基于锚定PCR技术对10种重要农业害虫微卫星DNA位点的筛选及其特征分析

摘要

Background Microsatellite DNA consists of both prokatryotic and eukaryotic genomes .The characteristics include rich polymorphism and easy detection .The technique has been applied in animal and plant genetics and breeding , for the construction for genetic maps .[Method]Using the 5′anchored PCR method , microsatellite DNA from 10 important agricultural pests were isola-ted and analyzed , including Carposina sasakii ( Lepidoptera: Carposinidae ) , Conogethes punctiferalis ( Lepidoptera: Pyralidae ) , Ostrinia nubilalis ( Lepidoptera: Pyralidae ) , Proxenus lepigone ( Lepidoptera: Noctuidae ) , Frankliniella intonsa ( Thysanoptera:Thripidae), Thrips hamaiiensis (Thysanoptera:Thripidae), Thrips palmi (Thysanoptera:Thripidae), Drosophila suzukii (Diptera:Drosophilidae ) , Lissorhoptrus oryzophilus ( Coleoptera:Curculionidae ) and Phenacoccus solenopsis ( Hemiptera: Pseudococcidae ) . The characteristics of microsatellite DNA in these insects were analyzed .[Result]The positive clone rates , microsatellite rates , clo-ning efficiency and redundancy rates of microsatellite DNA among the species were variable .Dinucleotide repeats were the most a-bundant repeat types (93.2%~100%), while trinucleotide and tetranucleotide were rare .Among the dinucleotide repeats , AC/GT is the most abundant type (89.2%~100%).This is closely related to the anchor primer;The average repeat number within these species was 6.7~8.9 and the highest repeat number (34) was detected in O.nubilalis;The types of microsatellite mainly consist of the perfect type ( 91 .0%~100%) .[Conclusion and significance]The anchored method can be used to reveal the microsatellite DNA in the agricultural pests , and can be used in monitoring and detection .%微卫星DNA广泛存在于真核和原核生物的基因组中，具有多态性丰富、易于检测等特点，在遗传图谱的构建、动植物遗传学育种等方面被广泛应用。利用5′锚定PCR技术对桃小食心虫、桃蛀螟、玉米螟、二点委夜蛾、花蓟马、黄胸蓟马、棕榈蓟马、斑翅果蝇、稻水象甲、扶桑绵粉蚧10种重要农业害虫进行微卫星DNA筛选，并分析各个物种的微卫星DNA的特点。不同物种的阳性克隆率、微卫星比率、克隆效率和冗余率存在不同程度的差异；在核苷酸碱基数目上，主要为二核苷酸重复序列，占重复位点总数的93．2％～100％，三、四核苷酸重复位点较少。在二核苷酸重复位点中， CA/TG重复位点最为丰富，占重复位点总数的89．2％～100％。这与使用的锚定引物密切相关；10种害虫的微卫星DNA平均重复次数为6．7～8．9次，其中玉米螟具有最高的微卫星DNA重复次数（34次）；在序列类型上，完全型序列占上述害虫序列总数的91．0％～100％。5′锚定PCR技术能够快速挖掘害虫的微卫星DNA位点，本研究结果为这些害虫微卫星位点的进一步利用奠定了基础。