黑龙江省一株红小豆锈病病原菌鉴定

Identification of a fungal isolate causing adzuki bean rust in Heilongjiang

【目的】为明确发生在黑龙江省大庆市红小豆田中的锈病病原物的分类地位。【方法】从大庆市采集红小豆锈病标样,采用单孢子堆分离法获得一株红小豆锈病菌纯培养物ZXL01。采用观测夏孢子芽孔数目、位置和冬孢子壁厚度等形态学特征结合ITS序列分析的方法,对其进行鉴定。【结果】ZXL01夏孢子发芽孔多为2个,位于孢子赤道部位较远之处,冬孢子壁厚度为2.9μm-3.3μm。在基于r DNA-ITS序列构建的系统发育树中,ZXL01菌株与两株豇豆单胞锈菌(Uromyces vignae)的参比菌株(Gen Bank登录号:AB115718和AB115731)在自举值99%相聚一群。用豇豆单胞锈菌的特异性引物UV-ITSF/R进行检测,ZXL01菌株可扩增出500 bp左右的特征片段。【结论】黑龙江省大庆市红小豆锈病病原菌ZXL01菌株为豇豆单胞锈菌,ZXL01菌株的Gen Bank登录号是KM461700。

[Objective]The aim of the study is to identify the pathogen causing adzuki bean（ Phaseolus angularis） rust in Daqing,Heilongjiang province. [Methods]Adzuki bean rust leaves were collected from Daqing,Heilongjiang province,China. A pure culture of rust isolate ZXL01 was obtained by single pustule isolation. Its taxonomic status was determined by observing the number of germ pores of urediniospores,germ pore location and the wall thickness of teliopores,and sequencing ribosomal DNA internal transcribed spacer（ r DNA-ITS）. [Results]Morphological studies showed that most of the urediospores of ZXL01 had two germ pores that were far from spores＇ equator area. The wall thickness of teliopores ranged from 2. 9 to 3. 3 μm. The r DNA-ITS sequence of ZXL01 was clustered in one clade with 2 reference isolates of Uromyces vignae（ Gen Bank accession numbers AB115718 and AB115731） at 99% bootstrap levels in the phylogenetic tree. A 500 bp amplified product was obtained by the specific primers UV-ITSF / R,which was specific for U. Vignae.[Conclusion]The morphological features and ITS analysis indicated that the rust fungus ZXL01 occurred on leaves of adzuki bean in Daqing was U. Vignae,and the accession number of Gen Bank was KM461700.