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从田间褐飞虱Nilaparvata lugens(Stl)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。
A strain of Metarhizium anisopliae was isolated from the pest of Nilaparvata lugens (Stl) in the field. The strain was used as the test strain, and the genomic DNA of the strain was extracted by benzyl chloride method, CTAB method and lysate method ITS1 and ITS4 were the common primers of Metarhizium anisopliae. The rDNAITS sequences of the tested strains were amplified by PCR, detected by agarose gel electrophoresis and sequenced, and the homologous sequences were compared in the nucleotide sequence database. The experimental results showed that the genomic DNA of Metarhizium anisopliae extracted by lysis method was of high purity and good quality, followed by benzyl chloride method, and CTAB method was not suitable for extracting genomic DNA of this strain. Molecular identification showed that the strain was Metarhizium anisopliae Metarhizium flavoviride. It is of great importance to utilize the biocontrol agent for the brown planthopper to identify the species of Metarhizium anisopliae that infect the BPH, and then understand its biological characteristics and host pathogenicity.