一种褐飞虱病原真菌的分子生物学鉴定

Molecular biological identification of entomogenous fungi isolated from the brown planthopper

从田间褐飞虱Nilaparvata lugens(Stl)罹病虫体分离得到一株绿僵菌,以该菌为供试菌体,采用氯化苄法、CTAB法以及裂解液法分别提取了该菌的基因组DNA;以ITS1和ITS4为绿僵菌通用引物,对供试菌株的rDNAITS序列进行PCR扩增、琼脂糖凝胶电泳检测和序列分析,并在核酸序列数据库中进行同源序列对比。实验结果表明裂解液法提取的该种绿僵菌的基因组DNA纯度高且质量好,氯化苄法次之,CTAB法不适合提取此菌的基因组DNA;分子鉴定结果显示该菌为绿僵菌属黄绿绿僵菌(Metarhizium flavoviride)。明确侵染褐飞虱的绿僵菌种类,进而掌握其生物学特征和对寄主的致病性,对于利用其对褐飞虱进行生物防治具有重要的意义。

Three different methods were used to extract genomic DNA from a strain of Metarhizium spp.isolated from the surface of infected Nilaparvata lugens（Stl） collected from rice fields;benzyl chloride,CTAB and cracking solution.The fungus＇ rDNA ITS sequence was amplified with the primers ITS1 and ITS4,and the amplified ITS sequence detected by agarose gel electrophoresis and analyzed in Genebank.The results show that the cracking solution method obtained the highest quantity of the purest DNA,the benzyl chloride method was the next best and the CTAB method was not suitable.Homologous sequence contrast conducted in the DNA sequence database identified the fungus as Metarhizium flavoviride.Identifying the entomogenous fungi that infect N.lugens is an important first step in determining their control potential with respect to this pest.