松墨天牛成虫标本保存及其DNA提取质量比较  

Comparison of the quality of genomic DNA extracted from adult specimens of Monochamus alternatus preserved by different methods

作　　者：曲良建[1] 王丽娟[2] 王青华[1] 王玉珠[1] 张永安[1] 

机构地区：[1]中国林业科学研究院森林生态环境与保护研究所,北京100091 [2]中国林业科学研究院林业研究所,北京100091

出　　处：《应用昆虫学报》2014年第3期741-748,共8页Chinese Journal of Applied Entomology

基　　金：国家自然科学基金项目(30972378);863课题(2012AAI01503)

摘　　要：【目的】探讨适合DNA提取的天牛成虫标本保存方法。【方法】采用SDS-蛋白酶K消化法对液氮中冷冻保存、无水乙醇-20℃冷冻保存、无水乙醇室温保存和干标本室温保存且保存时间在2年以上的松墨天牛Monochamus alternates Hope成虫标本基因组DNA进行提取,并对不同保存方式提取的DNA样本进行了质量比较和分析。【结果】在上述常见的松墨天牛成虫标本4种保存方式中,以液氮中冷冻保存效果最佳,其次为无水乙醇-20℃冷冻保存,插针干标本室温保藏效果最差。利用昆虫线粒体基因CO I和CO II的通用引物从上述DNA中均能够成功扩增出目的片段,测序结果证实扩增片段符合预期。【结论】液氮和无水乙醇-20℃冷冻保存适合松墨天牛成虫标本长期保存,且不影响后续的PCR扩增和测序。[Objectives] To find which method of preserving Monochamus alternatus specimens produced the best quality genomic DNA. [Methods] Genomic DNA from Monochamus alternatus adults that had been preserved using different methods （liquid nitrogen, 100% ethanol at -20℃, 100% ethanol at room temperature and museum specimens） for more than 2 years, was extracted using the SDS-proteinase K method and the quality of the extracted genomic DNA compared. [Results] The best quality genomic DNA was obtained from specimens that had been stored in liquid nitrogen, and the next best from those that had been preserved in 100% ethanol at -20℃. [Conclusion] The best quality DNA was extracted from specimens that had been frozen in liquid nitrogen or preserved in 100% ethanol at -20℃. The DNA samples obtained from such specimens are suitable for PCR amplification and sequencing.

关 键 词：松墨天牛 天牛成虫 DNA提取 保存方式 

分 类 号：S763.7[农业科学—森林保护学]