梨小食心虫微卫星标记的扩增稳定性及多态性分析

Amplification stability and polymorphism of microsatellite loci in the oriental fruit moth, Grapholita molesta (Lepidoptera: Tortricidae) in China

【目的】筛选适合我国梨小食心虫Grapholita molesta种群遗传学研究的微卫星位点,并依据所筛选的微卫星位点进行梨小食心虫地理种群的遗传多样性分析。【方法】利用欧洲梨小食心虫和苹果蠹蛾Cydia pomonella种群中已报道的11个微卫星位点,分析各位点在我国12个种群257头梨小食心虫样本中的扩增稳定性,再进行其多态性分析,筛选适合的位点,然后进行种群遗传多态性分析。【结果】在分析的11个微卫星位点中,位点Gm01,Gm03,Gm04和Cyd15无法稳定扩增;位点Gm05扩增成功率较低,位点Gm07遗传多态性较低;而位点Gm02,Gm06,Gm08,Gm09和Gm10等扩增效果稳定且遗传多态性丰富。这5个稳定扩增的微卫星位点平均等位基因数量（NA）为7.417~12.500,平均观察杂合度（Ho）为0.366~0.655,平均期望杂合度（He）为0.642~0.846,多态信息含量（PIC）为0.800~0.935。【结论】本研究成功筛选出位点Gm02,Gm06,Gm08,Gm09和Gm10等5个微卫星位点。基于这5个微卫星位点标记的结果显示,山东和陕西不同梨小食心虫地理种群均具有丰富的遗传多样性。这5个位点可以适用于我国梨小食心虫种群的进一步遗传分析研究。

Abstract： [ Aim] The objective of this research was to screen microsatellite loci which were available for population genetic research of the oriental fruit moth, Grapholita molesta in China. [ Methods ] The amplification stability and genetic polymorphism of 11 microsatellite loci previously reported in European populations of G. molesta and Cydia pomonella were characterized from 257 individuals from 12 geographic populations of China. The screened polymorphic microsatellite loci were then used for population genetic analysis of the G. molesta populations. [ Results] The results showed that loci Gin01, Gin03, Gin04 and Cydl5 could not be amplified from the samples. Gin05 could be amplified from a low proportion of individuals. Gin07 could be stably amplified, but was weakly polymorphic. Loci Gin02, Gm06, Gin08, Gm09 and Gin10 showed high polymorphism and could be amplified stably. For the five polymorphic loci, the mean number of alleles （ NA ） ranged from 7. 417 to 12. 500, the values of mean observed heterozygosity （Ho） and expected heterozygosity （He） were from 0. 366 to 0. 655 and from O. 642 to 0.846, respectively, and the polymorpbic information content （PIC） was between 0.800 and 0. 935. [Conclusion] We successfully selected five loci, i. e., Gin02, Gin06, Gm08, Gm09 and Gm10. High genetic diversity of G. molesta populations sampled in Shandong Province and Shaanxi Province was revealed by data from these five loci. These five loci can be used for further population genetics research of G. molesta populations from China.