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Trichothecium roseum Causes Fruit Rot of

Tomato, Orange, and Apple in Pakistan

Article in Plant Disease September 2014

Impact Factor: 3.02 DOI: 10.1094/PDIS-01-14-0051-PDN

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5 authors, including:

M. Imran Hamid Muzammil Hussain

Chinese Academy of Sciences Chinese Academy of Sciences
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Mubashar Raza Xingzhong Liu

Chinese Academy of Sciences Chinese Academy of Sciences
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Available from: Muzammil Hussain

Retrieved on: 28 April 2016
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September 2014

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Editor-in-Chief: Mark L. Gleason
Published by The American Phytopathological Society
Phytopathology
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Tomato, Orange, and Apple in Pakistan
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M. I. Hamid, Department of Plant Pathology, University C ollege of
View Most Agriculture, University of Sargodha, 40100, Pakistan, and State Key
Downloaded Articles Laboratory of Mycology, Institute of Microbiology, C hinese Academy of
Sciences, Beijing, 100101, C hina; M. Hussain, M. U. Ghazanfar, and
About Plant Disease M. Raza, Department of Plant Pathology, University C ollege of
Editorial Board Agriculture, University of Sargodha, 40100, Pakistan; and X. Z. Liu,
State Key Laboratory of Mycology, Institute of Microbiology, C hinese
Submit a Manuscript Academy of Sciences, Beijing, 100101, C hina

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Policies/Procedures During a field survey of greenhouses and fresh markets in 2013, fruits
of tomato, oranges, and apples exhibited rot symptoms with white
Online e-Xtras mycelial growth and salmon-color sporulation in the vicinity of
Sargodha city (3251 N, 724016 E), Pakistan. Diseased fruit
= "Open" Access samples were collected in plastic bags and taken to laboratory on ice
for further diagnosis. Diseased fruits were observed under a
STEREO MICROSCOPE and single spores were removed using an
inoculating needle. Isolation from single spores showed pink to white
colonies on potato dextrose agar (PDA) containing hyaline, 2-celled,
ellipsoid to pyriform conidia (17 to 24 7 to 11 m) with slanting and
truncate basal mark and produced in clusters. C onidiophores were
branched (105 to 254 2 to 4 m) and hyphae were hyaline (3 to 5
m in diameter). These characteristics of the fungus were similar to
Trichothecium roseum (Pers.) as reported by Incio et al. (1).
Genomic DNA was extracted by using C TAB buffer from a single pure
colony of one isolate of the fungus and PC R ANALYSIS was
performed for ITS region and part of the 5 end of the beta tubulin
(TUB) gene (2,3). Single fragments of 550 bp and 1.5 kb length from
ITS and TUB gene were amplified and sequenced (GenBank Accession
Nos. KF975702 and KJ607590, respectively). Sequence ANALYSIS
showed 99% similarity with T. roseum isolates from different regions of
the world. Phylogenetic ANALYSIS (MEGA version 5.2 with WAG
model) showed the close relatedness to the isolates of T. roseum from
Pakistan and isolates from other parts of the world that revealed the
low genetic variability of ITS region. TUB gene sequence analysis
 indicated 100% homology with isolates of T. roseum and to the other
species in Hypocreales. Pathogenicity tests were performed on tomato
cvs. Nova Mech and Rio Grande, orange cv. Kinnow, and on apple cv.
Golden Delicious by inoculating five fruits from each cultivar. Spore
suspensions (105 conidia/ml of sterilized distilled water) were
inoculated into all wounded fruits (9 wounds/fruit) of each cultivar and
incubated at 25C for the development of symptoms. Five wounded
fruits of each cultivar were inoculated with sterilized distilled water as a
control treatment. The fruits were kept in plastic boxes and incubated
in humid chambers for 5 days. The symptoms on apples were
observed as brown rot with pinkish spores on rotted tissue. The cross
section of apple fruits also showed the brown rotted tissues internally.
The fungus developed mycelium and spores on the surface and caused
severe rotting inside the tomato and citrus fruits. T. roseum was re-
isolated by picking a single spore from rotted tissues of fruits under a
STEREO MICROSCOPE , and culturing on PDA. The re-isolated
fungus was confirmed morphologically and by molecular techniques.
Tomato and apple has been reported as a host for T. roseum (1,4,5)
but oranges have not. To our knowledge, this is the first record of T.
roseum infecting tomato, oranges, and apples in Pakistan.

References: (1) C . A. Incio et al. Plant Dis. 95:1318. 2011. (2) K.

O'Donnell, and E. C igelnik. Mol. Phylogenet. Evol. 7:103, 1997. (3) T. J.
White et al. Page 315 in: PC R Protocols: A Guide to Methods and
Applications. Academic Press, San Diego, C A, 1990. (4) Y. H. Yun et al.
Afr. J. Microbiol. Res. 7:1128, 2013. (5) M. abka et al.
Mycopathologia. 162:65, 2006.

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Editor-in-Chief: Mark L. Gleason

Published by The American Phytopathological Society

ISSN: 0191-2917 Home > Plant Disease > Table of Contents > Supplemental Material Quick Links
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September 2014, Volume 98, Number 9
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Phytopathology http://dx.doi.org/10.1094/PDIS-01-14-0051-PDN Alert me when new
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About Plant Disease

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Author Instructions

Policies/Procedures

Online e-Xtras

= "Open" Access
Pathogenicity assay on fruits and leaves. A to C, Rotting and spore
production on citrus fruits. D to F, Rot symptoms and spore formation
on the surface of apple fruits and also cause lesions on the leaves of
apple. G to I, Formation of mycelia mass and spore on the surface of
tomato fruit and rotting symptoms on inside tissues. J to L,
Morphological characteristics of T. roseum on PDA for 10 days.
 Phylogenetic ANALYSIS of Trichothecium roseum isolates. A,
Phylogenetic tree by using ITS locus. B, Phylogenetic tree with beta
tubulin gene locus. Multiple alignments were performed by using
C lustalX PROGRAM . Maximum likelihood (ML) trees were
constructed using the program MEGA version 5.2 with the WAG model.
The scale indicates 0.02 substitutions per site.

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