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【背景】韭菜迟眼蕈蚊是我国重要的农业害虫,然而它的遗传资源有限。本研究旨在开发韭菜迟眼蕈蚊EST-SSR标记,为研究不同地区的韭菜迟眼蕈蚊种群结构和遗传多样性奠定基础。【方法】从韭菜迟眼蕈蚊的表达序列标签(EST序列)中设计16对简单重复序列(SSR)引物,进一步筛选出9对具有多态性的SSR引物。【结果】从42095条unigene中确定了3383个SSR位点。利用查找到的SSR位点共设计出16对引物,进一步检测筛选发现9对引物具有多态性,引物的每个位点平均有3.33个等位基因。利用9对引物对30头韭菜迟眼蕈蚊进行检测,共获得30个等位基因,观测杂合度和期望杂合度的范围分别为0.0000~0.6875和0.0370~0.6877;其中,9个位点中有5个位点显著偏离Hardy-Weinberg平衡。【结论与意义】本研究成功从迟眼蕈蚊EST序列中筛选出9个具有多态性的微卫星位点,这为进一步分析该害虫种群的遗传结构和遗传多样性奠定了基础。
【Background】 Leek Mycelia are important agricultural pests in our country, however, their genetic resources are limited. The aim of this study was to develop the EST-SSR marker of Lepidoptera brasiliensis, which laid the foundation for the study on the population structure and genetic diversity of Leishmania axyridis in different regions. 【Method】 Sixteen pairs of simple repeat (SSR) primers were designed from the expression sequence tags (EST sequences) of M. persicae and 9 pairs of SSR primers were further screened. 【Result】 3383 SSR loci were identified from 42095 unigene. A total of 16 pairs of primers were designed based on the SSR loci found, and 9 pairs of primers were further detected by screening. The average number of alleles per primer site was 3.33. Thirteen pairs of primers were used to detect 30 M. leungi, and 30 alleles were obtained. The range of observed heterozygosity and expected heterozygosity ranged from 0.0000 to 0.6875 and 0.0370 to 0.6877, respectively. Of the 9 loci Five sites deviated significantly from Hardy-Weinberg equilibrium. 【Conclusion and Significance】 Nine microsatellite loci were successfully screened from EST sequences of M. persicae, which laid the foundation for further analysis of the genetic structure and genetic diversity of the population.