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Fungal Diversity (2011) 46:171182

DOI 10.1007/s13225-010-0049-x

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 and


flower anthracnose of jasmine (Jasminum sambac) in the
Ho Chi Minh region of Vietnam are reported. The disease
of jasmine plantations was considered serious as it likely
reduced flower yield. Leaves were colonized by Colletotrichum species which formed chlorotic regions with light
brown necrotic centres, which eventually covered the whole

S. Wikee : N. Pairin : E. Chukeatirote : K. D. Hyde


School of Science, Mae Fah Luang University,
Chiang Rai 57100, Thailand
K. D. Hyde
e-mail: kdhyde3@gmail.com
L. Cai (*) : Y.-Y. Su
Key Laboratory of Systematic Mycology & Lichenology,
Institute of Microbiology, Chinese Academy of Sciences,
Beijing, Peoples Republic of China
e-mail: mrcailei@gmail.com
E. H. C. McKenzie
Landcare Research,
Private Bag 92170, Auckland, New Zealand
H. N. Thi
Post Entry Quarantine Center No. 2,
Plant Protection Department,
District 1,
Ho Chi Minh City, Vietnam
A. H. Bahkali : M. A. Moslem : K. D. Hyde
Botany and Microbiology Department, College of Science,
King Saud University,
P.O. Box: 2455, Riyadh 1145, Saudi Arabia
K. Abdelsalam
College of Science, Abdul Rahman A1-Jeraisy DNA Research
Chair, King Saud University,
P.O. Box 2455, Riyadh 1145, Saudi Arabia

leaf and subsequently caused defoliation and dieback and


whole flowers were blighted. Nine strains of Colletotrichum species were isolated from diseased leaves and
flowers and partial ITS rDNA sequences were analysed
and morphologies compared across similar species. Based
on ITS sequence analysis and morphological characters,
three strains were identified as C. truncatum, while one
strain was identified as C. siamense. The remaining five
strains did not cluster with any known species for which
type sequences are available and therefore partial actin
(ACT), -tubulin (TUB2), calmodulin (CAL), glutamine
synthetase (GS), glyceraldehyde-3-phosphate dehydrogenase (GPDH) genes of the isolates were sequenced. Based
on the reconstructed multiloci molecular phylogeny, two
taxa are formally introduced as new species. Another strain
was not well resolved in the phylogenetic tree and herein
described as Colletotrichum sp. Further studies are needed
to prove its distinctiveness. The morphology and growth
rate of all taxa are described and compared with similar
species.
Keywords Anthracnose . Multilocus phylogeny .
Plant disease . Systematics

Introduction
Jasmine (Jasminum sambac (L.) Aiton) is grown as an
ornamental throughout Asia where it is native and is used
for garments, bouquets and in massage oils. In China and
Vietnam it is used to produce the flowers used in jasmine
tea and is grown in plantations. There are relatively few
reports of disease of jasmine; these include witches broom
caused by phytoplasma (Al-Zadjali et al. 2007), tomato
mosaic virus infection (Kamenova et al. 2006), cercospor-

172

oid leaf spots (Braun and Sivapalan 1999) and several other
folicolous 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. dematium
from leaf spots.
Jasmine plantations have become a common and
important industry in Asia. In Vietnam Colletotrichum
species cause serious problems to the plantations, infecting
leaves and flowers, often resulting in defoliation and
dieback. The objective of this study was to examine and
describe Colletotrichum species that cause disease of
jasmine plants in Ho Chi Minh, Vietnam.

Materials and methods


A survey was made to jasmine plantations in Cu Chi
District, Hoc Mon District and the 12th District of Ho Chi
Minh 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 the
jasmine plants and returned to the laboratory in individual
snap lock plastic bags. In the laboratory they were
examined for visible sporulation and the fungi present were
confirmed to be Colletotrichum species. A single spore
isolation technique was applied to plant tissues where spore
masses were formed. Spore masses were picked off with a
sterilize wire loop or fine forceps and suspended in
sterilized water. The spore suspension was diluted to a

Fig. 1 a Jasmine plantation in Vietnam. b Se variety

Fungal Diversity (2011) 46:171182

reasonable concentration and spread onto the surface of


PDA agar, followed by incubation overnight at room
temperature (25C). Single germinating spores were picked
up with a sterilized needle and transferred to new PDA
plate for morphological and molecular study (Goh 1999).

Morphological studies of Colletotrichum from jasmine


Mycelial discs (0.5 cm diam.) were taken from 10 day old
cultures and transferred to potato dextrose agar (PDA), malt
extract agar (MEA) and oat agar (OA), and incubated at
27C following the methods of Cai et al. (2009). Colony
diameters of three replicate cultures of each species
growing on PDA were measured daily for 7 days. Growth
rate was calculated as the 7 day average of mean daily
growth (cm per day). After 7 days, colony size was
recorded. Colony characteristics on each type of agar were
also recorded.
Appressoria were produced using a slide culture technique in which a 1 cm2 square of a Colletotrichum culture
on PDA was placed in an empty Petri dish and a sterile
cover slip was placed over the agar (Cai et al. 2009). After
7 days, the shape and size of appressoria formed on the
underside of the cover slip were studied. Morphological
data were analyzed using analysis of variance (P<0.05)
with equal variance assumed (Duncans test).

DNA extraction
For production of mycelium for DNA extractions, isolates
were grown on PDA and incubated for 7 days. Mycelium
was obtained from the surface by scrapping. Genomic DNA
was extracted by using a Biospin Fungus Genomic DNA

Fungal Diversity (2011) 46:171182

173

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.
C.
C.
C.
C.

anthrisci
anthrisci
asianum
asianum
asianum

CBS 125334a
CBS 125335
MFU 090232a
MFU 090233
MFU 090234

GU227943
GU227944
FJ 903188
FJ 907424
FJ 907421

GU228139
GU228140
FJ 907434
FJ 907439
FJ 907436

FJ 917501
FJ 917506
FJ 917503

FJ 972586
FJ 972595
FJ 972598

GU228237
GU228238
FJ 972571
FJ 972576
FJ 972573

GU227845
GU227846
FJ 972605
FJ 972612
FJ 972615

C.
C.
C.
C.
C.
C.
C.
C.
C.
C.
C.
C.
C.
C.
C.
C.
C.
C.

chlorophyti
chlorophyti
circinans
circinans
curcumae
dematium
dematium
jasminigenum
jasmini-sambac
jasmini-sambac
jasmini-sambac
lineola
lineola
liriopes
liriopes
siamense
siamense
fructi

IMI 103806a
CBS 142.79
CBS 111.21
CBS 221.81a
IMI 288937a
CBS 125.25a
CBS 125340
LLTX-01a
LLTA-01a
HLTX-01
CLTA-01
CBS 125337a
CBS 125333
CBS 119444a
CBS 122747
MFU 090230a
MFU 090231
CBS 346.37a

GU227992
GU227993
GU227952
GU227953
GU227991
GU227917
GU227918
HM131508
HM131507

HM131510
GU227927
GU227930
GU227902
GU227903
FJ 907423
FJ 907422
GU227942

GU228188
GU228189
GU227952
GU228149
GU228187
GU228113
GU228114
HM153770
HM153768
HM153769
HM153772
GU228123
GU228126
GU228098
GU228099
FJ 907438
FJ 907437
GU228138

HM131494
HM131492
HM131493
HM131496

FJ 917505
FJ 917504

HM131504
HM131502
HM131503
HM131506

FJ 972596
FJ 972597

GU228286
GU228287
GU228246
GU228247
GU228285
GU228211
GU228212
HM131499
HM131497
HM131498
HM131501
GU228221
GU228224
GU228196
GU228197
FJ 972575
FJ 972574
GU228236

GU227894
GU227895
GU227854
GU227855
GU227893
GU227819
GU227820
HM131513
HM131511
HM131512
HM131515
GU227829
GU227832
GU227804
GU227805
FJ 972613
FJ 972614
GU227844

C.
C.
C.
C.
C.
C.
C.
C.
C.
C.
C.
C.
C.
C.
C.
C.
C.
C.

fructicola
fructicola
fructicola
simmondsii
simmondsii
gloeosporioides
gloeosporioides
gloeosporioides
horii
horii
kahawae
kahawae
rusci
spaethianum
spaethianum
trichellum
truncatum
truncatum

MFU 090226a
MFU 090227
MFU 090228
BRIP 28519
CBS 294.67
CBS 953.97
CORCG4a
CORCG5a
TSG001
TSG002
IMI 319418
IMI 363578
CBS 119206a
CBS 167.49
CBS 100063
HKUCC10378
CBS 151.35a
CBS 136.30

FJ 907427
FJ 907425
FJ 907426
FJ 907428
FJ 907429
FJ 907430
HM034800
HM034801
GU133374
GU133379
FJ 907432
FJ 907433
GU227916
GU227905
GU227906
GQ856786
GU227960
GU227974

FJ 907442
FJ 907440
FJ 907441
FJ 907443
FJ 907444
FJ 907445
HM034810
HM034811
GU133375
GU133380
FJ 907446
FJ 907447
GU228112
GU228101
GU228102
GQ849447
GU228156
GU228170

FJ 917509
FJ 917507
FJ 917508
FJ 917510
FJ 917511
FJ 917512
HM034802
HM034803
GU133376
GU133381
FJ 917514
FJ 917515

GQ849466

FJ 972592
FJ 972594
FJ 972593
FJ 972591
FJ 972590
FJ 972589

GU133377
GU133382
FJ 972588
FJ 972587

FJ 972579
FJ 972577
FJ 972578
FJ 972580
FJ 972581
FJ 972582
HM034806
HM034807
GQ329682
GQ329680
FJ 972583
FJ 972584
GU228210
GU228199
GU228200
GQ856749
GU228254
GU228268

FJ 972602
FJ 972611
FJ 972603
FJ 972601
FJ 972610
FJ 972609
HM034808
HM034809
AY787483
AY791890
FJ 972608
FJ 972607
GU227818
GU227807
GU227808
GQ485589
GU227862
GU227876

CBS 182.52
LLBM-04
IMI 45525a

GU227964
HM131509
GU227904

GU228160
HM153771
GU228100

HM131495

HM131505

GU228258
HM131500
GU228198

GU227866
HM131514
GU227806

C. truncatum
Colletotrichum sp.
C. verruculosum

ACT actin, TUB-2 partial -tubulin, CAL calmoudulin, GS glutamine synthetase, GDPH glyceraldehydes-3-phosphate dehydrogenase, ITS
complete rDNA-ITS region.
a

indicate the ex-type culures. The newly generated sequence in this study are shown in bold.

174

Extraction Kit (BioFlux) according to the instructions of


the manufacturer. DNA concentrations were estimated
visually in agarose gel by comparing band intensity with
a DNA ladder 100 bp (Transgen Biotech).

PCR, sequencing and phylogenetic analysis


Partial actin (ACT), -tubulin (TUB2), calmodulin
(CAL), glutamine synthetase (GS), glyceraldehyde-3phosphate dehydrogenase (GPDH) genes and the complete rDNA-ITS (ITS) region from five Colletotrichum
strains were amplified by PCR reaction, as described by
Prihastuti et al. (2009). DNA sequencing was performed at
the SinoGenoMax Company Limited, Beijing, China. The
accession numbers of all sequences are listed in Table 1.
Parsimony and Bayesian analyses were performed by
using PAUP* 4.0b10 (Swofford 2002) and MrBayes
3.0b4 (Huelsenbeck and Ronquist 2001), with details
outlined in Cai et al. (2006).

Results
Nine strains of Colletotrichum were isolated from jasmine
leaves and flowers and morphological and culture characteristics established using the methods suggested by Cai et al.
(2009). The entire ITS region of all strains were sequenced
and compared with known ex-type cultures of Colletotrichum. Three strains (LLBM09, LLTX05, LLNB06) were
identified as C. truncatum, while one strain (LLBM02) was
identified as C. siamense, based on ITS sequence analysis
and morphological characters (results not shown). The other
five strains did not cluster with any currently known species
based on molecular and morphological characters. A further
five gene regions of these five strains were therefore
sequenced and phylogenetic relationships reconstructed
using parsimonious and Bayesian methods (Figs. 6, 7).
These five strains did not group with any other species of
Colletotrichum in the phylogenetic tree (Figs. 6, 7). Strains
CLTA01, LLTA01 and HLTX01 were resolved in a well
separated lineage with C. siamense as its sister group. Strain
LLTX01, which is characterized by curved conidia,
clustered in a distinct lineage basal to C. curcumae and C.
truncatum. Strain LLBM04 however, was not well resolved
in the six gene tree and herein tentatively named as
Colletotrichum sp.

Taxonomy
Colletotrichum jasminigenum Wikee, K.D. Hyde, L. Cai
and McKenzie, sp. nov.

Fungal Diversity (2011) 46:171182


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. c
Ten days growth on MEA. d Ten days growth on OA. e, f Curved
conidia. Note the central guttule in (f). gh Long conidiophores with
attached conidia. ip Irregularly-shaped appressoria, some with
numerous lobes and very complicated, some with conical, pale
apical cells. Note the small light ocular regions which may be germ
pores. Bars=10 m

MycoBank: MB 518490 Fig. 2


Etymology: 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. Setae
absent. Conidiophores hyaline, 1-septate, not branched,
3980 m. Conidiogenous cells 1014 m long. Conidia
19.526.52.54 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. Appressoria
variable, ovoid, clavate or irregular, formed mostly from
mycelia, often becoming complex and lobed with age,
brown, with 16 small, ocular, paler regions, possibly germ
pores, 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 deeply
lobed, in reverse with brown spots and brown zones at the
centre, no pigment in medium; on OA, grey, with dark
spots from the centre to the margin, margin fimbriate,
reverse yellowish to dark brown at the centre, no pigment
produced 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, 12th
District, Thanh Xuan Ward, on living leaves of Jasminum
sambac, May 2009, Hoa Nguyen Thi, LLTX01 (MFLU
100212, 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. 3
Etymology: Named after its host, Jasminum sambac.
Colletotrichi siamensi simile, sed conidiis ad appessoriis
grande, in vitro (PDA), conidiis 13153.54 m, unicellularis, hyalinis, cylindricis, Appressoriis irregulariter
ovoidis, clavatis.
On PDA: Vegetative hyphae hyaline, septate, branched,
2028 m wide. Chlamydospores not observed. Setae absent.
Conidiophores hyaline, septate, branched, 2339 m long.
Conidiogenous cells hyaline, 1215 m long. Conidia
formed in grey to black areas, 13153.54 m, (x=14

Fungal Diversity (2011) 46:171182

175

176

0.73.80.2 m, n=20), cylindrical, straight with obtuse


ends, hyaline, aseptate, smooth-walled. Appressoria variable
in 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 cream
at 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 in
diam. after 7 days at 27C).
Material examined: Vietnam, Cu Chi District, Trung An
Ward, on living leaves of Jasmin sambac, January 2009,
Hoa Nguyen Thi, LLTA01 (MFLU 100214, holotype);
ex-type living cultures CGMCC 3.14193, MFLUCC 10
0277; ibid., Thanh Xuan Ward, on flower of Jasmin
sambac, September 2009, Hoa Nguyen Thi, HLTX01
(MFLU 100215); living cultures CGMCC 3.14191,
MFLUCC 100275; ibid., January 2009, Hoa Nguyen
Thi, CLTA01.
Notes: Strains CLTA01, LLTA01 and HLTX01 were
not statistically different in growth rate and spore morphology. LLTA01 is chosen for representative strain of the new
species (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 Chi
district, Binh My Ward, on living leaves of Jasmin sambac,
January 2009, Hoa Nguyen Thi, LLBM02 (MFLU 10
0216); 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 Chi
district, Binh My Ward, on living leaves of Jasmin sambac,
June 2009, Hoa Nguyen Thi LLBM09 (MFLU 100217);
living cultures (MFLUCC 100344); ibid., LLTX05, 12th
District, Thanh Xuan Ward, on living leaves of Jasmin
sambac, June 2009, Hoa Nguyen Thi LLTX05 (MFLU
100218); living cultures (MFLUCC 100345); ibid., Hoc
Mon district, Nhi Binh Ward, on living leaves of Jasmin
sambac, June 2009, Hoa Nguyen Thi LLNB06 (MFLU
100219); living cultures (MFLUCC 100346).
Colletotrichum sp. Fig. 5
On PDA: Vegetative hyphae hyaline, septate, branched,
1923 m wide. Chlamydospores not observed. Setae
absent. Conidiophores hyaline, septate, unbranched, up to
136144 m becoming long with age. Conidiogenous cells
hyaline, cylindrical 1314 m long. Conidia 11.5163.5
6.5 m (x=13.6150.7 m, n=20), cylindrical with
obtuse ends, aseptate, smooth-walled, one-celled, hyaline.
Appressoria variable, brown, ovoid, clavate or slightly
irregular with 16 small, ocular, lighter regions, possibly
germ pores.

Fungal Diversity (2011) 46:171182


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 growth
on PDA. c Ten days growth on MEA. d Ten days growth on OA. ef
Cylindrical conidia with rounded ends. gi Conidiophores producing
conidia. jl Germinating conidia. mp Irregularly shaped appressoria.
Bars=10 m

Cultural characteristics: Colonies on PDA, flat, with


grey concentric rings, black spots at margin, no pigment in
medium; in reverse grey with dark grey spots in the centre.
On MEA and OA, at first white, becoming grey to dark
grey with age, with dark circular area at the centre, margin
undulate, in reverse grey with dark grey spots, no pigment
in medium; growth rate on PDA 26.211 mm/day. (41
42 mm in diam. after 7 days at 27C)
Material examined: Vietnam, Cu Chi District, Binh My
Ward, from Thanh Xuan Ward, on living leaves of
Jasminum sambac, June 2009, Hoa Nguyen Thi LLBM
04 (MFLU 100213); living cultures CGMCC 3.14192,
MFLUCC 100274.

Discussion
Colletotrichum species on jasmine have been poorly
studied, with reports of only C. jasminicola and C.
dematium (Tilak 1960; Agarwal 1962; Agarwal and Sahni
1965). Colletotrichum jasminicola has smaller conidia (8
1135 m) than any of the species reported in the
present study (Tilak 1960). It is also unclear from the
description and diagnosis that C. jasminicola is actually a
Colletotrichum species. Colletotrichum dematium was
only recently epitypified, with CBS 125.25 as the exepitype (Damm et al. 2009). Therefore, it cannot be
confirmed if Agarwal and Sahni (1965) correctly identified their species as C. dematium. The current study
isolated nine strains representing five distinct taxa and
three of them have not been reported in previous studies.
Although this investigation is limited in sampling scale
and isolations obtained, it appears that jasmine may host a
relatively high diversity of Colletotrichum species.
Until recently most identifications of Colletotrichum
species have been based on morphological data and
although strides were made to incorporate sequence data
they generally proved confusing (Hyde et al. 2009a;
2009b). The difficulties arose as most species lacked type
cultures and many of the sequences deposited in GenBank
were wrongly named (Crouch et al. 2009; Cai et al. 2009).
Cannon et al. (2008) epitypified the ubiquitous species
Colletotrichum gloeosporioides and for the first time it
was possible to compare fresh collections against a living
ex-epitype culture. Recent rapid progress in molecular

Fungal Diversity (2011) 46:171182

177

178

Fungal Diversity (2011) 46:171182

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

phylogenetics has also made it possible to recognise


stable and well-resolved clades within Colletotrichum
species complexes (Crouch and Beirn 2009; Damm et al.
2009; Prihastuti et al. 2009; Shivas and Yu 2009). A
polyphasic approach has been recommended for study into
the systematics of Colletotrichum, which emphasizes
comparison with type specimens, and multilocus phylogeny in conjunction with recognisable polyphasic characters (Cai et al. 2009). Recently, Damm et al. (2009)
described 18 species with curved spores of which 10 were
epitypes and 4 were new species. Prihastuti et al. (2009)
and Yang et al. (2009) described six new species from
coffee berries and amarylarids, while Shivas and Yu
(2009) discussed epitypification of the acutatum complex. The establishment of two new species in this study,
together with above recently described new species agree
well with the above concepts for recognizing a distinct
species.

Among the three taxa described in this paper, two fit


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

Fig. 5 Colletotrichum sp. (from MFLU 100213). a Symptoms on


leaves. Note the chlorotic areas (arrow). bd Colonies on different
media. b Two weeks growth on PDA. c Ten days growth on MEA. d
Ten days growth on OA. (el). eg Cylindrical conidia with rounded
ends. h Germinating conidium. i Conidiophore with developing
conidium. jl Irregularly shaped appressoria. Bars=10 m

Fungal Diversity (2011) 46:171182

179

180

Fungal Diversity (2011) 46:171182

Fig. 6 Phylogram generated


from maximum parsimony analysis based on combined ACT,
GPDH, ITS, TUB2, GS and
CAL sequences, showing the
phylogenetic relationships of
Colletotrichum jasmini-sambac.
Values above the branches are
parsimony bootstrap (>50%).
Thickened branches represent
significant Bayesian posterior
probability (95%). The tree is
rooted with Colletotrichum trichellum.*indicate the ex-type
strains. Strains isolated in this
study are shown in bold

C jasmini-sambac MFLUCC 10
C.
100277*
0277
77

C. jjasmini-sambac MFLUCC 100275


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

99

C. siamense BPD I2*

100
Colletotrichum sp. MFLUCC 100274
C. hymenocallidis CSSN3
50

C. hymenocallidis CSSN2*
C ffructicola
C.
i l BPD I12
98

59

C
C. fructicola BPD I18
C. fruiticola BPD I16*
I16

73

C. asianum BML I14

99

97

C. asianum BML I3
C. asianum BPDI4*

92
100

C. gloeosporioides CORCG5
C. gloeosporioides CBS 953.97*
C. gloeosporioides CORCG4

100

100

C. horii TSG001
C hhorii
C.
ii TSG002

100
100

C
319418*
C. kahawae IMI 319418
C. kahawae IMI 363578*

100

C. jasminigenum
MFLUCC 100273*
j
g
100

C. simmondsii BRIP 28519*


C. simmondsii CBS 294.67

C. trichellum
10

Colletotrichum jasmini-sambac is most closely related to


C. siamense, which is a common pathogen that has been
reported from a wide range of hosts (Prihastuti et al. 2009;
Yang et al. 2009; Phoulivong et al. 2010). Colletotrichum
jasmini-sambac has an overlapping conidial size range with
C. siamense and both produce white and cottony colonies
on PDA. However, the mean conidial length of the
former is significantly longer (14.01 vs. 10.18 m), and
it 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 and
appressorial morphology and growth rates.
Colletotrichum sp. (LLBM04) is represented by only
one strain. Further investigations are suggested so as to

have a better understanding of its phylogenetic relationships


and infraspecific variation. In the phylogenetic tree, strain
LLBM04 appears as sister clade between C. siamense and
C. hymenocallidis. Colletotrichum sp. (LLBM04) produces slightly smaller conidia than C. hymenocallidis, but
they are similar to those of C. siamense. Further collections
and studies are required to clarify the phylogenetic relationships between these strains.
Colletotrichum jasminigenum (strain LLTX01) produced curved conidia (Fig 2), which have close affinity to
species in truncatum-complex. Colletotrichum jasminigenum is most closely related to C. truncatum and C.
curcumae (Fig. 7) (Damm et al. 2009). Colletotrichum
jasminigenum differs in producing significantly larger

Fungal Diversity (2011) 46:171182

181

Fig. 7 Phylogram generated


from maximum parsimony analysis based on combined ACT,
GPDH, ITS and TUB2 sequences, showing the phylogenetic
relationships of Colletotrichum
jasminigenum. Values above the
branches are parsimony bootstrap (>50%). Thickened
branches represent significant
Bayesian posterior probability
(95%). The tree is rooted with
Colletotrichum simmondsii.
*indicate the ex-type strains.
Strain isolated in this study is
shown in bold

C. fructicola BPD I12*

100

C. fructicola BPD I16*


68
94

C. fructicola BPD I18*


100 C. gloeosporioides CBS 953 97*
C. gloeosporioides CORCG5

100 100 C. horii TSG001


C. horii TSG002
100 C. kahawae IMI 319418*
C. kahawae IMI 363578*

100

64
67
85

C. truncatum CBS 151.35*


C. truncatum CBS 136.30
C. truncatum CBS 182.52

98

C. curcumae IMI 288937*


C. jasminigenum MFLUCC 100273*
100 C. chlorophyti IMI 103806*
C. chlorophyti CBS 142.79
87 C. dematium CBS 125.25*
98
68

C. dematium CBS 125340


C. lineola CBS 125337*
C. lineola CBS 125333

100
C. fructi CBS 346.37
100 C. anthrisci CBS 125334

70

50

C. anthrisci CBS 125335


100

C. circinans CBS 221.81


C. circinans CBS 111.21

96 C. liriopes CBS 119444*


100
C. liriopes CBS 122747

100
100

C. verruculosum IMI 45525*


100 C. spaethianum CBS 167.49*
C. spaethianum CBS 100063

98

C. trichellum HKUCC
C. rusci CBS 119206*

C. simmondsii CBS 294.67*


10

conidia and appressoria than both species and the appressoria of C. jasminigenum are very variable in shape.
Acknowledgements This research was funded by Chinese Academy
of Forestry and the Knowledge Innovation Program of the Chinese
Academy of Sciences, No. KSCX2-YW-Z-1026, and research grants
51101010029 and 52101010002 awarded by Mae Fah Luang
University Chiang Rai, Thailand.

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