两种书虱微卫星富集文库的构建及比较  

Construction and comparative analysis of enriched microsatellite library from Liposcelis bostrychophila and L. entomophila genome

作　　者：魏丹丹[1] 袁明龙[1] 王保军[1] 朱礼明[1] 王进军[1] 

机构地区：[1]西南大学植物保护学院,昆虫学及害虫控制工程重庆市市级重点实验室,重庆400716

出　　处：《生态学报》2011年第15期4182-4189,共8页Acta Ecologica Sinica

基　　金：国家自然科学基金资助项目(30871631,31000860);国家教育部博士点基金资助项目(200806350009);国家教育部新世纪优秀人才支持计划资助项目(NCET-04-0584)

摘　　要：利用链霉亲和素与生物素之间的强亲和性原理,将链霉亲和素偶联的磁珠与微卫星探针(AC)12、(TC)12、(ATC)8、(ATG)8、(AAC)8、(ATAC)6及(GATA)6退火结合后,再亲和捕捉含接头和微卫星序列的单链书虱基因组DNA限制性酶切目的片段,经PCR扩增形成双链后进行克隆、建库。成功构建了嗜卷书虱和嗜虫书虱共13个微卫星富集文库,包括6个嗜卷书虱文库,7个嗜虫书虱文库,其平均阳性克隆率为71.17%。经检测发现共得到两种书虱260个微卫星位点。这两种书虱微卫星富集文库的建立和高多态性微卫星位点的筛选将为嗜卷书虱和嗜虫书虱的种群遗传与进化、基因连锁图谱构建、分子系统发育研究等提供大量分子遗传标记,对其在实仓中的持续控制提供遗传学信息。The psocids Liposcelis bostrychophila and Liposcelis entomophila are major pests of stored grain and commonly occur on a wide range of stored products.Their world-wide prevalence and increasing importance as pests contaminating food and agricultural commodities have been documented in recent years.Intensive use of chemical insecticides for pest control has facilitated resistance development in psocids,especially,resistance to PH3.Control of these pests has proven difficult because they do not respond to management tactics that have been developed for other stored-product pests.Previous research has focused on psocids physiology,biochemistry,and basic biology in grain storage systems.However,the population genetic structure has not been well categorized,which may be useful to understand the formation of resistance mechanisms of psocids to insecticides.Therefore,a detailed understanding of L.bostrychophila and L.entomophila gene flow patterns,determining the geographical origins and dispersal of source populations and the resultant genetic structure among populations within China is needed.Microsatellites or simple sequence repeats（SSRs） are tandemly repeated motifs of 1-6 genetic base pairs.Because of their ubiquity,neutrality,hyper-variability,co-dominance,sensitivity and high polymorphism,microsatellite markers have become a powerful tool for revealing genetic variation and subsequently the population structure in recent decades.Up to now,only 11 microsatellite loci have been isolated from Liposcelis decolor（Pearman）. In the present paper,microsatellite-enriched libraries of L.bostrychophila and L.entomophila were constructed utilizing methodologies that exploit the strong affinity between biotin（vitamin B7） and the protein streptavidin.We propose a fast and easy protocol which is combination of two different published methods.Briefly,high-quality genomic DNA was digested by the restriction enzyme MseI and then ligated to designed adaptors.250-1000 bp microsatellite-containing DNA fragments were capt

关 键 词：嗜卷书虱 嗜虫书虱 啮虫目 微卫星 富集文库 

分 类 号：Q953[生物学—动物学] S379.5[农业科学—农产品加工]