浙江省大豆红冠腐病的病原菌鉴定  

Identification of Red Crown Rot of Soybean Caused by Calonectria ilicicola in Zhejiang Province,China

作　　者：盖云鹏[1] 黄子凌 陈好好 周乔俐 马海杰[1] 李红叶[1] GAI Yun-peng HUANG Zi-lin CHEN Hao-hao ZHOU Qiao-li MA Hai-jie LI Hong-ye(Institute of Biotechnology, Gollege of Agriculture of Biotechnology, Zhejiang University, Hangzhou 310058, China)

机构地区：[1]浙江大学农业与生物技术学院生物技术研究所,浙江杭州310058

出　　处：《大豆科学》2016年第6期986-991,共6页Soybean Science

基　　金：浙江大学大学生科研训练计划(2015032199)

摘　　要：大豆红冠腐病(red crown rot of soybean,RCR)是世界范围内的重要的大豆病害之一,病害症状为罹病植株茎基部变红,表面聚集了大量橙红色球状子囊壳。对采自浙江省桐庐县的疑似大豆红冠腐病的病原进行分离和培养,从分离获得的菌株中选择1个代表菌株ZJHZ01进行后续试验和分析,为浙江省大豆产区的红冠腐病的防控提供依据。致病性测定结果表明:菌株ZJHZ01接种大豆茎基部可引起发病,回接症状与田间自然发病症状较为相似。病原菌落初为白色,后变为鹅黄色,菌落中可产生大量红褐色微菌核和厚垣孢子,在PDA、V8和康乃馨叶片培养基中极易产生子囊孢子,与已报道的大豆红冠腐病菌的菌落形态特征相似,初步认为ZJHZ01为冬青丽赤壳(Calonectria ilicicola)。进一步在分子生物学水平上,对其TUB、HIS和EF-1α序列进行PCR扩增和测序,对比Gen Bank中已经录入的丽赤壳属相应序列。基于以上3种基因联合序列,通过采用邻接法构建了的系统发育树。结果表明:菌株ZJHZ01与Gen Bank中C.ilicicola菌株GDBL01存在着极高的同源性,序列相似度达到了99%以上。通过对浙江省桐庐地区大豆产区的大豆病株的病原菌进行致病性、形态学和分子生物学分析,明确了浙江省桐庐大豆产区的红冠腐病是由C.ilicicola侵染所致,这是该病害在浙江省的首次报道。Red crown rot （RCR） is one of worldwide fungal diseases in soybean [ Glycine max （L.） Merr]. It causes yield loss and seed quality deficiency seriously. The objective of this study is to identify the pathogen causing stem rot on soybean in Zhejiang province, and to provide an IPM strategy for disease control. Typical symptoms of the disease were red discoloration at the base of the soil line with numerous reddish perithecia on the lesion surface. Diseased samples were collected from Tonglu County, Zhejiang province and ascospore suspensions were placed on potato dextrose agar （PDA） plate. One repre- sentative isolate ZJHZ01 was chosen for morphological and molecular study. Pathogenicity was verified by conidial suspension inoculation on the basal stems of soybean cuhivars. The colonial morphology was basically white, and the colonies tend to change into brown. Numerous yellowish-brown microsclerotia and chlamydospores were consistently formed on colonies. Orange-to-reddish perithecia had formed in old cultures of PDA, V8 and carnation leaf agar. Based on the morphology and pathogenicity analysis, the strain ZJHZ01 was primarily identified as Calonectria ilicicola Boedijin ＆ Reitsma. In addition, the molecular identification of representative strain ZJHZ01 was conducted, and the TUB, HIS and EF-1α gene were amplified and sequenced and compared with sequences deposited in GenBank. The NJ phylogenetic trees were constructed by comparison with strain ZJHZ01 and reference strains. Comparing with other known Calonectria sp. in GenBank, the three genes of ZJHZ01 shared high sequences homology with GDBL01 at a level of 99% similarity. Identification of the pathogen was based on morphology of colony and conidial, pathogenicity and phylogenetic analysis. This is the first report of red crown rot of soybean caused by C. ilicicola in Zhejiang Province, China.

关 键 词：大豆 红冠腐病 冬青丽赤壳 病原鉴定 

分 类 号：S435.651[农业科学—农业昆虫与害虫防治]