金银花黑斑病病原菌番茄匍柄霉的分离与鉴定

林若竹; 淮稳霞; 杨金库; 赵文霞; 姚艳霞; 刘端冲

doi:10.12403/j.1001-1498.20230079

金银花黑斑病病原菌番茄匍柄霉的分离与鉴定

Isolation and Identification of Pathogenic Fungus Stemphylium lycopersici Causing the Black Spot Disease on Honeysuckle (Lonicera japonica Thunb.)

摘要

摘要:
目的分离并鉴定引起金银花黑斑病的病原菌，为该病害的有效防控提供理论依据。
方法采用常规组织分离法对发病金银花叶片进行分离，结合形态学、多基因序列分析（ITS、gpd和EF-1α序列）和致病性测定进行病原物种鉴定。
结果在形态学上，分离获得的菌株YP4在PDA培养基上的菌落呈灰白或黄白色棉毛状，与番茄匍柄霉的菌落特征相符；在基于ITS、 gpd和EF-1α序列联合构建的3种系统发育树中，供试菌株YP4均以较高支持率与番茄匍柄霉（Stemphylium lycopersici）聚在同一分支上；致病性接种实验结果表明，菌株YP4接种至金银花叶片上和番茄叶片上均可产生病斑，并可再分离获得其纯菌株；因此，结合形态学、分子系统发育学以及致病性测定，该病原真菌被鉴定为番茄匍柄霉。
结论本研究明确了引起河北省金银花黑斑病的病原菌为番茄匍柄霉，这是该病菌在我国危害金银花的首次报道。

Abstract:
Objective To clarify the pathogen causing black spot disease on Lonicera japonica, and to provide a theoretical basis for effective prevention and control of this disease.
Method The strains of the pathogen were isolated by conventional tissue plate method, and were identified by the morphological characters and phylogenetic analysis of ITS, gpd and EF-1α sequences. Its pathogenicity was tested through in vitro inoculating healthy leaves of L. japonica and Solanum lycopersicum.
Result The isolated fungal strain YP4 morphologically formed greyish to yellowish white, cottony colonies on Potato Dextrose Agar, which were consistent with published descriptions of Stemphylium lycopersici. In the three phylogenetic trees based on ITS, gpd and EF-1α sequences, the isolate YP4 fell into the S. lycopersici clade with strong support. Based on the pathogenicity measurement of Koch's rule, the strain YP4 was pathogenic to leaves of L. japonica and S. lycopersicum.
Conclusion Combined with the morphological characteristics, the pathogenicity determination and phylogenetic analysis of ITS, gpd and EF-1α sequences, the pathogen causing black spot disease on honeysuckle is identified as S. lycopersici. To our knowledge, this is the first report of S. lycopersici causing black spot of L. japonica in China.

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