[1]盖云鹏,黄子凌,陈好好,等.浙江省大豆红冠腐病的病原菌鉴定[J].大豆科学,2016,35(06):986-991.[doi:10.11861/j.issn.1000-9841.2016.06.0986]
　GAI Yun-peng,HUANG Zi-lin,CHEN Hao-hao,et al.Identification of Red Crown Rot of Soybean Caused by Calonectria ilicicola in Zhejiang Province, China[J].Soybean Science,2016,35(06):986-991.[doi:10.11861/j.issn.1000-9841.2016.06.0986]
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浙江省大豆红冠腐病的病原菌鉴定

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第35卷 期数: 2016年06期 页码: 986-991 栏目: 出版日期: 2016-11-20
Title:
Identification of Red Crown Rot of Soybean Caused by Calonectria ilicicola in Zhejiang Province, China
作者:
盖云鹏; 黄子凌; 陈好好; 周乔俐; 马海杰; 李红叶
（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）
Author(s):
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）
关键词:
大豆; 红冠腐病; 冬青丽赤壳; 病原鉴定
Keywords:
Soybean;  Red crown rot;  Calonectria ilicicola;  Pathogen identification
DOI:
10.11861/j.issn.1000-9841.2016.06.0986
文献标志码:
A
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GenBank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GenBank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。
Abstract:
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 representative isolate ZJHZ01 was chosen for morphological and molecular study.Pathogenicity was verified by conidial suspension inoculation on the basal stems of soybean cultivars. 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. Orangeto-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 EF1α 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.

参考文献/References:

［1］朱丹华. 2010年浙江省大豆生产调研报告［J］.大豆科技, 2010(6):25-26. (Zhu D H.Research report on soybean production in Zhejiang province in 2010［J］. Soybean Science and Technology,2010(6):25-26.)

［2］刘欣, 姚晗珺, 章强华, 等. 浙江省出口菜用大豆使用农药现状及风险分析［J］.大豆科学, 2011, 30(2):298-302 (Liu, X, Yao H J, Zhang Q H, et al. Status of pesticide and risk analysis of exporting vegetable soybeans in Zhejiang province［J］. Soybean Science,2011,30(2):298-302.)
［3］葛起新, 戎文治. 浙江植物病虫志病害篇［M］. 上海:上海科学技术出版社, 1991. (Ge Q X, Rong W Z. Diseases of plant diseases and insect pests in Zhejiang［M］. Shanghai: Shanghai Scientific and Technical Publishers, 1991)
［4］盖云鹏, 潘汝谦, 关铭芳, 等.一种值得关注的大豆病害—大豆红冠腐病［J］. 植物保护, 2014, (4):118-121. (Gai Y P, Pan R Q, Guan M F, et al. A notable soybean disease of red crown rot of soybean［J］. Plant Protection, 2014（4）:118-121.)
［5］Kuruppu P U, Schneider R W, Russin J S. Effects of soil temperature on microsclerotia of Calonectria ilicicola and soybean root colonization by this fungus［J］. Plant Disease, 2004, 88 (6): 620-624.
［6］潘汝谦, 关铭芳, 徐大高, 等. 花生黑腐病菌的生物学特性［J］. 华中农业大学学报, 2012, 30 (6):701-706 (Pan R Q, Guan M F, Xu D G, et al. Biological characteristics of Cylindrocladium parasiticum［J］. Journal of Huazhong Agricultural University, 2012, 30 (6): 701-706)
［7］Fisher N L, Burgess L W, Toussoun T A, et al. Carnation leaves as a substrate and for preserving cultures of Fusarium species［J］. Phytopathology, 1982, 8(2):1041–1049.
［8］刘少华, 陆金萍, 朱瑞良,等.一种快速简便的植物病原真菌基因组DNA提取方法［J］. 植物病理学报, 2005, 35(4):362-365. (Liu S H, Lu J P, Zhu R L, et al. A rapid and simple extraction method for plant pathogenic fungi［J］. Acta Phytopathologica Sinica, 2005, 35(4):362-365.)
［9］Crous P W, Groenewald J Z, Risède J M, et al.Calonectria species and their Cylindrocladium anamorphs: Species with sphaeropedunculate vesicles［J］. Studies in Mycology, 2004, 18:415-430.
[10］Tamura K, Dudley J, Nei M, et al. MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4-0［J］. Molecular Biology and Evolution, 2007, 24: 1596-1599
［11］Crous P W, Wingfield M J, Alfenas A C.Cylindrocladium parasiticum, sp- nov a new name for C.crotalariae［J］. Mycological Research, 1993, 97 (7):889-896.
［12］Pan R Q, Guan M F, Xu D G, et al. Cylindrocladium black rot caused by Cylindrocladium parasiticum, newly reported on peanut in China［J］. Plant Pathology, 2009, 58(6):1176-1176
［13］盖云鹏, 潘汝谦, 徐大高,等. 进境检疫性有害生物—寄生帚梗柱孢霉［J］. 植物检疫, 2014, 28 (4):76-81.(Gai Y P, Pan R Q, Xu D G, et al. Quarantine plant pathogen-Cylindrocladium parasiticum［J］. Plant Quarantine,2014, 28 (4):76-81.)
［14］Hawksworth D L, Crous P W, Redhead S A, et al. The amsterdam declaration on fungal nomenclature［J］. Mycotaxon, 2011, 116(1):105-112.
［15］Greer A M, Spurlock T N, Coker C M. First report of Neocosmospora stem rot of soybean caused by Neocosmospora vasinfecta in Arkansas［J］. Plant Disease, 2015, 99 (4):554-554.
［16］Sun S L, Kim M Y, Van K, et al. First report of Neocosmospora vasinfecta var vasinfecta causing soybean stem rot in South Korea［J］. Plant Disease, 2014, 98 (12):1744-1744.
［17］Ma Z H, Zhang Z D, Wang Y X, et al.Cylindrocladium crotalariae, causing red crown rot of soybean in China［J］. Plant Pathology, 2004, 53(4):537-537.
［18］Guan M F, Pan R Q , Gao X G, et al. First report of red crown rot caused by Cylindrocladium parasiticum on soybean in Guangdong, southern China［J］. Plant Disease, 2010, 94(4):485-485.
［19］Kuruppu P, Schneider R, Russin J. Factors affecting soybean root colonization by Calonectria ilicicola and development of red crown rot following delayed planting［J］. Plant disease, 2004, 88 (6):613-619.
［20］Sidebottom J R, Beute M K. Inducing soil suppression to Cylindrocladium black rot of peanut through crop rotations with soybean［J］. Plant Disease, 1989, 73 (8): 679-685.

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备注/Memo

基金项目：浙江大学大学生科研训练计划（2015032199）。

第一作者简介：盖云鹏（1986-），男，博士，主要从事植物病原真菌学与生物信息学研究。E-mail：gaiy@zju.edu.cn。
通讯作者：李红叶（1963-），女，博士，教授，主要从事植物病原真菌学与分子植物病理学研究工作。E-mail：hyli@zju.edu.cn。

更新日期/Last Update: 2016-12-09