蚕豆赤斑病病原菌分离鉴定及生物学特性研究

Isolation,Identification and Biological Characteristics of Broad Bean Chocolate Spot

采用组织分离法进行病原物的分离、柯赫氏法则进行致病性测定,并利用真菌通用引物ITS1/ITS4和蚕豆葡萄孢特异性引物G3PDHfor/G3PDHrev进行基因组DNA扩增与序列分析,对甘肃省渭源县蚕豆赤斑病的致病菌种类进行研究。结果表明:该病原菌为蚕豆葡萄孢(Botrytis fabae),系统发育树分析表明该病原菌与GenBank已报道的蚕豆葡萄孢属聚为一类。生物学特性测定表明,该病原菌菌丝生长最适培养基为PDA培养基,最适碳源和氮源分别为乳糖和酵母膏,最适温度为20℃,最适pH为8,对光照不敏感。

The tissue isolation was used to isolate pathogen from diseased tissues,Koch’s rule was used to test pathogenicity,and fungal universal primers ITS1/ITS4 and Botrytis fabae specific primers G3PDHfor/G3PDHrev primers were used to amplify genomic DNA and to analyz the sequence,the classification of the pathogenic bacteria in broad bean chocolate spot occurring in Weiyuan county was studied.The results showed that B.fabae was the causal pathogen of broad bean chocolate spot.Phylogenetic tree analysis indicated that sequences of the pathogen and B.fabae submitted on GenBank were clustered into the same clade.Biological tests showed that optimum medium for mycelial growth was PDA medium.Optimum carbon and nitrogen sources were lactose and yeast extract.Optimum temperature was 20℃,and optimum pH was 8.Mycelial growth of the pathogen was insensitive to light.