Cover and inside page (web) · CSSA Seed Health Symposium 55 Seed Testing InternationalNo. 125 April 2003 1 CONTENTS The 2nd Announcement of the Extraordinary Meeting 2003, to be

IN THIS ISSUESEED TECHNOLOGY

GMO 1st & 2nd Proficiency Test10

ISF Position on GMO12

Latest Developments in Seed Health20

ISHI - Industry Initiatives22

ISTA ACCREDITATION

Report from Workshop in Slovenia44

Kenyan Seed Trade45

TECHNICAL COMMITTEES

Flower Seed Testing32

Report on the Tetrazolium Workshop39

Report from the PDC Workshop 42

Update from the Proficiency TestCommittee

45

MEMBERSHIP

Seed Testing in Hungary48

100 Years of Seed Testing in Canada51

Calendar 200357

48Seed Testing in HungaryThe 27th ISTA Congress is to be held in theEastern European Country. Dr. Ertsey givesus insight into their Seed Testing System

Seed TestingINTERNATIONAL

ISTA News Bulletin No. 125 April 2003

EDITORIAL OFFICE

EDITOR-IN-CHIEFDr. Michael Muschick

CO-EDITORSMs. Sarah Anne Meier

Ms. Patricia Raubo

ADMINISTRATIONDr. Bettina Kahlert

(Technical Committees)

Ms. Martina Rösch(Accreditation)

Ms. Cannice Gubser(Membership)

International Seed Testing Association (ISTA)

ISTA SecretariatZürichstrasse 50

P.O. Box 3088303 BassersdorfCH - Switzerland

Phone: +41 1 838 6000Fax: +41 1 838 6001

E-mail: [email protected]: www.seedtest.org

Next IssueDeadline for article submission:

July 31, 2003Publication Date of Seed Testing

International No. 126:October 2003

Other ISTA PublicationsFor information and orders of other

ISTA Publications, please contact theISTA Seretariat at the above address, or

visit our website at

www.seedtest.org

Dear Reader,

What you now have in your hands isthe ISTA News Bulletin with a newconcept, a new cover and the new titleSeed Testing International. Seed Testing International should be amagazine by seed testers for seedtesters. Seed Testing International willprovide articles about and around seedtesting. Seed Testing International will mainlyaddress the international challengesand developments but also describeregional challenges and developmentsand hence provide a colourful pictureon all different regions and cultures ofour ISTA family. Seed Testing International will containarticles from our Executive Committeedealing with governance and politicalissues as well as from our TechnicalCommittees dealing with improve-ments and developments in the area ofseed science and technology. Moreoverall our partner organisations around theworld shall have a word in Seed TestingInternational describing challengesand developments from their point ofview or in their region. I am proud toannounce that in this first issue of SeedTesting International articles are pub-lished by ISF, APSA, ISHI and IPPC inorder to present to you an interestingand vivid picture of the seed testingworld and around it. A focal point of this issue is the GMdiscussion. A hot topic which wasalready addressed in the last ISTANews Bulletin (December 2002 issue /No. 124). ISF provide their views onthis very challenging issue, next to thedetailed information on all our ISTAactivities in that particular area, whichare numerous. With these contributions

we hope to provide you an overall pic-ture of the different international andregional developments in this area. Also plant health issues are an interna-tionally challenging topic. Please readan update on the developments of theISTA Plant Disease Committee as wellas articles about ISHI and the IPPC(International Plant ProtectionConvention) on this subject.I hope you are aware that the nextISTA Congress is coming up quitesoon. The first announcement for thisimportant event as well as an articleabout seed testing in Hungary can befound in this issue.We sincerely hope that you will enjoyreading this first issue of Seed TestingInternational and are looking forwardto receiving your comments/input onour new magazine as well as commentsand questions on the different articlesin this issue, which we would bepleased to publish in the next issue ofSeed Testing International. Your feed-back is important to us!Before closing, I would also like toremind you that the next membershipmeeting is coming up very shortly: theISTA Extraordinary Meeting 2003 tobe held in Zurich, Switzerland fromJune 30 to July 3, 2003 and it will bemy pleasure to welcome you personal-ly in Switzerland at this importantevent.

Yours sincerely,Michael Muschick

EditorialBy Michael Muschick,ISTA Secretary General

Seed TestingINTERNATIONAL

CONTENTS

PRESIDENT’S REPORT 2

MEMBERSHIP MEETINGSISTA Extraordinary Meeting 2003 Announcement 3EOM 2003 - The Venue: Zurich 427th ISTA Congress 2004 Announcement 5ISTA Seed Symposium 2004 - Call for Papers 6

ISSUES OF COMMON TECHNICAL INTERESTGenetically Modified Crops 8Summary of 1st ISTA GMO Proficiency Test 10Protocol for 2nd ISTA GMO Proficiency Test 11ISF Position on GMO 12APSA Marches Forward 16IPPC Global Situation 17Latest Developments in Seed Health 20ISHI - Industry Initiative in Seed Health 22

TECHNICAL COMMITTEESMoisture Content in Proficiency Test 25Quality Assurance - Temperature Control 28Purity Survey Overview 31The Making of Flower Seed Working Sheets 32Statistics 38Report on ISTA Tetrazolium Workshop 39Report on ISTA PDC Workshop in New Zealand 42Upcoming Workshops 43Update from the ISTA Proficiency Test Committee 45

ACCREDITATIONReport on ISTA QA Workshop in Slovenia 46Report on Kenyan Seed Trade Association Workshop 47Newly Accredited ISTA Member Laboratories 46

MEMBERSHIPSeed Testing in Hungary 48Prof. Leist’s 60th Birthday Celebration 50100 yrs Seed Testing in Canada 51India Applauds Danish Aid & Dr. Mathur 52Information on ISTA Membership 53New Members and Mutations 53Calendar 57

REPORTS FROM MEETINGSICPP Seed Health Symposium 54OECD Forest Meeting 55CSSA Seed Health Symposium 55

Seed Testing International No. 125 April 2003 1

CONTENTS

The 2nd Announcementof the ExtraordinaryMeeting 2003, to beheld in Zurich, June 30- July 3

Including RegistrationForm, and details aboutthe meeting and Zurich

Page 3

Dr. KatalinErtsey talksabout SeedTesting inHungary, wherethe 27th ISTACongress is tobe held in 2004

Page 48

The ISTA FlowerSeed Committeediscuss thepreparation oftheir ISTAFlower SeedHandbook,includingWorking Sheetsof Cyclamen andPetunia

The ISTAGMO TaskForce send outthe samplesfor the 2nd

ISTA GMOProficiencyTest

Page 11

Seed TradeAssociationin KenyaholdAccreditationWorkshop inNairobi

Page 47

Dear ISTA Members

We can rely on our Secretariat to be effi-cient at all times; they even reached meelectronically here at the workshop "onvarietal verification and GMO detection"at the ARC Roodeplaat Vegetable andOrnamental Plant Institute near Pretoria,South Africa, reminding me to write thisoutstanding letter.

Our workshop here is an example of thefruitful cooperation between FAO andISTA, whereby we provide the technicalexpertise and the organization, and FAO,the financial maintenance, i.e. travelexpenses for the participants and the lec-turers. Following the successful conceptthat had already been tested in Argentina in2001 for South America, the first part ofthis workshop deals with varietal verifica-tion and hybrid quality by electrophoresisof storage proteins in theory and practice.Thereby, the participants practiced the iso-electronical focusing with hybrid maize,barley and sunflower, and rice under theexperienced/approved instruction ofRainer Knoblauch, whereas Dr. EnricoNoli, leader of the GMO Task Force work-ing group, 'Exchange of Information',excellently presented and instructed thePCR analysis. The 18 participants fromseven African countries were highly moti-vated to learn the techniques and it wasintensively discussed. Often in workshops,there is an interesting mixture of partici-pants from governmental, company andother private seed testing laboratories,from regulatory officials and universities.This leads to very interesting discussionsregarding highly actual topics.

We were pleased that Dr. Michael Larindefrom FAO participated in this workshopnot only as an observer but also with apipette in hand. In this he was able to gaina direct insight into the efficiency of thisevent, which not only results into a net-work of communication, but leads to newfriendships. Dr. Bettina Kahlert, Head ofTechnical Committee Administration fromthe ISTA Secretariat, was the responsibleperson for all organization, and was mostsuccessful. We would like to thank Dr.Graham Thompson, the assistant directorof Plant Protection and BiotechnologyARC, and his colleague, Gurling Bothma,

for providing the excellent working andlecture rooms, and their practical supporton location. With this workshop SouthAfrica contributed to the further develop-ment of the African continent.

Dear ISTA Members, your ISTA ExecutiveCommittee will also meet in Africa March10 to 13, 2003. We are invited by ourECOM Member Jeffrey Luhanga to holdour ECOM Meeting in Malawi for theapproval of ISTA internal items as well asfor the preparation of the 2nd ExtraordinaryMeeting, Zurich, June 30 to July 3, 2003.Please find below a few topics selectedfrom the full agenda. (Two of the topicslisted below raised at the ExtraordinaryMeeting in Bolivia, were very remarkableand have to be continued):

1. The ISTA position paper regardingstronger restructuring, and reorientationbetween governmental dependency andindependency, as a free and autonomousassociation, with all consequences for vot-ing rights, methodology acceptance andfinancing. In Bolivia, the delegates clearlysupported innovation in careful, gradualchanges, as well as carefully monitor theexciting structures. This needs accurateconsiderations and decisions.2. The presentations of all TechnicalCommittee Chairs, at the ExtraordinaryMeeting 2002, were so excellent, brief andprecise all at the same time that theExecutive Committee would like, infuture, to keep this format of the meetingagenda for following ExtraordinaryMeetings. 3. One important issue to discuss is thefuture information and publication policy,including in particular the electronicmedia. Also the use of the ISTA Logo byour members needs a clear regulation.

As a matter of course, the ISTAAccreditation as well as the progress of ourTechnical Committee work will have theirown time in the meeting.

You will see that this full programme, dec-orated with many single items, fills theweek. On Friday, the ECOM will be invit-ed to a meeting with the national andregional representatives to intensify thecooperation between this region and ISTA,

in regards to our goals defined at the con-gress in Angers to support Africa, Asia andSouth Africa.

As Member, you might be interested toknow that the 2nd ISTA Proficiency Test onGMO Testing has begun and that the firstsamples have been sent to participatinglaboratories. Remarkable is also therestructuring of our ISTA web site due tothe intensive cooperation between PatriciaRaubo from the Secretariat and JimSheppard from Canada.

Finally, we would like to congratulate theInstitute of Plant Breeding, Seed Scienceand Population Genetics, from theUniversity of Hohenheim on their 125th

Birthday Celebration. ISTA is most appre-ciative of this Institute, which is currentlyunder the direction of Prof. Dr. MichaelKruse, whom we all hold in esteem as anenergetic leader and member of our com-mittees.

We think of Prof. Lakon who developedthe Tetrazolium Test, and incorporated itinto the ISTA Rules, also of Prof. Dr.Lindenbein, Dr. Bulat and Prof. Dr.Steiner, who took on important roles with-in ISTA, with great success. We haveexciting role models in our Association, aswell as many young, energetic SeedTesters, so that ISTA can reach it's goalsuccessfully: Uniformity in Seed Testing.

I look forward to seeing all of you inZurich at our 2nd Extraordinary Meeting,where I will be able to greet you personal-ly.

Norbert LeistPresident of ISTA

2 Seed Testing International No. 125 April 2003

PRESIDENTS REPORT

President’s ReportBy Norbert Leist, ISTA President


MEMBERSHIP MEETINGSISTA Extraordinary Meeting 2003

ISTA Extraordinary Meeting 2003Zürich, Glattbrugg, SwitzerlandJune 30 - July 3, 2003

Dear Colleagues,The International Seed Testing Association (ISTA) takes pleasure in invitingyou to participate in its Extraordinary Meeting 2003, to be held from June 30- July 3, in Glattbrugg, Zurich, Switzerland.Following a decision of the ISTA Membership during the last ExtraordinaryMeeting in Santa Cruz, Bolivia, July 2002, the 2nd Extraordinary Meeting willbe held in Switzerland. Through the Extraordinary Meetings, ISTA has beenable to speed up its activities - something not possible from the previous threeyear meeting cycle - therefore we expect you will be joining us!This meeting is aimed at discussing and deciding on proposals for changes tothe ISTA International Rules for Seed Testing and business items of theAssociation, with the international participation of ISTA delegates and repre-sentatives from both the seed industry and governments, including experts inseed technology, scientific research and laboratory accreditation.The main subjects of the meetings will be: Testing of GM Seeds - Chapter inthe ISTA Rules and international harmonisation; Generic Method ValidationProgramme; ISTA Quality Assurance Programme; Accreditation ofLaboratories world-wide - latest developments and international collabora-tion; Discussion on ISTA's relationship with governments.

THE OCCASION TO DISCUSS ALL YOUR TECHNICAL PROBLEMS AND CHALLENGES WITHTHE BEST EXPERTS IN SEED TESTING FROM ALL PARTS OF THE WORLD.

Preliminary Programme30 June - 1 JulyTechnical Committee SessionsSessions from all the ISTA TechnicalCommittees, As well as longer sessions onAccreditation, Proficiency Testing, theGeneric Method Validation Programme andthe ISTA Rules.

2 - 3 JulyOrdinary Meeting of the AssociationUpdates on invited partner organisations,consideration and adoption of activityreports of the Association’s Committees,consideration and adoption of all changes tothe International Rules for Seed Testing,consideration and adoption of current busi-ness matters of the Association, discussionon further activities and developments ofthe Association.A full Preliminary Programme is currentlyavailable in the 2nd Announcement.However it is subject to change. The finalprogramme will be published in the finalannouncement, once the ExecutiveCommittee has given its approval. For fur-

ther information, or to receive the finalannouncement, please contact the ISTASecretariat.

Travel Information

Regular scheduled flights from every conti-nent and most countries and major cities ofthe world land at Zurich's international air-port. The busy Hauptbahnhof, Zurich'smain railway station, dates from 1871 andis located on Bahnhofplatz, in the city cen-tre, and has excellent connections with bothtram and bus lines. Trains run frequentlyfrom major Swiss centres. High speed InterCity Expresses also connect with most ofthe major cities within Europe, makingtravelling by train a viable option.

ISTA has determined to provide compli-mentary transportation from the airport tothe hotel. The NOVOTEL Shuttle bus runsbetween the Airport and the hotel every 15minutes. For participants arriving atZurich's main railway station, there aredirect trains from the station to the airport(the S7 and S2), where the shuttle can thenbe taken to the hotel.

Public TransportA highly efficient and easy-to-use tram andbus network (known as Züri Linie) operateswithin the city, making sightseeing intoZurich city a pleasurable experience.Ferries also form part of Zurich's transportsystem. A boat trip is a must in summer.Short and extended cruises are available,and a sundowner on the Zurich Lake is theperfect way to spend a scenic afternoon.

Accompanying PersonsThe accompanying person fee includes par-ticipation of the official dinner, lunches andcoffee breaks. Please note that there is noofficial programme for accompanying per-sons, however guided tours and day tripscan be arranged at short notice through thehotel. For further information, contact thereception desk at the Novotel, Zurich.

To register, please fill out the registrationform enclosed herewith, and return to the

ISTA Secretariat. You may alsoregister online at

ww.seedtest.org

Zurich (or more familiarly, Züri) is situatedin the heart of German speakingSwitzerland, with a population of 363 000.Positioned at the northern tip of theZürichsee (Lake Zurich), lakeside prome-nades and expensive houses are prominentand can be spotted along both shores. Butthe city's most familiar sites are, without adoubt, the Fraumünster and GrossmünsterMinsters, which solemnly face each otheracross the River Limmat. The Old Townspans this river, and some of the most inter-esting lanes and buildings are clusteredalong its banks. In summer the view of thecity is beautiful, with the lake reflecting themountains and clear blue sky. The summertemperature averages at about 25oC andrarely rises above the 30oC mark. Thehumidity is at a comfortable level.

Zurich dates its origins from 15BC, whenthe Roman customs post of Turicum wasfounded. By the tenth century, the town hadacquired the status of a city. The modernZurich is a city of bankers in a country ofbanks. This concentration of wealth canmost readily be seen along theBahnhofstrasse, flanked by lime trees.

Other riches lie in the city's excellent uni-versities - Zurich is a powerhouse forresearch, with public-private partnershipsleading to innovations in design and thehigh-tech area. The city also has a strongcultural presence - over 30 museums, art

galleries, auction houses, the opera, orches-tras and the Schauspielhaus theatre.

The citizens enjoy a high standard of living,and this is evident in the many fashionableand enjoyable bars, cafés and restaurantsthat fill the Old Town. The ambience isheightened by the large swathes on eitherside of the River Limmat that are pedestri-an-only areas. But for those who find thecomfortable burgher lifestyle a little tootame, there are always alternative places toseek out. This is, after all, the city that sawthe birth of the artistic movement ofDadaism - the antithesis of conformity.

The Extraordinary Meeting 2003 will takeplace at the NOVOTEL, Zurich AirportMesse, which is situated only 5 minutesfrom the Zurich International Airport in aquiet green belt area close to Zurich. TheNovotel is only 12km from the city centreof Zurich, and easily reachable by publictransport.

Key AttractionsGrossmünster The twin towers of this attractive Minster -the largest in Zurich and the city's symbol -face onto the River Limmat and are bestseen from Rathausbrücke. Fraumünster Although this beautiful Minster dates fromthe ninth century (when it was aBenedictine Abbey), it is often the five

20th-century stained-glass windows in thechoir by Marc Chagall (1970) that attractvisitors. These glass works of art are bestseen in the morning light.

Kunsthaus Zürich (Zürich Art Gallery) This is the city's most important art gallery,boasting a collection of paintings and sculp-tures by Swiss and international masters,covering most periods from medieval timesbut predominately from the 19th and 20thcenturies. Two of Monet's Water Liliespaintings and the largest collection ofEduard Munch's works outside Norway canbe found here.

Day trips/Excursions

RapperswilThe beautiful little 'City of Roses' has amedieval character dating back to 1229,with romantic lanes and restaurants on thelakeside. It is located on the northern shoreof Lake Zurich.

JungfraujochFrom this mountainous location in theBernese Oberland one can see Switzerland'slargest glacier and the spectacular Alpinescenery of this region. The cogwheel moun-tain train from Lauterbrunnen climbs to thehighest railway station in Europe at 3454m(11,333ft), on the Jungfraujoch. Bus tripscan be arranged, or alternatively, a train tripwill take about 5 hours, however, the Alpsare always just outside the window.


MEMBERSHIP MEETINGSISTA Extraordinary Meeting 2003

ISTA Extraordinary Meeting 2003The Venue: Zurich, SwitzerlandZurich, Glattbrugg, SwitzerlandJune 30 - July 3, 2003In this part of the world, everything is that bit smaller, but with somany things to do, this simply means that you have all the more timefor an unforgettable visit. Although the largest city in Switzerland,Zurich aptly promotes itself as the 'little big city' and has a historiccentre compact enough to be explored on foot.

Budapest, the capital of Hungary, is an eco-nomic, financial and cultural center withtwo million inhabitants. The city which isbeautifully situated on both sides of theDanube river has a history dating back over2000 years. There are ruins from the time ofthe Roman Empire as well as from theMiddle Ages. Its main characteristicsreflect the atmosphere of the end of the 19thcentury when the millennium of theHungarian State was celebrated. It boasts anumber of museums (picture gallery of themuseums), theatres, concert halls, a lot ofrestaurants and other amenities. Severalbaths and thermal waters of various medic-inal springs are also at the disposal of visi-tors. Budapest can easily be reached by air,train or car

Peaceful and bustling, a big metropolis andyet friendly, it treasures the old andembraces the new. Here the historic blendswith the modern, the hills harmonize withthe river, that is, the Danube, which flowsthrough the city along a stretch of 28 kilo-metres.

Budapest has over 1,000 restaurants offer-ing Hungarian and international cuisine.You will not have any difficulty in finding aplace to eat which suits your taste andbudget.

Development of the today face of Budapestbetween 1870 and 1900 - So fast was thisgrowth that it earned the description of an'American tempo'. However, Budapestresembled Chicago only in the speed of itsgrowth. The development was carefullyplanned and the effect was delightful. In1870 the municipality set up the Council ofPublic Works, which elaborated a grandmaster plan, and the city had the power torealize it. Everything that marked the stan-dards of the age could be found in the mas-ter plan: there was a system of ring roadsand boulevards, and a network of urbanpublic transport: the height of the buildingswas set, green spaces were included, and soforth. Though a major part of the city wasbuilt within the space of twenty years, theresult was not monotony but a harmoniousuniform style.

Up to the first third of the eighteenth centu-

ry the left-bank settle-ment, thehistoric cen-tre of theformer townof Pest, consisted only of the district lyingbetween today's Liberty Bridge, ChainBridge, Múzeum körút and Károly körút,that is to say, it extended only as far astoday's Kiskörút (Little Boulevard). Thetown was completely rebuilt and it grewgradually; today, of the original town ofPest, there only remain some parts of thefifteenth-century town walls, and from theeighteenth century, only the churches, aswell as a few monasteries and public build-ings. In contrast to the Castle District ofBuda, which is by-passed by the main traf-fic of the city, the Inner City, with its shops,offices and important traffic arteries, is partof the city's everyday life.

The city is divided into two parts, the hillyside of Buda on the western bank and theflat plain of Pest on the eastern bank of theriver Danube. These two parts of the citywere once separate towns and were mergedtogether with Ancient Buda (Óbuda) only in1873. The name Buda Castle covers morethan a castle or the Royal Palace in the cap-ital city; it extends to the historical quarterfull of sites. On bright spring days peopleinvite friends for a "walk in the Castle", i.e.to wander around the Castle Hill quarter.The most exiting way of getting to theCastle is by taking the Funicular, a littlecable car up the Castle Hill.

Places of InterestPál Valley Cave (Budapest) This is a cave which was discovered in1904 in the Rózsadomb area of Budapest. Itis a hot spring cave and has been open to the

public since 1919. Though the Pál Valleycave became noted for its exceptional sta-lagmites and stalactites, in reality the nar-row, high, crevice-like passageways, thelarge differences in levels, and the sphereshaped forms resulting from hot water sed-imentation are more characteristic of theplace. The cave has 500 metres of built uppathway along which you see a variety ofstalagmites and stalactites, the hot watersedimentary spheres and ancient shellshaped depressions. A pleasantly land-scaped quarry area in front of the cave givesyou a chance to rest after your labours inthe cave.

Szemlohegy Cave (Budapest) This cave was discovered in 1930 but onlyopened to the public in 1986. This cave isalso the result of hot spring sedimentationand its most characteristic feature is thethick layer of mineral deposit covering thewalls in shapes reminiscent of cauliflowerand grapevine, which are known as peast-ones and are a wonderful sight. The specialclean, dust free air in the cave makes it pos-sible to operate cave therapy courses,thanks to which thousands of asthmaticsand other sufferers of respiratory ailmentsfind relief here.

Alcsút Arborétum (Alcsútdoboz) This is a classical palace belonging to thePrince Palatine and designed by MihályPollack, though sadly only the palacefaçade and palace chapel remain. The garden is an English landscape garden.There are several structures among the hugetrees contemporary with the house, likebeautiful bridges, the music pavilion, thechild's house, the bear house, the water fea-ture, and the Lourdes Cave. The recon-structed chapel has an exhibition entitledHistorical Gardens of Hungary and in thechild's house there is one on the Soldiers ofthe Archduke Joseph. Thanks to the work ofthe owners, the Palatine Joseph and theArchduke, the park today has a collection


MEMBERSHIP MEETINGS27th ISTA Congress

27th ISTA Congress 2004The Venue: Budapest, HungaryMay 13 - 24, 2004

Meier

Archduke, the park today has a collection

1. Papers will be selected according to theiroriginality, either of the methodology or ofthe results. They should be written concise-ly and with clarity.

2. Authors are reminded that acceptance ofa paper at the symposium does not meanautomatic submission to the journal 'SeedScience and Technology'. Authors whowish to submit a symposium paper to 'SeedScience and Technology' should follow theregulations for that journal and submit their

papers independently to the editor.3. For consideration for the symposium,authors should submit their papers in thefollowing format:

(a) Papers should be written in English.

(b) The summary should be headed by thepaper's title and the author's name(s) andaddress(es) and the number of the session tobe presented in. It should be written inEnglish and include information on the

author's aimand majorfindings. Itshould notcontain dis-cussion orr e f e r e n c e sand must not exceed 250 words. All sum-maries will be published prior to the sym-posium. Authors are kindly asked to submitthe summaries as attachment via e-mail oron computer disk either CD or floppy disk


MEMBERSHIP MEETINGSISTA Seed Symposium 2004 - Call for Papers

Intending participants are encouraged topresent papers and posters dealing withseed related topics. For the detailed scien-tific topics of the Sees Symposium, pleaserefer to the information on page 6.

The following should be noted:

1. Papers should be of a high standard andshould follow the usual format for scientif-ic manuscripts with the subject matterarranged under 5 main headings:

- Introduction- Materials and Methods- Results- Discussion- Acknowledgements

(For exact details see the "Instructions toAuthors" below.)

2. A summary of not more than one A4 pageshould be typed separately and should

accompany the manuscript. The summaryshould give a clear indication of the topicscovered and the main findings.

3. Please indicate the session (see topics ofthe Seed Symposium) in which the papershall be presented.

4. Papers will be presented orally or inposter form, both of which have equal sta-tus. As the number of oral presentations willbe limited by time constraints, oral presen-tation of your paper may not be possible,and you may be asked to present your paperas a poster. This decision will be made bythe symposium organisers, and you will beadvised accordingly by the end of January2004.

5. Authors who wish to present a paper inposter format only (i.e. not be consideredfor oral submission) are not required to sub-mit a full text of the paper. Instead theyshould submit a summary only, but indicate

clearly in a covering letter that the paper isfor poster presentation only.

6. Summaries of symposium papers will begiven to all participants at the Congress.Complete manuscripts will be required bysession leaders.

Deadline for Submission:September 30, 2003

Papers must be sent to:

ISTA SecretariatSeed Symposium 2004

Zürichstrasse 508303 BassersdorfCH-Switzerland

Tel: +41 1 838 60 00Fax: +41 1 838 60 01

E-Mail [email protected]

ISTA Seed Symposium 2004Budapest, Hungary, May 13 - 15, 2004

CALL FOR PAPERSThis is the first invitation to people interested in presenting a paper during

the Seed Symposium of the 27th ISTA Congress under the theme

"Towards the future in seed production, evaluation and improvement"

Instructions to AuthorsFor the ISTA Seed Syposium, 2004

31/2

. Suitable formats are as follows:

- Operating system: Windows X- Word processing systems:

Word, WordPerfect and ASCII;

(c) The paper (excluding the summary)should include the title, name(s) andaddress(es) of the author(s), the number ofthe session, the text, tables and figures. Thetext should contain the following:

Introduction should state the main reasonsfor undertaking the work, together with aclear definition of its purpose.

Methods used should be described withenough detail to show how the results wereobtained (i.e. experimental design, treat-ments, operating conditions, materialsused, methods of statistical interpretation).

Results should include statistical informa-tion (i.e. significance of results, coefficientof variation etc.).

Discussion should review the results inrelation to the existing literature, to indi-cate their scientific or practical signifi-cance, and to come to a brief conclusion.

Tables and figures; results may be present-ed either in tables or figures, but in no caseshould information be presented in bothways.

The list of References must contain onlythe papers cited in the text, arranged inalphabetical order of the authors.

4. Authors are reminded that the purpose ofsubmitting their paper is to allow the sym-posium organisers to decide:

(a) Whether the subject material and quali-ty of the paper is acceptable for inclusion inthe ISTA Seed Symposium.

(b) What subject area the paper should beassigned to.

(c) Whether the paper should be presentedorally or in poster form.

The decisions of the organisers are notrelated to publication of the paper, but onlyto its symposium presentation. Therefore,authors may choose to either submit theirpaper in a format which meets all of the'Seed Science and Technology' require-ments, or submit the paper in a formatwhich simply meets the requirements listedin 3(c). If the former option is chosen,authors are reminded that they must still

submit their paper to the editor of 'SeedScience and Technology' for considerationfor publication.

5. To give the symposium organisers timeto decide on the programme and form ofpresentation, and the ISTA Secretariat timeto preprint and distribute all the summaries(oral plus poster) and to prepare a sympo-sium programme (giving paper titles,authors and time of presentation), authorsare requested to meet the following dead-lines:

(a) The text plus a separate summary pagemust reach the ISTA Secretariat bySeptember. Papers received after this datewill not be considered for inclusion in thesymposium.

(b) Authors will be notified of the accept-ance of their paper, the subject area towhich it has been assigned, and whether itis to be presented in oral or poster form, byJanuary 31, 2004

(c) Authors invited to present their papersorally must send the text of the spoken ver-sion of the paper (i.e. what will be said inthe 10 minutes available) to their sessionchairperson and to the local organisers byFebruary 28, 2004. Note that this is the textwhich will be used by the translators.


MEMBERSHIP MEETINGSISTA Seed Symposium 2004 - Call for Papers

TOPICSof the ISTA Seed Symposium,

Budapest, May, 2004

Theme“Towards the future in seed produc-tion, evaluation and improvement"

Session 1:Application of advanced

technologies

Topics: Identification of GM, varietalidentification; diagnostics in plantpathology.

Session 2:Organic and conventional seed pro-

duction

Topics: Seed crop management; harvest;processing; post-harvest technology;seed certification.

Session 3:Viability and vigour: evaluation and

impact

Topics: Methods of evaluation; influenceof seed quality on seed performanceand/or emergence, storage potential,responses to stress.

Session 4:Seed systems in emerging and devel-

oping economies

Topics: Development and status of seedsystems; seed quality management; eval-uation of seed quality; role of informaland formal seed sector.

Session 5:Seed lot hygiene

Topics: Weed contamination, seed-bornepathogens, pests, endophytes, phytosani-tary standards.

Session 6:Seed improvement

Topics: Dormancy breaking; seed sort-ing; priming; chemical and biologicalcontrols; pre-storage treatments; pellet-ing.

Session 7:Physiological basis of

seed quality

Topics: Seed development; desiccation;storage; molecular, cellular and bio-chemical aspects of germination.

Poster Session

Host instituteNational Institute for AgriculturalQuality ControlLocal OrganisersDr. Károly NeszmélyiPresident of the Organising CommitteeDr. Katalin ErtseySecretary of the Organising CommitteeE-mail: [email protected] ConvenorDr. Alison PowellE-mail: [email protected] CorrespondenceISTA Secretariat - Seed SymposiumZürichstrasse 50, CH-8303 BassersdorfSwitzerlandTel: +41 1 838 60 00Fax: + 41 1 838 60 01E-mail: [email protected] SecretariatDiamond Congress Ltd.Fõ u.68., H-1027 Budapest, HungaryTel: +36-1-214-7701, +36 1 214 7698, Fax: +36 1 201 2680E-mail: [email protected]

To put 58.7 million hectares into context, itis more than 5% the total land area of Chinaor the US or almost two and half times theland area of the United Kingdom. Theincrease in area between 2001 and 2002 is12%, equivalent to 6.1 million hectares. Asustained rate of annual growth of morethan 10% per year has been achieved everyyear for the last six years, since their intro-duction in 1996. During the seven-year peri-od 1996 to 2002, global area of transgeniccrops increased 35-fold, from 1.7 millionhectares in 1996 to 58.7 million hectares in2002.

An increasing proportion of GM crops aregrown in developing countries. More thanone quarter (27%) of the global GM croparea of 58.7 million hectares in 2002, equiv-alent to about 16 million hectares, wasgrown in nine developing countries.

In 2002, four principal countries grew 99%of the global transgenic crop area. The USAgrew 39.0 million hectares (66% of globaltotal), followed by Argentina with 13.5 mil-lion hectares (23%) despite the economicsituation, Canada 3.5 million hectares (6%),and China 2.1 million hectares (4%).

Globally, the principal GM crops were GMsoybean occupying 36.5 million hectares in2002 (62% of global area), followed by GMcorn at 12.4 million hectares (21%), trans-genic cotton at 6.8 million hectares (12%),and GM canola at 3 million hectares (5%).

During the six-year period from 1996 to2002, herbicide tolerance has consistentlybeen the dominant trait with Bt insect resist-ance, particularly the European Corn Borer,

second. In 2002, herbicidetolerance, deployed in soy-bean, corn and cotton, occu-pied 75% or 44.2 millionhectares of the global GM58.7 million hectares, with10.1 million hectares (17%)planted to Bt crops, andstacked genes for herbicide tolerance andinsect resistance deployed in both cottonand corn occupying 8% or 4.4 millionhectares of the global transgenic area in2002.

In 2002 for the first time more than half ofthe world's population lived in countrieswhere GM crops are approved and grown.There is cautious optimism that global areaand the number of farmers planting GMcrops will continue to increase in 2003.Currently the development of transgeniccorn lines with new traits has become one ofthe main activities and research interests ofthe agro-industry (International Service forthe Acquisition of Agri-biotechApplications, ISAAA).

The application (import, use and cultiva-tion) of GMO crops as food, animal feedand seeds in Europe and Switzerlandrequires an authorization.

In Europe, the marketing and labelling ofGMO's is regulated by the Novel FoodDirective or by national Food and FeedOrdinances. The Swiss Ordinances, forexample, are based on the detectability ofthe modified gene sequences.

Once approved they have to be labelled ifthe GMO's exceed a fixed threshold. For

Switzerland these thresholds are: 0.5% forseeds, 1% for food, 3% for feed ingredients

and 2% for mixed feeds. In addition, the EUhas planned that additives and flavours con-taining GMO material have to be declaredon the product's label, regardless of theamount used (Amended proposal for aREGULATION OF THE EUROPEANPARLIAMENT AND OF THE COUNCILON GENETICALLY MODIFIED FOODAND FEED, COM (2002) 559 final). Alsolarge groups of consumers, especially inEurope are concerned about the use ofGMO food. In order to comply with manda-tory labelling regulations there is a strongneed to have access not only to specific butalso to quantitatively reliable methods,which are easy to handle.

Therefore, quality as well as safety of seed,food and feed is an issue of growing publicconcern. Public control institutions, privateservice laboratories, and commercial manu-facturers see an increasing need to test forgenetically modified organisms (GMO) inthese matrices.

To fulfil the duty of the Swiss legislation,the Swiss Federal Research Station forAnimal Production established its ownGMO laboratories in 1997. As the GMOlaboratories are accredited by ISO/IEC


ISSUES OF COMMON TECHNICAL INTERESTGenetically Modified Crops

Genetically Modified CropsCurrent situation and some considerations regarding detection techniquesBy Christoph Haldemann, ISTA GMO Task Force Member

Our technician with real-time PCR machines

The cultivation of genetically modified (GM) crops grewextensively all over the world in 2002. The estimatedglobal area of GM crops for 2002 is 58.7 millionhectares, grown by between 5.5 and 6.0 million farmersin sixteen countries - up from 5 million farmers and thir-teen countries in 2001.

Meier

laboratories are accredited by ISO/IEC

17025, all methods have to be validated bya well-defined procedure.

Table 1 shows the methods to detect GMO's,available in our laboratories.Basically we have two types of customers:- External customers, e.g. seed importers,

which send the samples straight to our labo-ratories.- Internal customers, i.e. samples derived

from the official seed and feed control inSwitzerland.

Real-time multiplex PCR (PolymeraseChain Reaction)Since in the last ISTA News Bulletin, No.124, an article about the most commonmethods for detecting GMO's was publishedby Bettina Kahlert, the author needn't gointo further detail in this issue. In addition tothis article there is the real-time multiplexPCR. The fluorescence-based real-timePCR (rt-PCR) is widely used nowadays forthe quantification of DNA and is a criticaltool for basic research, molecular medicineand biotechnology. Assays are relativelyeasy to perform, capable of high throughput,and can combine high sensitivity with reli-able specificity. The technology is evolvingrapidly with the introduction of newenzymes, chemistries and instrumentation.The simultaneous amplification of morethan one DNA sequence (in the same reac-tion tube) is called real-time multiplex PCR.This multiplex DNA-amplification is a veryuseful and powerful technique for the detec-tion of GMO contaminations. All Swissauthorized GMO's contain a 35S-promotorinsert. Therefore we use this technique inour laboratories as a quantitative screeningtest, i.e. to check quantitatively whetherGMO's are contained in the sample. For thisscreening we established a system withwhich we can amplify the 35S-promotorgene and the invertase-gene respectively,the lectine-gene, simultaneously. For moredetailed information visit our homepage:http://www.sar.admin.ch/rap/en/fodder/gvo_method_aug01.pdf.

If the customer wants to know what GMvariety is responsible for a positive finding,the laboratory has to search selectively forthe individual genetic engineering modifica-tions. Thus, further examinations are neces-sary. Table 1 shows all specific GM vari-eties, which can be detected in our laborato-ry at the moment.

Quality managementTaking into account possible DNA degrada-tion, due to food processing involving heator chemical treatments, UV-radiation, etc,we compute the ratio of the amplified trans-genic DNA to the amplified endogenous

DNA. Errors of the endogenous DNAanalysis can affect very much the final resultof the transgenic content of the unknownsample. This ratio is calculated only for seedand feed ingredients. For mixed seeds wetake only the 35S-promotor into account.

To prevent errors and to ensure high qualityof analysis we have to set stringent condi-tions for our results:- If the result of the endogenous DNA is

smaller than 1%, the final result will begiven as: no DNA could be detected.- If the result of the endogenous DNA is

higher than 1% and the result of the trans-genic DNA is higher than 0.02% the finalresult of the transgenic content is calculatedby the ratio of the transgenic DNA to thetotal DNA.- If the result of the endogenous DNA is

higher than 1% and the result of the trans-genic DNA is equal to or smaller than0.02% the final result will be given as: resultunder the limit of quantification.

Further requirements to fulfil the high qual-ity demands and to prevent contaminationmust be carried out by the GMO analysis infour spatial separated laboratories:- For sample grinding- For DNA extraction- For master mix preparation- For DNA amplification

Moreover we successfully take part in sev-eral different Proficiency Tests (PT), such asISTA PT on GMO testing, GeMMA PT(Genetically Modified Material AnalysisScheme), Bipea (Bureau Interprofessionneld'Etudes Analytiques).

Frequency of GMO analysis

In 2002 a total of more than 1500 sampleswere analysed by our laboratories.Approximately 10% of these samples con-

sisted of seeds. As seeds the followingspecies were found:- soybeans- corn- canola- sugar beet- wheat- tomato- barley- flax

The seeds were analysed qualitatively forthe 35S-promotor and for the NOS-termina-tor (terminator of nopaline synthase genederived from Agrobacterium tumefaciens).The 35S-promotor also exists in the cauli-flower mosaic virus, hence, in case of posi-tive results it must also be checked whetherthis might not possibly be the 35s-promotorfrom the virus, due to a contamination or avirus infection. For canola seeds, in additionto the previously mentioned were analysedfor the GT73/RT73 event as well. For allinvestigated seeds there was no result to berejected, i.e. there was no positive result.

Today's challenges for 'GMOLaboratories'

Since the analytical know-how regardingGMO detection is located in a rapidly devel-oping field, one of the main goals for GMOlaboratories is to efficiently develop andestablish further methods for detectingnewly designed GM crops launched on theworld market. In this context access to cer-tified reference material (CRM) is of para-mount importance. Without CRM methodcalibration and method comparison betweendifferent laboratories becomes enormouslydifficult.

Last but not at least I would like to stressanother relevant issue, the DNA extraction.Poor DNA quality, e.g. containing amplifi-cation inhibitors, such as polysaccha-


ISSUES OF COMMON TECHNICAL INTERESTGenetically Modified Crops

Qualitive Methods

Maize BT11 (Novartis)Bt176 (Novartis)MON810 (Monsanto)T25 (Bayer AG)CBH 351 (Aventis)GA21 (Monsanto)

Soybean

Canola GT73/RT73

Quantitative Methods

BT11 (Novartis)Bt176 (Novartis)MON810 (Monsanto)T25 (Bayer AG)Invertase (endogenous gene for maize)

GTS 40-3-2 (Monsanto)Lectine (endogenous gene for soybean)

35S-Promotor

Table 1: Qualitative and quantitative methods to detect specifically different GMspecies, available in our laboratories.

Meier

17025, all methods have to be validated by

Meier

a well-defined procedure.

Meier

inhibitors, such as polysaccha-

The aim of the 1st Proficiency Test was tocheck the ability of the participating labora-tories to detect the presence of GM seeds insamples of conventional seed of Zea maysL.

Each participating laboratory received a setof 30 maize samples. 12 samples were neg-ative (no GM seeds added) and 18 sampleswere positive. Each sample contained about300 seeds. The GMO content in the positivesamples was about 1% (3 in 300). 6 out ofthe 18 samples were positive due toMon810, 6 samples due to T25 and 6 sam-ples due to both (1 seed from Mon810 and2 seeds from T25).

A first communication of results was givenin the ISTA News Bulletin 124.

43 laboratories from 20 countries world-wide (Fig. 1a), from governmental, compa-ny and other private seed testing laborato-ries participated (Fig. 1b).

Each laboratory reported for the individualsample whether this is a negative sample or

a positive sample. So for a given sample theresult reported by the laboratory can beeither correct or false.

30 laboratories (70%) reported for all 30samples correct results and 13 reportedfalse results (30%, Fig. 2). On the laborato-ry basis more laboratories reported falsenegative results for T25 than for Mon810(Fig. 3a). On the sample basis also the per-centage of false negative results reportedwas higher for T25 than for Mon810 (Fig.3b).

The following methods were used: 31 labo-ratories used qualitative PCR; 9 with falseresults. 4 laboratories used real time PCR;1 with false results. 2 laboratories usedqualitative and real time PCR combinedwith correct results. 2 laboratories usedELISA; 1 with false positive results. 2 lab-oratories used Bioassay, both with correctresults. 1 laboratory used qualitative PCRand ELISA combined and reported falsenegative results. 1 laboratory used ELISAand Bioassay combined and reported falsepositive results.

The results of this proficiency test showedthat about 70% of the participants were ableto identify all 30 maize samples correctly.This is a clear indication of the high per-formance of the majority of the participantsto perform both accurate GMO tests for theevents T25 and Mon810 with a spikinglevel of 1% and accurate tests for negativesamples. The remaining 30% showed thewhole range from minor up to severe prob-lems with GMO testing. This clearly indi-cates, the need for further training and stan-dardisation efforts in these laboratories.Thus, in summary, the results provide a firstestimate for the great potential of the per-formance based approach in the field ofGMO testing as developed by ISTA in theGMO position paper. The ratio betweenlaboratories without and with false resultsof 70:30 indicates that, the level of difficul-ty of the 1st Proficiency Test was appropri-ate, and shows the potential for further pro-ficiency tests. In the view of ISTA, as anaim to provide uniform test results all overthe world, 70% correct results is not sufficient and calls for action for improvement.


ISSUES OF COMMON TECHNICAL INTEREST1st ISTA GMO Proficiency Test

Europe(66%)

America(30%)

Africa(2%)

Asia(2%)

Other private laboratories

(5%)

Seed company laboratories

(33%)

Governmental laboratories

(62%)

only correct results (69.8%) false results (30.2%)

false negative results (18.6%)

false positive results (7.0%)

both false negative and positive results

(4.6%)

9.3 5 50

20406080

100

T25 Mon810 T25 andMon810

Events of the sample

Perc

enta

ge o

f sam

ples

(%)

11.611.618.6

020406080

100

T25 Mon810 T25 andMon810

Events of the sample

Perc

enta

ge o

f lab

orat

orie

s (%

)

1st ISTA Proficiency Test on GMO Testing of Zea mays L.By Bettina Kahlert, Proficiency Test Working Group Leader of ISTA GMO Task Force and Michael Kruse, Rules Chapter Working Group Leader of ISTA GMO Task Force

Figure 1b. Status of participating laboratories. Figure 2. Percentage of laboratories reporting only correct resultsand false results.

Figure 3b. Percentage of positive sam-ples with T25, Mon810 and T25 andMon810 which were reported as falsenegative results by the laboratories.

Figure 3a. Percentage of laboratorieswhich reported false negative resultswith positive samples with T25, Mon810and T25 and Mon810.

Figure 1a: Region of participating labo-ratories:

Aim

The aim of this 2nd ISTA Proficiency Test onGMO Testing is to check the ability of indi-vidual laboratories to detect and, on a vol-untary basis, to quantify the presence ofGM seeds in samples of conventional seedof Zea mays L. The object of data analysis will not be toidentify deviating laboratories but to com-pile the performances in the laboratoriesand to provide data for the laboratory´sinternal performance data base.

Sample description

Each participating laboratory will receive10 maize samples. They will be labelledonly with an identification number provid-ed by ISTA. Samples will contain approxi-mately 3000 seeds. This number will not beexact as samples will be prepared on thebasis of an average 1000 seeds weight.Some of the samples will be positive (i.e.contain GM seeds) and the others will benegative (i.e. contain no GM seeds).Laboratories will have to prepare the floursfrom seed samples.

Obligatory qualitative test

A qualitative result is requested.Laboratories can use the method they thinkappropriate for this test. The result for thequalitative test, i.e. a sample is positive ornegative, must be sent back for each samplealong with the sample identification num-ber provided by ISTA. Participants are notexpected to identify events in the positivesamples.

Optional quantification of GMO in posi-tive samples

On a voluntary basis laboratories can do aquantification of the GMO level in the pos-itive samples by either a semi-quantitativetest (sub-sampling strategy) or by a quanti-tative test.

Semi-quantitative test (sub-sampling strate-gy)A semi-quantitative test using the sub-sam-

pling strategy is optional to the participants.The participants shall report as a result ofthis test whether the GMO level in the testsample is above the level of 1% or not.Laboratories can use the method they thinkappropriate for this test. The result for thesemi-quantitative test must be sent back foreach sample along with the sample identifi-cation number by ISTA.

Quantitative test

The true quantitative test is also optional.This quantitative test is for checking theability of the laboratories to quantify theGMO content in a sample. The participantsshall report the GMO level of the test sam-ple. Laboratories can use the method theythink appropriate for this test. The result forthe quantitative test must be sent back foreach positive sample as estimates, alongwith the sample identification number byISTA. Furthermore, a statement on theuncertainty of measurement should bereported. The procedures shall be clearlydocumented. The test procedures will notbe evaluated.

ImplementationParticipation feeThere is a participation fee for non ISTAmembers of 150 US$ for the 2nd ISTAProficiency Test on GMO Testing whichhave to be paid as pre-requirement for

receiving the samples The participation feeof ISTA Members is included in their ISTAmembership fee.

Material Transfer AgreementParticipating laboratories in the 2nd ISTAProficiency Test on GMO Testing are kind-ly requested to sign the Material TransferAgreement with ISTA. It will be not possi-ble to participate without signing it.

Shipping

The shipment you will receive will contain10 samples, each of about 3000 seeds ofZea mays, in total about 10kg. The seedsare not treated. The samples are packed inhermetically sealed plastic bags and theshipment will be labelled as 'seeds fordestructive laboratory tests'. If necessary, aphytosanitary certificate will be attached.

Time schedule

1. Registration February 05, 2003 toApril 18, 20032. Final submission of results to the ISTASecretariat May 16, 20033. Presentation of the overall results andconclusions July 01, 2003 (ISTAExtraordinary Meeting, Zürich)

Thank you for your interest, we are lookingforward to a successful proficiency test.


ISSUES OF COMMON TECHNICAL INTERESTAnnouncement of the 2nd ISTA GMO Proficiency Test

Announcement of the 2nd ISTA GMOProficiency Test GMO Testing of Zea mays L.By Michael Kruse, ISTA GMO Task Force MemberBettina Kahlert, ISTA GMO Task Force Member

GMOs today are in the "heart" of a passionate debate. When speak-ing about GMOs, there are in fact two different aspects that areoften confused: one is the development of genetically modified(GM) varieties and their commercialization and use, the other oneis the adventitious presence of GM material in non-GM varieties.

1. Development of GM varieties1.1. Historical backgroundThe history of plant transformation began with the discoveriesmade by phytopathologists working on soil bacteria responsible forcrown gall and "hairy root" syndrome in the 1960s(1). In 1974 , itwas demonstrated that these "natural" transformations were due tothe transfer of plasmids from bacteria to the plant(2). Using theseplasmids, the first stable expression of a "foreign" gene in a plantwas demonstrated for the first time in tobacco in 1983(3). It was thefirst GM plant.In 1987 the first field trials were planted in France and USA, andin 1994 the first authorization for commercial production of atransgenic variety, a Tomato, was obtained in the USA.1.2 The present situation of transgenic crops in the worldSince 1994 the acreage of transgenic varieties has increased drasti-cally, the technology being probably the one adopted the more rap-idly by farmers in the recent history of agriculture. The evolution isas follows(4),(5). Global Area of Transgenic Crops, from 1996 to 2002

During the seven-year period 1996 to 2002, the global area of trans-genic crops increased by 35 fold, reflecting the growing acceptanceof transgenic crops by farmers in both industrial and developingcountries.

By country, the situation is as follows:Global Area of transgenic crops from 1998 to 2002(millions of hectares)

By crop, the situation is as follows:Global area of transgenic crops from 2000 to 2002(millions of hectares)


ISSUES OF COMMON TECHNICAL INTERESTGenetically Modified Varieties and the Seed Industry

Genetically Modified Varieties and theSeed IndustryBy Bernard Le Buanec, ISF Secretary General

Genetic engineering started in the early 1970s and was at that timelimited to bacteria.But rapidly the technology was extended to the plant kingdom andthe first transgenic plants, tobacco plants, were published in 1983.Since then, the evolution has been very fast and the transgenic plantvarieties, known as GMOs, were grown on more than 60 millionhectares in the world in 2002.

Year

1996199719981999200020012002

Million hectares

1.711.027.839.945.254.260.7

Country

USAArgentinaCanadaChinaBrazilSouth AfricaAustraliaIndiaRomaniaSpainUruguayMexicoBulgariaIndonesiaColombiaHondurasGermanyFrance

Total

2000

30.310.03.00.51.00.20.2--

<0.1<0.1<0.1<0.1<0.1

--------

<0.1

45.2

%

6722712

<1<1--<1<1<1<1<1--------<1

100

2001

35.711.83.21.51.60.20.2--

<0.1<0.1<0.1<0.1<0.1<0.1

----

<0.1--

54.2

%

6622633

<1<1--<1<1<1<1<1<1----<1--

100

2002

39.013.53.52.12.00.30.1

<0.1<0.1<0.1<0.1<0.1<0.1<0.1<0.1<0.1<0.1

--

60.7

%

6422633

<1<1<1<1<1<1<1<1<1<1<1<1--

100

2000

26.810.35.32.8

<0.1<0.1<0.1

45.2

%

5923126

<1<1<1

100

2001

34.99.86.82.7

<0.1<0.1

--

54.2

%

6418135

<1<1--

100

2002

38.512.46.83.0

<0.1<0.1

--

60.7

%

6321115

<1<1--

100

Crop

SoybeanMaizeCottonCanolaSquashPapayaPotato

Total

In terms of value, the market of GM varieties seed was US$ 3.0billion in 2000, increasing to US$ 3.8 billion in 2001. It is estimat-ed at US$ 4.25 billion in 2002, representing almost 15 % of the 30billion value of the global commercial seed market. The marketvalue of the seed of GM varieties is based on the sale price of theseed, plus the technology fees that apply(6).

1.3 Regulatory Issues

1.3.1 Regulations at international level

Discussions are going on at international level on regulation ofGMO and products issued from GMO in several fora as CodexAlimentarius, OECD Seed Schemes, the Convention on BiologicalDiversity (CBD), but except the Biosafety protocol, which is a pro-tocol to the CBD, at present no agreement has been reached.

In summary, the Protocol on Biosafety, known as the CartagenaProtocol, adopted in February 2000, states that transboundarymovement of LMO (Living Modified Organisms) with the objec-tive of intentional introduction into the environment of the import-ing country must be subject to the advance informed agreement ofthat importing country.

Are excluded from that obligation:

- LMOs which are pharmaceuticals for humans that are addressedby other international agreements;- LMOs in transit or for contained use;- LMOs intended for direct use as food or feed, or for processing.

In fact, in the plant domain, the protocol will apply almost exclu-sively to seed for planting.

In addition, LMOs which are subject to intentional transboundarymovement within the scope of the Cartagena Protocol must fulfillspecial handling, transport, packaging and identification:

- LMOs that are intended for food, feed or for processing must beclearly identified as "may contain" LMOs.- LMOs that are intended for intentional introduction into the envi-ronment must be clearly identified as such. The accompanying doc-umentation must specify the identity and relevant traits and/or char-acteristics, any requirements for the safe handling, storage, trans-port and use, the contact point for further information and, asappropriate, the name and address of the importer and exporter; andcontain a declaration that the movement is in conformity with therequirements of the Cartagena Protocol applicable to the exporter.The practicalities of these requirements are presently under discus-sion, in which ISF is involved. The Protocol will enter into forcewhen ratified by 50 countries. Today, 44 have ratified and the entry

into force will likely be in 2004, impacting on seed companies fortheir seed shipment, in particular for the winter nurseries.

1.3.2 Regulations at national level

The situations are very diverse from country to country, not interms of the technical approach of safety assessment, but in termsof general regulations and acceptability.

As regards safety assessment, all the countries where appropriatelaws and committees do exist, the main focus is on environmentand food safety, and for food safety, more particularly on possibleallergenicity. The process is always very comprehensive. It is worthnoting that the procedures are very efficient as, since 1994, no acci-dent has been documented, although LMOs/GMOs are grown ontens of millions of hectares every year and products derived fromGMOs are eaten by hundreds of millions of people every day. Thedisadvantage is that the safety assessment is extremely costly,between 1 and 2 million dollars for crop/trait. The consequencesare that such costs exclude de facto from development small andmedium size companies and small crops. In fact we are here in a"Catch 22" situation: the opponents to the technology claim thatonly large multinational companies are developing new traits forfew major crops but, by their request to add new tests in the safetyassessment procedures they are reinforcing that trend. Whenenough experience is gained in those issues, and we have now 20years of experience, those constraints should be taken into accountwhen designing and implementing the safety assessment proce-dure.

The general regulations on GMOs differ drastically form country tocountry, from a total ban, as in Algeria(7), to a complete liberaliza-tion after the safety assessment has been proved satisfactory, as inUSA or Argentina, by moratorium or de facto moratorium, such asin the European Union. It seems however that there is a global trendto the acceptability of GMOs at world level. The European Unionhas put in place in 2002 legal instruments that should lead to the liftof the de facto moratorium even if some European countries arestill reluctant; most of the African countries are now more interest-ed in the transfer of technology than in the ban of the GMOs(7) andmuch of Asia is rushing forward with the development and cultiva-tion of GMOs(8).

However, the development of compulsory labeling of GMOs inseveral countries lead to a very difficult problem facing the SeedIndustry: the adventitious presence of GM material in non-GMvarieties.

The Adventitious Presence of GM material in non-GM varieties iscontinued on page 14.



20 kg450 gr

20 kg80 M units

50 kg50 pounds

Bt100 US$1400 Roupies

Bt100 US$103 US$

RR20 US$22 US$

Conventional30400

Conventional8080

Conventional1713

Technology fee70 70%

1000 71%

Technology fee20 20%23 22%

Technology fee3 15%9 41%

CottonArgentinaIndia

MaizeArgentinaUSA

SoybeanArgentinaUSA

Examples of Seed Prices for GMO’s and “conventional” Crops

2. The Adventitious Presence of GM material in non-GM varieties

2.1 Background

The adventitious presence of GM material in seed, food and feed isthe unintended occurrence of plant material from crops improvedthrough biotechnology. It is of course the logical and unavoidableconsequence of the development of GMOs at world level. Thatadventitious presence may occur through natural pollen flow orfrom commingling that occurs in the production/distribution sys-tem.

Some countries, that are labeling GMO food and food ingredients,have defined thresholds above which the products should belabeled as GMOs. For instance in Europe the trigger point is 1%, inJapan 5% and in Brazil 4%. (Incidentally it is interesting noting thehuge variations as regards the thresholds chosen, showing clearlythat they are not based on scientific information but on politicalposturing). As early as 1999, FIS and ASSINSEL realized thatlabeling of GMOs and consequent demand for separation of GMand non GM products would have impact on the seed trade. Duringtheir Melbourne Congress in June 1999, the following motion wasadopted:

FIS and ASSINSEL Urge Countries to Adopt Practical Seed GMO Thresholds

"FIS and ASSINSEL consider that when the environmental andnutritional safety of GMOs and GMO products have been positive-ly assessed, there is no scientific reason to separate these productsfrom the non-GMO ones.

However, for various reasons, some markets are demanding sepa-ration of GMO and non-GMO products.

Whereas there is a link between the quality of seed and productsderived therefrom; and

Whereas new genetic and analytical technologies (e.g., PCR ampli-fication of DNA) have made it possible to detect minute amountsof all off-types; and

Whereas the world's seed industry has established and observesstandards and management practices that result in seed products ofconsistently high quality in which the presence of off-types is lim-ited to an agreed upon standard;

FIS and ASSINSEL support seed quality standards with practical,defined seed GMO thresholds for adventitious genetic off-typesfrom any source, that are consistent with current seed productionpractices and industry standards on a crop by crop basis."

In August 1999, FIS launched an international initiative to dealwith this issue, which was known at the end of that year as theInternational Seed Network Initiative, ISNI, with FIS, OECD,ISTA and AOSA. That initiative had three main objectives:

- Review the production processes to minimize the adventitiouspresence by pollen flow and commingling and if necessary todesign new quality insurance charts.- Establish laboratory's standards for assessing the presence of GMmaterial in non GM product.- Fix an experimental threshold of 1%, globally recognized.

Unfortunately, it was not possible to reach an agreement on thatproposal and after 3 years of discussions, the principle of such an

experiment has been abandoned. Discussions are continuing with aless exhaustive approach on possible thresholds and seed testing.They have shown that the issue was technically complex and polit-ically sensitive.

The Adventitious Presence issue can be divided in 3 main cate-gories:1) Events that have been approved for feed.2) Events that have been/will be approved for pharmaceutical uses.3) Events that have been approved for food.

There is an agreement that Adventitious Presence of events of thefirst two categories should not be accepted at all. It is very unlike-ly that, after the Star Link experience, companies will put on themarket food crops events approved for feed only and the risk ofA.P. is almost virtual.

The events for pharmaceutical uses deserve a special attention andthe industry is taking the matter extremely seriously. There are dis-cussions at the moment within the agro-food industry on the possi-ble ban of "pharmaceutical events" in food crops.

As regards the events that have been approved for food, three mainsubcategories may be established:1) The adventitious presence of GM seed, of events that are domes-tically approved.2) The adventitious presence of GM seed, of events that are notdomestically approved but have been approved in another countrythat employs OECD safety assessment process.3) The adventitious presence of GM seed, of events which are stillin the research stage and have received approval for field tests inany country member of the OECD Seed Schemes.

Categories 1 and 2 are of primary importance with regard to adven-titious presence. However, adventitious presence of event of cate-gory 3 also need to be taken into account, even if it is extremelyunlikely.

Where are we at the moment ?

2.2 Thresholds

2.2.1 At international level

The discussions are going on at OECD level. The position of ISF isthat zero tolerance is unachievable in practice and that a 1%threshold for event approved, including approved field trials, in anyof the member countries of the OECD Seed Schemes is a reason-



Incidentally it is interesting noting the hugevariations as regards the thresholds chosenabove which the products should be labeledGMOs, showing clearly that they are notbased on scientific information but on politi-cal posturing. However, the lack of harmo-nization at international level is creating newtrade barriers. A science-based approachshould facilitate the development of harmo-nized international regulations.

able transitional solution taking into account seed production real-ities and the absence of documented risks. Despite several meet-ings, no agreement has been reached for two main reasons:

- All the countries involved in the discussion are reluctant toinclude events which have not been approved by themselves. Ofcourse that position is much less damaging in countries wheremany events are approved like for instance North America andJapan than in countries like Europe where very few events areapproved due to the current de facto moratorium.- There is no agreement on the threshold level, some countries sup-porting ISF position, as USA, Argentina, Canada, Australia, …,some asking for lower thresholds, 0%, 0.3% or 0.5%, as theEuropean Union.

It is very unlikely that an agreement can be reached in the nearfuture.

2.2.2 At national level

Few regulations have been put in place. In Switzerland, the provi-sional level is 0.5% for events approved in the country and in theEuropean Union. It is an interesting step towards the acceptance ofdecision from other countries but the threshold is rather low. InArgentina, the threshold is 1%. In New Zealand, for sweet corn itis 0%, tested according to an official protocol.

In the European Union the existing official proposal states that:

- Genetically modified seed not having authorization for the plac-ing on the market shall not be present- Genetically modified seed having received authorization shall notexceed the standards laid down for varietal purity and, in any case,not more than:

· 0.3% for seed of a cross-pollinating species or variety, other than maize.

· 0.5% for seed of maize.· 0.5% for seed of a self-pollinating species or variety

other than field peas and soybean.· 0.7% for seed of field peas, or soybean.

Internal discussions among countries member of the EuropeanUnion are not yet completed.

ISF stresses that in fixing thresholds countries have to take intoaccount the situation where almost all the crops would be GMOs asin some areas in the world at the moment. In that case a practicalthreshold should be the average percentage of off-types which arepresent at the moment in commercialized seed.

2.3 Seed testing

Seed testing comprises two different parts: seed sampling anddetection of presence of GM material. Scientifically it should notbe too difficult to resolve that problem. However, from a practicaland economical point of view, detection methods remain a majorissue. This point will have to be taken into consideration whenthreshold and detection methods are proposed, while maintainingreliable methods and providing valuable information.

- Seed Sampling

Several organizations are working on this issue, including the ISTAGMO Task Force, AFNOR in France, CEN and ISO and probablyothers. At the moment there is no rule globally adopted but thework done by CEN and ISO should be finalized shortly.

- Detection

A variety of methods, both qualitative and quantitative, are cur-rently used in government, company and third party labs.Unfortunately, for many of these methods overall performanceparameters have not yet been established and well-characterizedreference material, with documented purity measurements, are ingeneral not available for validation studies.

Several organizations are working on detection protocol such as,for example, GIPSA in USA and JRC in Europe.

For seed, a first ring test was run by ISF in the year 2000, but dueto the low level of participation, the results were not significant.

ISTA, in the year 2000, has established a "GMO Task Force" withthe participation of other actors of the international seed chain: ISF,AOSA and ISST.

In 2001, ISTA adopted a "Strategy regarding Methods for theDetection, Identification and Quantification of GeneticallyModified Seeds in Conventional Seed Lots"(9), with in particular anew performance-based approach. In the frame of this newapproach, proficiency tests are performed. In 2002, 43 test series by41 laboratories on the traits T25 and Mon810 have shows thatprogress has to be made, as 13 series reported false results, i.e.30%. A new test should be done in 2003.

It is important that only results obtained by laboratories duly rec-ognized/accredited by national authorities be taken into considera-tion for regulatory purposes.

3. Conclusion

GMOs are expanding rapidly at a global level, with differencesbetween countries. This development is creating new opportunitiesfor the seed industry.

However, the lack of harmonization at international level is creat-ing new trade barriers and, in particular, the possible adventitiouspresence of GM material in non-GM seed has already caused somedisruptions in the international seed trade. A science-basedapproach should facilitate the development of harmonized interna-tional regulations.

References (1) Goldman A, Tempé J, Morel G: Quelques particularités de diverses souch-es d'Agrobacterium tumefaciens, CR Soc. Biol. 1968; 162: 630-1(2) Van Larebeke N, Engler G, Holsters M, Van den Elsacker S, Zaenan I,Schilperoort RA, Schell J. Large plasmid in Agrobacterium tumefaciensessential fro crown gall-inducing ability. Nature 1974; 252: 169-70.(3) Herrera-Estrella L, Deblock M, Messens E, Hernalsteens JP, Van MontaguM, Schell J. Chimeric genes as dominant selectable markers in plant cells.EMBO J 1983; 2: 987-95.(4) James C: Global Status of Commercialized Transgenic Crops, 2002;Preview. ISAAA Briefs No. 27. ISAAA, Ithaca, NY.(5) For Brazil, estimation from the author based on information received fromBrazilian Scientists and Agronomists.(6) Estimation from the author based on information from the seed industry inthe relevant countries.(7) Information given by the governments during the 9th regular session of theFAO Commission on Genetic Resources for Food and Agriculture, Rome,Italy, 14-18 October 2002.(8) Genetically modified crops are sprouting throughout Asia, InternationalHerald Tribune, p. 1, 22-23 February 2003.(9) Position paper on ISTA's Strategy regarding Methods for the Detection,Identification and Quantification of Genetically Modified Seeds inConventional Seed Lots, Approved by the Executive Committee of ISTA, 14November 2001.




The Asia and Pacific Association(APSA) is the largest regional seedassociation in the world. Active mem-bership of over three hundred provideenough strength but diverse agricultur-al, socio-economic, and environmentalsituations prevailing in the 30 membercountries offer several challenges.While the seed industry is quite organ-ized in some developed countries suchas Australia, New Zealand and Japan, itis at various stages of development inothers It is important to understand thatAsia-Pacific region shares 23 percent ofglobal land area and nearly 56.6 per-cent of world population. Countrieswith large land area include China,Australia, India, Indonesia, Iran andMongolia.

China, Japan, India and Australia sharesubstantially world commercial seedmarket, while the seed industry is devel-oping fast in other countries of theregion. During past few years, APSAhelped the seed industry considerably inunderstanding market information andarranged trainings and visits for itsmembers. This has helped privatizationand the industry is slowly entering themainstream of global seed business.

APSA developed technical and marketrelated information and a number ofsuch reports are available with us. Theseinclude the following:· Technical reports on new tech-

nologies.

· Country reports of the seedindustry in the region.· Information on seed produc-

tion, conditioning, storage, qualityassurance and marketing.· Information on IPRs and other

related issues.· Seed import/export statistics

for different countries in the region.

The above information is shared withour members and a part of it is also pub-lished in our journal - Asian Seed andPlanting Material and can also beaccessed through our website:http://www.apsaseed.com

APSA has all the national seed associa-tions in the region as its members andwe interact with them regularly. In addi-tion, we also interact on issues of mutu-al interest with other regional seed asso-ciations such as AFSTA, ASTA, andFELAS.

The main issues facing the seed industryin the region include:

¨ Increasingly difficult access toavailable germplasm due to uncertaintyon IPR issues.¨ Phytosanitary regulations

being used as non-transfer barriers,often clogging free flow of seed attransnational level. ¨ Seed quality assurance need to

be improved and ISTA 's assistance iscrucial.

¨ PVP/patents are not yet inplace in several countries as a result ofwhich seed trade is not expanding at thedesired pace.¨ Inadequate investments in

agbiotech research and limited informa-tion on transgenics may be critical forthe adoption of GM seeds.

In the Asia and Pacific region, agricul-ture is the mainstay of the economies inthe majority of countries. It is a paradoxthat as to date a big gap still existsbetween demand and supply of qualityseeds in the entire region. There is alsoan urgent need to improve seed quality.In this effort, assistance of ISTA willundoubtedly be of great significance.

ISSUES OF COMMON TECHNICAL INTERESTAPSA Marches Forward

APSA Marches ForwardThe Asia & Pacific SeedAssociationBy J.S. Sindhu, APSA Director

In the Asia and Pacific region, agriculture is the mainstay of theeconomies in the majority of countries. It is a paradox that as todate a big gap still exists between demand and supply of qualityseeds in the entire region. There is also an urgent need to improveseed quality. In this effort, assistance of ISTA will undoubtedly be ofgreat significance.

The Asia and PacificSeed Association

Kasetsart Post OfficeP.O. Box 1030

Bangkok 10903TH-Thailand

Phone +66 2 940 5464Fax +66 2 940 5467

[email protected]


Presented at the 4th ISTA PDC Symposiumon Seed Health. Wageningen, TheNetherlands, 29 April - May 1 2002

Abstract

The International Plant ProtectionConvention (IPPC) is a legally bindinginternational agreement deposited withFAO. The purpose of the Convention is "tosecure common and effective action to pre-vent the spread and introduction of pests ofplants and plant products, and to promoteappropriate measures for their control".Although the IPPC clearly has strong rele-vance to the regulation of trade, theConvention is not limited in this respect.International cooperation in many forms ofplant protection may fall within the scopeof the Convention. Likewise, plants are notlimited to cultivated plants and protection isnot limited to direct damage from pests.Therefore, the scope of the Conventionextends to the protection of natural floraand includes indirect harm or damage frompests such as weeds. The 1997 revision ofthe IPPC, modernised the Convention andbrought it in line with the WTO Agreementon the Application of Sanitary andPhytosanitary Measures (SPS Agreement).This revision process included the estab-lishment of the Secretariat for the IPPC(based in FAO), the Commission onPhytosanitary Measures (CPM), and theformalisation of the standard settingprocess. The SPS Agreement specificallynominated the IPPC as the formal bodyresponsible for the setting of InternationalStandards for Phytosanitary Measures(ISPMs). ISPMs are used to harmonisephytosanitary measures that will facilitatetrade and minimise unjustified phytosani-tary barriers to trade. Should an internation-al standard not be available, phytosanitarymeasures must be based on a pest riskanalysis.

The entry into force of the SPS Agreementand the reorganisation and changes inemphasis in the New Revised Text of theIPPC (1997), have resulted in countriesneeding to significantly review nationallegislation and phytosanitary capacity tomeet domestic and international obliga-tions. Given the large discrepanciesbetween the developed and developingcountries developing countries are in needof substantial capacity building, includingthe development of institutional capacity,legislative frameworks, and technicalexpertise and capacity. A component of thisincludes the use of and training in theunderstanding and implementation of inter-national standards for phytosanitary meas-ures. In this context, the ISTA seed healthstandards could play a more meaningfulrole in future, if a way is found to formallyinclude them in the international process.Expertise within the framework of ISTAcould also be utilized more effectively inthis capacity building process.Historical BackgroundThe International Plant ProtectionConvention (IPPC) is a multilateral treaty

for cooperation in plant protection that hadits beginnings with the agreement by twelvecountries to regulatory measures forgrapevines under the PhylloxeraConvention of Berne in 1881. This repre-sented the first efforts at formalizing inter-national cooperation in plant protection andled to the recognition of the need to addressother plant pests and enlist cooperationamong all countries.

The first text of an international conventionwith broader objectives was drafted at theInternational Conference for PlantProtection held in Rome in 1929. After along lapse due to a world war, the draft textwas again brought to the attention of gov-ernments. This time, the forum was theyoung FAO in its Third and Fourth Sessionsof Conference in 1947 and 1948 respective-ly. In 1951, the Sixth Session of FAOConference adopted the Convention and itwas deposited with the Director General ofthe Organization shortly thereafter. TheConvention first came into force in 1952after ratification by three signatory govern-ments; Ceylon, Spain, and Chile.

Amendments to the IPPC were proposed in1973. After a series of consultations, mem-bers agreed upon modifications thatinvolved updating terminology and describ-ing certain changes in the model phytosani-tary certificates. The amendments wereadopted by FAO in 1979. The Revised Textof the Convention came into force in 1991following acceptance of the amendments bytwo-thirds of the contracting parties.

In 1986, the landscape began to change sig-nificantly as the General Agreement onTariffs and Trade (GATT) entered into aneighth round of multilateral trade negotia-tions known as the Uruguay Round. Prior tothe conclusion of these negotiations in1993, it was clear to IPPC Members and

ISSUES OF COMMON TECHNICAL INTERESTThe International Plant Protection Convention (IPPC)

The International Plant ProtectionConvention (IPPC)The Harmonisation Of International Phytosanitary MeasuresThrough The Use Of ISPMs And The Need For CapacityBuilding Under The IPPCBy D.C. Nowell, and R.L. Robert, IPPC Secretariat

The International PlantProtection Convention

(IPPC) is a legally bindinginternational agreementdeposited with FAO. The

purpose of the Convention is"to secure common and

effective action to prevent thespread and introduction ofpests of plants and plantproducts, and to promoteappropriate measures for

their


FAO that the IPPC would have a prominentposition in the Agreement on theApplication of Sanitary and PhytosanitaryMeasures (the SPS Agreement). The roleenvisioned for the IPPC was to encourageinternational harmonization and elaborateinternational standards to help ensure thatphytosanitary measures were not used asunjustified barriers to trade.

In response, FAO established a Secretariatfor the IPPC in 1992, followed by the for-mation of the Committee of Experts onPhytosanitary Measures (CEPM) in 1993.The Secretariat immediately began anambitious program of standard setting. Atthe same time, interest mounted amongIPPC Members to amend the Convention tomore accurately reflect the contemporaryrole of the Convention, particularly withrespect to the relationship of theConvention to the SPS Agreement.

Negotiations for revision started in 1995and were finalized in November, 1997when the 29th Session of FAO Conferenceapproved the New Revised Text (NRT) ofthe IPPC. The amended IPPC now requiresratification by two-thirds of its contractingparties to come into force.

The IPPC is an international treaty, deposit-ed with FAO and administered by FAO butimplemented through the cooperation ofmember governments and Regional PlantProtection Organizations (RPPOs). TheIPPC currently has 117 contracting parties(members), and 40 countries have alreadyadhered or accepted the NRT of the IPPC.Contracting parties to the IPPC includesmost major trading partners except Chinaand the European Community (the individ-ual Members of the European Communityare all Members and the EC will be eligiblefor membership when the NRT of the IPPCcomes into force.

Objectives and Scope

The purpose of the Convention is "to securecommon and effective action to prevent thespread and introduction of pests of plantsand plant products, and to promote appro-priate measures for their control".

Although the IPPC clearly has strong rele-vance to the regulation of trade, theConvention is not limited in this respect.International cooperation in many forms ofplant protection may fall within the scopeof the Convention. Likewise, plants are notlimited to cultivated plants and protection isnot limited to direct damage from pests.Therefore, the scope of the Conventionextends to the protection of natural flora

and includes indirect harm or damage frompests such as weeds.

The role of the Convention with respect totrade has changed significantly in recentyears as reflected in the substantial amend-ments found in the New Revised Textapproved in 1997. In addition to describingthe fundamental elements of national plantprotection organizations, the IPPC is nowalso an active organization stronglyinvolved with international standard set-ting.

Organizational Structure

From 1951 to 1992, the IPPC existed as aninternational agreement administeredthrough FAO. Beginning in 1986, activitiesunder the Convention began to be coordi-nated through annual TechnicalConsultations with RPPOs organized byFAO.

In 1992, FAO established a Secretariat forthe IPPC followed by the formation of theCommittee of Experts on PhytosanitaryMeasures (CEPM) in 1993. The Secretariatis charged specifically with the coordina-tion of the work programme of the IPPC,particularly the elaboration of InternationalStandards for Phytosanitary Measures(ISPMs). The work was supported by theCEPM, a group of international expertswhich met annually to review and commenton the suitability of documents prepared bythe Secretariat. This expert group has nowbeen superseded by the Interim Standards

Committee which meets biannually for thesame purpose.

The New Revised Text of the Conventionmakes provision for the formation of aCommission on Phytosanitary Measures. Inadopting the New Revised Text, the 29th

Session of FAO Conference approved sev-eral interim measures, including the estab-lishment of an Interim Commission onPhytosanitary Measures (ICPM) until theamendments come into force and a perma-nent Commission is in place. Because theICPM is authorized and organized under

provisions of the FAO Constitution andBasic Texts, it is open to all FAO Members.When the amended Convention comes intoforce, the permanent Commission will becomprised of the contracting parties of theIPPC.

Key National Obligations

In terms of the IPPC, National PlantProtection Organizations (NPPOs) must beestablished and administered by contractingparties. Additional, key obligations are therequirement to nominate an identify officialcontact point, conduct treatments and certi-fy exports, participate in the developmentof, and observe / adhere to adopted, ISPMs.Technical assistance / capacity building isalso a basic need in the IPPC and countriesare encouraged to participate within theircapacity and resources.

Key Activities Within the IPPC

The key activities within the IPPC frame-work are:i) the international phytosanitary standardsetting (ISPMs) process;ii) the development of pest risk analysis forvarious classes of pests e.g. quarantinepests and regulated non-quarantine pests;iii) the establishment of a technical disputeresolution process;iv) the facilitation of official phytosanitaryinformation sharing (including legislative /regulatory information, conduct surveil-lance of/for pests, ports of entry, NPPOstructure, pest distribution, pest lists, pestoutbreaks, official national contact points,regulations, non-compliance, disputes, andobligations under 1997 IPPC); andv) providing and facilitating technicalassistance / capacity building given thelarge needs of developing countries giventhe 1997 revision of the IPPC.

The International Standard SettingProcess

Draft standards may be developed by theSecretariat or submitted to the Secretariatfrom any source. Nomination of topics forISPM development may come from theICPM, NPPOs, RPPOs, the IPPCSecretariat, or from another source via theIPPC Secretariat. The topic are finalised,and priorities established, at the annualmeeting of the ICPM. In general, standardshave their origin in national or regional ini-tiatives, and/or are drafted by expert groupsorganized by the Secretariat based on thetopics and priorities identified by the ICPMand specifications agreed by the StandardsCommittee (SC). Draft standards that havebeen reviewed and accepted by the SC arecirculated to IPPC member governments


The ISTA seed health stan-dards could play a moremeaningful role in future, if away is found to formallyinclude them in the interna-tional process.

Meier

circulated to IPPC member governments


for consultation before being finalization ofthe draft standard by the SC for submissionto the Commission for possible adoption.IPPC standards fall into three categories:reference standards; concept standards; andspecific standards. The IPPC has producedprimarily reference and concept standardsthat provide the foundation for specificstandards that may follow. The number andfrequency of standards under developmenthas a direct relationship to the resourcesavailable to the Secretariat and the technicalcomplexity of the issues being addressed.

ISPMs completed and the year of theiradoption:

No. 1: Principles of plant quarantine asrelated to international trade, 1995No. 2: Guidelines for pest risk analysis,1995No. 3: Code of conduct for the import andrelease of exotic biological control agents,1995No. 4: Requirements for the establishmentof pest free areas, 1995No. 5: Glossary of Phytosanitary Terms,1996 (revised 2002)No. 6: Guidelines for surveillance, 1997No. 7: Export certification system, 1997No. 8: Determination of pest status in anarea, 1998No. 9: Guidelines for pest eradication pro-grammes, 1998No. 10: Requirements for the establish-ment of pest free places of production andpest free production sites, 1999No. 11: Pest risk analysis for quarantinepests, 2001No. 12: Guidelines for phytosanitary cer-tificates, 2001No. 13: Guidelines for the notification ofnon-compliance and emergency action,2001No. 14: The use of integrated measures ina systems approach for pest risk manage-ment, 2002No. 15: Guidelines for regulating woodpackaging in international trade, 2002No. 16: Regulated non-quarantine pests:concept and application, 2002No. 17: Pest reporting, 2002

Possible role for ISTA in the ISPM stan-dard setting process

In future it is possible that ISTA may beable to play a role in this formal IPPC inter-national standards setting process.However, any procedure or informationsubmitted, through the process outlinedabove, would be submitted to the normalfull review process of the IPPC i.e. the factthat it may have been approved by ISTAmembers will have no bearing on the

process or procedure within the IPPC stan-dard setting process. This also means thatISTA may want to consider ways ofincreased official government involvementand possibly expend their membership tofacilitate this process. In addition, ISTAprobably needs to find ways of improvingcooperation and coordination with RPPOs.The benefits to ISTA would be internation-al acceptance as an ISPM that has legal sta-tus within the WTO process.

To start the process of cooperation it wouldalso be advisable to concentrate of pests ofquarantine importance (e.g. Erwinia stew-artii) and regulated non-quarantine pests(e.g. Xanthomonas in beans). These wouldbe of particular interest to the IPPC con-tracting parties and are more likely to besupported by RPPOs.

Capacity building

Capacity building is crucial to the imple-mentation of the IPPC, particularly fordeveloping countries. For this reason thereis a large on-going technical assistance pro-gramme from FAO and others donors thatcovers:i) infrastructure and institutionalization(including policy and legislation, training,implementation of the ISPMs, and equip-ment / facilities);ii) dispute avoidance and resolution; andiii) pest emergencies.

Opportunities for ISTA in CapacityBuilding

There are also potential opportunities forISTA in the area of capacity building. Thisincludes possibly participating in the imple-mentation of relevant specific standards,and in relevant training programmes devel-oped by donor agencies. In addition, theISTA community could act as a valuablesource of technical experts and advice inrelevant projects. However, for this to mate-rialise there is a greater need for communi-cation, improved networks and feedback,and a possibly increased participation inprojects and project development. This is agoal that our oganizations can worktowards.

Conclusions

In conclusion it can be noted that the IPPCis the tool for harmonisation of internation-al phytosanitary measures through the useof ISPMs, there are significant capacitybuilding needs in developing countries; andISTA could probably play a more meaning-ful role in both these areas if they desired.However, should this be desirable, such

partnerships, cooperation and coordinationneed to be developed carefully in the nearfuture.References

FAO. (1995). Principles of plant quarantine asrelated to international trade. IPPC Secretariat,FAO, Rome, Italy.FAO. (1995). Guidelines for pest risk analysis,1995. IPPC Secretariat, FAO, Rome, Italy.FAO. (1995). Code of conduct for the importand release of exotic biological control agents,1995 IPPC Secretariat, FAO, Rome, Italy.FAO. (1995). Requirements for the establish-ment of pest free areas, 1995 IPPC Secretariat,FAO, Rome, Italy.FAO. (1996). Glossary of Phytosanitary Terms,1996 (revised 2002) IPPC Secretariat, FAO,Rome, Italy.FAO. (1997). Guidelines for surveillance, 1997IPPC Secretariat, FAO, Rome, Italy.FAO. (1997). Export certification system, 1997IPPC Secretariat, FAO, Rome, Italy.FAO. (1998). Determination of pest status in anarea, 1998 IPPC Secretariat, FAO, Rome, Italy.FAO. (1998). Guidelines for pest eradicationprogrammes, 1998 IPPC Secretariat, FAO,Rome, Italy.FAO. (1999). Requirements for the establish-ment of pest free places of production and pestfree production sites, 1999 IPPC Secretariat,FAO, Rome, Italy.FAO. (2001). Pest risk analysis for quarantinepests, 2001 IPPC Secretariat, FAO, Rome,Italy.FAO. (2001). Guidelines for phytosanitary cer-tificates, 2001 IPPC Secretariat, FAO, Rome,Italy.FAO. (2001). Guidelines for the notification ofnon-compliance and emergency action, 2001IPPC Secretariat, FAO, Rome, Italy.FAO. (2002). The use of integrated measures ina systems approach for pest risk management.IPPC Secretariat, FAO, Rome, Italy.FAO. (2002). Guidelines for regulating woodpackaging in international trade. IPPCSecretariat, FAO, Rome, Italy.FAO. (2002). Regulated non-quarantine pests:concept and application. IPPC Secretariat,

FAO, Rome, Italy.


IPPC Secretariat, Plant Protection Service,

Plant Protection and ProductionDivision,

Agriculture Department, Food and Agriculture Organization

of the United Nations, vialle Delle Terme di Caracalla,

00100 Rome, Italy. Correspondence:

[email protected]

Meier

for consultation before being finalization

Meier

the draft standard by the SC for submission

Meier

to the Commission for possible adoption.

Meier

finalization of

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need to be developed carefully in the near

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future.

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partnerships, cooperation and coordination

Meier

This end of the paragraph should be at the bottom of the 2nd collumn. The 3rd Collumn should start with references.


Seed is the basic unit of production for theworld's food crop. In recent years seed hasbecome an international commodity usedto exchange germplasm around the world.Seed is, however, also an efficient meansof introducing plant pathogens into newareas as well as providing a means of sur-vival for pathogens from one cropping sea-son to another. Testing seeds for germina-tion and purity as a measure of seed quali-ty has been a universal practice since thefirst seed testing station was established byNobbe in Thrant Saxony in 1869. Nobbementions smut balls and sclerotia in con-nection with seed in his 1876 publicationSamenkunde but does not describe anymethod for their detection except visualexamination of the seed. The publication ofthe Manual for Determination ofSeedborne Diseases in 1938 by Dr. L. C.Doyer was the first to describe repro-ducible methods for detection and identifi-cation of fungi, bacteria, nematodes andinsects associated with seed.

Today seed health testing is routinely car-ried out in most countries for domesticseed certification, quality assessment andplant quarantine. Methods for seed healthtesting often vary from one laboratory toanother. In 1957 the Plant DiseaseCommittee established a comparative seedhealth testing program aimed at standardiz-ing techniques for the detection of seed-borne pathogens. The basic principle ofthe comparative testing program involvedthe distribution of uniform samples to anumber of laboratories working independ-ently followed by annual workshops inwhich the results were compared. Theobjective was to develop and standardizesimple methods suitable for application onseed lots moving internationally.

Diagnostics in research and in routine seedhealth testing have similar objectives buthealth tests must be completed within alimited time span and standardisation isnecessary to achieve results, which areconsistent when applied in practice. Thereare six main requirements for seed healthtests.1. Specificity: the ability to distinguish thetarget pathogen from all organisms likelyto occur on seeds from field or store, i.e. toavoid false positives.2. Sensitivity: the ability to detect organ-isms, which are potentially significant infield crops at a low incidence in seedstocks. 3. Speed: in some cases, small concessionsto accuracy may be necessary to ensurerapid results, but such results should be fol-lowed by more definite testing.4. Simplicity: the methodology shouldminimize the number of stages to reduceroom for error and to enable tests to be per-formed by not necessarily highly qualifiedstaff.5. Cost effectiveness: test costs shouldform part of acceptable production marginsfor each crop.6. Reliability: test methods must be suffi-ciently robust so that results are repeatablewithin and between samples of the samestock regardless of who performs the test(within the bounds of statistical probabilityand sample variation).

Historically seed health tests have beenclassified into four distinct groups based onthe general techniques used to observe thetarget pathogen these are (1) DirectInspection of dry seed or examination ofsuspensions obtained from washings of theseed; (2) Incubation tests, Blotter or AgarPlate; (3) Staining and examination of theembryo; and (4) Immunoassays andMolecular Techniques.

Direct InspectionSeed samples may be examined dry for thepresence of ergots, other sclerotia and smutballs without or with a stereomicroscope.The sample may be immersed in water oranother liquid to make fungal fruiting bod-ies, for example pycnidia, or symptomsmore visible or to encourage the liberation

of spores. After immersion, the seeds areexamined by means of a stereomicroscope.Seeds may be immersed in water contain-ing a wetting agent or in alcohol and shak-en to remove fungal spores, hyphae, etc.attached to or carried with the seeds.Excess liquid is then removed and theextract is examined at a higher magnifica-tion using a compound microscope.Although seed inspection methods are use-ful for determining the incidence of easilyvisible and relatively superficial fungi orfungal bodies, they give no indication oftheir viability.

Incubation Tests: Agar PlateThe agar test gives an indication of theviable inoculum present in an infected seedsample. This is done by placing seeds ontosterile agar, potato dextrose or malt agarsare most commonly used to encourage thegrowth of seedborne fungi. Plates are incu-bated at 20°C in the dark for 7 days whenthe characteristic growth form (mycelium)of the fungus can be identified by eye orusing a low or high power microscope.Near-ultraviolet light (NUV) may be usedduring incubation to encourage the devel-opment of fruiting bodies. Seeds are spacedon the Petri dishes to avoid cross-contami-nation. There are many variations of theagar test. Acidic agars may be used toreduce bacterial contaminants agars maybe made semi-selective by the addition ofspecific chemicals and/or antibiotics and/orfungicides.

Incubation Tests: Blotter TestsThe blotter test gives an indication of theinfection of the seed, as shown by the pres-ence of mycelium and fruiting bodies, and,in some tests, infection of the germinatedseedlings as demonstrated by symptoms onthe young plants. Standard detection meth-ods vary depending on which fungus isbeing tested. For example, in Phaseolusbeans, 400 seeds per sample are placedbetween water-soaked sheets of paper tow-elling and incubated for 7 days at 20°C,

ISSUES OF COMMON TECHNICAL INTERESTSeed Health Testing

Seed Health TestingThe State of the Art of Seed Health Testing By Jim Sheppard, ISTA Plant Disease Committee Chair

Diagnostics in research and inroutine seed health testing havesimilar objectives but healthtests must be completed within alimited time span and standardi-sation is necessary to achieveresults, which are consistentwhen applied in practice.


dark spots on the cotyledons are sympto-matic of Colletotrichum lindemuthianuminfection. In other tests the germination ofseeds is deliberately suppressed to allowseedborne infection to develop. ThusBrassica seeds (1000 per sample) areplaced on blotters irrigated with the herbi-cide 2,4-dichlorophenoxyacetic acid (2,4-D) solution and incubated for 11 days at20°C in alternating cycles of light and darkto allow the pycnidia of seedborne Phomalingam to develop. The germination ofseeds is inhibited by 2,4-D solutions,allowing greater numbers of seeds to betested. Similarly, carrot seeds (400 persample) are tested for Alternaria dauci andA. radicina on triple-layer blotters soakedin sterile distilled water by incubating themfor 3 days at 20°C, then at -20°C overnightfollowed by 7 days at 20°C in alternatingcycles of light and dark for production ofconidia. Sterile media including sand, artificialcomposts, etc. can be used for the detectionof certain pathogens. Results are based onthe presence of symptoms typical of theorganisms.

Staining and examination of the embryoStaining methods are used for seedbornepathogens which cannot be detected bydirect inspection or incubation methods.The standard method used in seed healthtesting is that of staining barley embryos offor the presence of loose smut ( Ustilagosegetum var. tritici) mycelium. Theembryos of barley seeds (2000-4000 seedsper sample) are extracted over 24 h at 20°Cin a 5% solution of sodium hydroxide.After clearing in lactophenol, the embryosare examined under magnification for thepresence of the golden brown mycelium ofthe fungus in the scutellum of seeds. In cer-tain laboratories lactic acid is preferred tolactophenol as a clearing agent.

ImmunoassaysSerological tests for plant pathogenic bac-teria have been known for nearly a century.The methods are based on the immunolog-ical principle that foreign molecules inject-ed into the bloodstream of mammals stim-ulate the immune system of those mam-mals to produce specific antibodies whichwill recognize and bind to the antigens.Such antibodies recognize many chemicalsites, referred to as epitopes, on target anti-gens; these are polyclonal antibodies.Immunoassays utilizing antisera producedagainst purified pathogens or extracts ofpathogens have been effective in the detec-tion of viruses but have had a more limitedvalue for the detection of bacteria and par-ticularly fungi.

The development of monoclonal antibodies(MAbs) has greatly increased specificity.Monoclonal antibodies recognize onechemical site (epitope) on target antigensand in that respect are homogenous andfree from the variability common to poly-clonal antisera. They can be selected to actat generic, specific, pathovar or strain lev-els.

The ELISA technique, has had consider-able impact on the diagnosis of virus andbacterial diseases of vegetatively producedcrops with a lesser but nonetheless impor-tant role in the diagnosis of seedborne dis-eases. There are many different forms ofELISA. Three of the main types are thedirect (double antibody sandwich (DAS-ELISA)) method, the indirect method andthe competitive assay. The direct method ismost often used in seed health assays.Using the direct method, seed or seedlingextract (i.e. the antigen) is selectivelytrapped and immobilized by solid-phase-specific antibody in microtitre wells.Enzyme-labelled antibody is then reactedwith the immobilized antigen and, afterremoving unreacted enzyme-labelled anti-body, the retained enzyme is assayed byadding a suitable substrate. Qualitativeassays may be made by eye and visualiza-tion may he improved by the addition ofsubstrates that give coloured hydrolysates.Quantitative assays are made using colori-metric or spectrophotometric equipment.

Molecular (PCR) AssaysThe application of molecular nucleic acidtests in practical methods for the detectionof seedborne organisms has not been fullyexploited. The versatile nature of molecu-lar methods has resulted in their use to sup-plement or replace established morpholog-ical, biochemical and serological tech-niques. Their introduction has beenfavoured in part, by their relatively quickdevelopment and validation time asopposed to serological and biochemicalassays that take longer to implement. Oncedeveloped and validated, molecular assayscan provide simple, quick, high-throughputdiagnosis that is relatively cheap and easyto transfer to other laboratories. Initially,non-amplification assays wereused, but the need for pure cultures andpoor repeatability did not make them suit-able for routine diagnosis. Development of'in-vitro' amplification by PCR is leading tospecific, sensitive, robust assays for bothpure cultures and, more importantly, for thedetection and identification of organismsdirectly from seed and plant material. Nucleic acid-based methods tend to be rel-atively expensive to apply and the advan-

tages of these more rapid methods have tobe considered against the lesser cost, butgreater inconvenience to the grower, of thelonger period required achieve identifica-tion of a pathogen using isolation-basedmethods. At present, a major disadvantageof nucleic acid methods in seed health test-ing is that of quantification. The technolo-gy is available and under evaluation, how-ever, although the technology is relativelysimple to use, the cost of the necessaryequipment remains prohibitive for use inroutine seed health testing.

Meeting future challenges

The time taken to complete a diagnostic

test can be critical for the release of seed. Itis necessary to determine the number andfrequency of tests that can be completedwithin a limited time span, a constraintintroduced in certain circumstances by theneed to sow the seed bulk within a matterof weeks. The development of rapid andmore advanced methods for the detectionof seedborne organisms will increase thefrequency of some diagnostic tests and inthat respect will benefit commercial seedtesting. It is important that, as new andmore advanced detection methods areintroduced, their performance should besubjected to critical appraisal to establishthe limits of their analytical and diagnosticsensitivity and specificity, ideally todemonstrate that they are at least as effec-tive as the techniques, which they seek toreplace. The ISTA Plant DiseaseCommittee has recently implemented aprogramme of method validation. Thismethod validation programme is open, anymethod developer or test kit manufacturermay submit a method for validation.Details of the programme can be obtainedfrom the ISTA Secretariat or the ISTAonline Website.

ISSUES OF COMMON TECHNICAL INTERESTSeed Health Testing

It is important that, as newand more advanced detectionmethods are introduced,their performance should besubjected to critical appraisalto establish the limits of theiranalytical and diagnostic sen-sitivity and specificity, ideallyto demonstrate that they areat least as effective as thetechniques, which they seekto replace.

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IntroductionThe International Seed Federation (ISF)represents the mainstream of seed trade andthe plant breeding community in the world.To facilitate the free movement of qualityseed through fair and reasonable regula-tions is one among its many goals. Andtherein evolves the twofold responsibilitythe seed industry assumes in the area ofseed health: to deliver good quality andhealthy seed to farmers and seed producers,and to respect international phytosanitaryregulations.

ISF members fund the International SeedHealth Initiative (ISHI), which through itsexclusive focus on seed health is perhapsunique in the world. Under the umbrella ofISHI, seed companies exchange informa-tion on seed-borne diseases and jointlydevelop methods for their control. TheInternational Seed Health Initiative forVegetable Crops (ISHI-Veg) was charteredin 1994 by the vegetable seed industries inthe Netherlands and France and was soonjoined by the vegetable seed industry in theUnited States, Israel and Japan. These coun-tries together represent the production ofover 75% of the world's vegetable seed sup-ply. Public sector institutions Naktuinbouwand Plant Research International (NL),Volcani Centre (IL), Groupe d'Étude et decontrôle des Variétés et des Semences(GEVES) (FR), the Horticultural Research

Institute (UK) and the Canadian FoodInspection Agency (CFIA), and private lab-oratories such as STA Laboratories Inc.(US) collaborate with ISHI-Veg membercompanies adding significant value to thework done.

Working together as an international groupof more than 40 seed pathologists ISHI-Vegassesses, develops and disseminates infor-mation on test protocols for economicallyimportant seed-borne pathogens of veg-etable crops. The combination ofresearchers and practitioners presents ISHI-Veg with the opportunity to develop proto-cols using both well-tested and novelapproaches. This paper describes the practi-cal and innovative activities of ISHI-Veg insecuring the delivery of healthy seed.

ISHI-Veg Accepted Methods

Seed health testing is an integral tool for allvegetable seed companies in seed-bornedisease risk management and is a part of theentire disease control program. The meth-ods used are techniques either generatedinternally through a developmental processor those available for public use.

For a method to be accepted by ISHI and beconsidered as suitable for further develop-ment as a reference method, it must beeither described and available for public useor published in a peer-reviewed journal, and

meet the approval of the technical groupwithin ISHI-Veg responsible for the rele-vant vegetable commodity. The aim whendeveloping a reference method is that it bestmeets the needs of the laboratory conduct-ing the test, often the seed producer, and itsclients.

Most methods then go through a validationprocess that includes extensive comparativetesting of the protocol. The process consistsof checking the parameters of the protocol -sensitivity (of the assay in terms of percent-infected seed or target pathogen quantifica-tion, for instance), specificity (e.g. in iden-tifying a range of isolates of the pathogenfrom different geographical regions andrace), repeatability and reliability.Historical use, peer review, research in pub-lic institutions, and unbiased comparativetesting by other groups also contribute toaccomplishing the task.

Comparative Testing by ISHI-VegSeed health tests differ from country tocountry and the results from one countrymay not be accepted in another. A measureof ISHI-Veg's success lies in the interna-tional acceptance of its seed health tests.Comparative tests in many laboratories andcountries are essential components of ISHI-Veg's activities. And to ensure reliableresults the selection of participating labsthat are familiar with the tests is critical.

ISSUES OF COMMON TECHNICAL INTERESTIndustry Initiatives in Seed Health

Industry Initiatives in Seed Health: The Example of VegetablesBy Radha Ranganathan, International Seed Federation Ruud Scheffer, Chairman ISHI-Veg Technical Co-ordination Group and Harrie Koenraadt, Naktuinbouw, The Netherlands

Radha Ranganathan Ruud Scheffer Harrie Koenraadt

The twofold respon-sibility the seedindustry assumes inthe area of seedhealth: to delivergood quality andhealthy seed to farm-ers and seed produc-ers, and to respectinternational phy-tosanitary regula-


Comparative tests require the availability ofnaturally infected seed lots. For somecrop/pathogen combinations there is a lackof naturally infected or contaminated seedwith the result that comparative tests aredelayed. In such cases ISHI-Veg examinesthe use of artificial reference materials orhas artificially infected seed lots.

Artificial Reference MaterialsBacterial preparations containing a speci-fied quantity of viable bacterial cells weredeveloped and used as reference material atPlant Research International. Preservationwas optimised by freeze-drying bacteria(Clavibacter michiganensis subsp. michi-ganensis (Cmm), Pseudomonas savastanoipv. phaseolicola (Psp), Xanthomonasaxonopodis pv. phaseoli (Xap) andXanthomonas campestris pv. campestris(Xcc) among others) in paper discs. Theadvantages of this approach are that the via-bility in time, the genotype, and the numberof colony forming units of the targetpathogen are well known or predictable. Inaddition, provided these reference materialsare commercially available at a reasonableprice, an unlimited supply of referencematerial can easily be made available.

A disadvantage of the use of such referencematerials is that the 'real world' of seed test-ing cannot be simulated. For instance, theextraction kinetics of target pathogens fromnaturally contaminated seed lots crucial fordetection of Cmm from treated tomato seedlots and Xcc from cabbage cannot be eval-uated. Also the wide genetic variability ofCmm, as demonstrated by Giora Kritzmannof the Volcani Center (pers. comm.), showsthat such reference materials cannot be usedto answer all questions. Neither can a 'stan-dard' laboratory execute the production ofsuch bacterial reference materials, as spe-cial equipment is needed.

Artificially Contaminated Seed LotsInoculation of plants by a target pathogenwith the aim of obtaining artificially con-taminated seeds has not yet been widelyexploited. Biggerstaff et al. (2000) success-fully produced Cmm-infected seedsthrough inoculation of tomato plants andISHI-Veg is trying to produce tomato seedlots infected with Cmm. It is planned toproduce watermelon seeds infected withAcidovorax avenae subsp. citrulli (bacterialfruit blotch), an important crop/pathogencombination for the seed industry.

'Dilution' of Naturally Contaminated Seedwith Healthy Seed and the Use of Pre-ground FlourA not altogether rare practice is one wherecomparative tests are made to evaluate the

performance of participating laboratories.In the past equal numbers of sub samplesfrom one heavily contaminated sample, twoor three lightly contaminated samples, andone healthy sample were tested. The resultsof such tests were difficult to evaluate,especially when lightly contaminated sam-ples were used, due to variation in the num-ber of infected seeds per sample/partici-pant. Using a relatively limited number oflightly contaminated sub samples it was dif-ficult, if not impossible, to concludewhether laboratories missed infected sam-ples or whether they received samples thathad fewer infected sub samples.An alternative approach is currently underinvestigation in Naktuinbouw. Several largeseed lots infested by Lettuce MosaicPotyvirus (LMV) stored at Naktuinbouwwere used for a pilot. From four heavilycontaminated seed lots, large pools of 10-fold dilutions (1x, 10x, and 100x) were pre-pared using one large healthy seed lot.From each dilution of every seed lot 20 subsamples were prepared. All the sub sampleswere renumbered to obtain a completelyblind comparative test. In the pilot theresults were as expected. In all sub samplesof the 1x dilution LMV was detected.Depending on the infection rate of eachseed lot, the number of infected sub sam-ples decreased to zero in the 10x or 100xdilution. Even more information could beobtained when fewer sub samples from theundiluted positive samples and more subsamples from the 10-fold dilutions wereused.

The flour of pea and squash seeds infectedwith Pea Seed-borne Mosaic Potyvirus(PSbMV), Pea Early Browning Tobravirus(PEBV) and Squash Mosaic Comovirus(SqMV), respectively, have also been usedin comparative tests. Seeds were ground bythe test-organiser to fine flour and distrib-uted to participating laboratories to ensurethat all had identical sub samples of flour.Data from these comparative tests revealedthat almost all participants detected thevirus in the sub samples. A big advantage ofthis approach is that since all participantsreceive identical sub samples, outliers canbe easily identified. A limitation of thisapproach, however, is that it is best suitedfor large seeds that can be easily ground. Inaddition the virus must be stable and dis-tributed evenly in the flour.

Although these formats of comparative test-ing are laborious for the organiser, especial-ly when many laboratories participate, it iseasy to identify a method or a laboratorywith many false positive or false negativesub samples. The Use of Multiple Methods

The ISHI experience has shown that usingone method or a common methodology inall testing facilities is not always feasible;on the contrary ensuring consistent resultsfrom all testing facilities is the most impor-tant factor in seed health testing. Multiplemethods where the results obtained fromtwo or more methods are compared withthat obtained from using only the accredit-ed or standard method may provide analternative.

Multiple methods have become accepted inclinical laboratories where there are goodreasons to deviate from an existing standardmethod. For instance, labs may haveexpertise that allows for the implementationof another method or a very high or lownumbers of samples, which for reasons ofeconomy demand an alternative to the stan-dard, or they may want a method that ischeaper, more sensitive or specific. Any ofthese reasons may result in the implementa-tion of an alternative to the standardmethod.

Clinical laboratories have one major advan-tage over vegetable seed companies and thetest laboratories related to them: there aremany more clinical labs and referencematerials are more easily available. In theNetherlands for instance, the monitoringorganization SKZL organizes weekly refer-ence tests at all participating laboratories.This allows SKZL to observe even gradualdeviations of a specific test carried out by alab, deviations such as a loss of sensitivityattributable to a spectrophotometer requir-ing maintenance.

In the field of medicine multiple methodshave been shown to generate identicalresults in comparative tests where the per-formance of each laboratory and theirmethod is evaluated rather than just themethod. In the case of seed, the 'dilutionmethod' described in the previous sectionwas used to compare the performance oflaboratories and not methods. If the meth-ods used by the participating laboratoriesare not identical, it adds another importantfactor to the equation. Methods must befirst investigated carefully in one or a fewlaboratories using a larger number of sam-ples with different infections rates, morevariation in genotypes, etc. If more than onemethod is used in a comparative test, theinevitable noise of inter-laboratory variancewill most likely confound the results check-ing for equivalence of methods. This phe-nomenon has been seen in most, if not all,comparative tests in which the 'in-houseand well known' method was comparedwith 'new and therefore unknown' methods.The authors believe that the comparison ofdifferent methods through elaborate com-


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different methods through elaborate com-

Meier

The authors believe that the comparison of different methods through elaborate com-

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parative testing in several laboratories with-out prior and ample basic research at onelocation would not be very useful.

Despite the limited number of vegetableseed testing laboratories and the laborioustask of first comparing methods before theperformance of labs, there is good reasonnot to reject the idea of multiple tests.Relying exclusively on an older but 'stan-dard' test could prove risky particularly inview of the rapid developments in technol-ogy. If a seed company had say, a quick andsensitive DNA chip test in routine use and ifin a series of reference tests it was demon-strated that this method was superior orequivalent to the standard it should beaccepted as an alternative to the standard.

In practice multiple methods are used todayand differences between the protocols areminor. Examples are the use of differentselective media, or differences in theextraction of bacteria from seeds, likegrinding, shaking the seeds in a medium, orusing a stomacher. Multiple methods basedon radically different technologies likeimmuno-fluorescence vs. a DNA-based testare still unusual. An exception is the nowrarely used ISTA method in which LettuceMosaic Potyvirus could be detected througheither a grow-out of lettuce seeds orthrough a bioassay on Chenopodium plants(1981).

As part of its Performance Based Approachadopted during the Extraordinary Meetingin Bolivia in July 2002, ISTA also plans touse multiple methods to test for the pres-ence of genetically modified organisms inseed.

Accepting multiple methods still stressesthe need for generally accepted standardmethods. Such reference methods are the'standards' to which alternative test methodsare compared, and used to 'calibrate' refer-ence materials. The ISHI-Veg Methods Manual

The ISHI-Veg manual lists methods accept-ed by ISHI-Veg and serves as a focal pointand guide for the development of ISHI-Vegreference methods: methods that consistent-ly provide reliable and repeatable detectionof a specified seed-borne pathogen at a pre-determined level of infection. The manualis available to all labs on the ISF website(www.worldseed.org).

For the purpose of this manual, status rat-ings for ISHI seed health testing methodsare defined in the following manner. o Category 1 - An ISHI ReferenceMethod. The method has been through theISHI comparative testing process, has beenpeer reviewed and accepted by ISHI.o Category 2 - An ISHI Accepted Methodunder Review. The method has been pub-lished and/or is publicly available, and iscommonly used in the seed industry fordetermining seed health. It is undergoingcomparative testing or certain aspects of thetest are being tested for retention or deletionfrom the method.o Category 3 - An ISHI AcceptedMethod. The method has been publishedand/or is publicly available, and is com-monly used in the seed industry for deter-mining seed health. It is not undergoing acomparative test at this time.When a method has been validated by ISTAand/or any other body this is also indicated.

ConclusionIn the foregoing sections we have illustrat-ed how through international co-operationISHI-Veg continually strives for reliablestate-of-the-art test methods. Through thispractical goal ISHI-Veg realises its mission- the delivery of healthy seed to customerson a worldwide basis.

The development of methods to test forseed health is complex. The epidemiologyof diseases, different levels of infection inthe seed, differences between climaticzones and the effect of such zones on infec-

tion rates are some variables that must beconsidered in developing reliable test meth-ods, and particularly so in the field of bac-terial diseases. Comprehensive studies thatelucidate on all these aspects are often notpossible to find, as data are usually scat-tered and inconclusive. The combinedexperience, data and expertise within ISHI-Veg allows for test methods to be developedthat take into account the above-mentionedfactors.

Clearly, scientific developments, changes incultural practices of both seed companiesand growers and changing customer needsadd to the complexity. The comparative testprogram of ISHI-Veg and its aim to con-clude on generally accepted standard meth-ods are key efforts to make progresstowards its mission. The careful evaluationof alternative test protocols does result inmultiple methods being recognized andaccepted. However, using ISHI's referencemethods and the methods designated 'gen-erally accepted', companies are able tomake risk analyses specific to the condi-tions under which they operate, taking alsoother factors such as resistance in their vari-eties, seed production region, and theregion where the seeds will be sold intoconsideration, all again to secure the deliv-ery of healthy seed.

References

Biggerstaff, C.M., Gleason, M.L. and Braun, E.J. 2000.Refinement of a non-destructive tomato seed assay forClavibacter michiganensis subsp. michiganensis usingseed fiber. Seed Sci. & Technol., 28:261-269.

Rohloff, I. and Marrou, J. (Eds) 1981. ISTA handbookon seed health testing. Working sheet No. 9.

R. Ranganathan, International Seed Federation, 7Chemin du Reposoir, 1260 Nyon, Switzerland; RuudScheffer, Chairman ISHI-Veg Technical Co-ordinationGroup, 7 Chemin du Reposoir, 1260 Nyon, Switzerland.Harrie Koenraadt, Naktuinbouw, P O Box 40, 2370 AARoelofarendsveen, The Netherlands.


www.seedtest.org

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How to volunteer to become leader of an ISTA Proficiency TestIt was the summer of 2002. It had been a long week: first there hadbeen the 28 hour journey to Santa Cruz and that had been followedby 3 long days of Executive Committee Meetings. Sunday was myday off. Relaxing at the side of the pool of the Los Tajibos Hoteland Conference Centre after a 14 hour tour to the world heritagesite at Samaipata - I was at my most vulnerable. "Lovely evening Ronnie" was the greeting in unison from HarryNijënstein (ISTA Moisture Committee Chairman) and MartinaRösch (ISTA Secretariat, Head of Accreditation). I had to agree, it was. "Wouldn't it be a good idea to have Moisture Content testingincluded in the next round of proficiency tests?" asked Harry. Again I had to agree; moisture content testing is one of the origi-nal seed quality tests but moisture content has never been includedin the proficiency testing program. "Why are moisture content tests not part of the program?" Martinaenquired. I tried to explain as simply as I could - Unlike purity and germi-nation testing where results are based on a qualitative characteris-tic of seeds like pure seed versus not pure seed or normal seedlingversus not normal seedling, moisture content is a quantitative traitwhich is expressed as a percentage. Because of this the usualmethod of statistical evaluation based on Tattersfield can not beapplied and this has resulted in the omission of moisture contenttesting from the normal program of proficiency tests. In 1998 therehad been a pilot test using the species Phalaris aquatica to ascer-tain the most appropriate means of statistical evaluation and todetermine the feasibility of including moisture content determina-tions future proficiency testing programs. The pilot went well andDr Michael Kruse had developed a statistical evaluation procedurebased on the general principles of Tattersfield. Michael had alsotested the evaluation procedure on a moisture content comparativetest that I had organised in 1999 and again it had been successful inidentifying laboratories that may have had analytical problems withmoisture content testing."Mmm" was the rely from Martina, "So there is no reason for mois-ture content to be excluded from future testing programs?" Again I had to agree."What we need, Martina," said Harry "is someone with Ronnie'sexperience in preparing moisture content comparative tests to vol-

unteer". Again I had to agree. I evenstated that I would be glad tovolunteer but that the major-ity of seed production inScotland involves cerealsand that the first moistureproficiency test should be on a small seeded species that didn't needto be ground. This I thought was the perfect justification/excuse fornot taking on this piece of ISTA work. "That's not a problem", said Martina, "we have arranged for RitaZecchinelli, from the Proficiency Test Committee, to send yousamples of the clover seed that will be used for the Germination,Purity, and Other Seed Determination Proficiency test that will besent out in February 2003." But what about phytosanitary requirements?,….timing?,…..proce-dures….? Harry and Martina had answers to all my questions andthere was no way I could wriggle out of it; I was the volunteer whowould prepare the ISTA Moisture content proficiency test evenalthough I can't quite remember actually volunteering. It was astitch-up!

The Samples and Preparation InstructionsThe samples arrived from Rita Zecchinelli in November - 3 bags ofabout 5 kg each of the same lots that would be used for the otherparts of the clover proficiency test. We would have plenty time toprepare the samples and send them to ISTA for distribution by theend of February, or so it seemed! Around about the same timeMartina was kind enough to send me the Protocol to be used for byProficiency test leaders: "REFEREE SAMPLES PREPARATIONINSTRUCTIONS FOR REFEREE TEST LEADERS - PURITY,GERMINATION AND OTHER SEED DETERMINATION TESTS."These guidelines had be adopted by the Proficiency (Referee)Committee to ensure the ISTA Proficiency Program is incorporat-ing the basic principles for proficiency testing programs outlined inISO Guide 43. In particular they ensure that proficiency samples,sent to participating laboratories, are homogeneous.

The instructions, twelve pages in all, were very detailed and theprocedures looked much more complex than the ones I had used toprepare cereal samples for the moisture content comparative testinvolving 12 laboratories. In addition there was no mention ofpreparing samples for a moisture proficiency test. Martina told me

TECHNICAL COMMITTEESMoisture Content Proficiency Test

Moisture Content Proficiency TestBy Ronald Don, ISTA Moisture Committee Vice-chair

The sculptured rock atSamaipata bears outstand-ing witness to the exis-tence in this Andeanregion of a culture withhighly developed religioustraditions, illustrated dra-matically in the form ofimmense rock sculptures.

The pool at the centre ofthe Los Tajibos hoteland conference complexin Santa Cruz, Boliviawhere the firstExtraordinary Meetingof ISTA was held in July2002.


not to worry and just adapt the procedures given for purity, germi-nation and other seed determination tests. Adjusting Sample Moisture Contents

This was the easy bit. For the proficiency test it was decided that the samples sent to par-ticipating laboratories should have a range of different moisturecontents. To achieve this objective the three lots of clover seedwere subjected to the following treatments:

Lot A - sent to laboratories without any moisture content adjust-ment.

Lot B - placed in an oven at 30oC for 6 weeks to reduce mois-ture content.

Lot C - placed in a room at a temperature of 20oC and 95%Relative humidity for 7 days to increase moisture content.

Preparation of Proficiency Samples

This was the difficult bit!!!Monday, Tuesday and WednesdayTo prepare the 160 samples, required by the Secretariat, from eachlot was complex and time consuming. The laboratory was busy andthe job of preparing the samples was allocated to me with help fromMargaret Orr, who normally provides the laboratory with adminis-trative support. Mixing and dividing was achieved using aCentrifugal Divider and each set of samples took a full day to pre-pare in a process that involved 10 steps:

Step 1Sample is mixed three times and then divided into two.

Step 2Each half is mixed three times and then divided into two halvesgiving 4 samples halves.

Step 3Alternate quarters are combined to give two half samples.

Step 4This is a repeat of Step 2 - each half is mixed three times and thendivided into two halves giving 4 samples.

Step 5The four quarters are each mixed three times and then divided intotwo halves giving 8 samples.

Step 6The eighths are recombined to give 4 samples.

Step 7A repeat of Step 5 - The four quarters are each mixed three timesand then divided into two halves giving 8 samples.


5 Kg Sample of Clover Seed

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2.5Kg

1.25Kg 1.25Kg 1.25Kg


This may seem simple when written down like this but believe meit was a major operation involving almost every mixing bowl in theseed laboratory. In addition, all seed not in the process of beingmixed and divided had to be placed in moisture proof containers toprevent any changes to their moisture content that would haveresulted in differences in moisture content between samples of thesame seed lot.Each lot took 10 hours to divide down into the 160 samplesrequired for the proficiency test. During the period of preparationdaily contact was maintained with Gerhard in the ISTA Secretariat.Gerhard would be responsible for the dispatch of the samples toISTA Laboratories and he also gave Margaret and I endless encour-agement. We had started the preparation on Monday morning andat 18:00 on Wednesday the last sample (sample 480) had beensealed in its foil bag and labeled. I was exhausted and needed a dayoff. I E-mailed Gerhard to say we had finished and left Margaretinstructions to pack the samples and send by courier to Gerhard.

ThursdayWhilst I was enjoying my day off, Gerhard E-Mailed Margaret toask for the results of tests to prove that the samples we had pre-pared were homogenous. This was something I had forgottenabout. Margaret had to arrange for 10 samples from each lot, ran-domly chosen by ISTA, to be tested for moisture content. I musthave been popular, and it's a wonder my ears were not ringing. But

the staff in the lab are veryobliging. Thirty moisturecontent tests were com-pleted by 15:00, and theresults E-mailed to ISTA.

FridayI was informed of the pre-vious days developmentsand told that Annabellawas due a box of choco-lates for doing all themoisture content tests atsuch short notice.Looking at the results ofthe tests I thought it wasMargaret and I that weredue a box of chocolates -the 10 results from eachlot were within ±0.2% ofeach other. I was confi-dent that H-test analysisby Günter Müller, ViceChairman of theProficiency TestCommittee, would showthat the samples were fitfor purpose and could beused for the MoistureContent Proficiency test.I was correct, Günter con-firmed this within thehour and we could sendthe samples off to ISTA.

Margaret found a box bigenough to accommodateall the samples. She care-fully packed them by seedlot and sample number tomake the job of the

Secretariat easier when dis-patching samples to participating laboratories. We had used justover 9 kg of seed but the weight of the foil bags and packing meantthat the final parcel had a weight of 17 kg - too heavy for airmail!It was sent by courier - cost £112.

Dispatch of Proficiency Samples to ISTA Laboratories

Two weeks later

ISTA clover proficiency samples arrive in the laboratory for testing.Usually they wouldn't be given any special thought. But these sam-ples were special -we will never look at proficiency/referee sam-ples in the same way again. We now appreciate the work and effortrequired in providing laboratories with these samples.

Step 8The eight eighths are each mixed three times and then divided into two halves giving 16 samples.

Step 9

The sixteenths are recombined to give 8 samples.

Step 10

From each of the eighths 20 samples of at least 25 g are obtained using the spoon method of sampling. Each of these samples is placed in a foil bag, which is heat sealed and labelled according to instructions from the Secretariat.


0.63 Kg

0.63 Kg

0.63 Kg

0.63 Kg

0.63 Kg

0.63 Kg

0.63 Kg

0.63 Kg

A B C D E F G H I J K L M N O P

A B C D E F G H I J K L M N O P

0.63 Kg

0.63 Kg

0.63 Kg

0.63 Kg

0.63 Kg

0.63 Kg

0.63 Kg

0.63 Kg

0.63 Kg

0.63 Kg

20 Samples of at least 25g each

The sampleswere dis-p a t c h e dfrom theI S T ASecretariatOffice.Thephotographshows ISTASecretariatStaff hard at


TemperatureTemperature is one of the most commonlymeasured physical quantities but its basis isnot widely understood. Whereas the unitsof other quantities, such as mass and timeare based on real physical entities tempera-ture is founded on a theoretical set of con-ditions. So whilst the perfect kilogram is inParis and time is based on atomic transi-tions in a caesium atom, temperature isbased on the thermodynamics of perfectsystems, such as ideal gases. This results inthe thermodynamic temperature scalemeasured in Kelvins (K), which is unattain-able. To overcome this we do the next bestthing and use imperfect thermodynamicsystems to achieve a working temperaturescale as near to the theoretical one as wecan get. Temperature is an important factor in anumber of tests carried out by ISTALaboratories. It can affect the outcome ofGermination, Moisture Content,Tetrazolium and Vigour tests and laborato-ries must take measures to control and mon-itor temperature. This set of guidelines hasbeen drawn up to assist laboratories andauditors when they are considering thearrangement for temperature measurement.

Temperature MeasurementThe two most common methods of measur-ing temperature are using thermometers orthermocouplesTHERMOMETERSA thermometer is an instrument that meas-ures the temperature of a system in a quan-titative way. The most direct 'regular' way isa linear one. For example, the element mer-cury is liquid in the temperature range of -38.9° C to 356.7° C. As a liquid, mercuryexpands as it gets warmer; its expansionrate is linear and can be accurately calibrat-

ed. When wires of different metals are fused atone end and heated, a current flows fromone to the other. The electromotive forcegenerated can be quantitatively related tothe temperature and hence, the system canbe used as a thermometer - known as a ther-mocouple. The thermocouple is used inmany electronic/digital thermometers andmany different metals are used, forexample - platinum and platinum/rhodium;nickel- chromium and nickel-aluminum.

Temperature Measurement in ISTALaboratoriesA variety of different equipment is used tomeasure temperature in ISTA laboratories.Some use thermometers / temperatureprobes with accuracy of ± 0.1oC whereasothers use thermometers where it would bedifficult to achieve a temperature reading

with an accuracy of ± 2oC.

Recommendation for TemperatureMeasurement in ISTA Laboratories

- Laboratories should use temperaturemeasurement instruments can be read accu-rately to at least 0.5oC and conform to ISO386.- For Accelerated Ageing Vigour Tests the

instrument must be read accurately to atleast 0.1oC.Calibration and the ISTA StandardAppropriate calibration samples, referencematerials and reference standards of meas-urement shall be held by the laboratory, andshould be used for calibration and referencepurposes only. Examples include calibra-tion samples for seed blowers, standardbuffer solutions for pH meters, calibrationweights for balances, seed reference collec-tions for the identification of unknownseed.Calibration of temperatureSome laboratories have all probes calibrat-ed externally, others use externally calibrat-ed probes to calibrate their working probes,others calibrate their thermometers withsteam and ice and others do no calibrationwhat so ever.

Calibration of Thermometers - ExternalCalibrationRECOMMENDATIONSThe current working temperature scale isthe International Temperature Scale of 1990(ITS-90) and is measured in degreesCelsius (°C).- When a thermometer is calibrated exter-

nally it should be to IPTS-90 or to anagreed previous temperature scale such asIPTS-68 or even IPTS-48.- External calibration should be repeated

every 5 years and this should be supple-mented by annual in-house ice point cali-bration.- Any probe whose temperature reading

differs from the standard by more or lessthan ±0.5oC should be removed from serv-ice.- For Accelerated Ageing Vigour Tests the

limit is ±0.1oC

Calibration of Thermometers - Againstan External StandardRECOMMENDATIONS

- When Calibrating a batch of thermome-ters against an externally calibrated probethe calibration should be carried at temper-atures within the normal working range and

TECHNICAL COMMITTEESTemperature Measurement and Control in ISTA Laboratories

Temperature Measurement and Controlin ISTA LaboratoriesGuidelines for Laboratories and AuditorsBy Ronald Don, ISTA Moisture Committee Vice-chair

The mercury-in-glass thermometer contains abulb filled with mercury that is allowed toexpand into a capillary. Its rate of expansion iscalibrated on the glass scale.


in conditions where fluctuation in tempera-ture between readings is limited.

- Where working probes are calibratedagainst an externally calibrated probe thisshould take place at least once a year.- Any probe whose temperature reading

differs from the standard by more or lessthan ±0.5oC should be removed from serv-

ice (For Accelerated Ageing Vigour Teststhe limit is ±0.1oC)

Calibration of Thermometers -International Ice Point Method The ice point may be realised in an insulat-ed flask or vessel containing an ice-watermelting mixture. The ice particles shouldbe no more than a few millimetres in diam-eter and the water and ice should be pure orprepared from de-ionised water, which isair saturated.For high precision the thermometer shouldbe maintained in the mixture for 10 minutes

prior to reading. In theory accuracies of±0.001oC may be achieved but in practice±0.005oC is more likely.

RECOMMENDATIONS- International Methodology should be fol-

lowed.- Calibration should take place at least

every year.- Any probe whose temperature reading

differs from the ice point by more or lessthan ±0.5oC should be removed from serv-ice.1

Calibration of Thermometers -Steam/Boiling Water MethodThe boiling point of water varies withatmospheric pressure and this is dependenton both weather conditions and altitude. Atsea level with an atmospheric pressure of760mm Hg the boiling point may be 100oCbut in Kathmandu in Nepal at an altitude of1310m and atmospheric pressure of 650mmHg the boiling point is 95.5oC.

RECOMMENDATION- Calibration of temperature probes using

steam/boiling water should not be used byISTA Laboratories

Calibration of Thermocouples,Electronic Meters and Temperature Data

Loggers

RECOMMENDATIONS- The procedures for the calibration of ther-mocouples and temperature probes of elec-tronic meters and data loggers should be thesame as that adopted for thermometers. Inother words they can be calibrated external-ly, or in-house· against an externally calibrated probe orthermometer; or · using the ice point method. - As with thermometers, any probe that dif-fers by more than ±0.5oC from the standardshould be removed from service (±0.1oC in

the case of Accelerated Aging Vigour tests).

Records of Calibration Checks

RECOMMENDATIONS

- Calibration Checks should be recordedand be available for inspection by auditors.The easiest way to do this is in aThermometer/Probe Register.- Records should be archived for a period

of at least 5 years. - Adjustments should be made for differ-

ences between probe temperature and thatof the calibrated standard. For example if aprobe reads 19.6oC when the calibratedthermometer/probe reads 20oC, and theprobe is to be used to monitor a germinationroom which has a performance requirementof 20±2oC the limits using the probe will be17.6 - 21.6oC.

Total immersion and partial immersionthermometers

Total immersion thermometers are designedto be totally submerged in the media whosetemperature is being measured. They aredistinguished by the engraved suffix '/TOTAL' or '/ TOT IMM'.

Partial immersion thermometers shouldonly be immersed to the indicated depth.They have a suffix that indicates the immer-sion depth, e.g. '/ 100 MM IMM' should beimmersed to the depth of 10 cm. They mayalso have an engraved ring on the stemindicating the immersion depth.

Measurement of Temperature - Totalimmersion thermometers and thermo-couples

RECOMMENDATIONS

- Total immersion thermometers and ther-mocouples can be used to monitor the tem-perature of incubators, walk in germinators,ovens and fridges.- For ease of taking measurements and to

reduce dramatic temperature fluctuationsthat can result from the opening of doors ofapparatus thermometers and probes shouldbe submerged in glycerol or sand.

Measurement of Temperature - Partialimmersion thermometers

RECOMMENDATION

- Partial immersion thermometers can beused to monitor the temperature of incuba-tors, ovens and fridges through an externalaperture on the apparatus provided that theycan be immersed to the required depth with-


-1000 0

1000 2000 3000 4000 5000

400 500 600 700 800 900 Atmospheric Pressure (mm Hg)

(m)

82.0 84.0 86.0 88.0 90.0 92.0 94.0 96.0 98.0 100.0102.0

(°C)

Temperature Measurement and Control in ISTALaboratories is continued on page 30.


in the apparatus.

Measurement of Temperature -Frequency

Many laboratories now use data-loggers orother temperature monitoring equipmentthat constantly monitors temperature andrecords of readings are available on anhourly basis or less. Such records should bearchived for a period of at least 5 years andbe available for the inspection of auditorswho may want to ascertain the temperatureof a particular piece of apparatus on a spe-cific date.

RECOMMENDATIONS for MANUALRECORDING

- For constant temperature equipment atleast 3 readings must be recorded per day atregular pre-set times. If records show thatthe equipment is stable, in terms of temper-ature, with variations of less than 1oCbetween readings the recording frequencycan be reduced to once per day. However,if there is any indication of a change in per-formance recording frequency must beincreased. - For alternating temperature equipment at

least 3 readings must be recorded per day atpre-set times. The timing of these readingsmust be such that both temperature phasesare monitored. - Laboratories should plot readings from

individual pieces of apparatus against time,as this is the easiest way to notice anychanges in the performance. - For alternating temperature equipment

monthly checks are required on the changeover time between high and low phases.This should be no more than 3 hours.- Where records show that a piece of appa-

ratus is stable then measures to take read-ings at weekends and public holidays arenot required. Where apparatus if unstablemeasures should be taken to ensure read-

ings are taken. This might involve the use

of a max/ minimum thermometer of a smallindividual logger.

Measurement of Temperature - Numberof Measuring Points

RECOMMENDATION

- For Germination /Temperature Rooms orCabinets with an area of 20m2 or morethere must at least three measuring points.

Measurement of Temperature -Temperature Profiles

RECOMMENDATIONS

- Before accepting a piece of temperaturecontrolled equipment (ovens, incubators,fridges, Copenhagen tanks, germinators,propagators, germination rooms, etc.) intoservice a laboratory must ascertain the tem-perature profile of the equipment to ensurethat it is fit for purpose.- There should be a minimum of 9 check

points covering different combinations ofheight, depth and breadth, as appropriate.For example, for incubators, fridges andovens temperature should be recorded ontop, middle and bottom shelves and on eachshelve the temperature should be measuredat 3 points - front (next door), middle andback. - The temperatures of all points measured

must be within ± 2oC2 before a piece ofequipment is accepted into service.- Once in service profiles are only meas-

ured again if a piece of equipment under-goes repair or major service.

1 For Accelerated Ageing Vigour Tests the limit is±0.1oC2 The Ageing Chamber for Accelerated AgeingVigour Tests must have a profile of less than±0.3oC when operating at a temperature of 41oC


Total and Partial immersion thermometers canNOT be used to monitor the temperature ofCopenhagen Tanks or Germinators scince it is notpossible to meet immersion requirements.

Thermocouples with contact probes must be used.

Measurement of Temperature - Copenhagen Tanks

ISTA Handbook forSeedling Evaluation, 3rd Edition, 2003By J. Bekendam, R. Schmitt-Grob and R. Don

ISBN 3-906549-26-7

This handbook is a most valuable guidefor seed analysts the world over.Although the 'International Rules forSeed Testing' define normal and abnor-mal seedlings in considerable detail, theadditional help provided by this hand-book, with its detailed instructions andmany illustrations, is vital if the princi-ples of seedling evaluation are to beapplied uniformly. The 3rd edition alsoincludes many tropical, subtropical,flower and tree species.

What is new about the 'ISTAHandbook for Seedling Evaluation'?

The first section of the new addition isaimed mainly at the trainee seed analystand trainers and contains backgroundinformation on the elements of seedbiology, an understanding of which isconsidered essential for seed analysts. Alarge number of new diagrams as wellas the classic Dutch diagrams from the2nd edition give life to the text and helpexplain the finer details of physiologyand seedling evaluation. For the firsttime, general evaluation rules such asthe 50% rule for the evaluation ofcotyledons are considered in depth andexplained with the use of diagrams andcolour prints of seedlings. The evalua-tion of different seedling types is fullyillustrated by colour prints of normaland abnormal seedlings. Diagrams andplates are also used to explain in detailthe intricacies of the evaluation ofspecies, such as Zea and Phaseolus, anddefects, such as physiological necrosisand negative geotropism.

It is planned to regularly update the newedition of the Handbook and to facili-tate this in a binder format with individ-ual sections adopting a QA numberingsystem. Amendments and additions tothe handbook can be easily made with-out the expense of reprinting the entirepublication.

ANNOUNCEMENT

There is a need for ISTA to have a "univer-sal list" of crop and weed species that ana-lysts in all ISTA-accredited laboratoriesshould be able to identify. There are twomain reasons for this list:

1. The Proficiency Test Committee and TestLeaders could use the species on this listwhen choosing crops and inclusions for ref-erees. 2. Audit teams could expect all accreditedlaboratories to have the listed species, as aminimum requirement, in their referencecollections.

In developing such a list, the ISTA PurityCommittee felt that it was important to haveinput from a broad spectrum of ISTA mem-bers to ensure that the list is relevant to allregions and climate zones. It was especiallyimportant that the list not be based solely onfamiliar temperate crops and weeds. A sur-vey form was sent out to ISTA membersfrom the Secretariat asking them to ranktheir top 10 crop species and their top 25weed species.

Results werereceived from 65laboratories in 40countries, one inAfrica, two in theSouth Pacific, sixin Asia, 25 inEurope, three inNorth America andthree in SouthAmerica. As youcan see, the infor-mation wereceived is stillheavily weightedin favour of thetemperate zonecrops. It would bevaluable to havemore data fromISTA laboratoriesin tropical and sub-tropical countries,so that theseregions would bebetter represented.Please contact theISTA PurityC o m m i t t e eChairperson if youwould like to con-

tribute.The data on kinds have been summarized.Table 1 is a list of the top 30 species ofcrops reported by respondents. The score isbased on their ranking (1-10) by the labora-tories and the number ofsamples received eachyear. Table 2 is a list of thetop 50 species of weedsreported by respondents.The score is based on theirranking (1-25) by the labo-ratories. These are the shortlists; the next step will beto decide how many ofthese species should beincluded in the final list.For example, there doesn'tseem to be much point inincluding both species ofLolium and their hybrid,since they are so difficult toseparate from each other.Again, any suggestionswould be welcomed.

Thank you to all who contributed data forthis important project and to BettinaKahlert at the ISTA Secretariat who typedthe numbers into a spreadsheet.


TECHNICAL COMMITTEESISTA Crop and Weed Species Survey

ISTA Crop and Weed Species SurveyBy Ken Allison, ISTA Purity Committee Chair

Zea mays 4558 33Triticum aestivum 3541 47Hordeum vulgare 2480 43Lolium perenne 1431 25Glycine max 879 17Secale cereale 584 19Avena sativa 458 31Festuca arundinacea 243 9Pisum sativum 234 25Medicago sativa 151 20Festuca rubra 105 12Triticum durum 72 7Helianthus annuus 70 17Lolium multiflorum 58 15Phleum pratense 38 16Beta vulgaris 32 12Poa pratensis 25 10Sorghum xdrummondii 18 2Brachiaria brizantha 16 1Trifolium repens 14 9Brassica napus 14 12Festuca pratensis 13 11Panicum maximum 12 3Sorghum bicolor 12 13Trifolium pratense 12 12Gossypium spp. 10 5Allium cepa 9 8Lycopersicon esculentum 9 8Oryza sativa 7 7Dactylis glomerata 7 8

123456789101112131415161718192021222324252627282930

Crop Species Score Labs

1234567891011121314151617181920212223242526272829303132333435363738394041424344454647484950

Weed Species Score Labs

Chenopodium album 733 42Rumex crispus/obtusifolius 610 41Polygonum aviculare 457 28Elytrigia repens 441 27Galium aparine 399 23Echinochloa crus-galli 373 20Amaranthus spp. 332 23Bromus hordeaceus 306 14Tripleurospermum perforatum 296 12Stellaria media 288 19Avena fatua 281 18Viola sp. 246 19Poa annua 240 16Vulpia spp. 228 10Apera spica-venti 225 13Raphanus raphanistrum 213 19Rumex acetosella 207 17Lapsana communis 179 15Sinapis arvensis 179 13Alopecurus geniculatus 174 14Capsella bursa-pastoris 166 16Plantago lanceolata 155 10Fallopia convolvulus 150 25Convulvulus arvensis 147 11Thaspi arvense 136 14Setaria viridis 120 7Galeopsis tetrahit 119 8Solanum nigrum 116 9Bromus sterilis 113 7Centaurea cyanus 96 7Polygonum persicaria 96 5Cirsium arvense 95 10Ipomoea spp. 93 6Myosotis sp. 84 15Digitaria sanguinalis 81 7Alopecurus myosuroides 76 7Panicum spp. 76 5Cirsium vulgare 72 8Poa trivialis 71 5Dactylis glomarata 68 5Geranium sp. 68 9Silene sp. 68 7Spergula arvensis 68 7Vulpia bromoides 67 4Sonchus asper 64 7Erodium cicutarium 60 6Ammi majus 59 3Rapistrum rugosum 59 6Sida spp. 59 4Melilotus spp. 56 4

Meier

contribute.

Meier

should be at bottom of 1st collumn. Text seems to have got smaller on this sheet, as you can see that the green box is too low down now.


The main work of the ISTA Flower SeedCommittee (FSC) since 1998 has been toprepare material for a flower seed testinghandbook. It is a challenge since no similarhandbook was issued before. We are wellaware of the fact that flower seed testing isnot a highlighted area in the seed testingworld but it is an important matter for thosewho make their living from flower seedmultiplication and trade. So hopefully thefuture handbook will be a very useful guidefor laboratories testing flower seeds.

The handbook will consist of two mainparts. First the general part with chaptersIntroduction, Apparatus, Reference -andperhaps Method Validation Programme-which contain important but general knowl-edge about seed testing and will not berepeated in the working sheets later. Thesecond part contains the working sheetswith a series of procedures and detaileddescriptions, instruction and conditions ofthe most important laboratory seed tests ofa flower species or genera. The main parts of a work sheet are:- Title page: gives the botanical data of the

species -botanical and common name andfamily- Seed description: text and coloured pho-

tos in natural size and enlarged- 1000 seed weight: data for information- Purity test: pure seed definition, sample

size- Germination test: method, seedling

description, coloured photo of normalseedlings and of the most frequent types ofabnormal seedlings- Tetrazolium test: method description and

colour drawings to help evaluation- Seed-borne diseases: list of pathogens,

diseases, distribution, reference, CMI andtest method- Literature: list concerning the species

In the beginning of the work 6 genera werepicked as the first task. These are:Calendula, Gaillardia and Tagetes from theAsteraceae family, Dianthus fromCaryophyllaceae family, Impatiens fromBalsaminaceae family and Viola from

Violaceae family. The work had a uniformpattern: 1. Determine if the present method is cor-rect: questionnaires were circulated amongFSC members by working group leadersand from the evaluation of these it can betold if a method is correct or there are someneed to change the conditions of the germi-nation test or modify the seedling evalua-tion 2. Detect points where changes are need-ed: by comparative tests it can be detectedwhere exactly the changes are needed. Aworking group leader organises the com-parative tests. In the work of workinggroups 5-7 FSC members participate andsometimes there are members from otherISTA committees also from laboratorieswhere they often test the given species.3. Proposal for changes in the presentmethod: discussions among FSC workinggroup members and also with other ISTAcommittees and when it is needed organisa-tion of repeated comparative tests4. Draft text of the work sheet: by thereport of the working group leaders the FSCchair prepares a draft text and circulate itfor validation within FSC and after withinISTA5. Issue of handbook work sheet: thefinally accepted version.

To get the right answers to the professionalquestions we work together with the ISTAGermination and Tetrazolium Committeesto find accurate test methods for everyspecies. We also try to harmonize the testmethods of ISTA and AOSA during thiswork and AOSA members also take part inour comparative tests and their evaluation. We started consultations with members ofAOSA and SCST as well as experts fromseed industry, which revealed that at presentit is unlikely to fully harmonize the twomethods because of the economic consider-ations of SCST. (It means mainly the differ-ence in the evaluation of normal and abnor-mal seedlings -presence and number of pri-mary and secondary roots and physiologicalnecrosis.) Therefore at the present stage inthe handbook we cite the ISTA methods and

point out the AOSA Rules as a note forinformation to the analysts interested.

Experience of the first 6 work sheets:The most complicated and time consumingtasks were: the seed description part, to findthe most frequent types of abnormalseedlings, to prepare TZ test method andcollect seed-borne diseases.For Calendula, Dianthus and Viola spp. -there was no difficulty in the preparation ofthe work sheets.Gaillardia spp. -there was no difficultyonly TZ method was new so it had to bechecked.Tagetes spp. -there are some trouble withthe evaluation of seedlings. The question iswhether seedlings with defective primaryroot and well-developed secondary rootsshould be considered normal or abnormal.Impatiens spp. - for germination ofImpatiens walleriana the addition of lightwas suggested in the method. Based on thecomparative tests of FSC the new seedlingevaluation group of Impatiens should beA.2.1.1.2.

Since e-mailing is general today our workbecame much faster and more communica-tive than before but I feel that we couldimprove it by making InterNet conferenceson the ISTA website. (I mention it becauseit is the sort of work, which demands ratherpersonal discussions among several people.We can experience friendly but animatedprofessional discussions during ISTA work-shops. I miss this feeling via bilateral e-mails.)

All this work was started by Lea Mazor for-mer chair of FSC and completed by FSCmembers who are always willing to makethis extra work to contribute to the maingoal of ISTA: uniformity in seed testing.Working group leaders were for Impatienswalleriana and Gaillardia: Lea Mazor,Viola tricolor and Petunia: SylvieDucournau, for Tagetes: Aleta Meyr, forCalendula officinalis: Marcia Taylor, forDianthus: Zita Ripka and for Cyclamen:Rita Zecchinelli. FSC also owes a lot ofthanks to other members of ISTA who

TECHNICAL COMMITTEESFlower Seed Testing

Flower Seed TestingPreparation Work of the ISTA Handbook on Flower Seed TestingBy Zita Ripka, ISTA Flower Seed Committee Chair


helped our work with their kind and helpfulcooperation. They are mainly the membersof ISTA Germination Committee, in TZtesting the LUFA staff, in seed-borne dis-eases Prof. Wen-Shi Wu, Jim Sheppard inwebsite matters and Bettina Kahlert ineverything general. Dr. Norma B. Deginani

and Dr Raul Pozner, the Darwin Institute ofBotany, Argentina gave their assistance inseed description.

In the future work the preliminary question-naires will be the basis of the work sheetswhere the result shows no difficulty or wish

to change the present method. Comparativetests will be carried out only for genera orspecies where some changes are needed andfor new species. This way we hopefully canquicken up the preparation of workingsheets for the next species and genera.


Questionnaire on Cyclamen sp.for the ISTA Flower Seed Committee working group

Return it please before 15 October 2001to: Rita Zecchinelli - Laboratorio ENSE - Via Emilia km 307 26838, Tavazzano (LO)

Italye-mail: [email protected]

Fax: +39 0371 760812Tel.: +39 0371 761919-760135

Name: ………………………………………………………………………………………Laboratory:…………………………………………………………………………………Number of samples tested a year (max): 0-10 10-30 30-50 50-100 +100Average of the last 5 years:……………..Are you a practised analyst in germination test of Cyclamen sp.: yes no

Please answer the following questions:1/ Indicate the frequency of seedling abnormalities by your experience:

O none, x seldom, xx seldom to regular, xxx regular, xxxx regular to often, xxxxxoften

Type of abnormality FrequencyI. Primary rootsI.2. StubbyI.3. RetardedI.8. SpindlyI.10. With negative geotropismI.11. GlassyI.12. Decayed as a result of primary infectionI.13. Only secondary roots developedII. Hypocotyl, epycotyl, mesocotylII.2. Not forming a tuberII.3. Deeply cracked or brokenII.6. ConstrictedII.10. SpindlyII.11. GlassyII.12. Decayed as a result of primary infectionIII. CotyledonIII.3. Broken or othetwise damagedIII.8. Decayed as a result of primary infectionV. Terminal budV.1. DeformedV.3. AbsentVII. Seedling as a wholeVII.1. DeformedVII.2. FracturedVII.4. Two fused togetherVII.6. Yellow or whiteVII.7. SpindlyVII.8. GlassyVII.9. Decayed as a result of primary infection

2/ Is it neccessary to revise the germination methods in ISTA Rules Table 5A Part3? Please indicate your suggestions in the following table:

Substrate Temp(ºC) First count Final count Breaking (days) (days) dormancy

ISTA Rules TP;BP;S 20;15 14-21 35 KNO3-Soak in water 24 hours

Changes………………………………………………………………………………………

3/ Do you think that seedling evaluation group A 2.1.4.3. is correct? YesNo, I think …………………………would be more adequate, because….………………………………………………………………………………………………….………………………………………………………………………………………………….………………………………………………………………………………………………

4/ Do you have data for 1000 seed weight of Cyclamen sp.? …………….. g.

5/ Do you agree with the Pure Seed Definition (N° 10) given by the ISTA Rules1999 for Cyclamen sp? Yes

No, I will suggest……………………………………………………………………………………………………………………………………………………………………………………

6/Can you provide photographs of normal and abnormal seedlings for thehandbook?

Yes No

7/Can you provide photographs of growing stages of Cyclamen sp seedlingsand young plants for the handbook?

Yes No

8/ Do you have any suggestion regarding Cyclamen sp lab tests and/or thehandbook?

…………………………………………………………………………………………………...

…………………………………………………………………………………………………...

…………………………………………………………………………………………………...

…………………………………………………………………………………………………...

……………………………………………………………………………………………………

Figure 1. Questionnaire on Cyclamen sp.

Report of ISTA FSC Cyclamen Working GroupSummary of Questionnaire on Cyclamen (October 2001)By Rita Zecchinelli, ISTA FSC Cyclamen Working Group Leader

The questionnaire was sent to 16 membersof FSC and reported by 11 of 10 countries.The answers are: 1. Degree of experience on testingCyclamen seeds: 6 laboratories: no experi-ence, 3 laboratories: just a few samples (<10 samples/year), 2 laboratories: moreexperience (> 50 samples/year)2. Frequency of seedlings abnormalities:see the table below

3. Is it necessary to revise the germina-tion methods in ISTA Rules: 1 answer:substrate to be compost, count days 35-49 1answer: final count 42 days, 2 answers:darkness4. Do you think that seedling evaluationgroup A 2.1.4.3 is correct: All answersCORRECT5.Data for 1000 seed weight of Cyclamen:from 4 to 11g, from 5 to 10g, 12 g.

6. Do you think that pure seed definitionN° 10 is correct: All answers CORRECT7. Photographs for abnormal: YES (Italy,France)8. Photographs for growing stages: YES(Italy, France)

The Report of FSC ISTA Cyclamen WorkingGroup is continued on page 34.

Meier

move the box


Summary of Cyclamen ring test (March 2002)

2 samples of Cyclamen seeds -Germination and TZ test

1. Germination ring test7 participating laboratories in alphabeticalorder:- Agri Seed Testing, Oregon - USA

(Sharon Davidson) - GEVES, SNES - F (Sylvie Ducournau) - ENSE - Italy (Rita Zecchinelli)- LaRAS - Italy (Enrico Noli)- Naktuinbow - Netherland (Petra

Remeeus) - National Institute for Agricultural

Quality Control - Hungary (GyöngyiIvanovics)

- Official Seed Testing for England andWales - Great Britain (Linda Maile)

Two laboratories carried out the tests bothin light and in darkness condition; two lab-oratories in darkness and three in light only.

Observations Generally speaking, germination ofCyclamen seeds seems to be higher in dark-ness than in light and the results obtainedby the different laboratories are morehomogeneous.

The different value of germination betweenlight and dark is due to the number ofabnormal seedlings: the light seems toincrease the amount of abnormalities.Regarding the kind of abnormal seedlings(shown in Table 1), the ring test confirmsthe results of the questionnaire and showsthat in case of Cyclamen the classificationof abnormalities regards most of the struc-tures of the seedlings.The final count for some laboratories was35 days, but most of them (5) preferredlonger duration of the trial, according toISTA Rules 5.6.4.A Come believed that theduration of 35 days does not fit to themajority of seed lots of Cyclamen.ConclusionsISTA method for germination is correct. Bythe results obtained from the referee test therecommended method is TP, 20°C forCyclamen. ISTA method gives no specification aboutthe light conditions but according ourobservations germination in dark gives bet-ter results and less abnormal seedlings. It often occurs that normal seedlings cannot develop within the 35 days period sofinal count day can be put 7 days lateraccording to ISTA Rules 5.6.4.A. No statis-tical evaluation was calculated.


Type of Abnormality

I. Primary rootI.2 StubbyI.3 RetardedI.8 SpindlyI.12 DecayedII. HypocotylII.2 Not forming a tuberII.6 ConstrictedII.10 SpindlyII.12 DecayedIII CotyledonsIII.3 Broken or damagedIII.8 DecayedVII Seedling as a wholeVII.1 DeformedVII.4 Two fused togetherVII.7 SpindlyVII.9 Decayed

Frequency in the questionnaire

XXXX0

XXX

XXXXXX

XXX

XXXXX

XXX

XXXX

Frequency in the ring test

XXXXXXXXXXX

XX00X

0X

XXX0

XX

Table 1. The most frequent types of abnormal seedlings:

X seldom; XX seldom to regular; XXX regular; XXXX regular to often; XXXXX often

Substrate

TP, BP, S

Temperature(°C)

20, 15

First count (days)

14-21

Final count (days)

35

Additional directions (breaking dormancy etc.)

KNO3, soak in water 24 hours

Table 2. Germination test method according to ISTA Rules

Figure 1 - 3:Results of germination ring test

of Cyclamen

Figure 1. Rate of normal seedlings

Figure 2. Rate of abnormal seedlings(Note: laboratory No. 1 reported 0 abnor-mal seedlings in both tests)

Figure 3. Rate of ungerminated seeds

light dark

0

20

40

60

80

100

1 2 4 5 6 1 2 3 7

laboratories

norm

al s

eedl

ings

(%)

Sample 1 Sample 2

light dark

05

10152025

1 2 4 5 6 1 2 3 7

laboratories

abno

rmal

see

dlin

gs

(%)

Sample 1 Sample 2

light dark

0

20

40

60

80

100

1 2 4 5 6 1 2 3 7

laboratories

unge

rmin

ated

see

ds (%

)

Sample 1 Sample 2


2. Tetrazolium ring test6 participating laboratories in alphabeticalorder:- Agri Seed Testing, Oregon - USA

(Sharon Davidson) - GEVES, SNES - F (Sylvie Ducournau) - Laboratorio Sementi, MiPAF, ex ASFD-

Italy (Fabio Gorian)- Naktuinbow - Netherland (Petra

Remeeus) - National Institute for Agricultural

Quality Control - Hungary (GyöngyiIvanovics)

- Official Seed Testing For England andWales - Great Britain (Linda Maile)

ISTA method:Pretreatment:- Soak 24 hours in water at 20°C

Preparation for staining:- Cut longitudinally through the seed near

the hilumStaining:- 24 hours, 1% TZ-solution, 30°C

Preparation for evaluation:- Expose the embryo by slicing the

endosperm step by stepEvaluation (maximum area of unstained,flaccid and/or necrotic tissue permitted):- The position of the embryo inside the

endosperm can differConclusions:The recommended ISTA method is correct. No statistical evaluation was calculated.


05

101520

2530

1 2 3 4 5 6

laboratories

non

viab

le s

eeds

(%)

Sample 1 Sample 2

020406080

100120

1 2 3 4 5 6

laboratories

viab

le s

eeds

(%)

Sample 1 Sample 2

The questionnaire was sent to 7 personsfrom 7 countries. The answers are:

1. Frequency of seedling abnormali-ties. (see table hereafter)2. Is it necessary to revise the germi-nation methods in ISTA Rules. All theanswers are: No3. Do you think that seedling evalua-tion group A.2.1.1.1 is correct. All the

answers are: Yes4. Can you provide photographs ofnormal and abnormal seedlings for theHandbook.One laboratory can provide photographs,one can try if necessary.5. Data for 1000 seed weight ofPetunia. 4 answers quite similar : around0,1 g (minimum: 0,09 g ; maximum :0,16 g)

Report of ISTA FSC Petunia WorkingGroup Summary of Questionnaire on Petunia(October 2001)By Sylvie Ducournau, ISTA FSC Petunia Working Group Leader

Frequency in the questionnaire

XX0X

XXX0

XX

XXXX

XXXXX

XXX0

XXXX

Frequency in the ring test

XXXX

XX00

XXXXX

0X0XX

XX0

XXXXX

Type of Abnormality

I. Primary rootI.1 StuntedI.2 StubbyI.3 RetardedI.4 MissingI.8 SpindlyI.11 Glassy

II. HypocotylII.1 Short and thickII.11 Glassy

III CotyledonIII.2 DeformedIII.3 Broken or damagedIII.5 DiscolouredIII.6 NecroticIII.8 Decayed (primary infection)

VII Seedling as a wholeVII.1 DeformedVII.6 Yellow or whiteVII.7 SpindlyVII.8 GlassyVII.9 Decayed (primary infection)

Table 1. The most frequent types of abnormal seedlings

X seldom ; XX seldom to regular; XXX regular; XXXX regular to often; XXXXX often


(see Table 2. below)· Substrate: one laboratory uses top of blot-ter, medium wet· Temperature: one laboratory uses a nonISTA temperature (25°C)· Light: one laboratory does not use light forthe germination· Duration: one laboratory has a duration ofthe germination test of 21 days instead of14 days· Pre treatment: 3 laboratories use pre chill-ing (2 to 5 days) and 2 laboratories useKNO3.ObservationsPre chilling does not seem to increase thegermination rate of samples used in the ringtest. Absence of light during the test does notallow to evaluate defects of cotyledon. Temperature of 25°C (non ISTA) does notdecrease the germination rate of the sam-ples tested. Deviation of laboratory 06 is difficult toexplain. Perhaps it could be due to substrateand/or humidity of substrate.

ConclusionsISTA method for germination is correct. Bythe results obtained from the referee test therecommended method is TP, 20-30 °C forPetunia.

Statistical evaluation of test resultsGermination results have been analysed sta-tistically with a tool developed to assessrepeatability and reproducibility in inter-laboratory tests according to ISO 5725-2(Grégoire S., ISTA Web Site).· Results are reported sample by sample · Statistical analysis is reported in the tablesbelow for normal seedlings, abnormalseedlings and ungerminated seeds.

The h values show the tendency for a labo-ratory to give over estimations or underestimations, compared to the mean of all theresults available.The k values show thevariability of the repeats.In general results of normal seedlings are ingood accordance between laboratories,except one lab (n°06). If we analyse theresults of abnormal seedlings, laboratory 06reports high quantity of abnormal seedlingswith root and hypocotyl defects.

The tendency observed for laboratory 06 tofind less normal seedlings and more abnor-mal seedlings than the overall mean is high-ly significant.Laboratory 07 has tendency to find lessabnormal seedlings than the mean of alllabs, but it is not significant.For the ungerminated seeds, no significantdifference has been found between labora-tories.


Substrate

TP

Temperature(°C)

20-30; 20

First count(days)

5-7

Final count(days)

14

Additional directions(breaking dormancy etc.)

Prechill, KNO3

Table 2. Test method according to ISTA Rules

Significance level H value 1% 5%

Sample 1 Lab 06 -2.100 Sample 2 Lab 06 -2.001 Sample 3 Lab 06 -1.989

1.98 1.71

0.0

20.0

40.0

60.0

80.0

100.0

Sample 1 Sample 2 Sample 3N

orm

al s

eedl

ings

(%)

Lab 01Lab 02Lab 03Lab 04Lab 05Lab 06Lab 07

h values superior to h theoric

Summary of Petunia ring test (2002)3 samples of Petunia seeds -

Germination test

7 participating laboratories in alphabeticalorder:- ENSE- Italy (Rita Zecchinelli)- GEVES-SNES, France (Sylvie

Ducournau)- Naktuinbouw- Netherlands (Petra

Remeeus)- National Institute for Agricultural

Quality Control,- Hungary (GiziHorvath)

- National Seed Testing Laboratory, -Yugoslavia (Mirjana Milosevic)

- Norvegian Agricultural InspectionService, Seeds Department, -Norway(Hakon Tangeras)

- Ransom Seed Laboratory- U.S.A. (AletaMeyr)

Figure 1. Normal Seedlings



0

5

10

15

20

Sample 1 Sample 2 Sample 3

Unge

rmin

ated

see

ds (

%)


0.0

10.0

20.0

30.0

40.0

Sample 1 Sample 2 Sample 3

Abn

orm

al s

eedl

ings

(%)


Significance level K value 1% 5%

Lab 01 Sample 3 1.551 Lab 05 Sample 2 1.718 Lab 06 Sample 1 1.620 Lab 06 Sample 2 1.595

N.S. 1.55 Significance level H value 1% 5%

Lab 07 Sample 3 1.797 1.79 1.55


Sample 2 Lab 05 -1.955 1.55


Sample 1 Lab 06 2.180 1.98 1.71 Sample 2 Lab 06 2.036 1.98 1.71


k values superior to k theoric

Figure 2. Abnormal seedlings Figure 3. Ungerminated seeds

k values superior to k theoric


Two computer programs are available in theISTA statistical toolbox on the web site athttp://www.seedtest.org/STA/sta_toolbox.cfmThese programs can help you to:- check the efficiency of existing controlprocedures- select appropriate new control proceduresif your aim is to check if the number or pro-portion of seeds which are not of theexpected common type are within accept-able limits.

If you wish to understand the principlesused by Qualstat and Seedcalc, you canread the paper "Statistical considerations inseed purity testing for transgenic traits" inSeed Science Research (2001) 11, 101-119Remund K., et al.

Both programs work: if you check each individual seed or plantor if you are able to detectunexpected seeds in a group(or pool) of seeds as inELISA, biochemical orDNA analysis to detect spe-cific traits or bacteria.

In both programs you cansee the influence of falsepositives and false negativeson your decision schemes.The graphics provide an easyway to see the efficiency ofdifferent sampling schemes.The values of the buyer andseller risk probabilities areprovided if you need to putthem in the protocols orquality assurance manuals. Qualstat runs independent ofany other software program.Seedcalc is a spreadsheetthat runs in MicrosoftExcel®.

Both programs providedesigns for standard singleand double stage samplingschemes. Qualstat also

includes some more complex sequentialsampling schemes. Seedcalc allows theuser to incorporate some simple calcula-tions toe s t i m a t ethe expect-ed cost ofimplement-ing a givens a m p l i n gscheme. Q u a l s t a thas anaccompa-nying userm a n u a l .S e e d c a l chas ana t t a c h e dsheet withbrief user

instructions for each calculations sheet. Amore detailed users manual is forthcomingfor Seedcalc.


TECHNICAL COMMITTEESStatistics Programmes OnLine

Qualstat and SeedcalcTwo Programmes available in the ISTA Statistical Toolbox atwww.seedtest.org/STA/sta_toolbox.cfmBy Sylvain Grégoire, ISTA Statistics Committee Chair andKirk Remund, ISTA Statistics Committee Member

Figure 2. Seedcalc

Figure 1. Qualstat


7th ISTA Tetrazolium Workshop 2002

Nineteen seed analysts from 17 differentcountries attended the 7th ISTA TetrazoliumWorkshop 2002 hosted by the StateAgricultural Extension and ResearchStation Augustenberg (LUFA), Karlsruhe,Germany. In addition, there was:- the host Prof. Dr. Norbert Leist, ISTAPresident, former TEZ Chairman, lecturerand co-organiser; - the leader of the Workshop Ing. AugustoMartinelli, Chairman of the TEZ; - co-organisers and lecturers:

Ronald Don, Vice-Chairman of the TEZand Member of the Executive Committee;

Prof. Dr. Dr. h. c. Adolf Martin Steiner,former Vice-chairman of the TEZ,Honorary Lecturer; and

Dr. Jose Franca Neto, Member of theTEZ.- Instructors supporting the practical workMrs. Stephanie Krämer and Helga Werth.From Augustenberg Mrs. Andrea Jonitztook care of organisational matters togetherwith Mr. Gerhard Lindörfer and RainerKnoblauch, the latter also being responsiblefor transportation. There were also numer-ous other assistants from Augustenberg pro-viding for the needs of the participants andensuring that the workshop was a success.

The organisers, lecturers and instructorswith the experience of the 5th TEZWorkshop in Edinburgh and the 6th TEZWorkshop in Buenos Aires (cf. ISTA NewsBulletin 116, 6-9, 1997) had prepared aprogram designed to achieve the aims of theWorkshop:(i) Adding to uniformity in TEZ testing onan international basis by jointly carryingout tests and discussing their evaluation.(ii) Deepening the understanding of TEZtesting by increasing participants knowl-edge of the scientific background to TEZtesting.(iii) Providing participants the opportunityto comment on the amendments in the ISTARules of 1999 and 2002.(iv) Explaining the working program of theISTA TEZ Committee and drawing up a listof future activities.

In view of these aims it was a pleasure towelcome colleague Annette Miller the

Chair of the TEZ Committee of ISTA'snational sister organisation AOSA.

As working basis for the participants theProceedings of the ISTA TetrazoliumWorkshop held in 1997 at the OSTSEdinburgh (published by the ISTA) and afolder containing the contents of addressesand instructions for preparation and evalua-tion of the species being studied in the prac-tical sessions was distributed. In relation tothe lectures Prof. Steiner provided hand-outs that documented the progresssince1997, and Vice-Chairman Don provid-ed copies of his excellent photographs withexplanations on a CD-ROM. Thanks to thegenerous support of the Workshop by spon-sors, a complete set of tools for TEZ testingcontained in a smart leather wallet with theinsignia of the Workshop was presented toeveryone as a practical and lasting memen-to.

For practical work, a spacious laboratorywas provided together with necessary addi-tional rooms in the same building whichwas completely reserved for the Workshop.The Zeiss-binoculars and the video demon-

stration facilities were of the most moderntype. The lectures were presented in a hallin the historical Castle Augustenberg. The200 meters between venues pleasantlyallowed participants some time for free dis-cussion whilst on route.

The program of the Workshop as shown inthe first insert was compiled as a result of aquestionnaire sent to participants. Whentesting a species, an introduction and finaldiscussion was held by Ing. Martinelliassisted by respective specialists. The lec-tures, listed in the second insert, weretimetabled to fill the gaps between thestages of practical testing. The alternate ses-sions of practical and intellectual challengecreated a stimulating atmosphere.

Special mention has to be made of theunsurpassed hospitality of our host. Theparticipants lodging in two Hotels weretransported to and from the workshop andtransport was also provided for cultural andsocial events. This made commuting com-fortable and supported discussion andfriendship between participants. TheWorkshop building was situated in an

TECHNICAL COMMITTEES

7th ISTA Tetrazolium Workshop 2002Karlsruhe-Augustenberg, Germany, July 29 - August 2, 2002By Adolf Martin Steiner, ISTA Member,Norbert Leist, ISTA President andRonald Don, ISTA Tetrazlioum Committee Vice-chair

Nineteen seed analysts from 17 different countries attended the 7th ISTA TetrazoliumWorkshop 2002 hosted by the State Agricultural Extension and Research StationAugustenberg (LUFA), Karlsruhe, Germany.


TECHNICAL COMMITTEES7th ISTA Tetrazolium Workshop 2002

arboretum neighbouring orchards. The flo-ral displays in the buildings were admiredby all and tea/coffee breaks took place in alight filled lobby or under the blue sky inthe front of the building. Hot and cold bev-erages and biscuits were available and eachday different tasty local food specialitieswere presented. Lunch was served in animpressive vault of a historical building 10minutes walk from the LUFA and this wascelebrated every day as another enjoyableget-together. All this contributed to thefriendly atmosphere and gave renewedimpetus for an afternoon of concentratedand dedicated earnest work.

Several cultural and social highlightsenriched the working program. On Sunday,prior to the start of the workshop, a guided

walking tour revealed scenic views of theold village of Durlach which was foundedin 1161. At the subsequent "Meeting in theBiergarten Traube" those having arrived toolate for the walking tour joined the othersfor a typical meal accompanied by sometraditional refreshments. On Monday therewas an evening tour to Karlsruhe, whichwas the residence and capital of the formerGreat Dukedom Baden. There we saw theBotanical Garden, the Castle, the SupremeCourt Buildings, the shopping mall"Kaiserstraße" and the Market Place. Theday ended with a meal at one large table inan old hall of a brewery. An invitation toRainer's cottage at Grötzingen followed onTuesday. After a humorous address by Prof.Leist and a kind welcome by Mr. Rainer

Knoblauch, in his back garden, Mrs. AliceLeist and Mrs. Birgit Klauke were intro-duced as those being responsible for thisevent. The yard and an old barn had beenset out with tables and chairs. Tasty salads,well-seasoned baked or grilled sausages,fresh vegetables, local breads and sweetdesserts delightfully combined with softdrinks, beers and local wines providedeveryone with a meal and a night toremember. Some ate in the garden understarlight and some in the barn with candlelight and when the meal was over the ISTAfamily came together for an unforgettablesing song. Our most cordial warm thanksmust go to the generous hosts. Wednesdayevening or what was left of it after amarathon practical session was at own dis-posal.

On Thursday morning we set out for anexcursion to the University of Hohenheim.On arrival in the Institute of Plant Breeding,Seed Science and Population Genetics,after a refreshment and a snack, Prof.Steiner presented a lecture on "The Historyof Tetrazolium Testing". Thereafter partici-pants were treated to an exhibition of his-torical tools, equipment, drawings and pho-tographs. These were from the times ofProf. Friedrich Nobbe, who founded seedtesting in 1869, and of early TEZ testingfounded by Prof. Georg Lakon in 1942.Following a tour through the HohenheimGerman Agricultural Museum lunch wastaken at the students refectory. Participantswere then treated by Prof. Steiner to a guid-ed tour of the 35 hectare Hohenheim

Gardens. Part of these form the famousState Arboretum and contains about 2500different tree species and varieties. Abouttwo dozens of the trees are more than 230years old having been planted by Duke CarlEugen of Württemberg, the founder of theGardens in 1776. There was the unique pos-sibility to compare an established 230 yearsold garden with a recently planted enlarge-ment. Finally, in one of the conserved his-torical buildings, contained in the gardens,the Small Museum of the History ofHohenheim was visited. Hohenheim wasfirst mentioned in 1100.

On the route back to Karlsruhe, the romanstyle Cistercian Monastery of Maulbronn,founded in 1147, was visited. This is thebest preserved German Monastery and isregistered as an UNESCO World CulturalHeritage Monument. An excellent guidedemonstrated why this monastery receivesmore than 200.000 visitors a year. The tourended with a Festive Workshop Dinner inthe "Monastery tap-room" housed in one ofthe huge historic buildings.

At this dinner Prof. Steiner and Mrs. Werth,who were both going to retire this year,were honoured and said farewell. With it 60years of TEZ research at Hohenheim willend. It had started with Prof. Georg Lakonwho invented TEZ testing in 1942. He wasfollowed by Prof. Werner Lindenbein andDr. Helene Bulat who established Chapter 6in the ISTA Rules in 1965, and they werefollowed by Prof. Steiner who revisedChapter 6 in 1999. Now, in GermanyAugustenberg takes over the lead in TEZtesting and research and, hence, organisedthis 7th Tetrazolium Workshop to honourHohenheim's achievements.

After five days of: - earnest work and enthusiastic discussionon tetrazolium testing, of - delightful cultural and social events and - renewed and new friendships, every participant received an ISTACertificate of Participation.

Special thanks were expressed to the organ-isers and lecturers by presenting each one aunique small hand-painted porcelain plate.This showed a painting of the oldAugustenberg Castle, and was inscribedwith the recipients name, Seed TestingStation Augustenberg and ISTA-Tetrazolium-Workshop. The gifted artistwas Mrs. Gerda Jonitz, sen.

For sure, nobody will ever forget the expe-rience gained by the daily work striving foruniformity in TEZ testing and nobody willforget the friendly hours spent with the col-leagues. Hopefully, many more of such sci-

Participants were then treated by Prof. Steiner to a guided tour of the 35 hectareHohenheim Gardens. Part of these form the famous State Arboretum and containsabout 2500 different tree species and varieties. About two dozens of the trees are morethan 230 years old having been planted by Duke Carl Eugen of Württemberg, thefounder of the Gardens in 1776.

entifically and personally rewarding Workshops are to come.Wholeheartedly, warm thanks again to the hosts, the organisers andlecturers, thanks also to the participants.

Finally, many thanks to Prof. Dr. Friedel Timmermann, Director ofthe LUFA Augustenberg, for his continuous support of the activi-ties of the Section Seed Testing and Applied Botany. Without suchsupport the Workshop would not have been possible.


TECHNICAL COMMITTEES7th ISTA Tetrazolium Workshop 2002

Program of the 7th ISTA Tetrazolium Workshop 2002

1st day Welcome and introductionLolium sp., Hordeum vulgare; preparation, staining and evaluation; lectures

2nd day Tagetes sp., Zea mays, Lotus sp., Lycopersiconesculentum; preparation and staining; lectures; evaluation of parallel germination test Zea mays; germination test versus viability test in Zea mays

3rd day Tagetes sp., Zea mays, Lotus sp., Lycopersiconesculentum, Oryza sativa; evaluation; lectures; guided tour LUFA Augustenberg Seed Testing Station; Glycine max evaluation; lecture

4th day Excursion to the University of Hohenheim

5th day Finishing practical work, lectures,discussion forum; Farewell and workshop closure

Lectures at the 7th ISTA Tetrazolium Workshop 2002

Presentation of the International Seed Testing AssociationISTA. N. Leist Presentation of the ISTA Tetrazolium Committee 2001 - 2004. A. Martinelli Tetrazolium salts and biochemistry of tetrazolium reduction. A. M. Steiner Presentation of the 2001 and 2002 amendments to the 1999edition of the ISTA Rules in the Annexe to Chapter 6. A. Martinelli Relationship between germination testing and viability testing. A. M. Steiner Explanations of staining patterns caused by heat damage,water damage, mechanical damage, freeze injury, microorganisms etc. R. Don Presentation of the new ISTA Tetrazolium Handbook.N. Leist Quality assurance and health and safety in tetrazolium testing. R. Don Tetrazolium testing in Glycine max. J. Franca Neto History of topographical tetrazolium testing. A. M. Steiner Tetrazolium testing: sample size, statistical basis for assessingtolerances and use of sequential testing procedures. A. M. Steiner and R. Don Tetrazolium staining as an indicator of vigour. A. M. Steiner and R. Don

Are you a native Englishspeaker? With

experience in seed sci-ence?

Would you like to help our colleaguesfrom countries where English is not

the first language (or second) to pub-lish papers in Seed Science and

Technology (SST) in a correct English?

Please contact:Anne Bülow-Olsen

Chief [email protected]

A N N O U N C E M E N TISTA Working Sheets on Tetrazolium Testing

Volume IAgricultural, Vegetable and Horticultural Species

Volume IITree and Shrub Species

Edited byNorbert Leist and Stephanie KrämerIllustrated by Jochen PfäfflinThe Tetrazolium Working Sheets Part 1 and 2 include detailedand standardised description to conduct and evaluateTetrazolium tests for the determination of viability from agri-cultural, horticultural and forest seed.

The working sheets present 120 agricultural and 122 forestspecies and genera, respectively, in as much as the testing ofthe species took place similar. The description is illustratedwith pictures of the seed morphology, the cutting instructionsand the different stages of non-viable seeds. The workingsheets support the International Rules for Seed Testing by pro-viding detailed working plans to the seed testing laboratories.This publication is a result of the experiences of the daily rou-tine work of a seed testing laboratory and the optimisation ofcompetent members of the ISTA Tetrazolium Committee fromall over the world. The Tetrazolium Working Sheets contributeperfectly to one of the main aims of ISTA: Harmonization ininternational seed testing.

The 8th International Congress of Plantpathology held in Christchurch, NewZealand provided an opportunity for thePlant Disease Committee of ISTA to con-duct a seed pathology workshop followingthe Congress at Lincoln University. Thiswas the first ever ISTA workshop in NewZealand and the first ISTA gathering inNew Zealand since the Fifteenth ISTACongress in 1968.

Twenty-five people registered for the work-shop. Five were from Australia, 4 from UK,3 from USA, 3 from New Zealand, 2 fromDenmark, 2 from India, and one each fromNorway, Israel, The Netherlands, Spain,Canada and Switzerland. Eight currentmembers of the PDC (Jim Sheppard,Valerie Cockerell, Guro Brodal, SteveRoberts, Denis McGee, Kees Langerak,Rivka Hadas, and Rouke Bakker) were atthe workshop. Presenters were assisted bytwo postgraduate students (from Brazil andUruguay), Dr. Wadia Kandula and MrRouke Bakker who also participated. Theworkshop concentrated on detection ofseed-borne diseases of vegetables and othercrops of interest to the Asia/Pacific region.Participants had the opportunity for practi-cal "hands-on" training, as well as informa-tion transfer provided by ISTA PDC mem-bers and other seed health specialists.

Daily sessions were composed of laborato-ry exercises interspersed with oral presenta-tions on various topics pertaining to seedhealth testing. Methods for detection ofseveral host/pathogen combinations weredemonstrated. Xanthomonas campestris pvcampestris (blackrot) of Crucifers (SteveRoberts). This method has been the subjectof extensive development and review isproposed for inclusion into the InternationalRules for Seed Testing this year. JimSheppard demonstrated a recent compara-tive test for detection of Phoma lingam(blackleg) of Crucifers using the currentISTA method, 7-004, and proposed modifi-cations using freezing blotters or absic acid.The results from the various treatmentswere compared. Guro Brodal (Norway)presented the Osmotic blotter test for detec-tion of Drechslera spp. in barley and oats

used in the Nordiccountries for manyyears. Rouke Bakkerdemonstrated a methodroutinely used byBiolinc for detection ofPseduomonas syringaepv.pisi. The advan-tages and disadvangesof three agar methodsfor detection ofMichrodochium nivaleand various Fusarium species was exam-ined in a session led by Valerie Cockerellwho organized a recent comparative test onthese pathogens. Pea seed borne mosaicvirus (PSbMV) and lettuce mosaic virus(LMV) are important seed-borne diseasesaffecting seed production. There is no stan-dard testing method currently available forPSMV but an ELISA method has beenadopted by Biolinc. An ELISA method hasalso been the subject of recent comparativetesting. Demonstration of both methods inthe laboratory and a discussion of issuesrelated to the suitability of the two tests forroutine testing was led by Rouke Bakker.Wadia Kandula present methods describedfor detection of Ascochyta in chickpea andthe Ascochyta complex in pea and dis-cussed recent advances in molecular meth-ods for detection of these pathogens.Other topics discussed during the workshopincluded the role of ISTA in establishinguniformity in seed health testing, moleculartechniques in seed health testing, seedhealth issues for the Asia/Pacific region,and the ISTA quality assurance, accredita-tion, proficiency test and method validationprogrammes as they relate to seed healthtesting.The learning experiences and the socialinteraction that occurred made this a highlysuccessful workshop. It gave organisers andparticipants the opportunity to put faces tonames, and to reassure themselves that theirwork was up to international standards. Theopportunities taken for networking will beinvaluable.

On the behalf of the PDC I would like tothank the NZSTI, and particularly Director

Professor Murray Hill for supporting theworkshop in so many ways. John Hamptonfor his courage and perseverance in hostingthis workshop following a major event suchas the ICPP; the dedication and hard MrRouke Bakker (PDC member) and DrWadia Kandula and the post graduate stu-dents. Without their efforts (and long hours)we would not have had such a successfulworkshop. I would like also to thank myfellow PDC members for assisting with theprogramme development, oral presenta-tions and laboratory demonstrations.

A proceedings of the workshop will be pub-lished by ISTA and many of the presenta-tions can be found on the ISTA onlineWebsite at http://www.seedtest.org/pdc/works_p.cfm.


TECHNICAL COMMITTEESISTA/ICPP Seed Pathology Workshop 2003

ISTA/ICPP Seed Pathology Workshop 2003 Lincoln University, Canterbury, New Zealand Feb. 10-13, 2003By Jim Sheppard, ISTA Plant Disease Committee Chair

Comments from the participants indicatethat they found this workshop informative,interesting and interactive.

"The presentations made by the differentspeakers were excellent"."A great opportunity for me and I am glad

I came"."Good balance between theoretical infor-

mation and practical work"."It was a good idea to include fungi, bac-

teria and viruses in the same workshop"."BBQ and dinner were superb. No room

for improvement in your kind hospitality"."The workshop was interesting, well

organised, and speakers were well cho-sen".

"Excellent chance to interact; overall anextremely useful workshop".

"A very productive workshop"."An entirely enjoyable experience".

Objectives The workshop will deal with practical prob-lems related to tree seed testing of bothconifer and broadleaf species. The generalaim of the workshop is to create discussionand exchange of information in this area.Based on input from the preliminary regis-tration, the workshop will cover all fields ofseed testing such as Purity, Germination,Tetrazolium, Health, Excised embryo,Moisture content and X-ray tests; also,other fields of interest are Referee tests,Quality Management System, Precision oftest results, Retesting frequencies of storedseeds and Practical work in the laboratory.Tentative ProgrammeFrom Monday, October 20, to Wednesday,October 22nd - presentations, lectures,practical "training"; and a visit to theCentral Institute for Supervising andTesting in Agriculture in Prague (CZDL03).Registration for the workshop is scheduledfor Sunday evening, October 19, andMonday morning, October 20.

There will be two alternative post-meetingtrips (from Thursday, October 23, toSaturday, October 25): 1. Visit to the State Tree Seed Centre inTyniste nad Orlici (1day bus trip of about300 km).2. Visit to the Seed Testing Laboratory forForest Tree Seeds in Uherske Hradiste(CZDL02), southeastern Czech Republic,and then continue on to visit the ForestSeed Testing Laboratory (SKDL02) in

Liptovsky Hradok, Republic ofSlovakia (3 day bus trip of about 1 000km).VenueThe workshop will take place at theCongress Conference Centre Floretlocated in the peaceful villagePruhonice, a suburb of Prague, Czech.Pruhonice is situated in a quiet area onthe main eastern highway leading toBrno and Vienna, only a few minutesby car from the Prague city centre andwithin the Prague municipal bus and

metro (subway) systems. In Pruhonice thereis a beautiful castle "Pruhonice Chateau"which presides over the Botanical Park,now protected by UNESCO. Instructions for the paper presentationsParticipants wishing to make a presentationare asked to submit an abstract of theirpaper. The papers have to be relevant to theWorkshop objectives.The Abstract includes: Title, author(s),address and text. Title should be CAPI-TALISED and bold. Authors should be list-ed in upper and lower case, with initialsbefore the surname. The address shouldstart in a separate line; include E-mailaddress of the presenting author. The textshould not exceed 250 words. Use MSWord 95 or later edition; Paper size A4;Margins 3.0 cm (top and bottom) and 2.5cm (left and right); Font: Times NewRoman, Font size: 9pt. for the entireAbstract.

Please submit an Abstract for oral presenta-tions (not more than 250 words) before 30June 2003 via e-mail to ZdenkaProcházková ([email protected]).

For your registration, travel informationand hotel reservation please visit theISTA website or contact:Zdenka PROCHÁZKOVÁFGMRI RS Uherske Hradiste686 04 KunoviceCzech Republice-mail: [email protected]


TECHNICAL COMMITTEESUpcoming Workshops

1st Announcement - ISTA ForestTree and Shrub Seed WorkshopOctober 20 - 25, 2003Prague, Czech Republic Zdenka Procházková, ISTA Forest Tree & Shrub Seed Committee Chair

ISTA PurityWorkshop June 11 - 13, 2003Seattle, USAKen Allison,ISTA Purity Committee Chair

Workshop contentIdentification Sessions:* Poa, Bromus, Centaurea, Lolium andFestuca

Quality Assurance Topics:* Blind testing of analysts* Training personnel* Monitoring of purity equipment

Discussion Topics:* Purity concerns for harmonization

ISTA/AOSA Rules* New rules and proposals for the

future

Registration

The workshop will begin at 1.00 pm onWednesday 11th and finish at 12.00 pmon Friday 13th June. The cost is US$150per person. Registration deadline isMay 10th, 2003!

Hotel reservationsPlease make reservations directly atDouble Tree Guest Suites in Seattle,WA. Phone +1-206-575-8220, ask forin-house reservation. Be sure to tellthem you are with the AOSA/SCSTAnnual Meeting to take advantage ofthe group discount rate of 119 US$ +Tax per night.

To register online please go towww.seedtest.org

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e-mail: [email protected]

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missing the last line

GeneralThe ISTA Moisture Committee and the Danish Plant Directorate have the pleasure toinvite you to their Workshop on Moisture Testing.

ScopeThe aim of the workshop is to create discussion and exchange of information in the fieldof moisture testing.

ProgrammeLectures and discussions· Background to seed moisture · Quality Assurance with reference to moisture testing· Future of moisture testing· New species and methods (method validation)· Development of handbook for moisture testingDemonstrations and excursions· Moisture meters, Danish Grain Network· Excursion to seed company and to manufacturer of moisture metersPractical work· Oven test (soft wheat test)· Calibration of moisture meters

RegistrationThe number of participants is limited to 20. The provisional registration costs are EUR250, which includes participation, handouts, refreshments during breaks, lunches, dailytravel from hotel to laboratory. The costs for a single hotel room including breakfast willbe around EUR 100 per night. The deadline for registration is August 1, 2003.

Return this form toISTA SecretariatZürichstrasse 50P.O. Box 3088303 Bassersdorf, SwitzerlandEmail: [email protected]: +41 1 838 6001


TECHNICAL COMMITTEESUpcoming Workshops

stay up to date...The New ISTA Publications

Catalogue 2003 and the ISTAWorkshops Programme 2003 are both available from the

ISTA Secretariat, free of charge.

If you have not yet received these, please contact the

Secretariat, and have them forwarded to you.

Or visit ISTA OnLine at

www.seedtest.org

1st ISTA Moisture WorkshopLyngby, DenmarkNovember 3 - 7, 2003Harry Nijënstein, ISTA Moisture Committee Chair, andJette Nydam, ISTA Moisture Committe Member

Registration Form

Title ……………………………… Position …………………………………

Name …………………………….. First name ……………………………....

Company / Institute……………..... Address …………………….……………

Postal Code …………………........ City ………………………………………

Phone number ………………….... Country ……………………………..…...

Email …………………………….. Fax Number ……………………………..

The Proficiency Test Committee invites par-ticipation by anyone interested in becominga proficiency test leader. This includes anywho have previously been leaders as well asthose with no experience in preparation ofproficiency test samples.

The schedule for the proficiency testing pro-gramme is finalised during each triennialCongress. The three year plan includes thelist of species for nine rounds, the tests whichwill be conducted (purity, other seed deter-mination, germination), and the names of thetest leaders who have volunteered for therounds. Each round leader is responsible forfinding three seed lots having a range ofquality for germination and which are rea-sonably free from impurities. If the testsinclude other seed determination, the leaderdevelops a list of impurities to be added,which is approved by the Committee Chair.The test leader prepares approximately 150sub-samples from each seed lot and conducts

homogeneity tests before distribution. TheISTA Secretariat carries much of the burdenof preparing the written communications,distributing the samples, entering data intothe database and preparing the reports. Thetest leader reviews data summaries preparedby the Secretariat and writes recommenda-tions for those laboratories which havedemonstrated difficulties with the proficien-cy test. The leader advises the Secretariat onany questions concerning the round and mayreceive requests for assistance from partici-pating laboratories which experienced spe-cific problems. Test leaders are providedwith a detailed protocol for sample prepara-tion and are assured advice and support fromthe Secretariat and Committee. Some of thecosts associated with sample preparation,(e.g. purchase of seed lots, containers, ship-ping, etc.), are reimbursed by the Secretariat,but the cost of labour cannot be reimbursed.At this time, the proficiency testing pro-gramme is focused on purity and germina-

tion testing; however, pilot studies are under-way or being considered for vigour, mois-ture, seed health, tetrazolium and tree seeds,so it is likely that in the near future leaderswill also be needed for these fields of testing.

Although there is a significant time commit-ment, previous test leaders will confirm thatthis is a very rewarding experience because itprovides an insight into the complexities ofproficiency testing and gives a direct behind-the-scenes view of a very important ISTAactivity. If you are interested in becoming aproficiency test leader, or would like toreceive more information, please contactMartina Rösch at the Secretariat. The newCommittee will be formed prior to the nextCongress in Budapest in May 2004.


TECHNICAL COMMITTEESProficiency Test Committee

Proficiency Test CommitteeCall for Test LeadersBy Doug Ashton, ISTA Proficiency Test Committee Chair

ISTA's primary instrument in promoting uniformity in seed testing pro-cedures is the 'International Rules for Seed Testing', which lays downdetailed standard techniques and procedures. The publication includes17 Chapters and Appendices describing principles and definitions indetail, assisted by many tables and the methods to be used. The‘International Rules for Seed Testing’ is designed for the principal cropspecies of the world, but apply in general, if not in detail, to any speciesof crop plant, even including those not mentioned in the text. The ‘International Rules for Seed Testing’ is approved and amended atISTA Ordinary and Extraordinary Meetings on the basis of advice ten-dered by the ISTA Technical Committees. The Edition 2003 includesthe latest changes, which were voted on at the ISTA ExtraordinaryMeeting 2002, held in Santa Cruz (Bolivia) from July 3 - 6.Explanatory note: The complete set of the ‘International Rules for SeedTesting’ as per January 1, 2003, will include the following two separatepublications: International Rules for Seed Testing, Edition 2003 andAnnexe to Chapter 7, Seed Health Testing Methods.What is new about the 'International Rules for Seed Testing'?The Edition 2003 will cometo you in a binder. Each year, updates, inthe form of a 'single paper collection' including additions or replace-ments of existing pages will be published, and could then be separate-ly inserted into the binder. With this system ISTA will be able to pro-vide more flexibility, enabling faster updates and improvements of theInternational Rules for Seed Testing.

Valid from January 1, 2003ISBN:3-906549-38-0(Available in English)

The price, including theAnnexe to Chapter 7: CHF 350.-(Approx. US$ 215.-)

*As part of the ISTA membership services, ISTA Members will receiveone free copy of the Rules 2003.

ISTA International Rules for Seed TestingAnnouncement of the new Edition 2003


ISTA held its workshop on AdvancedQuality Management (QM) in November2002. This workshop primarily addressedexperienced quality system managers whohave already participated in one of theprevious workshops and/or have set up aquality management system in theirorganisation. The purpose of this work-shop was to deepen knowledge about QMand initiate lively discussions among theparticipants and the exchange of ideas.

The workshop was being held at theAgricultural Institute of Slovenia (AIS) inLjubljana, November 11-15, 2002. It waspresented by Prof. Dr. Norbert Leist,ISTA President and Heinz Schmid, TheEssential Quality Management.

27 participants attended the workshopfrom 16 different countries. The work-shop programme was made up of the pre-liminary programme and suggestionsbrought in by the participants prior to theworkshop. At the very beginning the par-ticipants were requested to formulate theiraims and questions so that their needscould be adequately met.

Through the lectures the participantslearnt more about theoretical aspects ofquality management like internal audit-ing, QM tools and their application, basicprinciples of communication and staffmotivation. For the technical part manypractical examples were presented con-cerning e.g. calibration of laboratoryequipment and sampling of seed lots. Theworkshop did not only focus on theory butthe lectures were supplemented by prac-ticing the newly acquired knowledge ingroup work. These group work sessionswere also a good opportunity to exchangeindividual experiences made with qualitymanagement in small groups.

Also due to the efforts put into the organ-isation of this workshop by RomanaRutar, head of the seed testing laboratoryof the AIS and her team, the workshopwas a great success and the participantswent home with a suitcase full of newideas.

ACCREDITATIONWorkshop Reports

Advanced Quality ManagementReport on the Workshop in Ljubljana,Slovenia, November 11 - 15, 2002By Martina Rösch, ISTA Accreditation Department

GRDL01Seed Testing Station of AthensMinistry of AgricultureAntheon 2, Marousi151 23 AthensGR-GreecePhone: +30 10 6830471Fax: +30 10 6830917E-mail: [email protected]

Re-accredited ISTA Member LaboratoriesUp until February 15, 2003

CH - Switzerland

CHDL01Eidgenössische Forschungsanstalt fürAgrarökologie und Landbau FALGruppe SaatgutqualitätReckenholzstrasse 191/211Postfach 4128046 ZürichCH-SwitzerlandPhone: +41 1 3777111Fax: +41 1 3777201E-mail: [email protected]

GR - Greece

Quality AssuranceThe following Quality AssuranceDocuments are available directly from theISTA Secretariat.

1. ISTA Seed Testing LaboratoryAccreditation Standard, Version 3.1,20022. Guidelines for becoming an ISTAAccredited Member laboratory, Version1.1, 20023. Guidelines for Monitoring ISTAAccredited Company Seed Samplersand Company Laboratories, Version 1.0,20024. Procedures for Termination,Suspension and Withdrawl of ISTAAccreditation, Version 1.0, 20025. Guidelines for developing QualityDocumention, Version 1.0, 2002

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Following an invitation from the SeedTrade Association of Kenya (STAK) I par-ticipated as second resource person, togeth-er with Mr Dagallier from OECD in theaccreditation workshop held at the NairobiHilton in Kenya. Participants included rep-resentatives from Ministries of Agriculture,Universities and Seed Trade from Kenya,Tanzania and Uganda. The purpose of theworkshop was twofold:

-To explore the possibilities of theDesignated Authority of Kenya, delegatingsome elements of its supervisory activitiesto the private sector, and -To look into requirements of quality man-agement systems as far as documentationand quality manuals are concerned.This second purpose of the workshop wasmerely touched upon, as the printout fromthe two previous sessions was distributedand it was felt that this issue had alreadybeen covered in depth.

In a number of presentations variousaspects of accreditation and certificationwere looked at. It soon became apparent,that there was some confusion concerningthe different concepts applied in conformi-ty assessments. During the workshop theseissues could be clarified and all participantsreached a common understanding.

Looking at the situation in Kenya, itappeared that there was some potential forpartly delegating specific tasks to the pri-vate sector. Field inspection and seed test-ing were identified as two major compo-nents that would qualify for being carriedout by the private sector under close super-vision by government authorities.

ISTA Accreditation pertains to the technicaland managerial competence in a seed test-ing laboratory. As a consequence, accredita-tion is granted to the laboratory, not to indi-viduals and, following changes to the ISTAConstitution in 1995, this applies to labora-tories from both public and private sector.Initially, only private independent laborato-ries were taken into consideration, but nowcompany laboratories, too, can be accredit-ed. One of the two ISTA member laborato-ries in Kenya is currently in the process ofbeing re-accredited. It operates under theMinistry of Agriculture and is part of KenyaPlant Health Inspectorate Service(KEPHIS). KEPHIS, which is also theDesignated Authority according to nationallegislation, is willing to support and recog-nize ISTAAccreditation of private laborato-ries in Kenya.

Similarly, field inspection along the lines ofthe OECD Seed Schemes could be carried

out by recognized individuals under con-tract of or employed by members of the pri-vate sector. The recognition would be to begoverned by certain educational, trainingand expertise requirements set up byKEPHIS in consultation with other interest-ed parties. A presentation from Dr Muasyafrom Moi University Nairobi outlined aprospective training programme for candi-dates. It had been developed together withgovernment representatives to be a startingpoint in further building up a comprehen-sive system to meet the requirements offarmers, industry and authorities. Detailsstill have to be worked out; especially whowould qualify for official recognition, howand to what extent monitoring will be exert-ed.

The workshop was a considerable success,bringing most players in the region togeth-er and indicating the potential for futuredevelopment. It shed light on both limita-tions and possibilities of forthcoming co-operation. Still, a lot of work lies ahead andonly future will show whether the privatesector will take up some of the activitiesunder governmental supervision and whichpitfalls the process will reveal.

ACCREDITATIONWorkshop Reports

Seed Trade Association of KenyaReport on the Accreditation Workshop in Nairobi, Kenya,December 9 - 10, 2002By Gerhard Schuon, ISTA Accreditation Department

KEPHIS,which is alsotheDesignatedAuthority inKenya, sup-ports and rec-ognizes ISTAAccreditationof private lab-oratories inKenya.


Hungary is situated in Central-EasternEurope, surrounded by the Carpathians andthe Alps. The neighbouring countries areAustria, Slovakia, Ukraine, Romania, for-mer Yugoslavia, Croatia and Slovenia.

67 % of Hungary's territory, about 4,7-4,9million ha, is suitable for agricultural pro-duction. The main crops are maize, winterwheat, winter and spring barley, sunflower,potato and other crops as large and smallseed of legumes, grasses, oilseed rape, veg-

etables and so on. The seed sector has animportant and valuable role in theHungarian plant production.

In the context of this activity the inspectedseed multiplication acreage and the quanti-ty of certified seed, Hungary has becomestable in the last 10 years.

The seed lots moving in the internationaltrade are, according to the last officialOECD report:USA 29 %Hungary 15 %France 8 %Chile 8 %Italy 6 %New Zealand 5 %Australia 4 %Czech Republic 4 %Argentina 3 %Others 18 %The value of the Hungarian seed export isbetween 58 - 65 million USD on averageper year.

Of course such a strong seed industryrequires excellent seed testing with highlyeducated experts and seed analysts.

Hungarian seed testing has a long history toit. Hungary has been a member of Europeanand international organisations either sincetheir foundation or since the demand for itincreased.

1921 - European Seed Testing Association1924 - International Seed Testing

Association

The institutional history of the Hungarianseed testing laboratory:

1895 Royal Seed Testing Station, Budapest

1949 National Institute for AgriculturalQuality Control

1951 National Institute for Seed Testing

1952 National Inspectorate for Seeds1976 National Inspectorate for Seeds

and Propagating Materials1983 Institute for Plant Production

Quality Control1988 Institute for Agricultural Quality

Control1994 National Institute for Agricultural

Quality Control

Now the Seed Certifying Agency is a partof the National Institute for AgriculturalQuality Control and its task, among others,the official seed testing in Hungary.

The only ISTA Accredited Seed TestingLaboratory is located in Budapest.

The Seed Testing Laboratory works accord-ing to a Quality Assurance System, and wasone of the first accredited labs under thenew ISTA system.

The times of ISTA accreditation were: 30thOctober 1998 and 20th November 2001.

The central ISTA Laboratory and the other6 local labs fulfil the "General requirementsfor the competence of testing and calibra-

MEMBERSHIPSeed Testing in Hungary

Seed Testing in HungaryBy Katalin Ertsey, ISTA 2nd Vice President

19921993199419951996199719981999200020012002

tons306.208300.535344.464323.273316.301354.009335.007314.512372.020340.253348.383

tons32.53328.67034.50235.34829.44319.98624.31425.15826.17622.38322.900

tons338.741329.205378.964358.621345.744373.995359.321339.670398.196362.636371.283

100,097,2118,7105,9102,1110,4106,1100,3117,6107,1109,6

YearSeed Potato Total

%certified quantities

Figure 1. Inspected Area

Figure 2. Certified Seed

19921993199419951996199719981999200020012002

ha

169.993158.944180,287181.975160.457155.775180.364169.142156.234173.405161.018

%

100,093,5106,1107,194,491,6106,199,591,9102,094,7

Inspected AreaYear

The next (27th) ISTA Congress will be held in Budapest inthe Capital and the economic, financial and cultural cen-tre of Hungary.


tions laboratories ISO/IEC 17025:1999" aswell. The tested samples included morethan 200 varieties from cereals to flowerand medical seeds.

The ISTA laboratory of Budapest handlesthe ISTA samples and most of vegetable,flowe and medicine seed tests, whichdemand a well developed technical envi-ronment.

Number of tested samples in year 2002:Analytical purity 25 748 samplesGenetic purity 1 377 samples Germination 14 701 samples Seed health 9 997 samplesMoisture content 270 samples(germination includes TTC and vigourtests)

The staff is highly educated with severalexperts holding master degrees or more.22 seed analysts work in the ISTA lab.The lab has different training programs forthe company specialists, and an organiser inpractical education for the university ofagriculture.

Our Hungarian predecessor colleaguesworked earlier in different ISTA TechnicalCommittees, obtaining good professionalappreciation, and this trend continues nowas we have members in the Flower Seedand Tetrazolium Committees, as well as inthe Executive Committee.

The ISTA Laboratory in Budapest issuesmore than 7000 ISTA Certificates yearly.The regional labs work for the Hungarianmarket, which are accredited by theHungarian National Accreditation Body.The number of ISTA samplers is 56.

We are looking forward to host a largenumber of seed experts in May 2004 on theoccasion of the 27th ISTA Congress. Thescientific program for the SeedSymposium, challenges of the new tenden-cies, cultural programs and the excursionspromise a nice time for all participants.

MEMBERSHIPSeed Testing in Hungary

Figure 3. The position of the seed testing and sampling as part of the Seed Division

On page 7 you willfind more informa-

tion on the 27th ISTACongress to be held

in Hungary, and the Call for

Papers for the SeedSymposium.

Or visit our website atwww.seedtest.org


Prof. Dr. Norbert Leist, Director of Biologyand Head of the Division of Seed Testingand Applied Botany at the StateAgricultural Experimental and ResearchStation Augustenberg (LUFA), Karlsruhe,Germany, celebrated his 60th birthday on31st of December 2002. Born and grown upin Bruchsal, a town near Karlsruhe, hegraduated from secondary school of classi-cal education and was awarded the ScheffelPrize for excellence in the German lan-guage in 1962. While in the military serviceas a 2nd Lieutenant of the reserve hebecame familiar with such areas as plan-ning, organization and leadership and, as aparatrooper, with decision making andobjective-oriented behaviour. Already in hisyounger years he was fascinated by the mir-acles of nature. Hence, he began studyingbiology, chemistry and physics at theUniversity of Heidelberg. After the stateexamination in these fields in 1968 he pre-pared a dissertation on "The ontogenesis ofthe reticular venation of fern leaves" underthe guidance of Prof. Dr. W. Hagemann. In1972 a doctor's degree of natural scienceswas conferred on him. Supported by theGerman Research Foundation (DFG) a one-year stay in Columbia for field research onthe ecology of tropical ferns and woodecosystems followed. After terms asresearch assistant at the Universities ofKarlsruhe and Heidelberg in the followingyears, he joined the staff of the LUFAAugustenberg in 1977 and, in 1992, becamethe successor of Dr. B. Schmidt as Head ofthe Division.

Prof. Leist started his career with a highdegree of competence, an admirableendurance and prudent sense of enforce-ment. He developed his division to the mostmodern, internationally competitiveGerman seed testing laboratory. With himseed matters in the State of Baden-Württemberg are in good hands. In addi-tion, he enhanced counselling and researchactivities. Main areas of research are quali-ty seed production in maize, wild oat, oathybrids and fatuoids population biology inseed production, cereal seed health assess-ment, and studies on toxicity effects of seeddressings. Furthermore, there are numerousstudies on the improvement of the method-ology in tetrazolium testing and for the

application of thistest to newspecies. Morerecently, a greatdeal of activitywas devoted toverification of cul-tivar by storageprotein markersdeveloping UTLIEF techniques, in particu-lar for testing hybrid purity in seeds ofmaize and vegetable species. These studieswere extended now by using molecularDNA techniques to test for adventitiousGM seeds in conventional seed lots. TheAssociation of the German AgriculturalExperimental and Research Stations (VDL-UFA) honoured his research activitiesalready in 1981 with the rarely awardedFriedrich-Nobbe Prize. He also continuedhis comprehensive teaching program at theUniversity of Karlsruhe to help fascinatestudents with applied botany and seed sci-ence and technology. There, his intellectualcuriosity about nature together with hisingenuous ability to marvel at wondershitherto attracted 3 post-graduate studentsand another 37 graduates preparing theirtheses under his guidance. For these out-standing achievements he was awarded anhonorary professorship in 1992. Besides, heorganised and was leader of many nationaland ISTA workshops on seed sampling,tetrazolium testing, cultivar purity testingby protein electrophoresis and ISTA qualitymanagement. Moreover, acknowledgingalso his singular merits in vocational train-ing, the Chamber of Commerce of theUpper Rhine Valley bestowed on him theCarl-Friedrich-Nebenius Medal.

Active participation in associations accom-panies Prof. Leist`s daily work in a true-to-life manner. Since 1978 he has been aMember of the Board of the Seeds and SeedScience Section of the VDLUFA and, in1990, was elected Vice Chairman. As of1980 he has been a Member of severalISTA Technical Committees: Purity, ForestTree and Shrub Seeds, Variety, Tetrazolium(Chairman 1990 to 2000) and RulesCommittees. In 1995 he was electedMember of the ISTA Executive Committeeand in 1998 became 1st Vice President ofISTA. Already in 2000 he took over the

ISTA presidency when President Kevin Boyce parted before his regular three yearswere up. In 2001 Prof. Leist was confirmedby the ISTA General Assembly in Angers,France, as ISTA President for the period2001 to 2004. In the now 79 years of ISTAhistory, he is the first German ever to beelected for that post. Besides his stronginvolvement in the VDLUFA and in partic-ular the ISTA, he is a member and active inseveral responsible positions in numerousother scientific associations in the fields ofcrop science, plant breeding, floristics, lim-nology and his beloved hobby arachnology.Also he is member of several state and fed-eral committees of experts in these fields.

It is not possible to describe here the sur-prisingly broad interests and activities ofProf. Leist in more detail. His multi-facetednatural gifts, his widespread and profoundscientific and practical knowledge andskills, his open-mindedness and readinessto listen, his politeness and well-balancedtemper, his co-operative attitude and cre-ative power combined with fairness and,above all, constantly positive attitudetowards life in general are characteristicsfor which he earned national and interna-tional acceptance and high regard.

May Prof. Leist, in the years to come, begranted the opportunity to harvest richcrops while borne by his understandingfamily and supported by his loyal team ofco-workers and well-tried colleagues. Heloves hiking in the mountains and roamingtropical forests, diving into the mysteriousunderwater world of old father Rhein aswell as into the magic of coral reefs. Mayall this give him the necessary "vigour". Forseed science and technology as well as seedindustry and trade, the VDLUFA and theISTA, and his colleagues: we all need him.Happy Birthday and many happy returns.

MEMBERSHIPProfessor Leist’s 60th Birthday Celebration

ISTA President, Prof. Dr. Norbert Leist60th Birthday CelebrationBy Adolf Martin Steiner, ISTA Member


On December 5th, 2002, the LaboratoryDirectorate of the Canadian FoodInspection Agency (CFIA) hosted a recep-tion to celebrate the hundredth anniversaryof the establishment of the Ottawa SeedLaboratory and the federal seed testing pro-gram. The establishment of the laboratorymarked the start of the seed program inCanada, which was aimed at improving thequality of domestic and exported seed, aswell as protecting the virgin agriculturalland that was being opened up for settle-ment in the Western part of Canada fromthe introduction of foreign weeds. Formore of the history, see the Ottawa SeedLaboratory's profile (CADL04) on theISTA web page section for member labora-tory profiles.

The reception was attended by the staff ofthe Ottawa Laboratory as well as formerstaff that had been involved with the labo-ratory and had contributed to the develop-ment of the science of seed testing inCanada. Guests included staff from theAgency's Programs Branch, the LaboratoryDirectorate and colleagues fromAgriculture and Agri-Food Canada'sResearch Branch that had collaborated withthe seed laboratory over the years.

The highlight of the reception was the pres-entation by Mr. Richard Fadden, thePresident of the Canadian Food InspectionAgency of a commemorative plaque fromCanada's Minister of Agriculture. In hispresentation speech, the President praisedthe dedication of CFIA staff, current andretired, on their long-standing contributionto the public good and its strong history ofinternational participation and co-opera-tion.

We were pleased to have with us a numberof special guests who brought greetings andcongratulatory remarks for the occasion.These included Dr. Michael Muschick,Secretary General of ISTA, representativesfrom the Canadian Seed Growers

Association, the CanadianSeed Trade Association,the Canadian Seed Instituteand the Commercial SeedAnalysts Association ofCanada. Commemorativeplaques were also receivedfrom the Association ofOfficial Seed Analysts (theOttawa laboratory has beena member since 1908) andthe Society of CommercialSeed Technologists.

Displays of seed testingequipment from the early1900s, archival photos, documents and pic-tures drew many exclamations and com-ments as guests revisited the past.Commemorative pins incorporating adesign by Ken Allison, seed biologist at theOttawa lab, were given to staff and gueststo mark the occasion. As part of the cente-nary celebration, Sandy Ednie presentedMr. Fadden with a beautiful water colorillustration of seeds that was one of manydone by W.H. Wright in the 1930s and1940s to help train seed analysts in seedidentification. The illustration now hangsin the lobby of the CFIA Headquartersbuilding.

The celebration held in Saskatoon com-memorating 100 years of seed testing inCanada was a huge success. There wereover 80 people who attended the December17th event, including guest speakers fromthe Agriculture and Agri-Food ResearchStation, Commercial Seed AnalystAsscociation of Canada, SaskatchewanAgriculture Minister's Office, AgWestBiotech and Saskatchewan Seed GrowersAssociation. Joanne Hinke, Purity LabSupervisor and Janine Maruschak, ActingHead of the Western Seed Laboratory gavea presentation on the history and currentstatus of seed testing in Canada. Many ofthe speakers focused on the close workingrelationships amongst the seed testing

groups and those that rely on their serviceswithin Canada. There was praise fromresearchers, associations and clients thathave worked closely with the SaskatoonSeed Lab and there was recognition of theexciting times in seed testing to come. Forthe many retired staff on hand it was achance to greet friends and former col-leagues. The Federal Agriculture Minister,Lyle Vanclief's commemorative plaque waspresented by Liz Singh, AssociateExecutive Director, LaboratoriesDirectorate to the seed lab staff. Guests andmedia took the opportunity to tour the newseed laboratory, which is located inInnovation Place, a research park adjacentto the University of Saskatchewan campus.The Saskatoon reception also provided anopportunity for the Western Seed lab toannounce its new name - the Seed Scienceand Technology Section of the SaskatoonLaboratory. Print and TV Reporters wereon hand to cover the event and take videofootage of the laboratory and analysts,resulting in an article in the local newspaperand a segment on the local news and on aweekend agricultural television program.

Several articles have appeared in the popu-lar agricultural press covering the centenaryand the role of the seed laboratories inCanada's agricultural system.

MEMBERSHIPCentenary of Seed Testing in Canada

Centenary of Seed Testing in CanadaCanadian Food Inspection Agency’s Seed Laboratories Celebrate100 Years of Seed TestingBy Alexander B. Ednie, Head of Seeds Section, OttawaLaboratory (Carling) and ISTA Member Janine Maruschak, Acting Head of Seed Science andTechnology Section, Saskatoon Laboratory and ISTA Member


Assistance provided by Danida (Ministry ofForeign Affairs, Denmark) to India in thefield of Agriculture was recently acknowl-edged by the University of Mysore, India,when Director Dr. S.B. Mathur of theDanish Government Institute of SeedPathology for Developing Countries(DGISP) was awarded Prof. K.M. SafeeullaGold Medal on December 2, 2002. Thegold medal was instituted for the first timeby the university in memory of K.M.Safeeulla's significant contributions in thefield of plant diseases, especially for inter-nationally known research on downymildew diseases of millets.

The medal was awarded for Dr. Mathur'sexpertise in seed-borne fungal diseases oftropical and subtropical crops as well as hisknowledge of technological developmentsin seed health that that have inspired a gen-eration of scientists all over the world. Dr.Mathur's contribution to increased foodproduction through the use of improvedseed that have been internationallyacknowledged. Considering his achieve-ments and contributions he was awardedwith the prestigious 'FIS World Seed Prize'in 1992. Dr. Mathur is responsible forestablishing a seed pathology research andtraining centre at the University of Mysore.

The Danish Institute of Seed Pathology inDenmark that have been Dr. Mathur's brain-child and he along with Dr. Paul Neergaardthat have been responsible for founding theInstitute. The Institute is located on thecampus of the Royal Veterinary and

Agricultural University of Denmark(KVL). The institute that have trained 75agricultural scientists and technologistsfrom all parts of India. The trainees came toDenmark from universities, research insti-tutes, seed production and seed certificationagencies. The main objective of the trainingthat have been to improve the health ofseed, with ultimate goal of increasing andimproving food production. Professionaland technical support received by theUniversity of Mysore for over 25 years washighlighted by Dr. Shekar Shetty, Professorof Applied Botany at the medal presenta-tion ceremony.

Danida is presently upgrading research andtraining facilities at Mysore and it is antici-pated that the University of Mysore willeventually become a Seed PathologyCentre in Asia where scientists and techni-cians from different countries can betrained and they should then fight seedtransmitted diseases which negativelyeffect food production.

Assistance to developing countries in thefield of seed pathology and seed health byDenmark is considered by Indian Scientistsas a success story of Danida.

Dr. V. Prakash, Director, Central FoodTechnology Research Institute, Mysore pre-senting the Professor K.M. Safeeulla GoldMedal and citation to Dr. S.B. Mathur(right) Director, Danish GovernmentInstitute of Seed Pathology for developingcountries, Denmark.

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India Applauds Danish AidUniversity of Mysore, Mysore, India

Press ReleaseMinutes of the ISTA ExtraordinaryMeeting 2002Cited and PublishedBy the ISTA Secretariat

This document summarizes and con-cludes the discussions and decisions ofthe ISTA Extraordinary Meeting 2002 inSanta Cruz, Bolivia, July 5 and 6.

As minutes of the subsequent meeting ofthe ISTA membership, this transcript pro-ceeds the Post-Congress Proceedings ofthe 26th ISTA Congress 2001, Angers,France, published as Volume 29,Supplement 3 of Seed Science andTechnology (SST).These minutes contain a complete list ofall participants of the meeting.

Available as of April 2003 from the ISTASecretariat.

Price: CHF 70.00 (approx. US$ / EUR49.00)

Note: This publication will be deliveredas a free service to all ISTA Members,Designated Authorities as well as to theparticipants of the ISTA ExtraordinaryMeeting 2002. It will also be distributedto all participants of the upcomingExtraordinary Meeting 2003 to be held inZurich, Glattbrugg, Switzerland (seemore details on upcoming ISTA meetingsfrom page 3 to page 7).


MEMBERSHIPISTA Membership

New ISTA Members

JP-JAPANJPML06 / JPPM06Quality Assurance Department, Takii & Co Ltd, 180 Umekoji, Inokuma, Shimogyo-KUC.P.O. Box 7, Kyoto 600-8686Phone: +81 75 365 0123 Fax: +81 75 3650110E-mail: [email protected] member: Mr. Masahiko Komaba

KE-KENYAKEDM01 (additional member of KEDL01)Mrs. Gladys. N. Maina, Kenya Plant HealthInspectorate Service (KEPHIS) AssistantDirector, QC Service, Waiyaki Way, P.O. Box49592, NairobiPhone: +254 2 440087 Fax: +254 2 448940E-mail: [email protected]

Membership cancellation

AR-ARGENTINAARDM01 Ing. Agr. Adelaida Harries de CrespoPresidencia Instituto Nacional de Semillas, PaseoColón 922, 3° Piso, 1063 Buenos Aires

CL-CHILECLDM01Dr. Ricardo Romero Alpe (additional member ofCLDL01)Corporación Nacional Forestal Special Advisorto the Executive DirectorP.O.Box 10346, Santiago

TW-Separate Customs Territory of Taiwan,Penghu, Kinmen and MatsuTWDM01Mr. Chia-Chi ChenSeed Testing Laboratory, Council of Agriculture, 37 Nanhai Road, Taipei 100-14

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AU-AUSTRALIAAUDM02New personal member : Dr. Kevin G. BoyceSeed Technology Institute, Australia Pty Ltd,P.O. Box 410, Blackwood SA 5051Phone: +61 8 8278 3356 Fax: +61 8 82783356E-mail: [email protected](formerly Mr. Colin Beavis)

DK-DENMARKDKPM07New personal member : Ms. Dorrit AndersenL. Dæhnfeldt A/S, Germination Laboratory,Breeding Station Danefeld, Odensevej 825290 MarslevPhone: +45 63 17 5506 Fax: +45 63 17 5505E-mail: [email protected](formerly Mr. Finn V. Povlsen)

GR-GREECE GRDM01New personal member : Ms. Eftychia AndritsouSeed Testing Station of Athens, Antheon 2,Marousi, 151 23 AthensPhone: +30 10 6830471 Fax: +30 10 6830917E-mail: [email protected](formerly Mr. Athanasios Kouis)

KE-KENYAKEDM01New personal member : Dr. Joseph OlooAhenda Kenya Plant Health InspectorateService, Nakuru Regional Office, P.O. Box 1679, NakuruPhone: +254 37 850106 Fax: +254 37 851268E-mail: [email protected](formerly Mr. Jeremiah K. B. Kiplagat)

KR-KOREA, REPUBLIC OFKRDM01New personal member : Mr. Jin-kug KimCentral Seed Testing Laboratory, InspectionDivision, Experiment Research Institute ofNAQS, 560, 3-Ga, Dangsan-Dong,Youngdeungpo-Gu, SeoulPhone: +82 2 2165 6041 Fax: +82 2 21656005E-mail: [email protected](formerly Mr. Kyu-won Choi)

RU-RUSSIAN FEDERATIONRUDM01New personal member : Mr. Viktor TrusovThe State Seed Inspection of the Ministry ofAgriculture of Russia, Orlikov pereulok 1/11107139 MoscowPhone: +7 095 2073883 Fax: +7 095 2073883(formerly Mrs. Tatyana Kokina)

RUDM02New personal member : Ms. Galina FilipenkoPlant Germoplasm Conservation Laboratory, N.I.Vavilov, Institute of Plant Industry 42, BolshayaMorskaya Street, 190000 St. PetersburgPhone: +7 812 3147714 Fax: +7 812 3118728E-mail: [email protected](formerly Dr. V. A. Zaitsev)

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Christchurch, New Zealand was the venuefor the 8th International Congress of PlantPathology (ICPP 2003) which was held dur-ing the week of 2 - 7 February 2003. Nearly1300 people from 72 countries participated,enjoying the scientific information transfer,the chance to meet fellow plant patholo-gists, and the famous Kiwi hospitality.Under the theme of "Solving problems inthe real world", three plenary sessions, fivekeynote sessions, 32 concurrent sessionsand two poster sessions provided delegateswith the opportunity to immerse themselvesin all areas of plant pathology, includingseed pathology.

Concurrent Session 22 (Seed Pathology)was organised by ICPP 2003 ProgrammeCommittee member Professor JohnHampton, with assistance from ISTA PDCchairperson and deputy chairperson JimSheppard and Valerie Cockerell. This pro-vided an opportunity to showcase the cur-rent work of the PDC, particularly in seedhealth method validation and quality assur-ance and proficiency testing. The sessionwas chaired by PDC member Dr GuroBrodel from Norway.Jim Sheppard (Canada) briefly explainedISTA's purpose to the 160 strong audience,before presenting the ISTA Seed HealthMethod Validation Programme. Jimdemonstrated the steps required before afinal product (a validated seed health testmethod) can be included in the ISTA Rules.He also outlined the progress made to date,and some of the problems encountered. Hispresentation was followed by one fromValerie Cockerell (Scotland) who explainedthe ISTA accreditation process and dis-cussed how seed health testing could bebrought into the ISTA proficiency testingprogramme. This will be no easy task, andthere are a number of issues still to beresolved, including the availability of suit-able seed lots, the application of perform-ance standards, and the difficulty of annual-

ly assessingi n d i v i d u a llabs whichmay not all bep e r f o r m i n gthe same seedhealth tests.The thirdspeaker wasPDC memberDr. SteveR o b e r t s( E n g l a n d ) ,whose topic was the use of reference mate-rials in seed health method validation andproficiency testing. Steve made the pointthat variability in the test material (i.e.infected seed lots) is a major limitation tothe success of comparative testing and theinterpretation of their results. He suggestedthat the use of Standardised ReferenceMaterials (RM) containing a stable quantityof the pathogen was one way to overcomethe problem. While initially costly to pro-duce, RMs will be cost effective in the longterm, and an EU supported project hasalready resulted in RMs for four seed trans-mitted bacterial pathogens. Inter-lab studiesusing these RMs have demonstrated theirvalue for accuracy and precision of seedhealth assays.

The session concluded with a demonstra-tion by former PDC Chairperson Dr. KeesLangerak (The Netherlands) of how a vali-dated seed health method can be used inpractice, using Alternaria radicina inorganic carrot seed production. Hedescribed laboratory and field studieswhich determined to what extent seedinfection can contribute to seedling death,storage black rot and seed to seed transferof the pathogen, the objective being todetermine pathogen thresholds for the vali-dated seed health test method.

As the session organiser, chairperson and

speakers were all ISTA members, the audi-ence left having broadened their knowledgeof ISTA and the work of the PDC. In addi-tion, the ISTA information available to theaudience (which included the PublicationsCatalogue and News Bulletin) disappearedwithin minutes!

There was also a Seed Pathology section inPoster Session Two in which 21 posters byauthors from 10 countries were displayed.Twelve of these authors then had the oppor-tunity to make a brief (5 minute) oral pres-entation during the poster discussion ses-sion. Jim Sheppard chaired this session,during which PDC Member Rivka Hadas(Israel) spoke about her work on the devel-opment of a PCR detection method forXanthomonas campestris pv. campestris inbrassica seeds.

Seed pathology received a much higherprofile at ICPP 2003 than it has at previousInternational Plant Pathology Congresses.Jim Sheppard made the point several timesthat ISTA Membership is not a requirementfor submission of a seed health testingmethod to ISTA for consideration for vali-dation. The opportunity to present ISTA andits PDC to a wider audience will encouragesuch submissions, as seed pathologists notalready working within ISTA are nowaware of this work.


REPORTS FROM MEETINGSICPP Seed Health Symposium

Seed Pathology at the 8th InternationalCongress of Plant PathologyChristchurch, Canterbury, New Zealand, February 2 - 7, 2003By John Hampton, ISTA Executive Committee Member-at-large

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The biennial meeting was attended by rep-resentatives from 19 Scheme membercountries and 4 other organizations includ-ing ISTA. Membership in the Scheme isvoluntary and open to OECD Membercountries as well as all members of theUnited Nations.

The prime focus of this meeting was to seeka means of approving a new Scheme. Thecurrent Scheme, which was adopted in1974, was viewed as having become some-what dated so work began on developing anew Scheme in the early 1990's. In 1996 theScheme was presented for approval bymember countries however, there was areservation regarding the inclusion ofGenetically Modified Organisms (GMO)which resulted in the inability for theScheme to be adopted by OECD. Severalattempts have been made to revise theScheme but the proposed changes were notaccepted by all member countries. Anotherconsideration was the European Union's(EU) new Directive on the marketing offorest reproductive material scheduled tocome into force January 1, 2003. ThisDirective, which was developed in parallelwith the new Scheme, contains referencesto GMO. The EU recognizes "equivalence"between their directive and the OECDScheme so it is important that the newScheme be approved to provide harmonywhen reproductive material is tradedbetween EU and non-EU countries.Following lengthy, productive discussions arecommendation was made to form anexpert group to revise those portions of theScheme referring to GMO and submit therevised Scheme to member countries fortheir approval. The expert group planned tomeet in December.

Additional agenda items covered at themeeting included a document summarizing,for each of the four categories, weights offorest seed certified, used, or traded by par-ticipating countries in the 2000-2001 timeperiod. Total weight of seed certified fordomestic use amounted to a record of 1 363tonnes. An updated list of approved basicmaterial was presented. It illustrated thatthe "Source-identified" category, the mostbasic level of genetic quality, was still veryimportant for a number of countries with atotal area of 13 million hectares of forestfrom which seed is collected. The"Selected" category consisted of 515 000ha. "Untested seed orchards" comprised 2600 ha while the "Tested" category, con-taining stands and seed orchards, had 3 400ha. Clones and clonal mixtures were alsopart of this category for a number of coun-tries. The Slovakian delegate provided aninteresting presentation about seed manage-ment and certification activities in Slovakiawhich are compatible with the OECDScheme. Presentations were also made byOECD staff on consensus documentsdeveloped for a number of forest treespecies useful for people dealing with reg-ulatory assessment of genetically modifiedmaterial and the creation of a web pagededicated to the forest seed Scheme.Statements were also presented by repre-sentatives from FAO, CPFUE (Committeefor Forest Nurseries of the EU), ISF, andISTA. The delegates were hosted bySwitzerland for a one day visit to the areaadjacent to Lausanne to visit stands ofDouglas fir and Norway spruce, field trialsof foreign tree species, and several otherstops.


REPORTS FROM MEETINGSCSSA/OECD

OECD Scheme for theControl of ForestReproductive MaterialMoving in InternationalTrade Paris, France, October 1 - 4, 2002By Dale Simpson, ISTA Forest Tree & Shrub Seed Committee Member

The CSSA Symposium on WorldwideSeed Health Test StandardizationSystems was held on 13 November2003. This symposium was part of themuch larger joint annual meeting of theAmerican Society of Agronomy (ASA),the Crop Science Society of America(CSSA), and the Soil Science Society ofAmerica (SSSA). The meeting attractssome 4,000 participants mainly fromNorth America, but also from around theworld, representing academia, govern-ment and private industry, and includinga large contingent of undergraduate andgraduate students. The scale of themeeting was quite awesome with manyconcurrent sessions/symposia and a cav-ernous exhibition area for many hun-dreds posters and trade displays. Thesymposium started with presentationsby Denis McGee from the Seed ScienceCentre of Iowa State University, on theUSA National Seed Health System andRudy Scheffer, the chairman of ISHI-Vegetables, on the industry-backedInternational Seed Health Initiative.Steven Roberts, leader of the ISTA-PDCBacteriology Working Group, then gavea presentation on the ISTA-PDCMethod Validation Programme coveringsuch topics as: the need for method val-idation; the development of the ISTAprogramme; the steps involved methodvalidation and the progress to date.Although the audience was disappoint-ingly small for such a large meeting, thisdid mean that those present were all themore enthusiastic leading to a lively dis-cussion after the formal presentations.

CSSA Symposiumon WorldwideSeed Health TestStandardizationSystemsIndianapolis, Indiana,USA, November 2002By Steve Roberts,ISTA Plant Disease CommitteeMember

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ISF Press Release2003 ISF Congress

The 2003 ISF Congress will take place in Bangalore, India on 9-11 June 2003.

Bangalore will host the first congress of the International Seed Federation (ISF)formed from the historic merger of FIS and ASSINSEL, two associations estab-lished in 1924 and 1938, respectively. With participants from over 65 countrtiesexpected at the biggest international gathering of seedsmen andseedswomen from all the world over, the congress provides a forum for:

- discussion on the main issues of interest or concern to the seed indus-try at a global level

- meeting colleagues for scientific, technical or commercial purposes- learning more about the Indian seed industry and market

India is credited with a strong seed industry and is one of the largest unifiednational markets in the world. It is endowed with strong skills and state of theart technology at all levels, from research to seed production.

The ISF Seed Treatment and Environment Committee (STEC) has organised aSeed Treatment Conference also to be held in Bangalore, after the congresson 12 and 13 June 2003.

The full information package onthecongress is available on-line atwww.worldseed2003.com.

For practical information contactthe congress organizers at:

Conferences & IncentivesManagement (I) Pvt. Ltd.C1/D, 1st FloorGreen Park ExtnNew Delhi 110 016IndiaTel: +91 (11) 2653 6075Fax: +91 (11) 2653 6086Email: [email protected]

For more information, please contact the ISF Secretariat:

Chemin du Reposoir 7, 1260 Nyon, SwitzerlandTel: +41 22 365 44 20, Fax: +41 22 365 44 21

E-mail: [email protected], Web: www.worldseed.org

13 Feb 2003


MEETINGS2003 ISF Congress

Main items on the agenda- Essential derived varieties and in partic-ular principles for a code of conduct inLettuce- Arbitration procedures for dispute settle-ment during trade in seeds for sowing andfor the management of intellectual prop-erty- Genetically modified varieties:

Impact on the seed industryPosition of ISF on Adventitious Presence and GMO testing

- Presentation of the Indian seed industryand seed regulations- FAO MTA on access to genetic resourcesMore information can be found in thedetailed program

CALENDAR2003

May12-16 ISTA Flower Seed Testing Workshop

(Budapest, Hungary)14-16 ISTA Seed Vigour Testing Workshop

(Parndorf, Austria)

June09-11 ISF Congress

(Bangalore, India)11-13 ISTA Purity Workshop

(Seattle, United States)

July30-03 ISTA Extraordinary Meeting 2003

(Zurich, Switzerland)07-11 OECD Annual Meeting of the Seed Schemes

(Paris, France)

October20-25 ISTA Forest Tree and Shrub Seed Workshop

(Prague, Czech Republic)

November03-07 ISTA Moisture Workshop

(Lyngby, Denmark)

2004May13-24 ISTA 27th International Seed Testing Congress

(Budapest, Hungary)24-26 ISF Congress

(Berlin, Germany)

CALENDAR

International Seed Testing Association (ISTA)ISTA Secretariat, Zürichstrasse 50, PO Box 308, 8303 Bassersdorf, CH - Switzerland

Phone: +41 1 838 6000, Fax: +41 1 838 6001, E-mail: [email protected]

www.seedtest.org