Mycosystema
菌 物 學(xué) 報   15 May 2008, 27(3): 325-334
[email protected]
ISSN1672-6472  CN11-5180Q
2008 Institute of Microbiology, CAS, all rights reserved.

稗草葉枯病病原尖角突臍孢菌的鑒定

楊敏1,2  朱書生1,2   李健強(qiáng)1*  倪漢文1   張文華1

1中國農(nóng)業(yè)大學(xué)農(nóng)學(xué)與生物技術(shù)學(xué)院 北京 100094

2云南農(nóng)業(yè)大學(xué) 教育部農(nóng)業(yè)生物多樣性與病害控制重點實驗室 昆明 650201

摘  要：采用形態(tài)學(xué)及分子生物學(xué)的方法對采集自湖南和北京的3株尖角突臍孢菌分離物進(jìn)行了鑒定,。結(jié)果表明,，3株分離物（KF-1,、HN-14和K-12）和保存于中國農(nóng)業(yè)大學(xué)已定名的尖角突臍孢菌菌株G-9和X-27之間在菌落形態(tài)、產(chǎn)孢量及孢子大小和分隔方面存在較大差異,。其中,，K-12在PDA培養(yǎng)基上生長緩慢、產(chǎn)孢量??；菌株G-9、KF-1,、X-27和HN-14生長迅速,，產(chǎn)孢豐富。對菌株進(jìn)行分子鑒定結(jié)果表明,，菌株間ITS區(qū)序列相似度達(dá)98%以上，聚類分析也表明,，種內(nèi)各菌株之間的遺傳距離明顯小于種間的遺傳距離,；基于ITS1及ITS2序列,，能將尖角突臍孢菌和突臍蠕孢屬中其它種分開。由此可確定分離自湖南水稻田及中國農(nóng)業(yè)大學(xué)科學(xué)園溫室中自然發(fā)病的稗草病樣上的3株病原真菌均為尖角突臍孢菌,。

關(guān)鍵詞：形態(tài)學(xué),，分子鑒定，ITS區(qū)序列分析,，雜草生物防治

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. 是危害水稻生產(chǎn)最為嚴(yán)重的雜草之一,。稗草與水稻的伴生性強(qiáng)，極難清除,，同時也可發(fā)生于潮濕旱地,，危害棉花、大豆等秋熟旱作物（Holm et al. 1977）,。尖角突臍孢菌Exserohilum monoceras (Drechsler) Leonard & Suggs是近年來研究較為系統(tǒng)的一種對稗草致病性強(qiáng)對水稻安全的生防因子,，其侵染稗草可在葉片上造成枯斑，嚴(yán)重時可引起整株死亡,。作者在承擔(dān)農(nóng)業(yè)部農(nóng)業(yè)結(jié)構(gòu)調(diào)整重大技術(shù)研究專項課題中,，采集了來自國內(nèi)水稻不同生態(tài)區(qū)的稗草病樣，進(jìn)行了病原物的分離,，并對分離獲得的病原物與實驗室已有的尖角突臍孢菌菌株G-9和X-27進(jìn)行形態(tài)學(xué)的比較,，進(jìn)一步對分離物進(jìn)行ITS分子鑒定，為后續(xù)的研究提供基礎(chǔ)材料和條件,。

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