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DOI 10.1007/s13225-010-0049-x
Received: 1 June 2010 / Accepted: 15 July 2010 / Published online: 31 July 2010
# Kevin D. Hyde 2010
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.
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
173
Table 1 Sources of isolates and GenBank accession numbers used in this study
Colletotrichum species
Culture collection
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
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.
175
176
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
177
178
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
179
180
C jasmini-sambac MFLUCC 10
C.
100277*
0277
77
99
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
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. trichellum
10
181
100
100
64
67
85
98
100
C. fructi CBS 346.37
100 C. anthrisci CBS 125334
70
50
100
100
98
C. trichellum HKUCC
C. rusci CBS 119206*
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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