Identification and biological characterization of the pathogens responsible for sclerotinia rot in Codonopsis pilosula

doi:10.11686/cyxb2021462

Abstract

Abstract:

The aim of this study was to isolate and characterize the causal pathogen and study the biological characteristics of sclerotinia rot of Codonopsis pilosula. Pathogen material was obtained by tissue isolation. The isolated fungal species were comprehensively evaluated in terms of their morphology， rDNA-ITS sequence， RPB2 gene of RNA polymerase Ⅱ， and biological characteristics. The pathogens were identified as Sclerotinia nivalis and Sclerotinia sclerotiorum， and of the two， S. nivalis was the dominant species. The optimum temperatures for mycelium growth and sclerotia formation of S. nivalis were 20 and 15 ℃， respectively， while corresponding values for S. sclerotiorum were 25 and 25 ℃. The optimum pH for mycelium growth was 6 for both species， while optimum pH for sclerotia formation was 6 for S. nivalis and 7 for S. sclerotiorum. The optimum culture medium was 100% potato dextrose agar （PDA）. The mycelial growth rate approach was applied to test the toxicity of adiacylamine， which exhibited inhibitory activity with EC₅₀ on S. nivalis at 0.2625 mg·L^-1and on and S. sclerotiorum at 0.4165 mg·L^-1. In summary， this is the first report of S. nivalis and S. sclerotiorum causing sclerotinia rot on C. pilosula in China， and the data presented provide a reliable scientific proof of the pathogen diagnosis and a theoretical basis for the development of prevention measures.

Key words: Codonopsis pilosula, sclerotinia rot, pathogen identification, biological characteristics

Jing-jing JIANG, Hui DU, Ai-chang CHEN, Xue-ping LI, Min-quan LI, Yong-hong QI. Identification and biological characterization of the pathogens responsible for sclerotinia rot in Codonopsis pilosula[J]. Acta Prataculturae Sinica, 2022, 31(12): 181-190.

Figures/Tables 9

Fig.1 Symptom of C. pilosula sclerotinia rot and pathogenicity testing

Fig.2 The morphological characteristics of the pathogens causing C. pilosula sclerotinia rot

Fig.3 Phylogenetic tree for pathogensrDNA-ITS gene sequences

Fig.4 Neighbor-Joining tree based on RPB2 genes

Fig.5 Effect of different temperature on mycelium growth and sclerotial fresh weight

Fig.6 Effect of different pH on mycelial growth and sclerotial fresh weight

Fig.7 Effect of different medium on mycelium growth and sclerotia fresh weight

Fig.8 Inhibition of different concentration boscalid

Table 1 Inhibition of boscalid against mycelium growth of S. nivalis and S. sclerotiorum

病原菌

Pathogens

回归方程

Regression equation

References 29

1	National Pharmacopoeia Commission. The Pharmacopoeia of the People’s Republic of China. Beijing： China Medical Science and Technology Press， 2015.
	国家药典委员会. 中华人民共和国药典. 北京：中国医药科技出版社， 2015.
2	Jin P B， Hu J D， Mao G， et al. Effects of planting density on yield and secondary metabolite content of Codonopsis pilosula. Acta Prataculturae Sinica， 2018， 27（3）： 164-172.
	靳鹏博，胡佳栋，毛歌，等. 栽培密度对党参产量和次生代谢物含量的影响. 草业学报， 2018， 27（3）： 164-172.
3	Bai Y E， Zhang S S， Zhang R M， et al. Quality equivalence of Codonopsis radix-Codonopsis radix pieces-rice fried Codonopsis radix. Chinese Traditional Patent Medicine， 2020， 42（12）： 3228-3235.
	白云娥，张沙沙，张芮铭，等. 党参药材-饮片-米炒党参的质量等同性探究. 中成药， 2020， 42（12）： 3228-3235.
4	Liu S B. Study on commercial grade specification and rational analysis correlation of Gansu Codonopsis pilosula. Lanzhou： Gansu University of Chinese Medicine， 2016.
	刘书斌. 甘肃地产党参商品等级划分合理性分析及相关性的研究. 兰州：甘肃中医药大学， 2016.
5	Qi Y H， Li M Q， Cao S F， et al. Identification of Codonopsis pilosula crown rot disease and its sensitivity to fungicides. Journal of Agriculture， 2021， 11（2）： 74-78.
	漆永红，李敏权，曹素芳，等. 党参茎基腐病镰孢菌鉴定及其对杀菌剂的敏感性研究. 农学学报， 2021， 11（2）： 74-78.
6	Qi Y H， Li M Q， Cao S F， et al. Identification of a new Codonopsis pilosula root rot disease caused by Fusarium spp. Gansu Science and Technology， 2021， 37（1）： 43-46.
	漆永红，李敏权，曹素芳，等. 一种新的党参根腐病害镰孢菌鉴定. 甘肃科技， 2021， 37（1）： 43-46.
7	Yang K， Shi J， Yuan Y T， et al. Pathogen identification and cell physiological changes of Trifolium repens leaves infected with powdery mildew. Acta Prataculturae Sinica， 2021， 30（10）： 92-104.
	杨凯，史娟，袁玉涛，等. 白三叶草叶片感染白粉病的细胞生理变化及其病原鉴定. 草业学报， 2021， 30（10）： 92-104.
8	Huang Y P. Studies on the pathogens causing Angelica sinensis root rot. Lanzhou： Gansu Agricultural University， 2011.
	黄亚萍. 当归根腐病病原物研究. 兰州：甘肃农业大学， 2011.
9	Staats M， Van Baarlen P， Van kan J A L. Molecular phylogeny of the plant pathogenic genus Botrytis and the evolution of host specificity. Molecular Biology and Evolution， 2005， 22（2）： 333-346.
10	Wang C H， Shang W J， Wang Q， et al. White rot of Panax quinquefolius caused by Sclerotinia nivalis. Plant Pathology， 2021， 70（9）： 2034-2045.
11	Huang G Y. Pesticide trial technology and evaluation method. Beijing： China Agricultural Press， 2000.
	黄国洋. 农药试验技术与评价方法. 北京：中国农业出版社， 2000.
12	Zhao C S， Li D W， Yang D， et al. Pathogenicity and biological characteristics of several pathogenic fungi isolated on Codonopsis pilosulain root rot disease-Codonopsis pilosulain root rot disease continue to report Ⅰ. Journal of Hubei Agricutural Science， 1989（6）： 23-25.
	赵纯森，李德望，扬斗，等. 党参根腐病病株上分离的几种病原真菌的致病性及其生物学特性-党参根腐病研究续报Ⅰ. 湖北农业科学， 1989（6）： 23-25.
13	Sun X R， Zhong C P， Zhang X M， et al. Identification and control of the root rot pathogen of Codonopsis pilosula in Dingxi of Gansu Province. Plant Protection， 2020， 46（5）： 290-297.
	孙新荣，仲彩萍，张西梅，等. 甘肃定西地区党参根腐病病原鉴定与防治研究. 植物保护， 2020， 46（5）： 290-297.
14	Chenault K D， Maas A L， Damicone J P， et al. Discovery and characterization of a molecular marker for Sclerotinia minor （Jagger） resistance in peanut. Euphytica， 2009， 166（3）： 357-365.
15	Saito I. Sclerotinia nivalis， sp. nov.， the pathogen of snow mold of herbaceous dicots in northern Japan. Mycoscience， 1997， 38（2）： 227-236.
16	Lee C K， Lee S H， Choi Y J， et al. Sclerotinia rot of Aralia elata caused by Sclerotinia nivalis in Korea. Plant Pathology， 2010， 26（4）： 426.
17	Zhou R J， Xu H J， Fu J F， et al. First report of sclerotinia rot of Chinese atractylodes caused by Sclerotinia nivalis in China. Plant Disease， 2012， 96（12）： 1823.
18	Fan X， Zhang J， Yang L， et al. First report of Sclerotinia nivalis causing white mold disease on Sedum sarmentosum in China. Journal of Phytopathology， 2012， 160（10）： 595-598.
19	Xu H J， Zhou R J， Fu J F， et al. Characterization of Sclerotinia nivalis causing sclerotinia rot of Pulsatilla koreana in China. European Journal of Plant Pathology， 2015， 143： 1-9.
20	Guan Y M， Ma Y Y， Zhang L L， et al. Occurrence of postharvest snow rot caused by Sclerotinia nivalis on Asian ginseng in China. Plant Disease， 2022， 106（1）： 322.
21	Liu C Y， Wang Y， Hao Y J， et al. Studies on the identification and the biological characteristic of the pathogen of bean sclerotinia rot in greenhouse. Chinese Agricultural Science Bulletin， 2007， 23（12）： 316-319.
	刘春艳，王勇，郝永娟，等. 保护地菜豆菌核病病原鉴定及其生物学特性. 中国农学通报， 2007， 23（12）： 316-319.
22	Liu T T， Gu Y L， Yang T S， et al. Identification and characterization of the pathogen of sclerotinia rot of Aeschynanthus pulche. Journal of Plant Protection， 2016， 43（4）： 576-579.
	刘婷婷，谷医林，杨铁顺，等. 口红吊兰菌核病病原鉴定及其生物学特性分析. 植物保护学报， 2016， 43（4）： 576-579.
23	Chen Y Q， Li M， Mou D K， et al. Studies on the identification and the major biological characteristic of the pathogen of sclerotinia rot on Salvia splendens. Journal of Anhui Agricutural Science， 2009， 37（13）： 6067-6071.
	陈永强，李梅，牟登科，等. 一串红菌核病病原鉴定及其主要生物学特性研究. 安徽农业科学， 2009， 37（13）： 6067-6071.
24	Huang H C， Kokko E G， Erickson R S. Infection of alfalfa pollen by Sclerotinia sclerotiorum. Phytoparasitica， 1997， 25（1）： 17-24.
25	Young C S， Werner C P. Infection routes for Sclerotinia sclerotiorum in apetalous and fully petalled winter oilseed rape. Plant Pathology， 2012， 61（4）： 730-738.
26	Liu Y L. Effects of continuous cropping years on the growth of Codonopsis pilosula， soil physical and chemical properties and enzyme activities. Lanzhou： Gansu Agricultural University， 2021.
	刘垠霖. 连作年限对党参生长、土壤理化性状及酶活性的影响研究. 兰州：甘肃农业大学， 2021.
27	Shi Z Q， Zhou M G， Ye Z Y. Study on resistance mechanism of Sclerotinia sclerotiorum to dimethachlon. Chinese Journal of Pesticide Science， 2000， 2（2）： 47-51.
	石志琦，周明国，叶钟音. 核盘菌对菌核净的抗药性机制初探. 农药学学报， 2000， 2（2）： 47-51.
28	Wang K B， Pu T， Luo Y， et al. The synthesis and antifungal activity of nicotinamide derivatives. Journal of Yunnan Agricultural University （Natural Science）， 2018， 33（2）： 218-223.
	王凯博，普特，罗艳，等. 烟酰胺类化合物的合成及其抑菌活性. 云南农业大学学报（自然科学）， 2018， 33（2）： 218-223.
29	Zhang H Y， Li H Y， Meng Q L， et al. Effect of biological activity of boscalid on Sclerotinia sclerotiorum of sunflower. Journal of Agriculture， 2021， 11（1）： 71-74， 90.
	张海洋，李海燕，孟庆林，等. 啶酰菌胺对向日葵核盘菌生物活性的影响. 农学学报， 2021， 11（1）： 71-74， 90.

[1]	Chen-qin LI, Jun-qiao LI, Xin-ci WANG, Yong-kun NIU, Jun-ru QU. Isolation， identification， and biological characteristics of Fusarium perseae isolated from Potentilla anserina roots [J]. Acta Prataculturae Sinica, 2022, 31(4): 113-123.
[2]	Kai YANG, Juan SHI, Yu-tao YUAN, Li-ting WANG. Pathogen identification and cell physiological changes of Trifolium repens leaves infected with powdery mildew [J]. Acta Prataculturae Sinica, 2021, 30(10): 92-104.
[3]	WANG Qiong, DUAN Ting-yu, NAN Zhi-biao. Isolation and identification of an anthracnose pathogen on Vicia sativa [J]. Acta Prataculturae Sinica, 2020, 29(6): 127-136.
[4]	WANG Chun-ming, YUAN Wei-wei, ZHANG Xiao-jie, ZHOU Tian-wang, GUO Cheng, JIN She-lin. Isolation, identification and biological characteristics of Alternaria brassicicola leaf spot on Orychophragums violaceus [J]. Acta Prataculturae Sinica, 2020, 29(5): 88-97.
[5]	SUN Hai-rong, CHE Zhao-bi, CHEN Yi-shi, LU Wei-hua, WANG Shu-lin, LI Na-na, XIN Huai-lu. Ecological adaptability of biological traits and population distribution patterns for the ephemeral plant Leontice incerta in desert habitats [J]. Acta Prataculturae Sinica, 2019, 28(7): 198-207.
[6]	LI Jian-hong, LI Xue-ping, LI Chang-ning, HAN Bing, XU Wan-li, YAO Tuo. Characterization of a plant-growth-promoting rhizosphere bacterium, Gnyt1, and determination of its taxonomic status [J]. Acta Prataculturae Sinica, 2019, 28(5): 55-67.
[7]	DING Ai-qiang, XÜ Xian-ying, ZHANG Wen, LIU Jiang, FU Li, FU Gui-quan. Soil physicochemical and biological characteristics of Tamarix ramosissima Nebkhas in different degradation degree [J]. Acta Prataculturae Sinica, 2019, 28(2): 1-11.
[8]	YANG Cheng-De, BIAN Jing, CHEN Tai-Xiang, CHEN Xiu-Rong, WANG Han-Qi, YANG Xiao-Li, WANG Yang. Biological characteristics of the Angelica sinensis anthracnose causal agent, Colletotrichum dematium [J]. Acta Prataculturae Sinica, 2017, 26(6): 139-144.
[9]	HE Chun-Gui, HE Zhen-Fu, WANG Fei. Efficient double cropping pattern of photoperiod-sensitive sorghum-sudangrass hybrids in summer after winter wheat [J]. Acta Prataculturae Sinica, 2017, 26(5): 70-80.
[10]	LI Jian, LI Mei, GAO Xing-Xiang, FANG Feng, DONG Lian-Hong. Biological characteristics of Lubao No.1 biological control agent (Colletotrichum gloeosporioides) and construction of a T-DNA insertional mutant library [J]. Acta Prataculturae Sinica, 2017, 26(1): 142-148.
[11]	LI Jian, LI Mei, GAO Xing-Xiang, FANG Feng, DONG Lian-Hong. Isolation and biological characteristics of the biological control fungi BC-1 for Echinochloa crusgalli [J]. Acta Prataculturae Sinica, 2016, 25(8): 164-171.
[12]	JIA Hui, CHEN Xiu-Rong, LU Guang-Xin, KONG Ya-Li, YANG Cheng-De. Isolation of cellulose-degrading bacteria and determination of their degradation activity [J]. Acta Prataculturae Sinica, 2016, 25(3): 60-66.
[13]	LI Jian, LI Yan, GAO Xing-Xiang, FANG Feng, LI Mei. The biological characteristics of Digitaria sanguinalis pathogen Fusarium chlamydosporum strain ZC201301 [J]. Acta Prataculturae Sinica, 2016, 25(3): 234-239.
[14]	PAN Long-Qi, ZHANG Li, YANG Cheng-De, YUAN Qing-Hua, WANG Yu, MIAO Li-Hong. Identification and biological characteristics of Fusarium sporotrichioide isolated from Medicago sativa root [J]. Acta Prataculturae Sinica, 2015, 24(10): 88-98.
[15]	HE Zhen-Fu, HE Chun-Gui, WEI Yu-Ming, LIU Long-Sheng. Biological characteristics and nutrients of the photoperiod-sensitive Sorghum×Sudangrass varieties in the arid land of east Gansu [J]. Acta Prataculturae Sinica, 2015, 24(10): 166-174.