米尔顿姬小蜂rDNA ITS1和ITS2的序列分析及其分子鉴定
Molecular identification of Anselmella miltoni Girault through analysis of rDNA ITS1 and ITS2 sequences
黄蓬英1**，廖富荣1，，林玲玲1，洪钦阳2，李雯琳2，林石明1
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作者单位：1厦门出入境检验检疫局厦门361026； 2 福建农林大学植物保护学院福州350002
中文关键词：米尔顿姬小蜂, rDNA, ITS1, ITS2, 特异引物, 分子鉴定
英文关键词：Anselmella miltoni, rDNA, ITS1, ITS2, diagnostic primers, molecular identification
中文摘要： 本研究测定了米尔顿姬小蜂Anselmella miltoni Girault的rDNA ITS1和ITS2序列，以探讨其分子鉴定方法。米尔顿姬小蜂的ITS1和ITS2侧翼区（18S 和58S）序列相对稳定，ITS1和ITS2序列存在种间差异。根据18S rDNA部分序列，利用DNAMAN的Maximum Likelihood方法构建了与膜翅目其它科的系统发育树。根据米尔顿姬小蜂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，扩增效果理想，采用上述特异性引物可从单头米尔顿姬小蜂稳定地扩增出明显的目的DNA条带。因此，可以采用ITS1和ITS2区的特异性对米尔顿姬小蜂进行快速的分子鉴定。
英文摘要：AbstractIn order to explore the potential for the molecular identification of Anselmella miltoni Girault complete sequences of its rDNA ITS1 and ITS2 were determined. Although the flanking regions of rDNA ITS1 and ITS2 showed only limited variation, the complete sequences differed significantly in different species. A phylogenetic tree was inferred from 18S rDNA part sequences using DNAMAN Maximum Likelihood. Speciesspecific primers of A. miltoni were designed based on the rDNA ITS1 and ITS2 sequences. The results of PCR amplification of rDNA ITS1 and ITS2 indicate that the speciesspecific primers were applicable. The results show that specific primers could produce clear, unique and reproducible target DNA bands from individual adult wasps. Molecular identification of A. miltoni using speciesspecific primers from rDNA ITS1 and ITS2 region is therefore feasible.