Collectotrichum Jasmine Sambac

Colletotrichum species from Jasmine (Jasminum sambac)

Saowanee Wikee & Lei Cai & Noireung Pairin & Eric H. C. McKenzie & Yuan-Ying Su &Ekachai Chukeatirote & Hoa Nguyen Thi & Ali Hassan Bahkali &Mohamed Abdo Moslem & Kamel Abdelsalam & Kevin D. Hyde

Received: 1 June 2010 /Accepted: 15 July 2010 /Published online: 31 July 2010# Kevin D. Hyde 2010

Abstract Colletotrichum species associated with leaf andflower anthracnose of jasmine (Jasminum sambac) in theHo Chi Minh region of Vietnam are reported. The diseaseof jasmine plantations was considered serious as it likelyreduced flower yield. Leaves were colonized by Colleto-trichum species which formed chlorotic regions with lightbrown necrotic centres, which eventually covered the whole

leaf and subsequently caused defoliation and dieback andwhole flowers were blighted. Nine strains of Colletotri-chum species were isolated from diseased leaves andflowers and partial ITS rDNA sequences were analysedand morphologies compared across similar species. Basedon ITS sequence analysis and morphological characters,three strains were identified as C. truncatum, while onestrain was identified as C. siamense. The remaining fivestrains did not cluster with any known species for whichtype sequences are available and therefore partial actin(ACT), -tubulin (TUB2), calmodulin (CAL), glutaminesynthetase (GS), glyceraldehyde-3-phosphate dehydroge-nase (GPDH) genes of the isolates were sequenced. Basedon the reconstructed multiloci molecular phylogeny, twotaxa are formally introduced as new species. Another strainwas not well resolved in the phylogenetic tree and hereindescribed as Colletotrichum sp. Further studies are neededto prove its distinctiveness. The morphology and growthrate of all taxa are described and compared with similarspecies.

Keywords Anthracnose .Multilocus phylogeny .

Plant disease . Systematics

Introduction

Jasmine (Jasminum sambac (L.) Aiton) is grown as anornamental throughout Asia where it is native and is usedfor garments, bouquets and in massage oils. In China andVietnam it is used to produce the flowers used in jasminetea and is grown in plantations. There are relatively fewreports of disease of jasmine; these include witches broomcaused by phytoplasma (Al-Zadjali et al. 2007), tomatomosaic virus infection (Kamenova et al. 2006), cercospor-

S. Wikee :N. Pairin : E. Chukeatirote :K. D. HydeSchool of Science, Mae Fah Luang University,Chiang Rai 57100, Thailand

K. D. Hydee-mail: [email protected]

L. Cai (*) :Y.-Y. SuKey Laboratory of Systematic Mycology & Lichenology,Institute of Microbiology, Chinese Academy of Sciences,Beijing, Peoples Republic of Chinae-mail: [email protected]

E. H. C. McKenzieLandcare Research,Private Bag 92170, Auckland, New Zealand

H. N. ThiPost Entry Quarantine Center No. 2,Plant Protection Department,District 1,Ho Chi Minh City, Vietnam

A. H. Bahkali :M. A. Moslem :K. D. HydeBotany and Microbiology Department, College of Science,King Saud University,P.O. Box: 2455, Riyadh 1145, Saudi Arabia

K. AbdelsalamCollege of Science, Abdul Rahman A1-Jeraisy DNA ResearchChair, King Saud University,P.O. Box 2455, Riyadh 1145, Saudi Arabia

Fungal Diversity (2011) 46:171182DOI 10.1007/s13225-010-0049-x

oid leaf spots (Braun and Sivapalan 1999) and several otherfolicolous taxa (Agarwal and Sahni 1965; Sahni 1966).Reports of Colletotrichum species on jasmine are rare.Agarwal (1962) introduced a new species, C. jasminicola,which caused severe blighting of leaves and shoots in India,while Agarwal and Sahni (1965) identified C. dematiumfrom leaf spots.

Jasmine plantations have become a common andimportant industry in Asia. In Vietnam Colletotrichumspecies cause serious problems to the plantations, infectingleaves and flowers, often resulting in defoliation anddieback. The objective of this study was to examine anddescribe Colletotrichum species that cause disease ofjasmine plants in Ho Chi Minh, Vietnam.

Materials and methods

A survey was made to jasmine plantations in Cu ChiDistrict, Hoc Mon District and the 12th District of Ho ChiMinh City, by staff of Post Entry Quarantine No. 2 (PEQ2)from January to September 2009 (Fig. 1).

Isolation of Colletotrichum strains

Diseased leaves and flowers were removed from thejasmine plants and returned to the laboratory in individualsnap lock plastic bags. In the laboratory they wereexamined for visible sporulation and the fungi present wereconfirmed to be Colletotrichum species. A single sporeisolation technique was applied to plant tissues where sporemasses were formed. Spore masses were picked off with asterilize wire loop or fine forceps and suspended insterilized water. The spore suspension was diluted to a

reasonable concentration and spread onto the surface ofPDA agar, followed by incubation overnight at roomtemperature (25C). Single germinating spores were pickedup with a sterilized needle and transferred to new PDAplate for morphological and molecular study (Goh 1999).

Morphological studies of Colletotrichum from jasmine

Mycelial discs (0.5 cm diam.) were taken from 10 day oldcultures and transferred to potato dextrose agar (PDA), maltextract agar (MEA) and oat agar (OA), and incubated at27C following the methods of Cai et al. (2009). Colonydiameters of three replicate cultures of each speciesgrowing on PDA were measured daily for 7 days. Growthrate was calculated as the 7 day average of mean dailygrowth (cm per day). After 7 days, colony size wasrecorded. Colony characteristics on each type of agar werealso recorded.

Appressoria were produced using a slide culture tech-nique in which a 1 cm2 square of a Colletotrichum cultureon PDA was placed in an empty Petri dish and a sterilecover slip was placed over the agar (Cai et al. 2009). After7 days, the shape and size of appressoria formed on theunderside of the cover slip were studied. Morphologicaldata were analyzed using analysis of variance (P

Table 1 Sources of isolates and GenBank accession numbers used in this study

Colletotrichum species Culture collection GenBank accession number

ACT TUB-2 CAL GS GPDH ITS

C. anthrisci CBS 125334a GU227943 GU228139 GU228237 GU227845

C. anthrisci CBS 125335 GU227944 GU228140 GU228238 GU227846

C. asianum MFU 090232a FJ 903188 FJ 907434 FJ 917501 FJ 972586 FJ 972571 FJ 972605

C. asianum MFU 090233 FJ 907424 FJ 907439 FJ 917506 FJ 972595 FJ 972576 FJ 972612

C. asianum MFU 090234 FJ 907421 FJ 907436 FJ 917503 FJ 972598 FJ 972573 FJ 972615

C. chlorophyti IMI 103806a GU227992 GU228188 GU228286 GU227894

C. chlorophyti CBS 142.79 GU227993 GU228189 GU228287 GU227895

C. circinans CBS 111.21 GU227952 GU227952 GU228246 GU227854

C. circinans CBS 221.81a GU227953 GU228149 GU228247 GU227855

C. curcumae IMI 288937a GU227991 GU228187 GU228285 GU227893

C. dematium CBS 125.25a GU227917 GU228113 GU228211 GU227819

C. dematium CBS 125340 GU227918 GU228114 GU228212 GU227820

C. jasminigenum LLTX-01a HM131508 HM153770 HM131494 HM131504 HM131499 HM131513

C. jasmini-sambac LLTA-01a HM131507 HM153768 HM131492 HM131502 HM131497 HM131511

C. jasmini-sambac HLTX-01 HM153769 HM131493 HM131503 HM131498 HM131512

C. jasmini-sambac CLTA-01 HM131510 HM153772 HM131496 HM131506 HM131501 HM131515

C. lineola CBS 125337a GU227927 GU228123 GU228221 GU227829

C. lineola CBS 125333 GU227930 GU228126 GU228224 GU227832

C. liriopes CBS 119444a GU227902 GU228098 GU228196 GU227804

C. liriopes CBS 122747 GU227903 GU228099 GU228197 GU227805

C. siamense MFU 090230a FJ 907423 FJ 907438 FJ 917505 FJ 972596 FJ 972575 FJ 972613

C. siamense MFU 090231 FJ 907422 FJ 907437 FJ 917504 FJ 972597 FJ 972574 FJ 972614

C. fructi CBS 346.37a GU227942 GU228138 GU228236 GU227844

C. fructicola MFU 090226a FJ 907427 FJ 907442 FJ 917509 FJ 972592 FJ 972579 FJ 972602

C. fructicola MFU 090227 FJ 907425 FJ 907440 FJ 917507 FJ 972594 FJ 972577 FJ 972611

C. fructicola MFU 090228 FJ 907426 FJ 907441 FJ 917508 FJ 972593 FJ 972578 FJ 972603

C. simmondsii BRIP 28519 FJ 907428 FJ 907443 FJ 917510 FJ 972591 FJ 972580 FJ 972601

C. simmondsii CBS 294.67 FJ 907429 FJ 907444 FJ 917511 FJ 972590 FJ 972581 FJ 972610

C. gloeosporioides CBS 953.97 FJ 907430 FJ 907445 FJ 917512 FJ 972589 FJ 972582 FJ 972609

C. gloeosporioides CORCG4a HM034800 HM034810 HM034802 HM034806 HM034808

C. gloeosporioides CORCG5a HM034801 HM034811 HM034803 HM034807 HM034809

C. horii TSG001 GU133374 GU133375 GU133376 GU133377 GQ329682 AY787483

C. horii TSG002 GU133379 GU133380 GU133381 GU133382 GQ329680 AY791890

C. kahawae IMI 319418 FJ 907432 FJ 907446 FJ 917514 FJ 972588 FJ 972583 FJ 972608

C. kahawae IMI 363578 FJ 907433 FJ 907447 FJ 917515 FJ 972587 FJ 972584 FJ 972607

C. rusci CBS 119206a GU227916 GU228112 GU228210 GU227818

C. spaethianum CBS 167.49 GU227905 GU228101 GU228199 GU227807

C. spaethianum CBS 100063 GU227906 GU228102 GU228200 GU227808

C. trichellum HKUCC10378 GQ856786 GQ849447 GQ849466 GQ856749 GQ485589

C. truncatum CBS 151.35a GU227960 GU228156 GU228254 GU227862

C. truncatum CBS 136.30 GU227974 GU228170 GU228268 GU227876

C. truncatum CBS 182.52 GU227964 GU228160 GU228258 GU227866

Colletotrichum sp. LLBM-04 HM131509 HM153771 HM131495 HM131505 HM131500 HM131514

C. verruculosum IMI 45525a GU227904 GU228100 GU228198 GU227806

ACT actin, TUB-2 partial -tubulin, CAL calmoudulin, GS glutamine synthetase, GDPH glyceraldehydes-3-phosphate dehydrogenase, ITScomplete rDNA-ITS region.a indicate the ex-type culures. The newly generated sequence in this study are shown in bold.

Fungal Diversity (2011) 46:171182 173

Extraction Kit (BioFlux) according to the instructions ofthe manufacturer. DNA concentrations were estimatedvisually in agarose gel by comparing band intensity witha DNA ladder 100 bp (Transgen Biotech).

PCR, sequencing and phylogenetic analysis

Partial actin (ACT), -tubulin (TUB2), calmodulin(CAL), glutamine synthetase (GS), glyceraldehyde-3-phosphate dehydrogenase (GPDH) genes and the com-plete rDNA-ITS (ITS) region from five Colletotrichumstrains were amplified by PCR reaction, as described byPrihastuti et al. (2009). DNA sequencing was performed atthe SinoGenoMax Company Limited, Beijing, China. Theaccession numbers of all sequences are listed in Table 1.Parsimony and Bayesian analyses were performed byusing PAUP* 4.0b10 (Swofford 2002) and MrBayes3.0b4 (Huelsenbeck and Ronquist 2001), with detailsoutlined in Cai et al. (2006).

Results

Nine strains of Colletotrichum were isolated from jasmineleaves and flowers and morphological and culture character-istics established using the methods suggested by Cai et al.(2009). The entire ITS region of all strains were sequencedand compared with known ex-type cultures of Colletotri-chum. Three strains (LLBM09, LLTX05, LLNB06) wereidentified as C. truncatum, while one strain (LLBM02) wasidentified as C. siamense, based on ITS sequence analysisand morphological characters (results not shown). The otherfive strains did not cluster with any currently known speciesbased on molecular and morphological characters. A furtherfive gene regions of these five strains were thereforesequenced and phylogenetic relationships reconstructedusing parsimonious and Bayesian methods (Figs. 6, 7).These five strains did not group with any other species ofColletotrichum in the phylogenetic tree (Figs. 6, 7). StrainsCLTA01, LLTA01 and HLTX01 were resolved in a wellseparated lineage with C. siamense as its sister group. StrainLLTX01, which is characterized by curved conidia,clustered in a distinct lineage basal to C. curcumae and C.truncatum. Strain LLBM04 however, was not well resolvedin the six gene tree and herein tentatively named asColletotrichum sp.

Taxonomy

Colletotrichum jasminigenum Wikee, K.D. Hyde, L. Caiand McKenzie, sp. nov.

MycoBank: MB 518490 Fig. 2Etymology: meaning produced on Jasminum.Colletotrichi truncati simile, sed conidiis ad appessoriis

grande, in vitro (PDA), conidiis 19.526.52.54 m,incurvo, hyalinis. appressoriis irregulariter ovoidis, clavatis.

On PDA: Vegetative hyphae hyaline, septate, branched,825 m wide. Chlamydospores not observed. Setaeabsent. Conidiophores hyaline, 1-septate, not branched,3980 m. Conidiogenous cells 1014 m long. Conidia19.526.52.54 m (x=23.523.50.4 m, n=20),lunate, one-celled, smooth-walled, widest in the centre,hyaline, guttulate, with a larger central guttule. Appressoriavariable, ovoid, clavate or irregular, formed mostly frommycelia, often becoming complex and lobed with age,brown, with 16 small, ocular, paler regions, possibly germpores, some with conical, pale apical cells.

Cultural characteristics: Colonies on PDA, raised, grey,with black spots near the margin, in reverse with zonations;on MEA orange to brown with the edge generally deeplylobed, in reverse with brown spots and brown zones at thecentre, no pigment in medium; on OA, grey, with darkspots from the centre to the margin, margin fimbriate,reverse yellowish to dark brown at the centre, no pigmentproduced in the media growth rate on PDA 24.210.7 mm/day (3944 mm in diam. after 7 days at 27C).

Material examined: Vietnam, Ho Chi Min City, 12thDistrict, Thanh Xuan Ward, on living leaves of Jasminumsambac, May 2009, Hoa Nguyen Thi, LLTX01 (MFLU100212, holotype); ex-type living cultures CGMCC,MFLUCC 100273.

Colletotrichum jasmini-sambac Wikee, K.D. Hyde, L.Cai and McKenzie, sp. nov.

MycoBank: MB 518491 Fig. 3Etymology: Named after its host, Jasminum sambac.Colletotrichi siamensi simile, sed conidiis ad appessoriis

grande, in vitro (PDA), conidiis 13153.54 m, uni-cellularis, hyalinis, cylindricis, Appressoriis irregulariterovoidis, clavatis.

On PDA: Vegetative hyphae hyaline, septate, branched,2028mwide.Chlamydospores not observed. Setae absent.Conidiophores hyaline, septate, branched, 2339 m long.Conidiogenous cells hyaline, 1215 m long. Conidiaformed in grey to black areas, 13153.54 m, (x=14

Fig. 2 Colletotrichum jasminigenum (from MFLU 100212, holo-type) a Symptoms on leaves. Note the chlorotic areas (arrowed).bd Colonies on different media. b Two weeks growth on PDA. cTen days growth on MEA. d Ten days growth on OA. e, f Curvedconidia. Note the central guttule in (f). gh Long conidiophores withattached conidia. ip Irregularly-shaped appressoria, some withnumerous lobes and very complicated, some with conical, paleapical cells. Note the small light ocular regions which may be germpores. Bars=10 m

174 Fungal Diversity (2011) 46:171182

0.73.80.2 m, n=20), cylindrical, straight with obtuseends, hyaline, aseptate, smooth-walled. Appressoria variablein shape, brown, ovoid, clavate or slightly irregular.

Cultural characteristics: Colonies on PDA crateriform,white aerial mycelium, cottony, with fimbriate margin,reverse cream, no pigment in medium; on MEA white,crateriform, cottony, aerial mycelium white, reverse creamat the edge, with fimbriate margin, no pigment in medium;and on OA raised, white, cottony, aerial mycelium white,growth rate on PDA 41.419.7 mm/day (6674 mm indiam. after 7 days at 27C).

Material examined: Vietnam, Cu Chi District, Trung AnWard, on living leaves of Jasmin sambac, January 2009,Hoa Nguyen Thi, LLTA01 (MFLU 100214, holotype);ex-type living cultures CGMCC 3.14193, MFLUCC 100277; ibid., Thanh Xuan Ward, on flower of Jasminsambac, September 2009, Hoa Nguyen Thi, HLTX01(MFLU 100215); living cultures CGMCC 3.14191,MFLUCC 100275; ibid., January 2009, Hoa NguyenThi, CLTA01.

Notes: Strains CLTA01, LLTA01 and HLTX01 werenot statistically different in growth rate and spore morphol-ogy. LLTA01 is chosen for representative strain of the newspecies (as ex-holotype).

Colletotrichum siamense Prihastuti, L. Cai & K.D.Hyde, Fungal Diversity 39: 98 (2009). Fig. 4

Material examined: Vietnam, Ho Chi Minh City, Cu Chidistrict, Binh My Ward, on living leaves of Jasmin sambac,January 2009, Hoa Nguyen Thi, LLBM02 (MFLU 100216); living cultures MFLUCC 100343.

Colletotrichum truncatum (Schwein.) Andrus & W.D.Moore, Phytopathology 25: 122 (1935) Fig. 4

Material examined: Vietnam, Ho Chi Minh City, Cu Chidistrict, Binh My Ward, on living leaves of Jasmin sambac,June 2009, Hoa Nguyen Thi LLBM09 (MFLU 100217);living cultures (MFLUCC 100344); ibid., LLTX05, 12thDistrict, Thanh Xuan Ward, on living leaves of Jasminsambac, June 2009, Hoa Nguyen Thi LLTX05 (MFLU100218); living cultures (MFLUCC 100345); ibid., HocMon district, Nhi Binh Ward, on living leaves of Jasminsambac, June 2009, Hoa Nguyen Thi LLNB06 (MFLU100219); living cultures (MFLUCC 100346).

Colletotrichum sp. Fig. 5On PDA: Vegetative hyphae hyaline, septate, branched,

1923 m wide. Chlamydospores not observed. Setaeabsent. Conidiophores hyaline, septate, unbranched, up to136144 m becoming long with age. Conidiogenous cellshyaline, cylindrical 1314 m long. Conidia 11.5163.56.5 m (x=13.6150.7 m, n=20), cylindrical withobtuse ends, aseptate, smooth-walled, one-celled, hyaline.Appressoria variable, brown, ovoid, clavate or slightlyirregular with 16 small, ocular, lighter regions, possiblygerm pores.

Cultural characteristics: Colonies on PDA, flat, withgrey concentric rings, black spots at margin, no pigment inmedium; in reverse grey with dark grey spots in the centre.On MEA and OA, at first white, becoming grey to darkgrey with age, with dark circular area at the centre, marginundulate, in reverse grey with dark grey spots, no pigmentin medium; growth rate on PDA 26.211 mm/day. (4142 mm in diam. after 7 days at 27C)

Material examined: Vietnam, Cu Chi District, Binh MyWard, from Thanh Xuan Ward, on living leaves ofJasminum sambac, June 2009, Hoa Nguyen Thi LLBM04 (MFLU 100213); living cultures CGMCC 3.14192,MFLUCC 100274.

Discussion

Colletotrichum species on jasmine have been poorlystudied, with reports of only C. jasminicola and C.dematium (Tilak 1960; Agarwal 1962; Agarwal and Sahni1965). Colletotrichum jasminicola has smaller conidia (81135 m) than any of the species reported in thepresent study (Tilak 1960). It is also unclear from thedescription and diagnosis that C. jasminicola is actually aColletotrichum species. Colletotrichum dematium wasonly recently epitypified, with CBS 125.25 as the ex-epitype (Damm et al. 2009). Therefore, it cannot beconfirmed if Agarwal and Sahni (1965) correctly identi-fied their species as C. dematium. The current studyisolated nine strains representing five distinct taxa andthree of them have not been reported in previous studies.Although this investigation is limited in sampling scaleand isolations obtained, it appears that jasmine may host arelatively high diversity of Colletotrichum species.

Until recently most identifications of Colletotrichumspecies have been based on morphological data andalthough strides were made to incorporate sequence datathey generally proved confusing (Hyde et al. 2009a;2009b). The difficulties arose as most species lacked typecultures and many of the sequences deposited in GenBankwere wrongly named (Crouch et al. 2009; Cai et al. 2009).Cannon et al. (2008) epitypified the ubiquitous speciesColletotrichum gloeosporioides and for the first time itwas possible to compare fresh collections against a livingex-epitype culture. Recent rapid progress in molecular

Fig. 3 Colletotrichum jasmini-sambac (from MFLU 100214, holo-type). a A3B2 Symptoms on leaves. Note the chlorotic areas(arrowed). bd Colonies on different media. b Two weeks growthon PDA. c Ten days growth on MEA. d Ten days growth on OA. efCylindrical conidia with rounded ends. gi Conidiophores producingconidia. jl Germinating conidia. mp Irregularly shaped appressoria.Bars=10 m


phylogenetics has also made it possible to recognisestable and well-resolved clades within Colletotrichumspecies complexes (Crouch and Beirn 2009; Damm et al.2009; Prihastuti et al. 2009; Shivas and Yu 2009). Apolyphasic approach has been recommended for study intothe systematics of Colletotrichum, which emphasizescomparison with type specimens, and multilocus phylog-eny in conjunction with recognisable polyphasic charac-ters (Cai et al. 2009). Recently, Damm et al. (2009)described 18 species with curved spores of which 10 wereepitypes and 4 were new species. Prihastuti et al. (2009)and Yang et al. (2009) described six new species fromcoffee berries and amarylarids, while Shivas and Yu(2009) discussed epitypification of the acutatum com-plex. The establishment of two new species in this study,together with above recently described new species agreewell with the above concepts for recognizing a distinctspecies.

Among the three taxa described in this paper, two fitin the gloeosporioides-complex (cylindrical conidia) andthe other fits in the truncatum-complex (curved conidia).These three taxa can be well differentiated by morphology(mainly conidial shape and size), cultural characteristicsand growth rates. The two taxa in the gloeosporioides-complex also differ by their cultural characteristics andgrowth rates. Colletotrichum jasmini-sambac grows sig-nificantly faster than C. jasminigenum (41.38 vs.24.24 mm/day).

Fig. 4 Colletotrichum siamense from MFLU 100216. a Symptoms on leaves. Note the chlorotic areas (arrow). b Colony on PDA.Colletotrichum truncatum from MFLU 100217. c Symptoms on leaves. Note the chlorotic areas (arrow). d Colony on PDA

Fig. 5 Colletotrichum sp. (from MFLU 100213). a Symptoms onleaves. Note the chlorotic areas (arrow). bd Colonies on differentmedia. b Two weeks growth on PDA. c Ten days growth on MEA. dTen days growth on OA. (el). eg Cylindrical conidia with roundedends. h Germinating conidium. i Conidiophore with developingconidium. jl Irregularly shaped appressoria. Bars=10 m


Colletotrichum jasmini-sambac is most closely related toC. siamense, which is a common pathogen that has beenreported from a wide range of hosts (Prihastuti et al. 2009;Yang et al. 2009; Phoulivong et al. 2010). Colletotrichumjasmini-sambac has an overlapping conidial size range withC. siamense and both produce white and cottony colonieson PDA. However, the mean conidial length of theformer is significantly longer (14.01 vs. 10.18 m), andit also grows significantly faster (41.38 vs. 9.12 mm/day)(Prihastuti et al. 2009). We obtained three strains of C.jasmini-sambac and they all present similar conidial andappressorial morphology and growth rates.

Colletotrichum sp. (LLBM04) is represented by onlyone strain. Further investigations are suggested so as to

have a better understanding of its phylogenetic relationshipsand infraspecific variation. In the phylogenetic tree, strainLLBM04 appears as sister clade between C. siamense andC. hymenocallidis. Colletotrichum sp. (LLBM04) produ-ces slightly smaller conidia than C. hymenocallidis, butthey are similar to those of C. siamense. Further collectionsand studies are required to clarify the phylogenetic relation-ships between these strains.

Colletotrichum jasminigenum (strain LLTX01) pro-duced curved conidia (Fig 2), which have close affinity tospecies in truncatum-complex. Colletotrichum jasminige-num is most closely related to C. truncatum and C.curcumae (Fig. 7) (Damm et al. 2009). Colletotrichumjasminigenum differs in producing significantly larger

C jasmini-sambac MFLUCC 100277*C. jasmini-sambac MFLUCC 10 0277C. jasmini-sambac MFLUCC 10027577 j

C. jasmini-sambac CLTA-01C. siamense BML I15

C. siamense BPD I2*99

100Colletotrichum sp. MFLUCC 100274

100

C. hymenocallidis CSSN350 C. hymenocallidis CSSN2*

C f i l BPD I12

50

C. fructicola BPD I12

C fructicola BPD I1898 C. fructicola BPD I18

C. fruiticola BPD I16*59

C. fruiticola BPD I16

C. asianum BML I1473

C. asianum BML I39997

73

C. asianum BPDI4*

C. gloeosporioides CORCG592

C. gloeosporioides CBS 953.97*100


100 C. horii TSG001

C h ii TSG002

100100

C. horii TSG002

C kahawae IMI 319418*100100

C. kahawae IMI 319418

C. kahawae IMI 363578*

100

C. jasminigenum MFLUCC 100273*100

j g

C. simmondsii BRIP 28519*100

C. simmondsii CBS 294.67

10C. trichellum

10

Fig. 6 Phylogram generatedfrom maximum parsimony anal-ysis based on combined ACT,GPDH, ITS, TUB2, GS andCAL sequences, showing thephylogenetic relationships ofColletotrichum jasmini-sambac.Values above the branches areparsimony bootstrap (>50%).Thickened branches representsignificant Bayesian posteriorprobability (95%). The tree isrooted with Colletotrichum tri-chellum.*indicate the ex-typestrains. Strains isolated in thisstudy are shown in bold


conidia and appressoria than both species and the appres-soria of C. jasminigenum are very variable in shape.

Acknowledgements This research was funded by Chinese Academyof Forestry and the Knowledge Innovation Program of the ChineseAcademy of Sciences, No. KSCX2-YW-Z-1026, and research grants51101010029 and 52101010002 awarded by Mae Fah LuangUniversity Chiang Rai, Thailand.

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Agarwal GP (1962) Fungi causing plant diseases at Jabalpur (M.P.)VIII. Mycopathol Mycol Appl 17:309314

Agarwal GP, Sahni VP (1965) Fungi causing plant diseases atJabalpur (M.P.) XI. Mycopathol Mycol Appl 27:136144

Al-Zadjali AD, Natsuaki T, Okuda S (2007) Detection, identificationand molecular characterization of a phytoplasma associated with

10

C. fructicola BPD I12*C. fructicola BPD I16*C. fructicola BPD I18*

C. gloeosporioides CBS 953 97*


C. horii TSG001

C. horii TSG002



C. truncatum CBS 151.35*

C. truncatum CBS 136.30

C. truncatum CBS 182.52

C. curcumae IMI 288937*

C. jasminigenum MFLUCC 100273*C. chlorophyti IMI 103806*

C. chlorophyti CBS 142.79

C. dematium CBS 125.25*

C. dematium CBS 125340

C. lineola CBS 125337*

C. lineola CBS 125333

C. fructi CBS 346.37C. anthrisci CBS 125334

C. anthrisci CBS 125335

C. circinans CBS 221.81

C. circinans CBS 111.21

C. liriopes CBS 119444*

C. liriopes CBS 122747

C. verruculosum IMI 45525*

C. spaethianum CBS 167.49*

C. spaethianum CBS 100063

C. trichellum HKUCC

C. rusci CBS 119206*

C. simmondsii CBS 294.67*

100

100

68

100

100

100

64

67

98

100

100

85

87

98

68

100

100

100

70

96100

100

100

100

98

50

94

Fig. 7 Phylogram generatedfrom maximum parsimony anal-ysis based on combined ACT,GPDH, ITS and TUB2 sequen-ces, showing the phylogeneticrelationships of Colletotrichumjasminigenum. Values above thebranches are parsimony boot-strap (>50%). Thickenedbranches represent significantBayesian posterior probability(95%). The tree is rooted withColletotrichum simmondsii.*indicate the ex-type strains.Strain isolated in this study isshown in bold


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Colletotrichum species from Jasmine (Jasminum sambac)AbstractIntroductionMaterials and methodsIsolation of Colletotrichum strainsMorphological studies of Colletotrichum from jasmineDNA extractionPCR, sequencing and phylogenetic analysisResultsTaxonomyDiscussionReferences

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Collectotrichum Jasmine Sambac