about 17.500 ha. Inthe last century, these forests have facedintensive human damages. The grassed

grounds of P. vera forests have encoun-tered overgrazing of livestock belongingto the different Iranian tribes who live inthis region and the wood of pistachiotrees has been used for their thermal re-quirements. For these reasons, the ex-tension of P. vera forests have drama-tically reduced and the distance betweenpistachio trees has enlarged.

P. vera is not found as forest in other re-gions of Iran. Thus it seems that thesetrees have been adapted to the ecologiccondition of this region. P. vera is the ori-gin of all cultivated Iranian domestic culti-vars. The Gazvini cultivars, regarding nut

appearance, are similar to wild P. vera, inwhich both of them have small nuts. Inaddition, the nuts of P. vera are similar tothose of small Badamy cultivars.

DISTRIBUTION OF P. veraACCORDING TO ECOLOGICALFACTORSP. vera species is extended from north-west to south-east of Sarakhs region at300-1500 m high. The average yearlyrainfall in these regions is about 200-300mm and the average daily temperature isbetween 10-20 ºC. Considering resistan-ce of P. vera trees regarding the changeof day and night temperatures, its lowwater requirement, and the absence ofGummous disease for wild P. vera, it ispossible to use it as resistant rootstockfor pistachio or to cross it for hybridizationto improve the quality of cultivated pista-chios in the future.

REFERENCES

Abrishami M., 1995. Persian Pistachio,a comparative history. University press,Tehran, Iran.

Behboodi B. Sh., 2003. Ecological dis-tribution study of wild pistachios for se-lection of rootstocks. XIII GREMPAMeeting on Pistachios and Almond, 1-5June, Mirandela, Portugal.

Rechinger K. H. (1969) Flora Des Ira-nischen Hochlandes Und Der Umara-menden Gebirge. No. 63. AkademischeDruck-u Verlagsanstalt, Graz (Austria).

Behrooz Sh. Behboodi Department of Biology, Faculty of Science,

University of Tehran, Tehran, Iran.E-mail: [email protected]

Leaves of P.vera Nuts of P.vera

Distribution of Pistacia vera in Iran

Pistacia atlanticaDESF. 1800 IN IRAN

INTRODUCTIONThree important Pistacia species P. vera,P. khinjuk and P. atlantica are found inIran. For this reason Iran is regarded aspistachio origin and rising. The synonymspecies of P. atlantica is P. mutica Fischand C.A. May 1838. This species hasthree subspecies in Iran, which are diffe-rent in their botanical appearance.

BOTANICAL CHARACTERISTICSOF P. atlantica

Trees up to 7 m high. Leaves deci-duous, imparipinnate, leaflets 2-4 (-5)pairs, form and size very variable, rotun-date-ovate, oblong, or lanceolate, (2-)


P. atlantica sub-sp. mutica trees Forest of P. atlantica sub-sp. cabulica

Leaves of P. atlantica sub-sp. cabulica

2.5-8 cm x 0.7-2 (-2.5) cm, obtuse orblunt at apex, never mucronate, at marginciliate, becoming glabrous, nerves in-conspicuous or slightly conspicuous. Ra-chis marginate or narrowly winged. Maleinflorescence paniculate, 2-8.5 cm x 6-9.5cm, different in subspecies. Floweringtime early spring. Fruit drupe, with aroma-tic pericarp, seed oily (Rechinger 1969).

WORLD GEOGRAPHIC DISTRIBUTIONOF P. atlanticaP. atlantica is located in the Atlantic re-gion, Turkey, Iran, Caucus, Afghanistan,Pakistan, Iraq, Syria, north Africa, and Al-geria. However its origin is Iran, but it hasexpanded from east to southwest of Euro-pe and from southeast to North of Africaand the Canary islands.

Distribution of P. atlantica sub-sp. mutica

P. atlantica subspecies in IranRechinger classified P. atlantica intothree subspecies: mutica, kurdica, cabuli-ca (Rechinger 1969). However non-Ira-nian botanists have not accepted thisclassification.

P. atlantica subspecies mutica (Fishand May) Rech. 1969Rachis winged up to terminal leaflets,leaflets narrowly oblong, 3-8 cm x 1.5-3cm, at margin ciliate. Drupe slightlybroader than long. This subspecies is lo-cated in Turkey, Iran, and the Caucus. InIran, it is found in northwest, northeast,center, east and south including Azerbai-jan, Esfahan, Kohkiloyeh Va Boyer-Ah-mad, Fars, Kerman, Khorasan, Yazd,Semnan and Tehran provinces.

P. atlantica sub-sp. kurdica trees

Leaves of P. atlantica sub-sp. mutica

Leaves of P. atlantica sub-sp. kurdica


P. atlantica subspecies kurdica (Zoha-ry) Rech. 1969Rachises winged up to terminal leaflets,leaflets ovate, 4-9 x 2-5 cm, glabrous orrarely ciliate at margin. Fruits depressed-globouse, 5-8 mm x 8-10 mm. This sub-species, apart from Iran, is distributed inIraq, Syria, and Turkey. In Iran it is nativein Azarbaygan, Kurdestan, Kermansha-han, Kohkiloyeh Va Boyer-Ahmad, andTehran provinces.

P. atlantica subspecies cabulica (Sto-cks) Rech. 1969Leaflets (2) 3-4 pairs, which are lanceola-te, attenuated at the ends, 3-4 times lon-ger than broad. Drupe globous, withequal long and broad. This subspecies isfound in Afghanistan, Pakistan, and Iran.Its distribution in Iran is mainly limited tosoutheast and central parts of the countrywhere Hormozgan, Kerman and Balot-chestan provinces are placed.

Ecological distribution of P. atlanticasubspeciesP. atlantica subspecies mutica is foundbetween 900-2800 m of hight. The meanyearly rainfall is between 200-400 mm in

these regions and the mean of yearly daytemperature is between 15-20 ºC.

P. atlantica subspecies kurdica is distri-buted in hights between 900-2800 mwhere the yearly rain mean is about 500-600 mm and the average of yearly daytemperature is flanked by 5-15 ºC. Thedistribution of P. atlantica subspecies ca-bulica is essentially restricted to Iransoutheast where the hight of lands altersfrom 50 m to 2500 m. The subspecies ca-bulica is able to grow in poor soils. Themean rainfall in its growing places isyearly lower than 100 mm and increasesto 200 mm as maximum. The averageday temperatures are 15-25 ºC yearly. Itis the most tolerant subspecies regardinglow water requirements and standing hightemperatures (Behboodi 2003).

CONCLUSIONSThe subspecies cabulica’s tolerance ishigher than in other P. atlantica subspe-cies regarding enduring of xerothermicand warm weather. Thus to improve do-mestic pistachios we suggest to usesubspecies cabulica rootstocks for bree-ding with cultivated rootstocks or to pro-

duce new rootstocks using P. vera x P.atlantica subspecies cabulica or P. khin-juk x P. atlantica subspecies cabulica hy-brids (Behboodi 2003).

REFERENCES

Behboodi B. Sh., 2003. Ecological dis-tribution study of wild pistachios for se-lection of rootstocks. CHEAM JournalOptions Mediterraneennes, in press.

Behboodi B. Sh., 2003. Ecological dis-tribution study of wild pistachios for se-lection of rootstocks. XIII GREMPAMeeting on Pistachios and Almond, 1-5June, Mirandela, Portugal, AbstractBook.

Rechinger K. H. (1969) Flora Des Ira-nischen Hochlandes Und Der Umara-menden Gebirge. No.63. AkademischeDruck-u Verlagsanstalt, Graz (Austria)

Behrooz Sh. BehboodiDepartment of Biology, Faculty of Science,


Distribution of P. atlantica sub-sp. kurdica Distribution of P. atlantica sub-sp. cabulica

Nuts of P. atlantica sub-sp. mutica Nuts of P. atlantica sub-sp. kurdica Nuts of P. atlantica sub-sp. cabulica


Pistacia khinjuk STOCKS 1852.IN IRAN

INTRODUCTIONThe scientific name of Pistacia khinjukderives from the Persian word “Khenjok”and the origin of this word is the Korasanprovince of Iran. This species is the sy-nonym of the species P. acuminata Boissand Bushe 1860. P. khinjuk is describedas “Lentisque de Bombay” in French lan-guage, as “Östindisher Mastix” in Ger-many, and in English as “East IndianMastiche”. The P. khinjuk trees are widelydistributed in several mountainous regi-ons of Iran. It is resistant against xero-thermic conditions and hot and cold wea-ther. These properties suit it for graftingof pistachio trees (Behboodi, 2003).

BOTANICAL CHARACTERISTICSOF P. khinjukTrees are dioecious, 3-7 m high, cortexsmooth, grey coloured. Leaves deciduous,imparipinnate, sometimes (var. populifoliaBoiss.) reduced only to one terminal le-aflet, pubescent, velvety, becoming gla-brous in age, petiole terete, leaflets 1-2(3) pairs, 3-10 cm x 1.5-7 (-8) cm, broadovate, oblique, narrowed suddenly into a± acuminate apex, on the upper surfaceshiny. Fruiting panicle straight. Drupe 4-6mm x 4-5 mm, globose, subcompressed,suboblique, apiculate (Rechinger 1969).

DISTRIBUTION OF P. khinjukFORESTSP. khinjuk is reported to spread in Iran,Turkey, Pakistan, Iraq, Syria, North Afri-ca, and Palestine.

DISTRIBUTION OF P. khinjuk IN IRANACCORDING TO ECOLOGICALFACTORSP. khinjuk is distributed in west, center,east, south, and east-south of Iran, inKurdestan, Lorestan, Eshfahan, Kohkilo-yeh Va Boyer-Ahmad, Farse, Hormoze-gan, Kerman, Balotchestan, Khorasan,Semnan and Tehran provinces. Only a

thin north band in Gillan and Mazandaranprovinces, on the coast of Caspian sea isblank of this species. P. khinjuk spread at700-2000 m heights on hills, flats and onmedium heights of Alborze and Zagrosemountains. This species withstands tomost trouble weather situations. The ave-rage rainfall of its growing areas altersfrom 100 mm to 600 mm and the meantemperature varied from 10 ºC to 25 ºC.

P. khinjuk is the most interesting pista-chio species in Iran as it grows in diffe-

P. khinjuk tree

Leaves of P. khinjuk The forest of P. khinjuk tree

rent regions and diverse climates. It is ex-clusively distributed as continuos popula-tions or accompanying with subspecies ofP. atlantica. Distribution of this species isnot affected by wide ranging changes inday and night temperatures, height, andlow rain. It is adapted to xerothermic wea-thers, soil sorts, and difficult environmen-tal situations (Behboodi 2003).

Using P. khinjuk rootstocks alone, or utili-zing its hybrids such as P. khinjuk x P. at-lantica subspecies mutica or P. khinjuk x


Distribution of P. khinjuk

Nuts of P. khinjuk

P. atlantica subspecies cabulica or P.khinjuk x P. vera may assist breeding pro-grammes to obtain resistant pistachiorootstocks.

REFERENCES

Behboodi B. Sh., 2003. Ecological dis-tribution study of wild pistachios for se-lection of rootstocks. CHEAM JournalOptions Mediterraneennes, in press.

Behboodi B. Sh., 2003. Ecological dis-tribution study of wild pistachios for se-

lection of rootstocks. XIII GREMPAMeeting on Pistachios and Almond, 1-5June, Mirandela, Portugal, Abstract Book.

Rechinger K. H. 1969, Flora Des Iranis-chen Hochlandes Und Der Umaramen-den Gebirge. No.63. AkademischeDruck-u Verlagsanstalt, Graz (Austria)

Behrooz Sh. BehboodiDepartment of Biology, Faculty of Science,


GERMPLASM COLLECTIONOF Pistacia AT THE JACOB

BLAUSTEIN INSTITUTEFOR DESERT RESEARCH.

PHENOTYPIC TRAITSAND MOLECULAR MARKERS

Natural habitats are shrinking in theworld, causing reduction in genetic varia-bility of plants with important economicalvalue. Shift from traditional agriculture ofPistacia vera L. (pistachio nut tree, Ana-cardiaceae) to modern agriculture, ‘dis-turb’ the genetic pool of the existing natu-ral population of this species by introduc-tion of selected cultivars. The lengthyprocess of cultivars selection leads onone hand to yield producing crops, but onthe other hand to a loss of importanttraits. Wild relatives of crop species arethus useful as potential genotypes inbreeding programmes. Gene banks andlive germplasm collections are importantin the conservation of the genetic variabi-lity, which still exists at a rather limitedscale in the wild.

At the Jacob Blaustein Institute for DesertResearch (BIDR), Ben-Gurion Universityof the Negev, Israel, a live germplasm co-llection of Pistacia species was establis-hed (Golan-Goldhirsh and Kostiukovsky,1998). The collection consists of ca. 600accessions of P. vera, P. atlantica, P.khinjuk, P. chinensis, P. lentiscus, P. te-rebinthus, and P. palaestina. All plantsgrowing in the field collection developedfrom seeds collected from natural popula-tions of Pistacia spp. around the Mediter-ranean basin, and Asia (Table 1). TheBIDR germplasm collection at its currentstatus serves as a small-scale gene bank,representing the genetic variability thatoccurs in nature. Thus, offering an oppor-tunity to study the genetic relations amongand within Pistacia species. Young trees (6to 8 years old) are growing under similargrowth conditions and therefore, any phe-notypic variability can be related to gene-tic difference (see Fig. 1). During the lastfew years we characterized the germ-plasm by assessing phenotypic traits, fol-lowing plant growth rate and related the-se to molecular markers.

Molecular marker techniques, amplifiedfragment length polymorphism (AFLP)and random amplified polymorphic DNA(RAPD) as well as morphological descrip-tors and plant growth parameters (follo-wing IPGRI descriptors for P. vera, 1997)were used to assess the relationshipsamong Pistacia species and amongaccessions of P. vera (Barazani et al.,2003; Golan-Goldhirsh et al., 2004).


Zohary (1952), in his famous monographon Pistacia spp. divided the eleven spe-cies of the genus into four different sec-tions (Fig. 2). The three evergreen spe-cies of the genus: P. lentiscus, P. wein-manifolia and P. saportae were includedin the section EU Lentiscus; the two Ame-rican species were clustered in the sec-tion Lenticella; in the section EU Terebin-thus he included the five species P. tere-binthus, P. palaestina, P. vera, P. khinjuk,and P. chinesis, while P. atlantica wasseparated into a single section Butmela(Zohary, 1952). Studies conducted atBIDR germplasm collection on the rela-tionship among Pistacia species usinggenetic marker techniques (Golan-Gold-hirsh et al., 2004) and morphological des-criptors, support Zohary’s division of thegenus (Fig. 2). However, while parsimonyanalysis of the morphological descriptorsclustered the six studied species accor-ding to Zohary’s division, molecular tech-niques analyses (AFLP and RAPD) inclu-ded P. atlantica, P. vera and P. khinjuk inthe same genetic subgroup. The geneticclustering indicated that the genetic dis-tance between P. atlantica and P. vera-P.khinjuk was smaller than that of the lattertwo species to P. terebinthus-P. palaesti-na (Fig. 2). We assume that the same

center of distribution of P. vera, P. khin-juk and P. atlantica in central Asia is thecause to the close genetic relationsamong these species. However, a muchdeeper study, such as alignment of geno-mic nucleotide sequences of conservedgenes, might assist in deciphering thephylogenetic tree of Pistacia sp.

The germplasm collection at BIDR inclu-des accessions of P. vera that were de-veloped from seeds collected from Bad-ghiz, Turkmenistan. There, natural standsof P. vera still exist in two populations,Kepele and Agachli, close to the Iranianand Afghan borders. Plant growth para-meters (plant height and trunk diameter)over two years of measurements at thecollection site indicated that accessionsfrom Agachli had higher growth rate thanthese of Kepele. Statistical analysisshowed that these differences were signi-ficant at 96% and 94% levels for plantheight and trunk diameter, respectively(Barazani et al., 2003). Agachli acces-sions can therefore be considered for fu-ture breeding programmes of P. vera.Moreover, significant statistical relationwas found between plant growth rate andthe genetic matrix as was assessed byRAPD (Barazani et al., 2003). Trees inthe two populations, Kepele and Agachli,although geographically close, grow indifferent habitats, which probably led tothe genotypical differences.

The pistachio tree is an important nutcrop in many countries. However, P. at-lantica, P. khinjuk, P. palaestina and P.terebinthus are used by local farmers inthe Middle East as rootstocks for graftingof P. vera. Therefore, wild relatives ofhorticultural cultivars may be useful stockmaterial for breeding programmes (Zoha-ry, 1995), pointing to the significance ofpreservation of the genetic variability.

Increasing concern on loss of genetictraits has led, over the last decades, toconservation plans and more effort togene banks management, as well as es-tablishment of germplasm collections ofwild relatives of economically importantcrops. We destined the Pistacia spp. live

germplasm for this purpose and for selec-tion of suitable genotypes for future bree-ding programmes.

ACKNOWLEDGMENTS

This research was supported in part un-der Grant No. TA-MOU-98-CA17-028 fun-ded by the U.S. – Israel Cooperative De-velopment Research Program, Bureau forEconomic Growth, Agriculture, and Tra-de, U.S. Agency for International Deve-lopment.

REFERENCES

Barazani, O., Atayev A., Yakubov B.,Kostiukovsky, V, Popov, K., Golan-Gold-hirsh, A. 2003. Genetic variability inTurkmen populations of Pistacia vera L.Genet. Resour. Crop Evol. 50: 383-389.

Golan-Goldhirsh, A., Kostiukovsky, V.1998. Mediterranean Pistacia genusgermplasm collection at Sede Boker Is-rael. Acta Hort. 470: 131-137.

Golan-Goldhirsh, A., Barazani, O.,Wang, Z.S, Khadka, D.K., Saunders,J.A., Kostiukovsky, V., Rowland, L.J.2004. Genetic relationships among Me-diterranean Pistacia species evaluatedBy RAPD and AFLP markers. PlantSyst. Evol. (in press).

IPGRI. 1997. Descriptors for Pistachio(Pistacia vera L.). International PlantGenetic Resources Institute, Rome,Italy.

Zohary D. 1995. The genus Pistacia L.In : Padulosi S., Caruso T., Barone E.(eds.) Taxonomy, distribution, conser-vation and uses of Pistacia genetic re-sources. IPGRI, Italy, pp. 1-11.

Zohary M. 1952. A monographical studyof the genus Pistacia. Palestine J. Bot.(Jerusalem Series) 5: 187-228.

O. Barazani and A. Golan-GoldhirshBen-Gurion University of The Negev, The

Jacob Blaustein Institute for Desert Research,Albert Katz Department of Dryland Biotechno-logies, Desert Plant Biotechnology Laboratory,

Sede Boqer Campus 84990, IsraelE-mail: [email protected]

Fig. 2. Genetic markers and morphological descriptors clustering of six Pistacia species (bottom) incomparison to the classical division of the genus as was proposed in 1952 by Zohary (top)

Table 1. Pistacia species growing inthe germplasm collection at BIDR and

their geographical origin.

Pistacia species Origin

P. atlantica CyprusIsraelSpainSyria

P. chinensis USA

P. khinjuk Syria

P. lentiscus CyprusIsraelTunisiaSpain

P. palaestina IsraelSyria

P. terebinthus Syria

P. vera TurkmenistanSyria

Fig.1. View of accessions of Pistacia atlantica(front) and Pistacia lentiscus (back) growing in

the germplasm collection at BIDR


WHERE IN A PISTACHIO TREEIS Xanthomonas AND HOW DID

IT GET THERE?

BACKGROUNDXanthomonas translucens has been iden-tified as the probable causal agent of die-back of pistachio (Pistacia vera) in Aus-tralia. The bacteria have been regularlyisolated from twigs with internal stainingfrom diseased trees. The location of Xan-thomonas in other parts of the tree andthe identification of possible entry pointsare the focus of research in progress atthe Sunraysia Horticultural Centre (Vic)and the University of Adelaide (SA).

INVESTIGATIONIn spring 2001 we performed various sur-veys to determine the tissues in whichbacteria are most likely to be found. Hun-dreds of inflorescences and individualmale and female flowers, leaves, vegeta-tive buds and current season growth wereexamined for the presence of the bacte-ria. Bacteria were present in male inflo-rescences, leaves and current seasongrowth from highly diseased trees.We also felled diseased male and femaletrees and healthy male trees at Kyalite(NSW) to locate the bacteria, through la-boratory isolations, in different parts ofthe wood. Samples of wood from therootstocks up to one-year old twigs, andtissues from young and old sapwood andheartwood were also examined.

X. translucens was located mainly in theyoung, unstained or stained sapwood ofthe main trunk (Figure 1), primary bran-ches and younger branches. The bacteriawere rarely found in discoloured heart-wood (dark heartwood is normal in matu-re trees) (Figure 1), and incidence in lea-ves was very low. Root and soil samplesdid not yield Xanthomonas. Bacteria orpathogenic fungi were not found in thebark and cortex associated with lesionswhereas the stained young xylem benea-th the lesions yielded Xanthomonas (Fi-gure 2). Scanning electron microscopy isbeing used to confirm these results.Experiments were initiated in the glass-house or shade house to determine if pru-ning wounds and/or scars are possibleentry points for the bacteria. We inocula-ted pruned branches (Figure 3) and leafscars (Figure 4) on young trees with sus-pensions of bacteria. Also, attempts aremade to introduce the bacteria into thetrees by injection (Figure 5) to simulateentry via an insect vector or wound, andto produce symptoms similar to those ob-served in the field. Results from these ex-periments will also indicate how fast thebacteria move in the trees.In the coming months, roots and flowerswill be inoculated with the Xanthomonasbacteria to determine if those are alsopossible entry points.

SUMMARYWe have determined where Xanthomo-nas translucens is located in the trees.Results from the ongoing experiments willclarify how the bacteria got into the trees.

Figure 1. Section of main trunk of a diseasedtree. Heartwood and young sapwood indicated

with arrows

Figure 2. Lesion from a branch of a diseasedtree. Unstained and stained sapwood indicated

with arrows

Figure 3. Pruned branch of a young potted treebeing inoculated with a drop of bacterial

suspension

Figure 4. Leaf scar of a young potted tree beinginoculated with bacterial suspension

Figure 5. Young potted tree being inoculated withan injection of bacterial suspension

More research is needed to determine theways of dispersion of the bacteria in theorchard and to elucidate the cycle of thedisease.Other experiments will be set up to studypossible control measures.

E. Facelli1, C. Taylor2, E. Scott1, R. Emmett2

and M. Sedgley1

1School of Agriculture and Wine, Universityof Adelaide, Waite Campus, South Australia,Australia. 2 Department of Primary Industries,

Mildura, Victoria Australia.E-mail: [email protected]

CHESTNUT PRODUCTIONIN ANDALUSIA

INTRODUCTIONLast data about Spanish chestnut produc-tion estimates 18.259 t in Spain, half ofthe amount achieved in the seventies.That production corresponds to approxi-mately 50.000 ha out of the 128.537 haexisting in Spain. Though chestnut pro-duction is located mainly in North Spain,an interesting area for chestnut produc-tion is localised in Andalusia, South Spain(Pereira et al., 2001). Production in Anda-lusia is considered to be only 2.000 t butit could be underestimated according tomarketing data and it could reach 6.000 tin 6.000 ha. There are two main areas: i)Sierra de Huelva in Huelva province (Fi-gure 1), and ii) Serranía de Ronda (Figu-re 2) and Sierra de Las Nieves, Málagaprovince. Other chestnut areas are loca-ted in Granada, where new orchards are


being established (Figure 3) though tradi-tionally chestnut production came fromseedlings. Chestnut is an interesting al-ternative in Andalusia, mainly in Málaga,because of the precocity of two cultivars,‘Temprana’ and ‘Pilonga’, harvested atthe end of September. Most of the pro-duction is commercialised by cooperati-ves where big quantities of chestnuts are

cleaned and classified. Chestnuts aresold in Spain and in the international mar-ket. Recently, an important industry fortransforming chestnuts has been establi-shed in Huelva. Both in Huelva and Mála-ga, production is based on local cultivars.Growers graft them over seedlings produ-ced in the same area. In Málaga, due tothe high prices of the nuts, trees are plan-

ted so close as 5 m x 5 m, though it is ne-cessary to prune severely or remove partof the trees later. In Granada, no localcultivars have been found and new or-chards are based on the excellent culti-vars of Málaga (Figure 3). Weather condi-tions for chestnut areas in Spain are quitesimilar (Table 1), with a rainfall around1.000 mm per year and mild temperatu-

Table 1. Climatic characteristics of chestnut areas in Asturias (www.inm.es), El Bierzo (www.inm.es), Extremadura (www.inm.es),Tenerife (www.inm.es), Andalusia (www.inm.es) and Galicia (Carballeira, 1983).

Region Province Location Annual average Annual rainfall Average temperature Annual temperaturetemperature (°C) (mm) in January (°C) range (°C)

Asturias Asturias Gijón 13,8 971 4,7 —Oviedo-El Cristo 12,9 973 4,2 —Aeropuerto Ranón 13,2 1140 5,4 —

Bierzo León Ponferrada 13,0 675 1,1 17,5Extremadura Cáceres Guadalupe 14,7 755 3,3 17,6Canarias Tenerife Aeropuerto Tenerife

Norte-Los Rodeos 16,5 557 12,8 —Andalucía Málaga Gaucín 15,0 1161 6,2 35,0

Huelva Almonaster La Real 16,0 1137 4,9 34,9Aracena 14,0 1107 3,1 35,8

Galicia Lugo Outeiro de Rei 12,0 1004 2,0 23,3Vilar do Caurel 7,9 1679 -1,3 24,5Ferreira 3,2 1086 2,4 25,7Fonsagrada 8,8 1754 -1,0 23,3

Ourense Mesón de Pentes 10,6 1460 0,8 26,0Poboa de Trives 8,9 874 0,1 26,6A Rúa-Petín 15,6 763 5,1 26,7

Table 2. Main characteristics of the cultivars of Andalusia (adapted from Pereira and Ramos, 2003).

Cultivar Origin Harvesting time Use Male Nuts/kg Lateral Poly-embrionic Splitcatkinz nut (g) nuts (%) nuts (%)

Capilla Málaga 20 - 26 Oct Industry, fresh, pollinizer L 66 17,00 11,00 16,00Comisaria Huelva 20 Oct - 2 Nov Industry, fresh, pollinizer L 80 13,37 6,00 0,50Corriente Málaga 18 Sep - 4 Oct Fresh, pollinizer L 65 18,58 15,50 4,50Dieguina Huelva 20 Oct - 1 Nov Marrón al natural, fresh L and M 87 13,61 8,00 2,50Helechal Huelva 22 - 29 Oct Industry, fresh B and L 80 14,14 7,00 1,50Pelón Huelva 19 Oct - 1 Nov Marmalades and purees, L 113 10,22 3,54 7,81

pollinizerPilonga Málaga 17 Sep - 4 Oct Industry, fresh, pollinizer L 63 18,32 7,61 8,67Planta Alájar Huelva 20 - 29 Oct Fresh M 71 16,06 10,50 2,25Rubia Tardía Málaga 25 Oct Industry, fresh, pollinizer L 49 20,76 10,00 2,50Temprana Málaga 15 Sep - 4 Oct Fresh, pollinizer L 61 18,76 11,67 8,53Tomasa Málaga 20 - 29 Oct Fresh, pollinizer L 44 24,86 14,38 23,13Vázquez Huelva 20 Oct - 5 Nov Fresh, industry, pollinizer L 73 16,05 4,00 1,50

zL = longuistaminate; M = mesostaminate; B = braquistaminate

Figure 2. New orchard with irrigation in Güimar-Sierra, Granada

Figure 1. High density spacing in new orchardsof Serranía de Ronda, Málaga

Figure 3. Old chestnut orchard in Sierra deHuelva, Huelva


res. Chestnut orchards in Tenerife sho-wed lowest rainfall with 700 mm, followedby Castilla-León and Extremadura, around700 mm. In Andalusia, orchards are loca-ted in the most humid areas of SouthSpain.

In 1995 a study was started to localiseand characterise cultivars of South Spain.An inventory was published in 2001 (Pe-reira et al., 2001) and the results foundwere published in a guideline book (Pe-reira and Ramos, 2003) and in a workwhere cultivars from Andalusia were com-pared with previous studies (Ramos andPereira, 2004). The paper is a summary ofthe main traits of the Andalusian cultivars.

CULTIVARSUp to now 12 cultivars have been studiedin Andalusia which are used locally fornut production and propagated by graf-ting (Figures 4 to 15): ‘Capilla’, ‘Comisaria’,‘Corriente’, ‘Dieguina’, ‘Helechal’, ‘Pelón’,‘Pilonga’, ‘Planta Alájar’ ‘Rubia Tardía’,‘Temprana’, ‘Tomasa’ and ‘Vázquez’.

Cultivars from Andalusia are differentfrom cultivars from other areas of Spain(Ramos and Pereira, 2004). In compari-son with those from Galicia (Pereira andFernández, 1997) with better characteris-tics in nut size, but with higher percenta-ge of poly-embryonic and split nuts, andmore intrusions of the inner coat in co-

tyledons what makes it more difficult topeel them. Nuts coming from Andalusiancultivars showed less worms and disea-ses what could be related with a drier en-vironment.

The best cultivar in Andalusia is ‘Pilonga’(Figure 10) due to its early ripening (endof September), big nut size and less per-centage of poly-embryonic nuts (Table 2).The worst feature of ‘Pilonga’ is the highpercentage of split nuts, over 8 %. ‘Tem-prana’ (Figure 13), cultivated in the samearea of ‘Pilonga’ is a little worse due to ahigher percentage of poly-embryonic nutsand intrusions of the inner coat in cotyle-dons. ‘Capilla’, ‘Corriente’, ‘Rubia Tardía’

Figure 4.- Capilla Figure 5.- Comisaria Figure 6.- Corriente

Figure 7.- Dieguina Figure 8.- Helechal Figure 9.- Pelón

Figure 10.- Pilonga Figure 11.- Planta Alájar Figure 12.- Rubia Tardía

Figure 13.- Temprana Figure 14.- Tomasa Figure 15.- Vázquez


and ‘Tomasa’, (Figures 4, 6, 12 and 14)are three excellent cultivars of Málaga,but they are harvested later than ‘Pilonga’and ‘Temprana’, at the end of October,when most of the Spanish chestnut pro-duction is concentrated and prices areusually lower. Moreover, ‘Capilla’ and‘Tomasa’ showed the highest percenta-ges of split nuts and ‘Corriente’ the high-est percentage of poly-embryonic nuts. InHuelva four main cultivars should be pro-moted, ‘Comisaria’, ‘Helechal’, ‘PlantaAlájar’ and ‘Vázquez’ (Figures 5, 8, 11and 15) due to their big nut size and lowpercentage of split nuts. ‘Planta Alájar’showed a higher percentage of poly-em-bryonic nuts. In comparison with cultivarsof Málaga, all cultivars in Huelva are har-vested at the end of October, main periodfor chestnut harvesting in Spain, whenprices are lower. Most of the cultivarsfrom Andalusia produce pollen, since onlyone accession studied of ‘Helechal’ wassterile. The most frequent type of malecatkin found was longuistaminate andonly ‘Planta Alájar’ and one accession of‘Dieguina’ showed mesostaminate cat-kins. ‘Pelón’ (Figure 9) is a very spreadcultivar in Huelva that produces the smal-lest nuts in Andalusia what makes it lessinteresting for present market. These cul-tivars are being collected and introducedin the Universidad de Santiago de Com-postela to establish a core collection.

REFERENCES

Carballeira, A., 1983. Bioclimatologíade Galicia. Fundación Pedro Barrié dela Maza. ISBN: 84-85728-27.

Pereira, S.; Fernandez, J. 1997. Loscultivares autóctonos de castaño (Cas-tanea sativa Mill.) en Galicia. Monogra-fías I.N.I.A., 99: 1-533. ISBN: 84-7498-461-0

Pereira, S.; B. Diaz, B.; Ciordia, M.; As-casibar, J; Ramos, A.M.; Sau, F. 2001.Spanish chestnut cultivars. HortScien-ce, 36 (2): 344-347.

Pereira, S.; Ramos, A.M. 2003. Carac-terísticas morfológicas e isoenzimáticasde los cultivares de castaño (Castaneasativa Mill.) de Andalucía. Monografíasdel INIA, 13, 160 pages.

Ramos, A.M.; Pereira, S. 2004. Geneticrelationship between Castanea sativaMill. cultivars from North-western toSouth Spain based on morphologicaltraits and isoenzymes. Genetic Resour-ces and Crop Evolution (in press).

S. Pereira, A.M. RamosUniversidad de Santiago de Compostela,

Escola Politécnica Superior, Departament ofProducción Vegetal, Campus de Lugo, 27002

Lugo, SpainE-mail: [email protected]

THE WORLD MACADAMIAINDUSTRY AND RESEARCH

ACTIVITIES

INTRODUCTIONMacadamia is one of the world’s finestnuts. The kernel is consumed as salted orunsalted snack food, as chocolate enro-bed confectionary, and as an ingredientin products such as biscuits or ice-cream.The kernel has a high oil content (70-80%) and most of the oil is mono-unsatu-rated lipids with 65.0 and 18.5% oleic andpalmitoleic acid respectively (Cavaletto etal., 1966). Oil content has been shown tobe an important component of quality;kernels with a specific gravity of 1.0 (oilcontent of more than 72%) or less wereidentified as having superior eating androasting qualities and are classed as firstgrade kernels or the commercial stan-dard. Total sugar content of raw M. inte-grifolia kernel ranges from between 3 to5% (Riperton et al. 1938), the majority ofwhich is sucrose.

The Macadamia genus is a member ofthe Proteaceace, which is a characteristicGondwana family (Johnson and Briggs,1983). Of the nine species, seven are en-demic to Australia, with the other two oc-curring on the islands of Indonesia. Onlytwo of these species, M. integrifolia andM. tetraphylla produce edible kernel. The-se species occur as a medium sizedevergreen tree in the lowland subtropicalrainforests of eastern Australia. The fruitconsists of a fleshy husk (pericarp) sur-rounding the hard shell (testa) and theedible kernel (embryo and cotyledons)(Gross 1995).

WORLD PRODUCTIONMacadamias represent less than 2% ofthe world tree nut market. The market ischaracterised by undersupply and pricefluctuations (Hargreaves 2003).

AustraliaAustralia is the main macadamia produ-cer with approximately 40% of the worldproduction or 10.000 t of kernel (Table 1).Despite recent surveys indicating thatabout 1/3 of the industry is new plantings,

industry growth that was forecast at 15%yearly has not been reached (Hargreaves2003) due to poor seasons in 2002 and2003. The major growing regions arearound Alstonville (28o 51’S, 153o21’E) inNorthern NSW and Bundaberg (24o 33’ S,152o00’ E) in South Eastern Queenslandwith approximately 45 and 35 % of theAustralian crop produced in these areasrespectively. There is renewed interest toidentify new growing areas in Australia formacadamia production. The major mar-kets for Australian production are USA,Japan, Germany and China.

USAMacadamias were first commercialized inHawaii and most of the USA productioncontinues to originate from these islands.The Hawaiian industry is characterised bymature orchards with new plantings atmuch smaller levels than for the otherproducing countries (Vidgen 2003). Do-mestic sales, including duty-free, are amajor market of USA production. Exportsare shipped predominately to Japan.

South AfricaOver 45% of the trees in South Africa areless than 5 years of age and there is thepotential for significant growth of this in-dustry (Lee 2003). Production of kernelhas grown by about 30% between 2000and 2003 to approximately 3000 t despitepoor production seasons in 2002 and2003. Half its production is exported toUSA, with Europe being the next majormarket (25%).

OthersMacadamias are also produced in Kenya(1500 t), Malawi (1000 t), Brazil (500 t),New Zealand (35 t), Thailand and CostaRica.

HORTICULTUREPropagationMacadamias are generally clonally propa-gated. Seedling rootstocks are commonin Australia using open pollinated seedfrom the cultivar H2. Seedlings from othercultivars have also been used to a smal-ler extent. Practical experience suggeststhat the roots system is not well deve-loped in cuttings of some varieties in Aus-tralia. There is little information to guide

Table 1. Macadamia worldwide kernel production (t) (www.inc.treenuts.org)

Region 2002 Import Production Domestic Export 2003Carry-over Carry-over

Australia 800 0 9.900 2.000 7.900 800Africa 43 0 6.440 190 6.250 30USA 2.000 7.466 5.500 12.216 770 2.000Latin America 170 0 3.370 300 3.070 170

Total 3.013 7.466 25.210 14.706 17.990 3.000


rootstock choice, although recently an ex-tensive rootstock trial of own rooted cut-ting, seedling and clonal rootstocks using12 cultivars was established in Australia(Hardner and McConchie, 2003). Suitablescion material is selected from the desi-red cultivar and girdled to increase itscarbohydrate content. Scions are then ei-ther grafted or budded onto the rootstock.Initial results show that budding successrate is affected by the rootstock and scioncombination, which is consistent withanecdotal evidence from macadamia pro-pagators.

Orchard designOrchards are established at varying den-sities from 10mx10m (100 trees per ha) to7mx4m (357 trees/ha). Tree size variesamong cultivars (Hardner et al. 2002) andmay influence planting density. There hasbeen trend in recent years to establish or-chards at higher densities to offset highinitial capital investments at an earlierage. In addition, there has been somepractical experimentation with high densi-ty planting (5mx2m) (Bell and Bell, 2001)that produced high yields of between 8and 10 t/ha before canopy management,but there is little knowledge of the laterage performance of trees to evaluate thissystem.

IrrigationOrchards located in Queensland are ge-nerally irrigated, especially in the Bunda-berg region where there is a significantlylower annual rainfall than in NorthernNSW where the majority of orchards arenot irrigated. Average annual rainfall forAlstonville in Northern NSW and Bunda-berg is 1600 mm and 1140mm respecti-vely. Stephenson et al. (2003) reportedthat water stress at different phenologicalstages including floral development, im-mature nut drop and nut maturation hadadverse effect on yield through decreasein nut number and nut size. Adverseeffects of irrigation in Northern NSW havealso been observed. Trochoulias andJohns (1992) found that NIS yield per unittrunk area decreased with irrigation by upto 10% at excessive rates of irrigation.Variable yield, water use and quality res-ponses between cultivars to irrigationhave been reported for macadamiasgrown in Bundaberg (Searle and Lu,2003).

NutritionAn extensive account of macadamia nu-trition was undertaken as part of a reviewof macadamia by Nagao and Hirae(1992). Orchards in Australia have beenestablished on a range of soil types thathave differing chemical and physical pro-perties, Stephenson et al. (1986) obser-ved the soil nutrient levels and fertiliserapplication on three common soil types

used to grow macadamias; red podsolic,krasnozem and sandy red earth, andfound varied nutrient levels and fertiliserapplications. Macronutrients are general-ly applied as granular fertilizers althoughin regions where irrigation is used fertiga-tion may also be used. Micronutrients areoften recommended as foliar sprays forrapid amendments of nutrient deficien-cies. The methods of determining nutrientrequirements throughout the industryvary; however they generally involve oneor a combination of soil nutrient analysis,leaf nutrient analysis, crop removal repla-cement or crop logging. Foliar nutrientcontent varies with leaf age, leaf position,time of year and recent work by Huett etal. (2001) showed that content of severalleaf parameters including nitrogen andphosphorus increased with decreasedshading and concluded that the currentsampling protocols for determination ofleaf nitrogen and phosphorus levels werenot reliable.

Flowering and fruit developmentInitial flowering generally occurs betweenthe 3rd and 5th year after planting. Flowersare hermaphroditic and borne on race-mes that contain between 100 to 300 flo-wers each (Figure 1), between 6 to 35%of these flowers develop into immaturefruit (Urata 1954) and only 0.3% will re-main at maturity (Ito 1980). It is traditio-nally accepted that flowers are located onwood that is at least 2 to 3 years of ageand there appears to be variation betwe-en cultivars. The major pollinator appearsto be the European honeybee, althoughthe Australian native species of Trigona

has been observed. Flowering in Austra-lian orchards occurs in September, thereare significant differences in timing of flo-wering between cultivars although thereis still high potential for cross-pollinationbetween cultivars (Boyton and Hardner,2002).

Abscission of immature fruit occursthroughout fruit development, howeverthe maximum rates are observed from 3to 8 weeks after anthesis (Sakai and Na-gao 1984). Increases in fruit diameter andfruit fresh weight follow a single sigmoidalpattern in which growth continues until 14to 15 weeks after anthesis and 15 to 16weeks after anthesis for fruit diameterand fresh weight respectively. Truemanand Turnbull (1994) found that shell har-dening occurred between 12 and 15weeks after anthesis and effectively cea-sed increases in fruit diameter althoughtotal fruit dry weight per branch increaseduntil 20 weeks post anthesis where moni-toring ceased. Increases in dry weight af-ter shell hardening are due to increasesin kernel mass, the majority of which is oilaccumulation (McConchie, et al. 1996). InAustralia, mature fruit drop begins fromMarch to April and can extend until Octo-ber for some cultivars.

HarvestingIn most Australian orchards, nut-in-huskis mechanically harvested from theground (Figure 2), although hand harves-ting may be used in excessively wet con-ditions and in very steep areas of the or-chard. Nuts are harvested every 4-6weeks to prevent deterioration of kernelon the ground (Mason and Wells, 1993).Tree shaking and Ethephon have beenused both experimentally and commer-cially to promote nut drop and so reducethe number of harvests, however theyhave not been widely adopted to date.Ethephon has been shown to be veryeffective but the response of leaf and nutdrop can vary with cultivar, time and me-thod of application and growing region.There are also concerns about the effectof Ethephon application on return andlong-term yields as variable responseshave been reported (Trueman, 2003,Penter et al., 2003).

Figure 1. Flowering raceme in M. integrifoliaFigure 2. Mechanical harvesting of fallen

macadamia nuts in Australia


Pests and DiseasesThe two major insect pests in Australiaare the larval macadamia nut-borer(Cryptophlebia viridula) and the fruit-spot-ting bug (Amblypelta spp.) (Huwer andMaddox, 2003). Nut-borer can cause pre-mature fruit drop or damage to the kernel,damage can be caused throughout nutdevelopment (Figure 3). Fruit spottingbug damages the kernel through surfacepits and also causes premature nut drop,the majority of damage being done beforeshell hardening occurs. Research on IPMstrategies includes the use of a nut-borerparasite, improved monitoring, trap crop-ping, and the use of IPM compatible pes-ticides (Huwer and Maddox, 2003). Fur-ther research is also needed to quantifythe impact of pest damage on the indus-try.

Husk spot (pseudocercospera) is the ma-jor pathogen in Australia. Considerableanecdotal evidence suggests that this di-sease infects the husk of the developingfruit and leads to premature fruit drop.Currently the major defence againsthusk-spot is prescription application ofchemical as infection occurs long beforethe symptoms are apparent. However,considerably more work is required to do-cument the lifecycle of the fungus, deve-lop robust chemical and genetic defencesand estimate the economic impact of thedisease.

shown to increase with canopy area andlight interception, so open systems suchas this will need to be tested and compa-red with the economics of the system.

Long term assessment of mature or-chards have found that, although maca-damias possess one of the most densetree-crop canopies, there seems smalldecline in yield due to crowding (McFa-dyen et al., 2003) but there was largebetween year variation. However, hed-ging of later age trees to maintain may re-duce the risk of crop losses due to pestand disease.

Post harvestThe husk is removed on farm as soon aspossible after harvesting, as heat of res-piration from the husk can be large. Themoisture content of wet nut-in-shell har-vested from the orchard may range from10 to 25% depending on environmentalconditions before harvesting (Cavaletto1996) but is generally dried to below 10%moisture content on farm where kernel isconsidered safe from mould develop-ment. However, kernel remains suscepti-ble to both hydrolytic and oxidative ranci-dity at this level of moisture content.

The nut-in-shell is transported to proces-sing companies where it is slowly dried toapproximately 3.5% (1-1.5% kernel mois-ture content), cracked and then sorted toremove reject kernel (immature, discolou-red, germinating, insect damaged ormould) and separate kernel into differentstyles defined by percentage of wholekernel and kernel size. Kernel is thenpackaged into foil vacuum-sealed bagsand stored in cool stores to reduce thedevelopment of oxidative rancidity. Cur-rently post-harvest research in Australiafocuses on development of in-line sortingtechnology for reject kernel, control of af-ter roast-darkening and identification oftraits linked to consumer preference.

Genetic improvementMost cultivars grown for production weredeveloped in Hawaii. Breeding and selec-tion programmes have also been under-taken in Australia, South Africa, Califor-nia, Brazil and New Zealand. To date the-re are approximately 700 names repre-senting about 500 genotypes (Hardnerand McConchie, 2003). However, there issome evidence that cultivar performanceis not stable across countries.

Australian industry is currently investingheavily in macadamia genetic improve-ment. The major focus of this programmeis the development of improved cultivarsfor the Australian industry however theprogramme also encompasses conserva-tion of wild macadamia germplasm, se-lection of elite rootstocks and scion bree-

Figure 3. Fruit damage due to nut-borer (courtesyof Gus Cambell NSW Agriculture)

Canopy managementMacadamia is a vigorous tree and thereare concerns about balance between ve-getative and reproductive growth andshading on yield. Canopies are very den-se, LAI measured in 11 year old macada-mia cultivar ‘Haes’ 344 was between 14and 16, which are among the highest re-ported for fruit and nut crops (Meyers etal., 1999). Generally trees are trained intoa single vertical leader with laterals to carrythe crop in the first 3 years after plantingto maintain an open canopy (O’Hare etal., 1996). Experience suggests that na-rrow crotch angles are susceptible tobreakage. New trials are being developedfor high-density plantings with intensivetree training and external tree supportsuch as trellises (Bell and Bell, 2001),trellis systems that promote irradiance tofruit are used for crops such as apples,however in macadamias yield has been

ding. The native origin of macadamias gi-ves the Australian industry a significantadvantage over other countries. Threeex-situ conservations plantations havebeen established in eastern Australia withcuttings collected from over 300 wildtrees across the natural range of M. inte-grifolia and M. tetraphylla. The industryhas also supported in-situ recovery of se-veral degraded wild populations.

The Australian macadamia breeding pro-gramme aims to efficiently produce newcultivars for the Australian macadamia in-dustry using a structured quantitative ge-netic approach. Breeding objectives aredeveloped with the industry and throughwhole of chain economic modelling toquantitatively assess the impact on profi-tability of changes in different traits. Thecurrent traits used in selection are treesize, cumulative yield, commercial kernelrecovery, proportion of whole kernel andkernel size. A breeding population of5.000 individuals has been establishedon 16 sites over 3 major growing regions(Bundaberg, Gympie and NNSW). Thefirst selection from these trials of candida-tes cultivars will be made in 2004 with re-lease to the industry expected from 2012.

ACKNOWLEDGEMENTSWe wish to thank the macadamia indus-tries of the different countries discussed,for their continued support of the rese-arch programmes that have developed toinformation included in this review.

REFERENCES

Bell D.J.D. and H.F.D. Bell. 2001. Expe-riences with intensively managed high-density orchards in macadamia. Austra-lian Macadamia Society News Bulletin28 (5):61-66.

Boyton S. J. and C. M. Hardner. 2002.Phenology of flowering and nut produc-tion in macadamia. Acta Horticulturae575: 381-387.

Cavaletto C. G., Dela Cruz A., Ross E.and H. Y. Yamamoto. 1966. Factorsaffecting macadamia nut stability. I.Raw kernels. Food Technology 20(8):108-111.

Gross C.L. 1995. Macadamia. Flora ofAustralia 16:419-425.

Hardner C. M., and C. A. McConchie.2003. The future of macadamia geneticimprovement in Australia. In: Procee-dings of the 2nd International Macada-mia Symposium 2003. Tweed Heads.Australian Macadamia Society, LismoreAustralia, pp. 85-89.

Hardner C., Winks C., Stephenson R.,Gallagher E. and C. McConchie. 2002.Genetic parameters for yield traits inmacadamia. Euphytica 225: 255-264.


Hargreaves G. 2003. World round-up –Australia. In: Proceedings of the 2nd In-ternational Macadamia Symposium2003. Tweed Heads. Australian Maca-damia Society, Lismore Australia, pp. 6.

Huett D. O., Gogel B. J., Meyers N. M.,McConchie C. A., McFadyen L. M. andS. C. Morris. 2001. Leaf nitrogen andphosphorus levels in macadamia inresponse to canopy position and lightexposure, their potential as leaf-basedshading indicators, and implications fordiagnostic leaf sampling protocols.Australian Journal of Agricultural Re-search 52: 513-522.

Huwer R. and C. Maddox. 2003. Inte-grated pest management in macada-mias – opportunities and challenges.In: Proceedings of the 2nd InternationalMacadamia Symposium 2003. TweedHeads. Australian Macadamia Society,Lismore Australia, pp. 106-110.

Ito P J. 1980. Effect of style removal onfruit set in macadamia. Hort Science15: 520-521.

Johnson L. A. S. and B. G. Briggs.1983. Proteales. IN: B. D. Morley, B. D.and H. R. Toelken (Eds.). FloweringPlants of Australia. Rigby, Adelaide.Pp. 238-244.

Lee, P. 2003. World round-up – SouthAfrica (and Africa). IN: Proceedings ofthe 2nd International Macadamia Sym-posium 2003. Tweed Heads. AustralianMacadamia Society, Lismore Australia,pp. 7-11.

Mason R. L. and I. A. Wells. 1983. Theeffect of harvest interval on the qualityof ground harvested macadamia nuts.Food technology in Australia 36: 373-378.

McConchie C. A., Meyers N. M., Ander-son K., Vivian-Smith A., O’Brien S. andS. Richards. 1996. Development andmaturation of macadamia nuts in Aus-tralia. IN: R. A. Stephenson and C. W.Winks (Eds). Challenges for Horticultu-re in the Tropics, Proceedings of theThird Australian Society of HorticulturalScience and the First Australian Maca-damia Society Research Conference.18-22 August 1996 Pan Pacific HotelBroadbeach, Gold Coast, Australia, pp.234-238.

McFadyen L., Morris S., Oldman M.,McConchie C., Huett D., Meyers N. andJ. Wood. 2003. Canopy management inmacadamia. In: Proceedings of the 2nd

International Macadamia Symposium2003. Tweed Heads. Australian Maca-damia Society, Lismore Australia, pp.139-142.

Meyers N. M., McFadyen L. M., HuettD. and C. A. McConchie. 1999. A com-parison of direct and indirect estimatesof leaf area index of Macadamia inte-grifolia (Maiden and Betcshe). IN: C. A.McConchie, L. M. McFadyen, N.M. Me-yers and D. Huett (Eds.). Canopy ma-nagement for consistent yield in maca-damia. Final Report MC 95008, Horti-cultural Research and DevelopmentCorporation, Gordon, Australia.

Nagao M. A. and H. H. Hirae. 1992. Ma-cadamia: cultivation and physiology.Critical Reviews in Plant Science. 10:441-470.

O’Hare P., Loebel R. and I. Skinner.1996. Growing macadamias. Queens-land Horticultural Institute, Departmentof Primary Industries, Queensland,Brisbane.

Penter M., Snijder B. and F. Kruger.2003. The use of ethephon to facilitateharvesting in the South African maca-damia industry. In: Proceedings of the2nd International Macadamia Sympo-sium 2003. Tweed Heads. AustralianMacadamia Society, Lismore Australia,pp. 133-139.

Ripperton J. C., Moltzau R. H. and D.W. Edwards. 1938. Methods of evalua-ting the macadamia nut for commercialuse and the variation occurring amongseedling plantings in Hawaii. HawaiiAgricultural Experimental Station bulle-tin 79.

Sakai W. S. and M. A. Nagao. 1985.Fruit growth and abscission in Macada-mia integrifolia. Physiologia Plantarum64: 455-460.

Searle C. and P. Lu. 2003. Whole-treewater use and irrigation scheduling inmacadamia. In: Proceedings of the 2nd

International Macadamia Symposium2003. Tweed Heads. Australian Maca-damia Society, Lismore Australia, pp.125-130.

Stephenson R A., Cull B.W., Price G.and J. Stock. 1986. Scientia Horticultu-rae. 30: 83-95.

Stephenson R. A., Gallagher E. C. andV. J. Doogan. 2003. Macadamia res-ponses to mild water stress at differentphenological stages. Australian Journalof Experimental Agriculture 54: 67-75.

Trochoulias T. and G. G. Johns. 1992.Poor response of macadamia (Maca-damia integrifolia Maiden and Betche)to irrigation in a high rainfall area ofsubtropical Australia. Australian Jour-nal of Experimental Agriculture 32:507-512.

Trueman S. J. 2003. Yield responses toethephon for unshaken and mechanica-lly shaken macadamia. Australian Jour-nal of Experimental Agriculture 43:1143-1150.

Trueman S. J. and C. G. N. Turnbull.1994. Fruit set, abscission and dry mat-ter accumulation on girdled branches ofmacadamia. Annals of Botany 74: 667-674.

Urata U. 1954. Pollination requirementsof macadamias. Hawaii agricultural Ex-perimental Station Technical Bulletin22: 5-40.

Vidgen, R. 2003. World round-up –Hawaii. In: Proceedings of the 2nd Inter-national Macadamia Symposium 2003.Tweed Heads. Australian MacadamiaSociety, Lismore Australia, pp. 11-12.

C. Hardner, J. Wilkie and C. McConchieCSIRO Plant Industry

Queensland Bioscience Precinct306 Carmody Road

St Lucia, 4067, AustraliaE-mail: [email protected]

CAROB TREE DEVELOPMENTIN CHINA

INTRODUCTIONChina is the world’s largest producer ofrice, wheat, cotton, peanuts and rape-seed. It ranks second in corn and fourthin soybean production. Subtropical fruitssuch as citrus, litchi and loquat grow to-gether with apples, peaches, apricotsand pears of cool temperate climates.The recent interest for the carob tree isframed in the possibility of growing asagroforestry species and for the restora-tion of degraded areas, mainly, of threeSouthern Chinese provinces with subtro-pical climates: Sichuan (Yibasan andTongAn), Yunnan (Lufeng and YuanJiang) and Guangxi (Shangsi and Pin-gGu) (Fig.1).

The project “Introducing of nursery andcultivation techniques of Ceratonia siliquain China” is funded by the Chinese gover-nment. One of the aims of this project isto find a tree species suitable to dry areasof southwest of China that will contributeto the ecological rehabilitation. The pro-ject was started in 2001 and will be com-pleted by 2005. During the year 2002 theChinese Academy of Forestry (CAF) con-tacted IRTA-Mas Bové (Catalonia, Spain)to start a possible collaboration on thisspecies between both institutions. Duringtwo years 2002 and 2004, IRTA sent toCAF more than 100.000 seeds, from im-portant carob cultivars: five of Spain (‘Ne-gra’, ‘Duraió’, ‘Rojal’, ‘Banya de Cabra’and ‘Matalafera’) and two of Portugal(‘Mulata’ and ‘Galhosa’), with the purposeof introducing them in the nursery and, la-ter be planted in the field (Fig 2). In thenew plantations two methods of planting

Figure 1. -Areas where the carob tree has beenintroduced in China •


were used: direct sowing in the field andsmall plants with containers. The last me-thod would be the most suitable and to beused in the future.

The necessity to know the possibilities ofdevelopment of the carob tree in China,as an answer to the new expectations ge-nerated by this species, fostered the rea-lization of this technical collaborationbetween China and Spain (Fig. 3).

ECOLOGYThe edaphoclimatic conditions of the Chi-nese areas where the carob has been in-troduced, Sichuan, Yunnan and Guangxiprovinces placed in the south, are diffe-rent from those of the Mediterranean Ba-sin to which carob is well adapted. It high-lights the great climatic variability, with al-titudes of 800-1000 m above sea level,minimum temperatures of 0 or -1ºC andmaximum of 40ºC, although there is a biggap of temperature between day and night(about 10 ºC), and rainfall of 700-800 mmin the county of Xichang (Sichuan) and,higher in the other two provinces (Yunnanand Guangxi), about 1000 mm, distribu-ted between June and September. Thecarob was planted on hilly lands and thesoils in overall are not very deep, clay-loam, grey-brown colours, with neutral orslightly acid pH (6-7) and low levels ofphosphorus and calcium (Fig. 4).

In the Mediterranean countries (Spain,Italy, Portugal, Greece, Morocco, Tunisia,etc.), the carob grows well in warm tem-perature and subtropical areas, with alti-tudes below 600 m above sea level, lowrainfall in summer (300-500 mm/year),and tolerates hot and humid coastal zo-nes. It can only withstand light frost; tem-peratures below –4ºC can damage youngtrees and flowers of mature trees. Carobcan adapt to a wide range of soil typesfrom poor sandy soils and rocky hillsidesto deep soils, but they cannot withstandwaterlogging. In the Mediterranean Basincarob generally grows in marginal, calca-reous and basic soils.

CROP DEVELOPMENT RESTRICTIONSThe areas where the carob tree has beenintroduced in China, in the case of the Si-

chuan province, the following remarkscan be highlighted:

The areas where the carob seeds havebeen planted are hilly, long away from thecoast, with high altitudes above sea level,high rainfall (800-1000 mm and concen-trated on three summer months) and withvery poor soils, neutral or slightly acidpH, and these conditions are quite diffe-rent from the Mediterranean countries.Temperatures of the Yibasan and TongAnareas are warm and more similar to caroboriginal areas of Mediterranean Basin.

Pests: Severe damage is caused inyoung trees by rabbits and some nativeants that gnaw trunks and new shoots.Diseases: High rainfall, concentrated onsummer months, can cause waterloggingproblems in young carobs and later pro-mote fungus attacks of Phytophtora, inthe neck of the tree, and Oidium in theleaves of the most sensitive carob seedtrees.

Pollination: the carob is a trioecious spe-cies with male, female and hermaphroditeflowers borne on different trees. It bloomsin summer-autumn and it seems to bemainly pollinated by insects, but also bywind. High rainfall and relative humidity inthe rainy summer season, could increasepoor fruit set during the pollination time(mainly the period that overlaps withblooming time).

Mechanical harvesting: currently this fac-tor is not very important because there islarge manpower and its cost is low.

RESEARCH AND DEVELOPMENTNEEDSResearch and development carob worksin the Chinese Academy of Forestry isvery limited, outstanding some nurseryworks (seed germination, planting me-thods, etc.). Future work would have toassess the performance of the seedlingsplanted in the three Chinese provinces(about 100.000 seedlings) that adapt bet-ter to the new environmental conditions,and select seedlings from more vigorouscultivars, female and hermaphrodite se-xes, tolerant to diseases (Oidium), early

bearing and fruits with good quality. Laterit would be necessary to bud the besttrees on seedlings and to introduce themin collections and comparative trials to fi-nish their agronomic and commercialevaluation. These works should be car-ried out within the new agroforestry sys-tems framework in which the carob couldsolve reforestation problems in degradedareas and its bean production used bothfor human and animal feeding (sheep,goats, pigs, cows, horses, etc.). A clima-tic and soil characterization of the possi-ble locations for this specie would also beinteresting, with the purpose of beingable to define its potential expansionarea in China, avoiding future problemsof lack of crop adaptation.

CONCLUDING REMARKSThere are some favourable aspects onthe development of the carob cultivationin China, large surfaces, large manpowerfor harvesting, potential agroforestryuses, being able to use their productionboth for human and animal feeding. Onthe other hand, it is necessary to choosewell the growing areas, which should bewindy, with little risk of frost, and the cul-tivars to be recommended should adaptwell to Chinese growing conditions be-cause they are different to those of theiroriginal areas (Spain, Italy, Portugal,etc.). Based on the previously described,the carob tree in China it can be consideras an alternative crop in dry land areaswith warm or subtropical climates for di-versification on the agroforestry farms.

ACKNOWLEDGEMENTSThese results emerged from a technicalvisit to China by the first author in Dec-ember 2003, which was supported by theChinese Academy of Forestry, Beijing(China).

J. Tous1 , and J. Chunqian2

1Institut de Recerca i Tecnologia Agroalimen-tàries (IRTA), Dept. d’Arboricultura Mediterrà-

nia, Centre Mas Bové. Apartat 415. 43280Reus, Spain.

2International Farm Forestry Training Center.Chinese Academy of Forestry, Beijing 100091,

China.E-mail: [email protected]

Figure 2.- New carob tree planting in Yibasancounty (Sichuan province)

Figure 3.- Research group that visits the carobseed plantations in several areas of the Xichang

county (Sichuan province)

Figure 4.- Carob tree reforestation in degradedareas of Sichuan province


IN MEMORIAM:MY PROFESSOR DALE KESTER

A few weeks before Dale’s death the 21st

of November 2003 in Davis, California, Iwas talking to a group of technicians ofseveral growers’ associations and I star-ted my talk recognising that I had beenfortunate in my professional life, and oneof the reasons was that I had been a stu-dent with Dale Kester. His influence hasbeen extremely positive for my work,even if we did not always agree on somesubjects, but his personal view of al-monds was in any case a clear exampleof dedication and interest. As most al-mond researchers, he had a passionatefor this species, as well as for his stu-dents, and was, as a consequence, ableto convey his passion for almond growingto us.

It is now quite difficult to find somebodywith this ability to give such a generalperspective of almond. Dale, as well asthe other great figures of almond rese-arch in the past century, Charles Grasse-lly and Antonio Felipe, was an expert bothon rootstocks and varieties, and this com-bined view of the tree complexity wasalso the way to obtain a complex over-view of all problems related to almondgrowing. Until recently, when we probablyfind more articles on almond molecularmarkers than on the almond itself, it washardly impossible to find any article on al-monds without his name cited several ti-mes in the bibliography list. As a matterof fact, his most cited article (Kester andGriggs, 1959. Fruit setting in almond: theeffect of cross-pollinating various percen-tages of flowers. Proc. Amer. Soc. Hort.Sci. 74: 214-219) was one of the pointsabout which we sometimes had disagreed.

Just after my arrival in Davis in December1972, he suggested four points upon whichI could start my research work. I only re-member one on leafing time and frost re-sistance, later taken by MustafaBüyükyilmaz, and the one I selected, onalmond self-compatibility. Once I hadmade my choice, he asked me if I couldread Portuguese, because he had an arti-cle in that language he was unable toread and that could be important on al-mond self-compatibility. This was the pio-neer work of Almeida (1945), so often re-ferred later after my review. This hasbeen definitive for my future work in al-mond, because I continue in the samefield of work.

NOTES AND NEWS

Although after my stay in California weonly met on a few occasions, we conti-nued to talk on the same subject. I re-member a trip to Davis in 1985 where heshowed me some pollination results diffi-cult to explain according to the S allelerelationships of the different cultivars. Wecommented several possibilities and fi-nally I suggested a mutation from an Sallele to a nul allele. This hypothesis waslater confirmed (Kester et al., 1994. Amutation of ‘Nonpareil’ almond conferringunilateral incompatibility. J. Amer. Soc.Hort. Sci. 119: 1289-1292).

During my stay there were other studentsworking under his guidance. They werethe mentioned Mustafa Büyükyilmaz fromTurkey, Juan Negueroles from Spain andMustafa Lasram from Tunisia. Also Ra-chid Hellali from Tunisia was partly wor-king with him. They are all now recogni-sed professionals in fruit growing, rese-arch, education and administration, and Iguess that I am the only one who conti-nues to work in almond, but I do not re-gret it. On the contrary, it makes me feelthat probably I could boast of being morehis heir than the others.

I cannot talk about Dale without mentio-ning his wife Daphne. They were alwaystogether and she was a real support forhim. They opened their home to me bothwhen I was a student, as on Thanksgivingday, and when I visited Davis later. Bothwere friendly and made other people feelat ease with them. They visited Spain se-veral times, and very much enjoyed Spa-nish cuisine. I remember that on their se-cond visit to Spain, they mentioned thatthey had attended meetings of the WeightWatchers in order to reduce weight asthey knew that in Spain they would eatvery well. They were delighted with the cro-quettes of Pili Royo, Antonio Felipe’s wife.

His work always had the support of ano-ther good man that I would also like tobring to mind now, his technician DickAsay. They formed a real team and Dalerecognised it when Dick appeared so of-ten in the scientific publications. This isanother example of the traits defining areal researcher: humble, adaptable andopen. He was humble, conscious of howmuch he did not know. Adaptable, in hisplace both in a scientific meeting and in agrowers’ talk. Open, willing to share hisknowledge.

I saw him for the last time during the In-ternational Horticultural Congress in To-ronto in August 2002, and now I wouldhave preferred not to have seen him be-cause he was not the same Dale. He hadsuffered Daphne’s loss and, although in-terested as always in his work and all theproblems related to almond bud failure,he seemed a little lost to me. However,now I find him more clearly than ever inour daily work.

R. Socias i Company

Professor Dale Kester at California, USA,in August 1997

XIII GREMPA MEETINGON PISTACHIOSAND ALMONDS

The XIII GREMPA Meeting on Pistachiosand Almonds took place in Portugal inMirandela, the Garden-City of Terra-Quente (Hot land), from 1 to 5 June2003. Mirandela, is located in the North-east of Portugal (Trás-os-Montes region).It has a Roman origin, but it was King D.Dinis who, in the XIV century, named thecity as Mirandela and ordered the cons-truction of a fortress wall surrounding it,part of which is still visible. The city stillhas other historical-architectonic structu-res, like the Roman bridge (“Ponte Vel-ha”) with 20 asymmetrical arches, the an-cient palace of the Távoras (the noble fa-mily accused of attempting regicide in1759), the church of “Misericórdia” andseveral pillories of medieval origin,among other interesting features.

The meeting was organized by the Direc-ção Regional de Agricultura de Trás-os-Montes (DRATM), the Instituto de Biolo-gia Experimental e Tecnológica (IBET)

CONGRESSESAND MEETINGS


and the Universidade de Trás-os-Montese Alto Douro (UTAD), with the collabora-tion of the Inter-Regional FAO-CIHEAMCooperative Research Network on Nuts,and having Vitor Cordeiro and M. Marga-rida Oliveira as the conveners.

The meeting was attended by more than80 participants from 14 countries: Algeria(1), Argentina (1), Australia (10), Belgium(1), France (3), Greece (1), Iran (5), Italy(8), Morocco (1), Portugal (22), Spain(17), Tunisia (3), Turkey (3) and USA (6).A Chinese delegation with 7 participantscould not attend the meeting because ofthe severe acute respiratory syndrome(SARS), although China was representedwith two abstracts. Also one participantfrom Macedonia and two other partici-pants from Algeria did not succeed to bepresent but managed to have their pos-ters presented.

The participants represented several sec-tors of activity linked to almond and pista-chio, including farmers, industrials andresearchers. Important scientific advan-ces have been reported and the meetingwas a good opportunity to exchangeknowledge and strategies among teamsfrom all around the world where almondand pistachio are grown. Besides thescientific sessions, numerous opportuni-ties for discussion and establishment ofnew collaborations were possible duringcoffee-breaks, meals or social events.

In total, 75 communications were presen-ted (39 oral communications and 36 pos-ters), unequally distributed by the diffe-rent sessions, which included:“Cultivars, Rootstocks and Breeding” (10oral communications and 12 posters),“Flowering, Pollination and Fruit Set” (5

orals, 5 posters), “Pests and Diseases” (3orals, 3 posters), “Harvesting, Marketingand Industry” (2 orals, 2 posters), “Or-chard Management” (4 orals, 4 posters)and finally “Physiology, Biology and Bio-technology” (15 orals, 10 posters). All theabstracts, including those of the partici-pants that finally did not attend the mee-ting (in total 50 orals and 40 posters weresubmitted) were included in the Book ofAbstracts and distributed by the partici-pants. Before closing the ceremony, aRound table was organized for discussionof common problems and strategies ai-ming to solve them. The proceedings ofthis meeting will be edited in a volume ofCahiers Options Méditerranéenes.

In the first working day, the participantswere transported to the city Hall Audito-rium by the touristic train of Mirandela. Inthe afternoon of the second working day,a visit was organized to an almond privatecompany “Amendouro”, and to the villageof Moncorvo where the secular churchwas visited, as well as a traditional storewhere almonds are processed in a secretway!!! The day ended with a prosperousmeal offered by the Town-hall Presidentof Alfândega da Fé.

On Wednesday, a full day was dedicatedto a long tour. The day started with a visitto almond orchards (at the ExperimentalCentre of Terra Quente), followed by a vi-sit to old wine presses in a Douro farm(Quinta da Pacheca) and a travel by boat,up the Douro river, until Pinhão.

On the last day, after the sessions andclosing ceremony, a farewell eveningbanquet organized at “Estalagem da Sen-hora das Neves” gathered most of theparticipants in a happy event where sin-

VI INTERNATIONALCONGRESS ON HAZELNUT

The 6th International Congress on Hazel-nut was held in Tarragona, North-East ofSpain, on June 14-18, 2004. The Con-gress offered an international forum tomeet each other and to exchange expe-riences related to the latest results con-cerning agronomic, industrial, health be-nefits and commercial aspects of hazel-nut growing. The Congress was organi-zed by the Institut de Recerca i Tecnolo-gia Agroalimentàries (IRTA) – CentreMas Bové in collaboration with the Inter-national Society for Horticultural Science(ISHS) and the FAO-CIHEAM Nut Net-work. This event was financially suppor-ted by several institutions: Diputació deTarragona, Ministerio de Ciencia y Tec-nología, INIA, European Union - projectPORTA, Generalitat de Catalunya –Agència de Gestió d’Ajuts Universitaris ide Recerca (AGAUR), MediterraneanAgronomic Institute of Zaragoza (IAMZ -CIHEAM), FAO-CIHEAM Nut Network,Caixa Tarragona, Town councils of Tarra-gona, Reus and Constantí; and the priva-te sponsors: Borges, Morella Nuts, Indús-tries Garriga, Denomination of Origin“Avellana de Reus”, Unió Agrària Coope-rativa and Coselva.

ging and dancing were also included andwhere the natural abilities of the partici-pants could be demonstrated.

Because of their contribution to this mee-ting special thanks are due to FAO-CI-HEAM, the Director of the Regional Agri-cultural Services of Trás-os-Montes andIBET, for their encouragement and sup-port during all the organization. Also thePresidents of Câmara Municipal de Mi-randela and Alfândega da Fé, as well asAmendouro Company, Mr. António Mar-tins and Mss. Maria de Lurdes Lopes aremuch acknowledged for their help andgranted availability.

Finally, we would like to express ourthanks to the Scientific and OrganizingCommittee of the Meeting and to all parti-cipants and contributors of the large al-mond and pistachio family. We hope thismeeting may have contributed for the ex-change of scientific knowledge and tostrengthen the collaboration of the partici-pants within the network or in connectionwith it.

M. Margarida OliveiraVitor Cordeiro

Participants of the XIII Grempa Meeting on Pistachios and Almonds held at Mirandela, Portugal


Several authorities were represented inthe opening session: the Councillor ofAgriculture (Mr. Antoni Siurana) of theGeneralitat of Catalonia, the President ofthe Diputació of Tarragona (Mr. Joan Are-gio), the Director of IRTA Dr. Josep Tar-ragó, the Chairman of the Nuts Sectionand Mediterranean Climate Fruits of TheInternational Society of HorticulturalScience (ISHS) Prof. Carlo Fideghelli, the

representative of The Centre Internatio-nal de Hautes Etudes Agronomiques Mé-diterranéennes (CIHEAM), Dr. Dunixi Ga-biña, and other local authorities.

The meeting was opened with a welco-ming of the convener, Dr. Joan Tous, whoreferred to the main economic characte-ristics of the Spanish hazelnut sector,with an average production of 9.000 t of

kernel per year and 23.000 ha, 95 % ofthis surface was concentrated in Catalo-nia and, mainly in Tarragona province(18.000 ha), where the hazelnut is, in di-verse districts, an important source of in-come. In Catalonia region, there are7.000 small hazelnut farms with an ave-rage orchard size of about 1 to 5 ha. Inaddition, most of these small farms needto be modernized. In this scenario, one ofthe most important aims is the reductionof the management cost and improve nutquality, to ensure greater returns forgrowers. The hazelnut sector is groupedin six OPA’s (Nut Growers Associations),all of them located in Catalonia. In Spain,hazelnut commercialization is mainly fo-cused on the chocolate and “snacks” in-dustries.

Dr. J. Tous also explained the history ofthe hazelnut Congresses in the world andnoted that the first Colloquium on Hazel-nut in Spain was held in Reus in 1862, or-ganised by the Institut Agrícola of SantIsidre, and that there was discussed tointroduce the hazelnut tree as possiblealternative to grapevine in the coastalareas of Tarragona, due to ‘powdery mil-dew’ fungus problems. Later, in the XXthcentury, in the year 1976, the 1st Interna-tional Hazelnut Congress was held inReus (in this case together with the al-mond). Since then, different cities all overthe world have hosted the following:Avellino, Italy (1983); Alba, Italy (1992);Ordu, Turkey (1996), Corvallis, USA

Participants of the VI International Congress on Hazelnut held at the Palace of the Diputació of Tarragona, Spain

Opening session (sitting from left to right): Dr. J. Tous (Convener), Dr. J. Tarragó (Director of IRTA),Mr. Antoni Siurana (Conseller of Agriculture of the Generalitat of Catalonia), Mr. J. Aregio (President

of Diputació of Tarragona), Prof. Carlo Fideghelli (Chairman, ISHS-WG Nuts and Mediterranean Fruits),and Dr. D. Gabiña (CIHEAM)


Table 1. Main Scientific conclusions of the VI International Congress on Hazelnut.Tarragona (Spain), 14-18 June, 2004.

Session Main aspects Remarks and conclusions(Nr. of contributions)

1. Germplasm and Genetic • Behaviour and adaptations of hazelnut cultivars • Current collections should be reviewed with Improvement (30) to new zones molecular markers.

• Behaviour of several breeding obtentions • International data base of molecular markers (limited advances until today) is useful.• Molecular markers• Many presentations focussed on table varieties

2.- Biology and Physiology (12) • Particular problems vary per country and zone • Phenological data of cultivars differ according (examples). sites and climatic conditions.

• Nut fall in Australia • The knowledge of incompatibility (S)• Selection of pollinizers in Chile alleles in hazelnut cultivars is required.• Studies on root distribution in Portugal• Shedding effects on flowering in Italy

3.- Propagation • Achievement of hardwood cuttings’ method • Rootstocks could be of great interest to solve and Rootstocks (6) specific problems, such as:

• Improvement of agronomic characteristics of ‘Negret’ cultivar with rootstocks (IRTA, Spain).

4.- Orchard Management (24) • Nutrition and pruning • High N fertilizer doses could have a yield• Improvement of harvest facility depressing effect. (machinery, pruning, green covers) • Reference of P in leaves should be revised• Organic production in some areas.

• The irrigation should be adapted to the physiologic phases of the crop.• The Spanish strategy is to develop Organised Technicians Groups (ADV) to help farmers with new EU policy regulations (IPM)*.

5.- Pest and Diseases (16) • Solution for specific problems in specific zones • New technologies of Teledetection (GPS or GIS)• General pest and diseases studies to study wide growing areas and their problems. for widespread areas • “Eastern Filbert Blight” fungus problem,• Increasing problems with traditional pests increases in Oregon hazelnut orchards. (bug species) which affect kernel quality • Possibilities for biological control will increase,• Biological control such as control of “Zeuzera” in Spain.

6.- Post Harvest • Chemical composition • More research is needed on food security and Quality (7) (old as well as new cultivars) (aflatoxin).

• Drying to avoid aflatoxin problems, in Turkey

7.- Health and nuts (2) • Nut composition and healthy effects • Most marketing efforts use the healthy strategy.• Scientific evidences of disease prevention • The nut kernels improve health and do not by the consumption of nuts seem to increase body weight.

8.- Industry, Marketing • Crop and international price prediction • The Turkey policy could reduce surface by 25%. and Economics (10) • Consequences of national policies, • Market for organic production seems

mainly for the main producing country (Turkey) on the increase.• Organic production • High alternate bearing in the crops

of the main producing countries.• Consumer studies can give relevant information to enhance strategies of producers, industries, distribution and research.• Other alternative nut species (Gevuina avellana, in Chile).

* IPM (Integrated Pest Management)


(2000) and finally Tarragona, Spain(2004), twenty eight years after the firstinternational event.

Then, Prof. Carlo Fideghelli (Chair of theISHS Section for Nuts and MediterraneanClimate Fruits) gave the “ISHS MedalAward” to the convener to acknowledge hiseffort for the organisation of the Congress.

The Congress was attended by 125 parti-cipants from 20 countries: Turkey, Italy,USA (Oregón), Spain (Catalonia), France,Portugal, Australia, China, New Zealand,Poland, South America (Chile and Argen-tina), Ukraine, India, Albania, Austria,Georgia, Switzerland, The Netherlandsand Slovenia. A total amount of 107scientific papers were presented in thisCongress, 43 as oral communicationsand 64 as posters. The topics of this Con-gress were related to the following issues:Plant material (30 papers), Biology andPhysiology (12), Propagation and Root-stocks (6), Orchard management (24),Pest and Diseases (16), Post harvest andQuality (7), Health benefits (2) and Indus-try, Marketing and Economics (10). In Ta-ble 1 the main scientific conclusions ofthis Congress are summarized.

Researchers had the opportunity to visitthe experimental fields of IRTA-Mas BovéResearch Center (collection of hazelnutgermplasm, rootstock and cultivar trialsand seedlings collection of Gevuina ave-llana specie), commercial hazelnut or-chards, an exhibition of several harvestmachines and nut industries (Borges andCoselva). An Open day for the sector wasorganised the last journey with two work-shops: “World hazelnut situation andperspectives in France, USA, Italy, Tur-key and Spain” and “Production and com-mercial aspects on hazelnut”. Also, mainscientific conclusions of the Congresswere presented to the sector.

After a ‘proposal’ of Italian researchers,the next meeting on the 7th ISHS Interna-tional Congress on Hazelnut, was deci-ded to be held in Viterbo (Italy), probablyin summer 2008.

The five days of the congress were afriendly course of excellent scientific pre-sentations, relaxed discussions and al-lowed to learn new aspects about our re-gion and local habits. Papers will be pu-blished in a volume of Acta Horticulturaein the near future.

J. Tous (convener) and M. Rovira (secretariat)IRTA-Centre Mas Bové. Apartat 415. 43280

Reus (Spain)E-mails: [email protected];

[email protected]

TO BE HELD:

Almond and Pistachio

IV ISHS International Symposium on Pistachios and AlmondsDate: May 22-26, 2005Place: Tehran, IranConvener: Dr. A. JavanshahAddress: Iranian Pistachio Research InstitutePO Box 77175 / 435Rafsanjan, Islamic Republic of IranTel: 98 391 422 52 02Fax: 98 391 422 52 08E-mail: [email protected]://www.pri.ir

Chestnut

III ISHS International Symposium on ChestnutDate: October 20-23, 2004Place: Chaves (Vila Real), PortugalConvener: C. AbreuAddress: University of Tras os Montes e Alto DouroP.O. Box 2025000-911 Vila Real – PortugalTel: 351 259 350 508Fax: 351 259 350 480Email: [email protected]/eventos/chestnutcongress

Walnut

V ISHS International Symposium on WalnutDate: November, 7-14, 2004Place: Sorrento, Naples, ItalyConveners: D. Avanzato (MiPAF) and M.E. Malvolti (CNR)Addresses: Istituto Sperimentale per la Frutticoltura di Roma (MiPAF)Via di Fioranello, 52, 00134 Roma (Italy)Tel: 39 06 79348186Fax: 39 06 79340158E-mail: [email protected]

Istituto di Biología Agro-ambientale e Forestale (CNR)Viale Marconi 2, 05010 Porano, ItalyTel: 390763374688Fax: 390763374330E-mail: [email protected]

Almond orchard at Almería, Spain

Bunch of pistachio nuts


ALMOND

AbdelFattah, H.M.; Shackel, K.A.;Slaughter, D.C., 2003. Methodology fordetermining almond shaker displacementand frequency. Applied Engineering inAgriculture, 19 ( 2): 141-144.

AbdelFattah, H.M.; Shackel, K.A.;Slaughter, D.C., 2003. Substantial verti-cal tree displacements occur during al-mond shaker harvesting. Applied Engi-neering in Agriculture, 19, (2): 145-150.

AbouJawdah, Y.; Dakhil, H.; ElMehtar, S.;Lee, I.M., 2003. Almond witches’-broomphytoplasma: a potential threat to al-mond, peach, and nectarine. CanadianJournal of Plant Pathology - Revue Cana-dienne de Phytopathologie, 25 (1): 28-32.

Al-Ababneh, S.S.; Shibli, R.A.; Karam,N.S.; Shatnawi, M.A., 2003. Cryopreser-vation of bitter almond (Amygdalus com-munis L.) shoot tips by encapsulation-de-hydration and vitrification. Advances inHorticultural Science, 17 (1): 15-20.

Alonso, J.M.; Socias, R. 2003. Lack ofmale-sterility allelism between peach andalmond. Genetics and Breeding of TreeFruits and Nuts. Acta Horticulturae, 622:257-260.

Aydin, C., 2003. Physical properties of al-mond nut and kernel. Journal of Food En-gineering, 60 (3): 315-320.

Basile, B.; Reidel, E.J.; Weinbaum, S.A.;De Jong, T.M., 2003. Leaf potassiumconcentration, CO2 exchange and light in-terception in almond trees (Prunus dulcis(Mill) D.A. Webb). Scientia Horticulturae,98 (2): 185-194.

Bi, G.; Scagel, C.F.; Cheng, L.; Dong, S.;Fuchigami, L.H., 2003. Spring growth ofalmond nursery trees depends upon nitro-gen from both plant reserves and springfertilizer application. Journal of Horticultu-ral Science & Biotechnology, 78 (6): 853-858.

Bi, G.; Scagel, C.F.; Cheng, L.; Fuchiga-mi, L.H., 2004. Soil and foliar nitrogen su-pply affects the composition of nitrogenand carbohydrates in young almondtrees. Journal of Horticultural Science &Biotechnology, 79 (2): 175-181.

Boskovic, R.; Tobutt, K.R.; Batlle, I.; Du-val, H.; Martinez Gomez, P.; Gradziel,

T.M., 2003. Stylar ribonucleases in al-mond: correlation with and prediction ofincompatibility genotypes. Plant Bree-ding, 122 (1): 70-76.

Boulila, M., 2002. Characterization of analmond isolate of Prunus necrotic ring-spot ilarvirus. Advances in HorticulturalScience, 16 (1): 30-37.

Channuntapipat, C.; Collins, G.; Sedgley,M. 2002. Self-compatibility of almond va-rieties. Australian Nutgrower, Sept- Nov2002, 40-41.

Channuntapipat, C.; Sedgley, M.; Collins,G., 2003. Changes in methyation andstructure of DNA from almond tissues du-ring in vitro culture and cryopreservation.Journal of the American Society of Horti-cultural Science, 128(6): 890-897.

Channuntapipat, C.; Sedgley, M.; Collins,G., 2003. Micropropagation of almondcultivars Nonpareil and Ne Plus Ultraand the hybrid rootstock Titan x Nema-guard. Scientia Horticulturae, 98 (4):473-484.

Channuntapipat, C.; Wirthensohn, M.;Ramesh, S.A.; Batlle, I.; Arus, P.; Sedg-ley, M.; Collins, G., 2003. Identification ofincompatibility genotypes in almond (Pru-nus dulcis Mill.) using specific primers ba-sed on the introns of the S-alleles. PlantBreeding, 122 (2): 164-168.

Corredor, E.; Roman, M.; Garcia, E.; Pe-rera, E.; Arus, P.; Naranjo, T., 2004. Phy-sical mapping of rDNA genes establishesthe karyotype of almond. Annals ofApplied Biology, 144 (2): 219-222.

De Herralde, F.; Biel, C.; Save, R., 2003.Leaf photosynthesis in eight almond treecultivars. Biologia Plantarum, 46 (4): 557-561.

Dicenta, F.; Martinez Gomez, P.; Marti-nez Pato, E.; Gradziel, T.M., 2003. Scre-ening for Aspergillus flavus resistance inalmond. Hortscience, 38 (2): 266-268.

Dirlewanger, E; Cosson, P; Howad, W;Capdeville, G; Bosselut, N; Claverie, M;Voisin, R; Poizat, C; Lafargue, B; Baron,O; Laigret, F; Kleinhentz, M; Arus, P; Es-menjaud, D., 2004. Microsatellite geneticlinkage maps of myrobalan plum and analmond-peach hybrid - location of root-knot nematode resistance genes. Theore-tical And Applied Genetics, 109 (4): 827-838.

Egea, J.; Ortega, E.; Martinez Gomez, P.;Dicenta, F., 2003. Chilling and heat requi-rements of almond cultivars for flowering.Environmental and Experimental Botany,50 (1): 79-85.

Garcia, M.; Martinez, P.; Dicenta, F.,2004. Breaking seed dormancy in almond(Prunus dulcis (Mill.) D.A. Webb). Scien-tia Horticulturae, 99 (3-4): 363-370.

Goldhamer, D.A.; Fereres, E.; Salinas,M., 2003. Can almond trees directly dicta-te their irrigation needs? California Agri-culture, 57 (4): 138-144.

Goldhamer, D.A.; Fereres, E., 2004. Irri-gation scheduling of almond trees withtrunk diameter sensors. Irrigation Scien-ce, 23 (1): 11-19.

Gradziel, T.M., 2003. Interspecific hybri-dizations and subsequent gene introgres-sion within Prunus subgenus amygdalus.Genetics and Breeding of Tree Fruits andNuts (Acta Horticulturae), 622: 249-255.

Gradziel, T.M.; Kester, D.E.; Martínez, P.,2002. A Development Based Classifica-tion for Branch Architecture in Almond.Journal American Pomological Society,56 (2): 106-112.

Gradziel, T.M.; Martínez, P., 2002. ShellSeal Breakdown in Almond is Associatedwith the Site of Secondary Ovule Abor-tion. Journal of the American Society forHorticultural Science, 127 (1): 69-74.

Ikeda, T.; Ishikawa, H.; Sasaki, T., 2003.Regulation of Wolbachia density in theMediterranean flour moth, Ephestia kueh-niella, and the almond moth, Cadra cau-tella. Zoological Science, 20 (2): 153-157.

Jiang, Y.Q.;Ma, R.C., 2003. Generationand analysis of expressed sequence tagsfrom almond (Prunus dulcis Mill.) pistils.Sexual Plant Reproduction, 16 (4): 197-207.

Kazantzis, I.; Nanos, G.D.; Stavroulakis,G.G., 2003. Effect of harvest time andstorage conditions on almond kernel oiland sugar composition. Journal of theScience of Food and Agriculture, 83 (4):354-359.

Koukourikou Petridou, M.A., 2003. Therelation between the levels of extractableand diffusible IAA in almond fruits andtheir ‘’June drop’’. Plant Growth Regula-tion, 39 (2): 107-112.

López, M.; Mnejja, M.; Rovira, M.; Collins,G.; Vargas, F.J.; Arús, P.; Batlle, I., 2004.Self-incompatibility genotypes in almondre-evaluated by PCR, stylar ribonuclea-ses, sequencing análisis and controlled po-llinations. Theor Appl Genet, 109: 954-964.

LondonShafir, I.; Shafir, S.; Eisikowitch,D., 2003. Amygdalin in almond nectarand pollen - facts and possible roles.

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Martins, M.; Tenreiro, R.; Oliveira, M.M.,2003. Genetic relatedness of Portuguesealmond cultivars assessed by RAPD andISSR markers. Plant Cell Reports, 22 (1):71-78.

MirAli, N.; Nabulsi, I., 2003. Genetic di-versity of almonds (Prunus dulcis) usingRAPD technique. Scientia Horticulturae,98 (4): 461-471.

Misaki, R.; Fujiyama, K.; Yokoyama, H.;Ido, Y.; Miyauchi, K.; Yoshida, T.; Seki,T., 2003. Characterization of almond al-pha-mannosidase and its application forstructure analysis of sugar chain. Journalof Bioscience and Bioengineering, 96 (2):187-192.

Ortega, E. ; Dicenta, F., 2003. Inheritanceof self-compatibility in almond breeding

strategies to assure self-compatibility inthe progeny. Theoretical and Applied Ge-netics, 106 (5) : 904-911.

Oukabli, A.; Lansari, A.; Wallali L.D.;Abousalim, A., 2001. Conséquences en-dogamiques sur la germination des grai-nes, la croissance et le développementdes semis de la variété d’amandier auto-compatible Tuono. Fruits 56 (3): 1-18.

Oukabli A., Lansari, A.; Wallali, L.D.;Abousalim A.; Egea, J.; et Michaux Fer-riere, N., 2000. Self and cross pollinationon pollen tube growth and fertilization inself-compatible almond Prunus dulcis‘Tuono’. Journal of Horticultural Scien-ces and Biotechnlogy. 75 (6): 739-744.

Oukabli A.; Wallali, L.D.; Lansari, A.;Abousalim, A.; Michaux-Ferriere, N.;Egea, J., 2001. Développement du sacembryonnaire et événements de la fécon-dation chez l’amandier autocompatiblePrunus dulcis (Mill.) D. A. cv ‘Tuono’. Re-vue Fruits 56 (2) : 93-99.

Romero, P.; Navarro, J.M.; García, F.;Botía, P., 2004. Effects of regulated defi-cit irrigation during the pre-harvest periodon gas exchange, leaf development andcrop yield of mature almond trees. TreePhysiology, 24: 303-312.

Rubio, M.; Martinez, P.; Dicenta, F.,2003. Resistance of almond cultivars toPlum pox virus (Sharka). Plant Breeding,122 (5): 462-464.

Sharma, A.; Gupta, M.N., 2004. Oil ex-traction from almond, apricot and ricebran by three-phase partitioning after ul-trasonication. European Journal of LipidScience and Technology, 106 (3): 183-186.

Shibli, R.A.; Shatnawi, M.A.; IQ Swaidat,I.Q., 2003. Growth, osmotic adjustment,and nutrient acquisition of bitter almondunder induced sodium chloride salinity invitro. Communications in Soil Scienceand Plant Analysis, 34 (13-14): 1969-1979.

Silva, C.; Tereso, S.; Nolasco, G.; Olivei-ra, M.M., 2003. Cellular location of Prunedwarf virus in almond sections by in situreverse transcription-polymerase chainreaction. Phytopathology, 93 (3): 278-285.

Socias i Company, R.; Alonso, J.M.,2004. Cross-incompatibility of ‘Ferragnes’and ‘Ferralise’ and pollination efficiencyfor self-compatibility transmission in al-mond. Euphytica, 135 (3): 333-338.

Socias i Company, R.; Felipe, A.J.; Apa-risi, J.G., 2003. Almond bloom in a chan-

ging climate. Journal American Pomologi-cal Society, 57 (2): 89-92.

Testolin, R; Messina, R; Lain, O; Marraz-zo, MT; Huang, WG; Cipriani, G., 2004.Microsatellites isolated in almond from anAC-repeat enriched library. MolecularEcology Notes, 4 (3): 459-461.

Takeoka, G.R.; Dao, L.T., 2003. Antioxi-dant constituents of almond [Prunus dulis(Mill.) D.A. Webb] hulls. Journal of Agri-cultural and Food Chemistry, 51 (2): 496-501.

Traynor, J., 2003. Almond pollinationcontractor uses microchips to discouragehive thefts. American Bee Journal, 143(12): 921-922.

Tsipouridis, C; Thomidis, T, 2004.Rooting of ‘GF677’ (almond x peach hy-brid) hardwood cuttings in relation tohydrogen hyperoxide, moisture content,oxygen concentration, temperature andpH of substrate. Australian Journal OfExperimental Agriculture, 44 (8): 801-805.

Vezvaei, A., 2003. Isozyme diversity inIranian almond. Genetics and Breeding ofTree Fruits and Nuts (Acta Horticulturae),622: 451-456.

Yi, W.G.; Law, S.E.; Wetzstein, H.Y.,2003. Fungicide sprays can injure thestigmatic surface during receptivity in al-mond flowers. Annals of Botany, 91 (3):335-341.

Yi, W.G.; Law, S.E.; Wetzstein, H.Y.,2003. An In Vitro Study of FungicideEffects on Pollen Germination and TubeGrowth in Almond. HortScience, 38 (6):1086-1088.

Young, C.T.; Schadel, W.E.; Pattee, H.E.;Sanders, T.H., 2004. The microstructureof almond (Prunus dulcis (Mill.) D.A.Webb cv. ‘Nonpareil’) cotyledon. Lebens-mittel - Wissenschaft und - Technologie -Food Science and Technology, 37 (3):317-322.

Zanetto, A.; Maggioni, L.; Tobutt, K.R.;Dosba, F., 2002. Prunus genetic resour-ces in Europe: Achievement and pers-pectives of a networking activity. GeneticResources and Crop Evolution, 49: 331-337.

CAROB

Burés, P.; Pavlicek, T.; Horova, L.; Nevo,E., 2004. Microgeographic genome sizedifferentiation of the carob tree, Cerato-nia siliqua, at ‘Evolution canyon’, Israel.Annals of Botany, 93 (5): 529-535.


Correia, P.J.; Pestana, M.; Martins, M.A.,2003. Nutrient deficiencies in carob (Ce-ratonia siliqua L.) grown in solution cultu-re. Journal of Horticultural Science & Bio-technology, 78 (6): 847-852.

Correia, P.J.; Martins-Louçâo, M.A.,2004. Effect of nitrogen and potassiumfertilization on vegetative growth and flo-wering of mature carob trees (Ceratoniasiliqua): variations in leaf area index andwater use indices. Australian Journal ofExperimental Agriculture, 44: 83-89.

Custodio, L; Martins-Loucao, MA; Roma-no, A., 2004. Influence of sugars on in vi-tro rooting and acclimatization of carobtree. Biologia Plantarum, 48 (3): 469-472.

Jahns, S., 2003. A late Holocene pollendiagram from the Megaris, Greece, givingpossible evidence for cultivation of Cera-tonia siliqua L. during the last 2000 years.Veget. Hist. Archaeobot, 12: 127-130.

Lo Gullo, M.A.; Nardini, A.; Trifilo, P.; Sal-leo, S., 2003. Changes in leaf hydraulicsand stomatal conductance following droug-ht stress and irrigation in Ceratonia sili-qua (Carob tree). Physiologia Plantarum,117 (2): 186-194.

Mediouni, J; Fukova, I; Frydrychova, R; etal., 2004. Karyotype, sex chromatin andsex chromosome differentiation in the ca-rob moth, Ectomyelois ceratoniae (Lepi-doptera: Pyralidae). Caryologia, 57 (2):184-194.

Ramon, L.; Mabberley, D.J., 2004. Theecological status of the carob-tree (Cera-tonia siliqua, Leguminosae) in the Medite-rranean. Botanical Journal of the LinneanSociety, 144 (4): 431-436.

Rhizopoulou, S, 2004. Aspects of cellwall extensibility in Ceratonia siliqua L.Flora, 199 (4): 327-333.

Von Haselberg, C; Ludders, P; Stosser,R, 2004. Pollen tube growth, fertilizationand ovule longevity in the carob tree (Ce-ratonia siliqua L.). J Appl Bot Food Qual,78 (1): 32-40.

Wang, F.; Wang, Y.J.; Sun, Z., 2002.Conformational Role of Xanthan in its In-teraction with Locust Bean Gum. Journalof Food Science, 67 (7): 2609-2614.

Zohary, D., 2002. Domestication of the ca-rob (Ceratonia siliqua L.). Israel Journal ofPlant Sciences, 50, Suppl. S: S141-S145.

Zunft, H.J.F.; Luder, W.; Harde, A.; Ha-ber, B.; Graubaum, H.J.; Koebnick, C.;Grunwald, J., 2003. Carob pulp prepara-tion rich in insoluble fibre lowers total andLDL cholesterol in hypercholesterolemic

patients. European Journal of Nutrition,42 (5): 235-242.

CHESTNUT

Bolvansky, M., 2002. Variability and grou-ping of chestnuts (Castanea sativa mill.)grown in old orchards at the locality Mo-dry Kamen. Folia oecologica, 29: 95-106.

Bolvansky, M.; Uzik, M., 2003. Phenoty-pic variation of nut characteristics in half-sib families of European chestnut (Casta-nea sativa). Biologia, Bratislava, 58/Supl.12: 27-34.

Bounous, G.; Botta, R.; Beccaro, G., 2001.Valore nutritivo e pregi alimentary dellecastagne. Frutticoltura, LXIII (10): 37-44.

Dane, F.; Lang, P.; Huang, H.; Fu, Y.,2003. Intercontinental genetic divergenceof Castanea species in eastern Asia andeastern North America. Heredity, 91 (3):314-321.

Fonti, P.; Sell, J., 2003. Radial split resis-tance of chestnut earlywood and its rela-tion to ring with. Wood and Fiber Science,35 (2): 201-208.

Lopez-Matas, MA; Nunez, P; Soto, A; etal., 2004. Protein cryoprotective activityof a cytosolic small heat shock proteinthat accumulates constitutively in chest-nut stems and is up-regulated by low andhigh temperatures. Plant Physiol, 134 (4):1708-1717.

Pereira, S.; Ramos, A.M., 2003. Caracte-rísticas morfológicas e isoenzimáticas delos cultivares de castaño. (Castanea sati-va Mill.) de Andalucía. Monografías INIA,Agrícola, 13: 160 pp. ISBN: 84-7498-486-6.

Portela, E.; Roboredo, M.; Louzada, J.,2003. Assessment and description ofmagnesium deficiences in chestnut gro-ves. Journal of Plant Nutrition, 26 (3):503-523.

Rhoades, C.C.; Brosi, S.L.; Dattilo, A.J.;Vincelli, P., 2003. Effect of soil compac-tion and moisture on incidence of phyto-phthora root rot on American chestnut(Castanea dentata) seedlings. Forest Eco-logy and Management, 184 (1-3): 47-54.

HAZELNUT

De Salvador, F. R., 2001. La situazionevarietale della corilicoltura italiana. Frutti-coltura, LXIII (10): 23-28.

Del Castillo, M.L.R.; Flores, G.; Herraiz,M.; Blanch, G.P., 2003. Solid-phase mi-croextraction for studies on the enan-

tiomeric composition of filbertone in ha-zelnut Oils. Journal of Agricultural andFood Chemistry, 51 (9): 2496-2500.

Moure, A.; Franco, D.; Sineiro, J.; Domin-guez, H.; Núñez, M.J., 2003. Simulationof multistage extraction of antioxidantsfrom Chilean hazelnut (Gevuina avellana)hulls. Journal of the American Oil Che-mists Society, 80 (4): 389-396.

Nas, M.N.; Read, P.E., 2004. A hypothe-sis for the development of a defined tis-sue culture medium of higher plants andmicropropagation of hazelnuts. ScientiaHorticulturae, 101 (1-2): 189-200.

Okay, Y.; Köksal, A.I.; Artik, N., 2003.Seasonal changes in fructose, glucoseand sucrose content of bark tissues of ha-zelnut grown in the black sea region. Gar-tenbauwissenschaft, 67 (6): 225-233.

Olsen, J.L., 2002. Oregon State Univer-sity’s Integrated Pest Management Pro-gram for the Oregon Hazelnut Industry.HortTechnology, 12 (4): 623-625.

Romero, A.; Tous, J.; Durfort, M.; Rius,M., 2003. Histology of hazelnut (Corylusavellana L.) kernel affected by brownspots in kernel cavity physiopathy. Spa-nish Journal of Agricultural Research, 1(3): 47-53.

Sariyar, L.; Heperkan, D., 2003. The roleof Aspergillus flavus and Aspergillus ni-ger in the hydrolysis of hazelnut fat. Inter-national Journal of Food Science and Te-chnology, 38 (4): 487-492.

Silva, A.P.; Rosa, E.; Haneklaus, S.H.,2003. Influence of foliar boron applicationon fruit set and yield of hazelnut. Journalof Plant Nutrition, 26 (3): 561-569.

Valentín, N.; Marinoni, D.; Botta, R.; Me,G., 2001. Cloni della cultivar di noccioloTonda Gentile delle Langhe a confronto:risultati di cinque anni di osservazioni.Frutticoltura, LXIII (10): 67-71.

NUTS

Bansode, R.R.; Losso, J.N.; Marshall,W.E.; Rao, R.M.; Portier, R.J., 2003. A d -sorption of volatile organic compounds bypecan shell- and almond shell-basedgranular activated carbons. BioresourceTechnology, 90 (2): 175-184.

Bayman, P.; Baker, J.L.; Mahoney, N.E.,2002. Aspergillus on tree nuts: incidenceand associations. Mycopathologia, 155:161-169.

GanMor, S.; Bechar, A.; Ronen, B.; Ei-sikowitch, D.; Vaknin, Y., 2003. Electro-


static pollen applicator development andtests for almond, kiwi, date, and pistachio- An overview. Applied Engineering inAgriculture, 19 (2): 119-124.

Mehlenbacher, S.A., 2003. Progress andprospects in nut breeding. Genetics andBreeding of Tree Fruits and Nuts. ActaHorticulturae, 622: 57-79.

Trullols, E; Ruisanchez, I; Rius, FX; et al.,2004. Qualitative method for determina-tion of aflatoxin B-1 in nuts. J AOAC Int,87 (2): 417-423.

Wang, SJ; Tang, JM, 2004. Radio fre-quency post-harvest quarantine andphytosanitary treatments to control insectpest in fruits and nuts. Prod Pract QualAsse, 4: 17-53.

Wiersma, P.A., 2003. Reproductive bar-riers in tree fruit crops and nuts. Geneticsand Breeding of Tree Fruits and Nuts.Acta Horticulturae, 622: 369-377.

PECAN

Acuña Maldonado, L.E.; Smith, M.W.;Maness, N.O.; Cheary, B.S.; Carroll, B.L.;Johnson, G.V., 2003. Influence of nitro-gen application time on nitrogen absorp-tion, partitioning, and yield of pecan.Journal of the American Society for Horti-cultural Science, 128 (2): 155-162.

Kraimer, R.A.; Lindemann, W.C.; Herrera,E.A., 2004. Recovery of late-season N-15-labeled fertilizer applied to pecan.Hortscience, 39 (2): 256-260.

Ng, C.; Marshall,W.E.; Rao, R.M.; Ban-sode, R.R.; Losso, J.N., 2003. Activatedcarbon from pecan shell: process descrip-tion and economic analysis. IndustrialCrops and Products, 17 (3): 209-217.

Nyczepir, AP; Shapiro-Ilan, DI; Lewis, EE,et al., 2004. Effect of entomopathogenicnematodes on Mesocriconema xenoplaxpopulations in peach and pecan. J Nema-tol, 36 (2): 181-185.

Partridge, E.C.; MorganJones, G., 2003.Notes on hyphomycetes. XC. Fusiclados-porium, a new genus for cladosporium-like anamorphs of Venturia, and the pe-can scab-inducing fungus. Mycotaxon,85: 357-370.

Reid, W., 2002. Current Pest Manage-ment Systems for Pecan. HortTechnolo-gy, 12 (4): 633-639.

Sparks, D., 2003. Pecan Tree DiebackFollowing 1999-2002 Drought Associatedwith September Rainfall. Journal Ameri-can Pomological Society, 57 (4): 142-146.

Thompson, T.E.; Grauke, L.J., 2003. Pe-can Tree Growth and Precocity. J. Amer.Soc. Hort. Sci., 128 (1): 63-66.

Thompson, T.E.; Grauke, L.J., 2003. Pe-can Nut and Kernel Traits Are Related toShelling Efficiency. HortScience, 38 (4):586-590.

Wetzstein, H.Y.; Richardson, E.A.; He,Y., 2002. Alterations in Anatomy and Ul-trastructure of Pecan Leaves Treatedwith Propiconazole during Shoot Expan-sion. Journal of the American Society forHorticultural Science, 127 (1): 08-12.

Wood, B.W.; Conner, P.J.; Worley, R.E.,2003. Relationship of Alternate BearingIntensity in Pecan to Fruit and CanopyCharacteristics. HortScience, 38 (3): 361-366.

PISTACHIO

Ahmad, R.; Ferguson, L.; Southwick,S.M., 2003. Identification of pistachio(Pistacia vera L.) nuts with microsatellitemarkers. Journal of the American Societyfor Horticultural Science, 128 (6): 898-903.

Attia, A.A.; Girgis, B.S.; Khedr, S.A.,2003. Capacity of activated carbon deri-ved from pistachio shells by H3PO4 inthe removal of dyes and phenolics. Jour-nal of Chemical Technology and Biotech-nology, 78 (6): 611-619.

Avanzato, D.; Quarta, R., 2004. Monoe-cious Pistacia terebinthus found in Bulga-ria. Crop wild relative, (2): 14-16.

Barazani, O., Atayev A., Yakubov B.,Kostiukovsky V., Popov K.P. and Golan-Goldhirsh, A. 2003. Genetic variability inTurkmen populations of Pistacia vera L.Genetic Resources and Crop Evolution,50 (4): 383-389.

Barazani, O., Dudai, N. and Golan-Gold-hirsh, A. 2003. Comparison of Mediterra-nean Pistacia lentiscus genotypes byrandom amplified polymorphic DNA, che-mical and morphological analyses. Jour-nal of Chemical Ecology, 29 (8): 1939-1952.

Caravaca, F.; Diaz, E.; Alguacil, M.M.;Roldan, A., 2003. Photosynthetic andstomatal responses of mycorrhizal Pista-cia lentiscus and Retama sphaerocarpaseedlings to water deficit. Agrochimica,47 (5-6): 188-194.

Cetin, AE; Pearson, TC; Tewfik, AH.,2004. Classification of closed- and open-shell pistachio nuts using voice-recogni-tion technology. T Asae, 47 (2): 659-664.

Chao, C.T.; Parfitt, D.E., 2003. Geneticanalyses of phenological traits of pista-chio (Pistacia vera L.). Euphytica, 129(3): 345-349.

Chen, W.Q.; Morgan, D.P.; Felts, D.; Mi-chailides, T.J., 2003. Antagonism of Pae-nibacillus lentimorbus to Botryosphaeriadothidea and biological control of panicleand shoot blight of pistachio. Plant Disea-se, 87 (4): 359-365.

Couladis, M.; Ozcan, M. ; Tzakou, O.;Akgul, A., 2003. Comparative essential oilcomposition of various parts of the tur-pentine tree (Pistacia terebinthus L)growing wild in Turkey. Journal of theScience of Food and Agriculture, 83 (2)136-138.

Elena, K.; Vlahoyiannis, D., 2003. Firstreport of Botrytis cinerea causing shootblight of pistachio in Greece. Plant Patho-logy, 52 (6): 810.

Epstein, L.; Beede, R.; Kaur, S.; Fergu-son, L., 2004. Rootstock effects on pista-chio trees grown in Verticillium dahliae-in-fested soil. Phytopathology, 94 (4): 388-395.

Fooladi, M.H.; Farahnaky, A., 2003. Afla-toxin removal from pistachio nuts by natu-ral natrolite. Journal of Food Science, 68(4): 1225-1228.

Golan, A.; Barazani, O.; Wang, Z.S.;Khadka, D.K.; Saunders, J.A.; Kostiuko-vsky, V.; Rowland, L.J., 2004. Genetic re-lationships among Mediterranean Pista-cia species evaluated by RAPD andAFLP markers. Plant Systematics andEvolution, 246 (1-2): 9-18.

Haddad, CR; Louw, SV; Dippenaar-Schoeman, AS., 2004. An assessment ofthe biological control potential of Helioph-anus pistaciae (Araneae: Salticidae) onNysius natalensis (Hemiptera: Lygaei-dae), a pest of pistachio nuts. Biol Con-trol, 31 (1): 83-90.

Katsiotis, A.; Hagidimitriou, M.; Drossou,A.; Pontikis, C.; Loukas, M., 2003. Gene-tic relationships among species and culti-vars of Pistacia using RAPDs and AFLPs.Euphytica, 132 (3): 279-286.

Kucukoner, E.; Yurt, B., 2003. Some che-mical characteristics of Pistacia vera va-rieties produced in Turkey. EuropeanFood Research and Technology, 217 (4):308-310.

Ila, H.B.; Kafkas, S.; Topaktas, M., 2003.Chromosome numbers of four Pistacia(Anacardiaceae) species. Journal of Hor-ticultural Science & Biotechnology, 78 (1)35-38.


Lee, N.A.; Wang, S.; Allan, R.D.; Kenne-dy, I.R., 2004. A rapid aflatoxin B-1 ELI-SA: Development and validation with re-duced matrix effects for peanuts, corn,pistachio, and soybeans. Journal of Agri-cultural and Food Chemistry, 52 (10):2746-2755.

Ma, Z.H.; Felts, D.; Michailides, T.J.,2003. Resistance to azoxystrobin in Alter-naria isolates from pistachio in California.Pesticide Biochemistry and Physiology,77 (2): 66-74.

Ma, Z.; Luo, Y.; Michailides, T.J., 2003.Nested PCR assays for detection of Mo-nilinia fructicola in stone fruit orchardsand Botryosphaeria dothidea from pista-chios in California. Journal of Phytopa-thology - Phytopathologische Zeitschrift,151 (6): 312-322.

Ma, Z.H.; Luo, Y.; Michailides, T.J., 2004.Spatiotemporal changes in the populationstructure of Botryosphaeria dothidea fromCalifornia pistachio orchards. Phytopa-thology, 94 (4): 326-332.

Martinez, J.J.Y.; Wool, D., 2003. Diffe-rential response of trees and shrubs tobrowsing and pruning: the effects on Pis-tacia growth and gall-inducing aphids.Plant Ecology, 169 (2): 285-294.

MirAli, N.; Nabulsi, I., 2003. Genetic di-versity of Syrian grown pistachio cultivars(Pistacia vera L.) using RAPD technique.Adv. Hort. Sci., 17 (4): 215-222.

MunneBosch, S.; Peñuelas, J., 2003.Photo- and antioxidative protection du-ring summer leaf senescence in Pistacialentiscus L. grown under Mediterraneanfield conditions. Annals of Botany, 92 (3):385-391.

Onay, A., 2003. Micropropagation of Pis-tachio. Micropropagation of Woody Treesand Fruits. Forestry Sciences, 75: 565-588.

Onay, A.; Pirinc, V.; Yildirim, H.; Basaran,D., 2004. In vitro micrografting of maturepistachio (Pistacia vera var. Siirt). PlantCell Tissue and Organ Culture, 77 (2):215-219.

Ozcan, M., 2004. Characteristics of fruitand oil of terebinth (Pistacia terebinthusL) growing wild in Turkey. Journal of theScience of Food and Agriculture, 84 (6):517-520.

Reig-Arminana, J; Calatayud, V; Cervero,J; Garcia-Breijo, FJ; Ibars, A; Sanz, MJ,2004. Effects of ozone on the foliar histo-logy of the mastic plant (Pistacia lentis-cus L.). Environmental Pollution, 132 (2):321-331.

Parfitt, D.E., 2003. ‘Bonsai’ OrnamentalPistachio. HortScience, 38 (6): 1260-1261.

Parfitt, D.E.; Arjmand, N.; Michailides,T.J., 2003. Resistance to Botryosphaeriadothidea in pistachio. Hortscience, 38 (4):529-531.

Sepaskhah, A.R.; Karimi Goghary, S.,2003. Growth and chemical compositionof pistachio affected by salinities anddepths of water table. Communications inSoil Science and Plant Analysis, 34 (3-4):343-355.

Vidrich, V.; Fusi, P.; Graziano, A.; Silves-trini, E.; Michelozzi, M.; Marco, F., 2004.Chemical composition of the essential oilof Pistacia lentiscus L. Journal of Essen-tial Oil Research, 16 (3): 223-226.

STONEPINE

Alvarez, JB; Toledo, MJ; Abellanas, B, etal., 2004. Use of megagametophyte sto-rage proteins as markers of the geneticdiversity in stone pine (Pinus pinea L.) inAndalucia, Spain. Genet Resour Crop Ev,51 (6): 621-627.

Mutke, S.M.; Sada, B.; Iglesias, S.; Gil,L., 2003. Evaluación de la producción in-dividual de piña en un banco clonal depino piñonero (Pinus pinea L.) en Madrid(in Spanish). Invest. Agrar.: Sist. Recur.For., 12 (1): 149-157.

Mutke, S.; Gordo, J.; Climent, J.; Gil, L.,2003. Shoot growth and phenology mo-delling of grafted stone pine (Pinus pineaL.) in Inner Spain. Annals of ForestScience, 60 (6): 527-537.

Ranaldi, F.; Giachetti, E.; Guerin, E.;Bacci, S.; Paoletti, E.; Boddi, V.; Vanni,

P., 2003. Gravitational stress on germina-ting Pinus pinea seeds. Comptes RendusBiologies (France). 326 (6): 553-564.

WALNUT

Aletà, N.; Ninot, A.; Voltas, J., 2003. Ca-racterización del comportamiento agrofo-restal de doce genotipos de nogal (Ju-glans sp.) en dos localidades de Cataluña(in Spanish). Invest. Agrar.: Sist. Recur.For., 12 (1): 39-50.

Amaral, J.S.; Casal, S.; Pereira, J.A.; Se-abra, R.M.; Oliveira, B.P.P., 2003. Deter-mination of sterol and fatty acid compositio-ns, oxidative stability, and nutritional valueof six walnut (Juglans regia L.) cultivarsgrown in Portugal. Journal of Agriculturaland Food Chemistry, 51 (26): 7698-7702.

Breton, C; Cornu, D; Chriqui, D; et al.,2004. Somatic embryogenesis, micropro-pagation and plant regeneration of “EarlyMature” walnut trees (Juglans regia) thatflower in vitro. Tree Physiol, 24 (4): 425-435.

Fady, B.; Ducci, F.; Aletà, N.; Becquey,J.; Díaz, R.; Fernández, F.; Jay-Alle-mand, C.; Lefèvre, F.; Ninot, A.; Panet-sos, K.; Paris, P.; Pisanelli, A.; Rumpf, H.,2003. New Forests, 25: 211-225.

Fukuda, T.; Ito, H.; Yoshida, T., 2003. An-tioxidative polyphenols from walnuts (Ju-glans regia L.). Phytochemistry, 63 (7):795-801.

Jensen, P.N.; Sorensen, G.; Brockhoff,P.; Bertelsen, G., 2003. Investigation ofpackaging systems for shelled walnutsbased on oxygen absorbers. Journal ofAgricultural and Food Chemistry, 51 (17):4941-4947.


Kevers, C; Bisbis, B; Crevecoeur, M; etal., 2004. Wood formation in in vitro pro-pagated walnut shoots in relation withroot formation and development. ActaBot Gallica, 151 (1): 45-53.

Koyuncu, M.A.; Ekinci, K.; Sabrán, E.,2004. Cracking characteristics of walnut.Biosystems Engineering, 87 (3): 305-311.

Mahoney, N.; Molyneux, R.J., 2004. Phy-tochemical inhibition of aflatoxigenicity inAspergillus flavus by constituents of wal-nut (Juglans regia). Journal of Agriculturaland Food Chemistry, 52 (7): 1882-1889.

McKenna, J.R.; Epstein, L., 2003. Sus-ceptibility of Juglans Species and Inter-specific Hybrids to Agrobacterium tume-faciens. HortScience, 38 (3): 435-439.

Menard, M.; Baudry, A.; LeSaux, M.,2004. First report of bacterial canker ofwalnut caused by Brenneria nigrifluens inFrance. Plant Disease, 88 (2): 220.

Ninot, A.; Aletà, N., 2003. Identificationand genetic relationship of Persian wal-nut genotypes using isozyme markers.Journal American Pomological Society,57(3): 106-114.

Orel, G.; Marchant, A.D.; McLeod, J.A.;Richards, G.D., 2003. Characterization of11 Juglandaceae genotypes based onmorphology, cpDNA, and RAPD. Hort-science, 38 (6): 1178-1183.

Sabatier, S.; Barthelemy, D.; Ducousso,I., 2003. Periods of organogenesis inmono- and bicyclic annual shoots of Ju-glans regia L. (Juglandaceae). Annals ofBotany, 92 (2): 231-238.

Sakr, S.; Alves, G.; Morillon, R.L.; Mau-rel, K.; Decourteix, M.; Guilliot, A. Fleurat

Lessard, P.; Julien, J.L.; Chrispeels, M.J.,2003. Plasma membrane aquaporins areinvolved in winter embolism recovery inwalnut tree. Plant Physiology, 133 (2):630-641.

Solar, A.; Ivancic, A.; Stampar, F.; Hudi-na, M., 2002. Genetic resources for wal-nut (Juglans regia L.) improvement in Slo-venia. Evaluation of the largest collectionof local genotypes. Genetic Resourcesand Crop Evolution, 45: 491-501.

Solar, A.; Stampar, F., 2003. Genotypicdifferences in branching pattern and frui-ting habit in common walnut (Juglans re-gia L.). Annals of Botany, 92 (2): 317-325.

Vahdati, K; Leslie, C; Zamani, Z; et al.,2004. Rooting and acclimatization of in vi-tro-grown shoots from mature trees ofthree Persian walnut cultivars. Hortscien-ce, 39 (2): 324-327.

Vettraino, A.M.; Belisario, A.; Maccaroni,M.; Vannini, A., 2003. Evaluation of rootdamage to English walnut caused by fivePhytophthora species. Plant Pathology,52 (4): 491-495.

Vyas, D.; Sharma, S.K.; Sharma, D.R.,2003. Genetic structure of walnut genotypeusing leaf isozymes as variability measure.Scientia Horticulturae, 97 (2): 141-152.

Yildiz, K.; Yilmaz, H., 2003. Effect ofTransplanting Rootstocks Before Graftingon Xylem Exudation and Graft Success inWalnut. Journal American PomologicalSociety, 57 (4): 146-148.

THESIS

Alonso, J.M., 2004. Efecto de la consan-guinidad sobre la expresión y la transmi-

sión de la autocompatibilidad en el al-mendro (Prunus amygdalus Batsch). PhDthesis. (in Spanish with summary in En-glish). Lleida University. Escuela TécnicaSuperior de Ingeniería Agraria, 227 pages.

Georges, A., 2003. Vessel-associated ce-lls in walnut trees (Juglans regia): Role ofplasma membrane H+-ATPase (in Frenchwith summary in English). Blaize PascalUniversity, France: 279 pages.

Gharnit, N., 2003. Characterization of thecarob tree (Ceratonia siliqua L.) from theProvince of Chefchaouen (NW of Moroc-co). PhD thesis. (in French with summaryin English). Faculty of Sciences and Te-chniques of Tangier, Université Abdelma-lek Essaâdi (Morocco).

López, M., 2004. Mejora del almendropara autocompatibilidad utilizando técni-cas biológicas y moleculares. PhD thesis.(in Spanish with summary in English).Lleida University. Escuela Técnica Supe-rior de Ingeniería Agraria, 270 pages.

Martins, M. 2003. Análise molecular emamendoeira: da diversidade e estabilida-de genómicas à identificaçao de resistên-cias. PhD thesis. (in Portuguese withsummary in English). Universidad de Lis-boa, 241 pages.

MASTERS

Türker, S. 2003., The effect of somerootstock x cultivar combinations of pista-chio to the phonological and pomologicaltraits. Master Thesis. University of Har-ran, Graduate School of Natural andApplied Sciences. Department of Horti-culture, 86 pages.


THE FAO-CIHEAM INTER-REGIONAL COOPERATIVE RESEARCH NETWORK ON NUTS

Network Coordinator: F.J. VargasEditor: I. BatlleEditorial staff: M. LannoyeTypeset by: Carácter Gráfico, S.L.E-mail: [email protected] 1020-0797

IRTA- Mas BovéDepartament d'Arboricultura Mediterrània.Apartat, 415.E- 43280 REUS (Spain)Tel.: +34-977 32 84 24Fax: +34-977 34 40 55E-mail: [email protected]

Network Coordination Centre Network Coordinator

Nut tree crops F. J. VargasAlmondStone Pine

Subnetworks Liaison Centre Liaison Officer

Hazelnut A.I.Köksal

Walnut and Pecan M. Lafargue

Pistachio B. E. Ak

Chestnut G. Bounous

Genetic Resources I. Batlle

Economics L.M. Albisu

FAO Jutta Krause

CIHEAM Dunixi Gabiña

Ankara University. Faculty of Agriculture.Department of Horticulture.06110 - Ankara (Turkey).Tel: 90 - 312 3170550. Fax: 90 - 312 3179119E-mail: [email protected]

Institut de Recerca i Tecnologia Agroalimentàries IRTA.Centre de Mas BovéDepartament d’Arboricultura MediterràniaApartat 415. E 43280 - Reus (Spain)Tel: 34- 977 328424. Fax: 34- 977 344055E-mail: [email protected]

University of Harran, Faculty of AgricultureDepartement of Horticulture63200 - Sanliurfa (Turkey)Tel: 90 - 414 2472697. Fax: 90 - 414 2474480E-mail: [email protected]

Institut National de la Recherche Agronomique INRA.Unité de Recherches sur les Espèces Fruitières et la Vigne.B.P. 81 - 33883 - Villenave d’Ornon (France)Tel: 33 - 556 843277. Fax: 33 - 556 843274E-mail: [email protected]

Università degli Studi di Torino.Dipartamento di Colture Arboree. Cattedra di ArboriculturaVia Leonardo Da Vinci, 44. 10095 - Grugliasco (TO) (Italy)Tel. 39 - 011 6708653. Fax: 39 - 011 6708658.E-mail: [email protected]

Institut de Recerca i Tecnologia Agroalimentàries IRTA.Centre de Mas Bové.Departament d’Arboricultura MediterràniaApartat 415. E 43280 Reus (Spain)Tel: 34 - 977 328424. Fax: 34 - 977 344055E-mail: [email protected]

Servicio de Investigación Agraria.Diputación General de Aragón. Apartado 72750080 - Zaragoza (Spain)Tel. 34 - 976 576361. Fax: 34- 976 575501E-mail: [email protected]

Regional Office for Europe REU:Viale delle Terme di Caracalla.00100 Roma (Italy)Tel: 39 - 06 57054405 Fax: 39 - 06 57055634E-mail: [email protected]

Instituto Agronómico Mediterráneo de Zaragoza IAMZ.Apartado 202, 50080 Zaragoza (Spain)Tel: 34 - 976 71 60 00 Fax: 34- 976 71 60 01E-mail: [email protected]

Documents

NUCISnetworks.iamz.ciheam.org/nuts/pdfs/NUCIS 12_2004.pdf · 2015. 11. 25. · FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004 1 NUCIS NEWSLETTER Information Bulletin of the