西花蓟马ISSR-PCR多态性引物及退火温度筛选

Screening of ISSR-PCR Polymorphism Primers and Their Annealing Temperature in Frankliniella occidentalis

西花蓟马是近年来在我国局部地区爆发成灾的重要外来入侵害虫,为进一步研究西花蓟马的遗传多样性和种群遗传结构,对100条ISSR分子标记引物对西花蓟马基因组DNA的扩增情况进行了初步筛选,并利用4个地区的西花蓟马种群研究了其扩增多态性和最佳退火温度。结果表明:100条ISSR引物中有51条引物在西花蓟马基因组DNA中有扩增产物,其中,多态性引物12条,分别为UBC815、UBC834、UBC835、UBC840、UBC844、UBC845、UBC853、UBC864、UBC868、UBC874、UBC879和UBC899,其最佳退火温度依次为48℃、56℃、52℃、50℃、54℃、58℃、50℃、52℃、48℃、58℃、44℃和56℃。12条ISSR引物在4个地区的西花蓟马种群共检测到68个扩增位点,其中,多态位点31个,多态性比率(P)为45.59%;其遗传距离变化范围为0.132～0.338;在4个地区中,昆明和玉溪的西花蓟马亲缘关系最近。

F. occidentalis is one of the invasive insect pests, which currently occurs in several provinces in China and causes serious economic loss in invaded regions. In order to study on the genetic diversity and population genetic structure in further, 100 inter-simple sequence repeat （ISSR） primers were screened in F. occidentalis and polymorphism primers were selected and the annealing temperatures were tested using four different populations of F. occidentalis. The results showed that 51 ISSR primers produced bands with F. occidentalis genome DNA, including 12 polymorphism primers, which were UBC815, UBC 834, UBC 835, UBC 840, UBC 844, UBC 845, UBC 853, UBC 864, UBC 868, UBC 874, UBC 879 and UBC 899, and the annealing temperatures of these polymorphism primers tested on F. occidentaliswere 48℃, 56℃, 52℃, 50℃, 54℃, 58℃, 50℃, 52℃, 48℃, 58℃, 44℃ and 56℃, respectively. Twelve ISSR primers were used to amplify four populations of F. occidentalis, and 68 discernible DNA fragments were produced, of which, 31 were polymorphic loci, and the percentage of polymorphic loci （P） was 45.59%. Genetic distance of the four F. occidentalis populations ranged from 0. 132 to 0. 338, and the population collected in Kunming and Yuxi have the closest genetic relationship.