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以采集自中国杭州和菲律宾的稻虱缨小蜂Anagrus nilaparvatae为研究对象,采用巢式PCR扩增Wolbachia的16S rDNA和wsp基因片段,并用DGGE分析稻虱缨小蜂体内Wolbachia的多样性。基于16S rDNA基因的分析结果准确地检测到稻虱缨小蜂体内细菌主要是Acinetobacter sp.,Methylophilus sp.,Acidovorax sp.,Burkholderia sp.和Wolbachia sp.。基于wsp基因的分析结果显示,杭州种群感染的Wolbachia属于A组的Mors亚组,菲律宾种群感染的Wolbachia属于A组的Dro亚组。结果说明,巢式PCR-DGGE是寄生蜂体内Wolbachia检测和多样性分析的有效方法,其中16S rDNA基因是检测Wolbachia存在的较佳分子标记,wsp基因是Wolbachia多样性分析以及种属鉴定和分型的较佳分子标记。
Anaerus nilaparvatae collected from Hangzhou, China and the Philippines were used to amplify the 16S rDNA and wsp gene fragments of Wolbachia by nested PCR. The diversity of Wolbachia in Wolbachia was analyzed by DGGE. Based on the analysis of the 16S rDNA gene, it was found that Acinetobacter sp., Methylophilus sp., Acidovorax sp., Burkholderia sp. And Wolbachia sp. Based on the analysis of the wsp gene, Wolbachia infected with Hangzhou population belonged to Mors subgroup A, and Wolbachia infected with Philippine population belonged to subgroup Dro. The results showed that nested PCR-DGGE is an effective method for Wolbachia detection and diversity analysis in parasitoid. 16S rDNA gene is a better molecular marker to detect the presence of Wolbachia, wsp gene is Wolbachia diversity analysis and species identification and typing Of the better molecular markers.