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Journal of Integrative Agriculture  2021, Vol. 20 Issue (11): 2957-2965    DOI: 10.1016/S2095-3119(21)63629-8
Special Issue: 植物病理合辑Plant Protection—Plant Pathology 植物病毒合辑Plant Virus
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Three Sclerotinia species as the cause of white mold on pea in Chongqing and Sichuan of China
DENG Dong1, SUN Su-li1, DU Chen-zhang2, XIANG Chao3, LONG Jue-chen2, CHEN Wei-dong4, ZHU Zhen-dong1 
 
1 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China
2 Chongqing Academy of Agricultural Sciences, Chongqing 402160, P.R.China
3 Crop Research Institute, Sichuan Academy of Agricultural Sciences, Chengdu 610066, P.R.China
4 United States Department of Agriculture-Agricultural Research Service, Washington State University, Pullman, WA 99164, USA
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摘要   

由核盘菌(Sclerotinia sclerotiorum)引起的豌豆菌核病是我国常见的豌豆病害。然而,我们近期在重庆和四川调查中发现,引起豌豆菌核病的病原菌可有多个核盘菌种。为此,我们对引起重庆和四川豌豆菌核病的病原菌种进行鉴定,以期为病害防控及抗病育种奠定基础。利用常规组织分离方法对采自重庆永川与合川、四川仪陇的病株及菌核进行病原菌分离,对获得的类似核盘菌的真菌分离物用PDA(potato dextrose agar)培养基培养,观察并测量菌落形态、生长速率与菌核大小。利用核糖体基因内转录区(rDNA-ITS)序列和种特异性分子标记进行分离物的分子鉴定。rDNA-ITS序列采用真菌通用引物ITS4/ITS5对分离物基因组DNA进行PCR扩增,种特异性分子标记鉴定分别利用核盘菌、小核盘菌(S. minor)和三叶草核盘菌(S. trifoliorum)特异性引物SMLcc2F/R、SSasprF/R与STCadF/ R对分离物基因组DNA进行PCR扩增,系统发育分析选取10个代表分离物进行ITS序列测序和采用Mega X软件中非加权组平均法(UPGMA)构建系统发育树。致病性测定采用菌丝块接种豌豆植株茎秆方法进行。菌落生长速率和菌核大小差异可将30个分离物区分为“快速生长-中菌核”、“中速生长-小菌核”和“慢速生长-大菌核”3中类型,分别有6个、7个和17个分离物,与已报道的核盘菌属中的核盘菌、小核盘菌和三叶草核盘菌的类似。ITS序列扩增片段将30个分离物分为2个带型,10个代表分离物进一步的系统发育分析表明,2个分离物与小核盘菌(KY550019、MN421822)聚在同一分支,2个分离物与核盘菌(MN216247、MK734068和MF776031)聚在同一分支,6个分离物与三叶草核盘菌(AY187070、AY547267)聚在同一分支。核盘菌种特异性引物扩增结果表明,30个分离物中分别有6、7和17个分离物扩增出小核盘菌、核盘菌和三叶草核盘菌的目标条带。综合形态和分子特征结果,我们确定30个分离物中有6为小核盘菌、7个为核盘菌和17为三叶草核盘菌。致病性测定结果表明,30个分离物对2个豌豆品种均具有致病性,病害症状与田间自然发病症状相似,且从感病植物中分离出病原菌分离物与接种分离物相同。本研究证明重庆市和四川省部分地区的豌豆菌核病有小核盘菌、核盘菌和三叶草核盘菌3种病原菌,其中小核盘菌和三叶草核盘菌引起豌豆菌核病是在我国西南地区首次报道。




Abstract   
White mold of pea caused by Sclerotinia sclerotiorum is a common disease in China.  However, we discovered that the diverse Sclerotinia species could cause white mold on pea plants in Chongqing and Sichuan of China during recent disease surveys.  Thus, the objective of this study was to confirm the causal agents from diseased pea plants.  The obtained isolates of white mold from Chongqing and Sichuan were identified by morphological characters and molecular characterization to determine the pathogen species, and their pathogenicity was confirmed on pea through completing Koch’s postulates.  Fungal isolates of Sclerotinia-like were obtained from diseased plants or sclerotia.  Based on morphological characteristics and molecular characterization, 30 isolates were identified to three species, six isolates as S. minor, seven as S. sclerotiorum, and 17 as S. trifoliorum.  In pathogenicity tests on pea cultivars Zhongwan 4 and Longwan 1, all 30 isolates caused typical symptoms of white mold on the inoculated plants, and the inoculated pathogens were re-isolated from the diseased plants.  This study confirmed that white mold of pea was caused by three Sclerotinia species, S. sclerotiorum, S. minor and S. trifoliorum in Chongqing and Sichuan. It is the first report that S. minor and S. trifoliorum cause white mold of pea in Southwest China.
 
Keywords:   Pisum sativum        white mold        Sclerotinia sclerotiorum        Sclerotinia minor        Sclerotinia trifoliorum  
Received: 26 August 2020   Accepted:
CLC Number:     
Fund:  This study was supported by the China Agriculture Research System of MOF and MARA (CARS-08), the National Crop Germplasm Resources Center of China (NCGRC-2020-09) and the Scientific Innovation Program of the Chinese Academy of Agricultural Sciences.
Corresponding Authors:  Correspondence ZHU Zhen-dong, Tel: +86-10-82109609, Fax: +86-10-82109608, E-mail: zhuzhendong@caas.cn   
About author:  DENG Dong, E-mail: 2499363366@qq.com;

Cite this article: 

DENG Dong, SUN Su-li, DU Chen-zhang, XIANG Chao, LONG Jue-chen, CHEN Wei-dong, ZHU Zhen-dong. 2021. Three Sclerotinia species as the cause of white mold on pea in Chongqing and Sichuan of China. Journal of Integrative Agriculture, 20(11): 2957-2965.

 
[1] WU Hong-qu, SUN Li-li, LIU Fang, Wang Zhi-ying, CAO Chuan-wang. Preparation of dry flowable formulations of Clonostachys rosea by spray drying and application for Sclerotinia sclerotiorum control[J]. >Journal of Integrative Agriculture, 2018, 17(03): 613-620.
[2] ZHAO Xu, SHAN Ya-nan, ZHAO Yan, WANG Ai-rong, WANG Zong-hua. A novel Arabidopsis miRNA, ath-miR38-3P, is involved in response to Sclerotinia sclerotiorum infection[J]. >Journal of Integrative Agriculture, 2016, 15(11): 2556-2562.
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