Mycosystema
菌 物 学 报 15 May 2008, 27(3): 325-334
jwxt@im.ac.cn
ISSN1672-6472 CN11-5180Q
2008 Institute of Microbiology, CAS, all rights reserved.
稗草叶枯病病原尖角突脐孢菌的鉴定
杨敏1,2 朱书生1,2 李健强1* 倪汉文1 张文华1
1中国农业大学农学与生物技术学院 北京 100094
2云南农业大学 教育部农业生物多样性与病害控制重点实验室 昆明 650201
摘 要:采用形态学及分子生物学的方法对采集自湖南和北京的3株尖角突脐孢菌分离物进行了鉴定。结果表明,3株分离物(KF-1、HN-14和K-12)和保存于中国农业大学已定名的尖角突脐孢菌菌株G-9和X-27之间在菌落形态、产孢量及孢子大小和分隔方面存在较大差异。其中,K-12在PDA培养基上生长缓慢、产孢量小;菌株G-9、KF-1、X-27和HN-14生长迅速,产孢丰富。对菌株进行分子鉴定结果表明,菌株间ITS区序列相似度达98%以上,聚类分析也表明,种内各菌株之间的遗传距离明显小于种间的遗传距离;基于ITS1及ITS2序列,能将尖角突脐孢菌和突脐蠕孢属中其它种分开。由此可确定分离自湖南水稻田及中国农业大学科学园温室中自然发病的稗草病样上的3株病原真菌均为尖角突脐孢菌。
关键词:形态学,分子鉴定,ITS区序列分析,杂草生物防治
The identification of barnyardgrass leaf spot caused by Exserohilum monoceras
YANG Min1, 2 ZHU Shu-Sheng1, 2 LI Jian-Qiang1* NI Han-Wen1 ZHANG Wen-Hua1
1College of Agriculture and Biotechnology, China Agricultural University, Beijing 100094, China
2The Ministry of Education Key Laboratory for Agricultural Biodiversity and Pest Management, Yunnan Agricultural University, Kunming 650201, China
Abstract: Three fungal isolates, KF-1, HN-14 and K-12, from barnyardgrass leaf blight, which were collected from Hunan and Beijing, were studied morphologically and molecularly. The results showed that there were differences between the three isolates and the two Exserohilum monoceras strains G-9 and X-27 which were accurately identified and deposited in culture collections of China Agricultural University in colonial morphology, sporulation and spore morphology especially septa of spore. G-9, KF-1, X-27 and HN-14 grow quickly and produce abundant spores on PDA plate, but K-12 grows slowly and produced less spores. To further identify the relationship of 5 isolates, 5.8S-ITS sequence were compared. Results indicated that the similarity of ITS sequence of five isolates were over 98% and the phylogenetic tree based on ITS1 and ITS2 sequence also revealed that G-9, KF-1, K-12, X-27, and HN-14 were all clustered into one group and distinct from the other outgroup and suboutgroup. Based on the above data, these three isolates were proved to be E. monoceras.
Key words: morphology, molecular identification, ITS sequence analysis, biocontrol of weed
稗草Echinochloa crusgalli (L.) Beauv. 是危害水稻生产最为严重的杂草之一。稗草与水稻的伴生性强,极难清除,同时也可发生于潮湿旱地,危害棉花、大豆等秋熟旱作物(Holm et al. 1977)。尖角突脐孢菌Exserohilum monoceras (Drechsler) Leonard & Suggs是近年来研究较为系统的一种对稗草致病性强对水稻安全的生防因子,其侵染稗草可在叶片上造成枯斑,严重时可引起整株死亡。作者在承担农业部农业结构调整重大技术研究专项课题中,采集了来自国内水稻不同生态区的稗草病样,进行了病原物的分离,并对分离获得的病原物与实验室已有的尖角突脐孢菌菌株G-9和X-27进行形态学的比较,进一步对分离物进行ITS分子鉴定,为后续的研究提供基础材料和条件。
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