Disease Note
The rDNA internal transcribed spacer (ITS), translation elongation factor
Diseases Caused by Fungi and Fungus-Like Organisms
First Report of Pestalotiopsis trachicarpicola Causing Leaf Blight on
Panax notoginseng in China
Xianjing Lin,1 Qingtao Wu,1 Juan Chen,1 Zhenting Liang,1
Yuhang Zhong,1 Qi Qin,1 Haihua Wang,1,2,3 and Ting Tang1,2,3,†
1
School of Life and Health Sciences, Hunan University of Science
and Technology, Xiangtan 411201, Hunan, China
2
Key Laboratory of Genetic Improvement and Multiple Utilization of
Economic Crops in Hunan Province, Xiangtan 411201, Hunan, China
3
Key Laboratory of Ecological Remediation and Safe Utilization of Heavy
Metal-Polluted Soils, College of Hunan Province, Xiangtan 411201,
Hunan, China
Funding: Funding was provided by the Natural Science Foundation of
Hunan Province, China (2020JJ4293), and Scientific Research Project of
the Hunan Education Department (21A0312). Plant Dis. XXX:XX,
XXXX; published online as https://doi.org/10.1094/PDIS-04-23-0681-
PDN. Accepted for publication 6 June 2023.
Panax notoginseng (Burkill) F. H. Chen ex C. Y. Wu & K. M. Feng is a
Chinese herbal medicinal plant for treating diseases of the central nervous
system and cardiovascular system, widely used as a medicine and health-
care product. In May 2022, leaf blight disease was found on leaves of
1-year-old P. notoginseng in plantings (27.904°N, 112.928°E) of Xiangtan
City (Hunan) with an area of 104 m2. Over 400 plants were investigated, and
up to 25% of the plants were symptomatic. From the margin of the leaf, the
initial symptoms of water-soaked chlorosis followed by drying and yel-
lowing with slight shrinkage appeared. Later, leaf shrinkage became severe,
and chlorosis enlarged gradually, leading to leaf death and abscission. To
identify the causal agent, 20 leaf lesions (4 mm2) collected from 20 indi-
vidual 1-year-old plants were sterilized with 75% ethanol for 10 s and 5%
NaOCl for 10 s, rinsed in sterilized water three times, placed on potato
dextrose agar (PDA) with lactic acid (0.125%) for inhibiting the growth of
bacteria, and incubated at 28°C for 7 days (Fang 1998). Five isolates were
obtained from 20 leaf lesions of different plants with an isolation rate of 25%
and purified by single sporing, which had similar colony and conidia mor-
phology characteristics. An isolate, PB2-a, was selected randomly for further
identification. Colonies of PB2-a on PDA were white with cottony myce-
lium, developing concentric circles (top view), and were light yellow (bot-
tom view) in color. Conidia (23.1 ± 2.1 × 5.7 ± 0.8 µm, n = 30) were
fusiform, straight or slightly curved and contained a conical basal cell, three
light brown median cells, and a hyaline conic apical cell with appendages.
†
Indicates the corresponding author.
T. Tang; tangting@hnust.edu.cn
© 2023 The American Phytopathological Society
1-alpha (tef1), and b-tubulin (TUB2) genes were amplified from genomic
DNA of PB2-a using the primers ITS4/ITS5 (White et al. 1990), EF1-
526F/EF1-1567R (Maharachchikumbura et al. 2012), and Bt2a/Bt2b
(Glass and Donaldson 1995; O’Donnell and Cigelnik 1997), respectively.
BLAST search of sequenced ITS (OP615100), tef1 (OP681464), and
TUB2 (OP681465) exhibited >99% identity with the type strain of
Pestalotiopsis trachicarpicola OP068 (JQ845947, JQ845946, and
JQ845945). Phylogenetic tree of the concatenated sequences was
constructed based on the maximum-likelihood method using MEGA-X.
The isolate PB2-a was identified as P. trachicarpicola based on mor-
phological and molecular data (Maharachchikumbura et al. 2011; Qi
et al. 2021). PB2-a was tested for pathogenicity three times, to fulfill
Kochʼs postulates. Twenty healthy leaves on twenty 1-year-old plants
were punctured with sterile needles and inoculated with 50 µl of co-
nidial suspension (1 × 10 6 conidia/ml). The controls were inoculated
with sterile water. All plants were placed in a greenhouse at 25°C under
80% relative humidity. After 7 days, all inoculated leaves developed
leaf blight symptoms identical to those described above, whereas the
control plants remained healthy. P. trachicarpicola was reisolated from
the infected leaves and was identical to the original isolates based on
the colony characteristics and the sequenced data of ITS, tef1, and
TUB2. P. trachicarpicola was reported as a pathogen of leaf blight on
Photinia fraseri (Xu et al. 2022). To our knowledge, this is the first
report of P. trachicarpicola causing leaf blight on P. notoginseng in
Hunan, China. Leaf blight is one of the destructive diseases in
P. notoginseng production. Thus, identification of the pathogen will be
useful to develop effective disease management and protect P. notoginseng,
a medical plant with economic value.
References:
Fang, Z. 1998. Page 122 in: Plant Disease Research Methods. China Agriculture Press,
Beijing, China.
Glass, N. L., and Donaldson, G. C. 1995. Appl. Environ. Microbiol. 61:1323.
Maharachchikumbura, S. S. N., et al. 2011. Fungal Divers. 50:167.
Maharachchikumbura, S. S. N., et al. 2012. Fungal Divers. 56:95.
O’Donnell, K., and Cigelnik, E. 1997. Mol. Phylogenet. Evol. 7:103.
Qi, M., et al. 2021. Antonie van Leeuwenhoek 114:1.
White, T. J., et al. 1990. Page 315 in: PCR Protocols: A Guide to Methods and
Applications. Academic Press, San Diego, CA.
Xu, X. L., et al. 2022. Plant Dis. 106:1520.
The author(s) declare no conflict of interest.
e-Xtra
Keywords: leaf blight, Panax notoginseng, Pestalotiopsis trachicarpicola
Plant Disease / 1