›› 2009, Vol. 52 ›› Issue (11): 1266-1272.

• 研究论文 • 上一篇    下一篇

基于rDNA ITS1和ITS2序列的褐飞虱、白背飞虱和灰飞虱的分子鉴定

刘玉娣, 林克剑, 韩兰芝, 侯茂林   

  • 出版日期:2009-12-18 发布日期:2009-11-20

Molecular identification of Nilaparvata lugens (Stål), Sogatella furcifera (Horvath) and Laodelphax striatellus (Fallén) (Homoptera: Delphacidae) based on rDNA ITS1 and ITS2 sequences

  • Online:2009-12-18 Published:2009-11-20

摘要: 本研究测定了褐飞虱Nilaparvata lugens、白背飞虱Sogatella furcifera和灰飞虱Laodelphax striatellus的rDNA ITS1和ITS2的序列, 以探讨这3种稻飞虱的分子鉴定方法。3种飞虱的ITS1和ITS2侧翼区(18S, 5.8S和28S)序列相对稳定, 但ITS1和ITS2序列在3种飞虱中变异较大。 ITS1在所分析的438个位点中可变位点达294个, ITS2在分析的403个位点中可变位点为177个。根据3种飞虱rDNA的ITS1和ITS2序列设计了特异性引物, 应用特异性引物对样品进行了PCR扩增, 分析发现3种飞虱ITS1区的特异性引物扩增效果不理想, 而ITS2区的特异性引物可以稳定地扩增出明显的目的DNA条带. 因此, 采用ITS2区的特异性引物可以对3种飞虱进行快速的分子鉴定。

关键词: 褐飞虱, 白背飞虱, 灰飞虱, rDNA, ITS1, ITS2, 特异引物, 分子鉴定

Abstract: The complete sequences of rDNA ITS1 and ITS2 were determined for the brown planthopper Nilaparvata lugens, the white-backed planthopper Sogatella furcifera and the small brown planthopper Laodelphax striatellus in order to explore the molecular identification method for them. The flanking regions of rDNA-ITS1 and ITS2 of the three planthoppers showed only limited variation, but the sequences of rDNA-ITS1 and ITS2 differed significantly. There are 294 variable sites in the 438 analyzed sites for the ITS1 region, and 177 variable sites in the 403 analyzed sites for the ITS2 region. Species-specific primers of N. lugens, S. furcifera, and L. striatellus were designed based on their rDNA-ITS1 and ITS2 sequences. The results of PCR amplification of rDNA-ITS1 in the three species indicated that the species-specific primers were not applicable. However, the species-specific primers based on the rDNA-ITS2 sequences proved to be useful diagnostic primers for the three planthoppers. It is so concluded that molecular identification of N. lugens, S. furcifera and L. striatellus using the species-specific primers from the rDNA-ITS2 region is feasible.

Key words: Nilaparvata lugens, Sogatella furcifera, Laodelphax striatellus, rDNA, ITS1, ITS2, diagnostic primers, molecular identification