大猿叶虫四地理种群的PCR-RFLP方法鉴别及遗传多样性分析

为了明确大猿叶虫Colaphellus bowringi Baly的遗传多样性水平及探索其种群快速鉴定的方法, 利用 7种限制性内切酶对其4个地理种群（江西龙南、江西修水、山东泰安、黑龙江哈尔滨）的线粒体COⅠ基因进行PCR-RFLP分析。结果表明: 利用AseⅠ, MboⅠ, NlaⅢ和RsaⅠ在大猿叶虫4个地理种群中均没检测到多态性; 利用AluⅠ和DraⅠ只检测到种群间的多态性; 利用HaeⅢ既检测到了种群间的多态性, 又在山东种群中检测到种群内多态性。根据酶切图谱, 共发现4种单倍型, 单倍型的特异性可以作为种群鉴别的标志。根据限制性片段共享度, 利用POPGEN 3.2计算4个种群的遗传距离, 并利用MEGA3.1进行聚类分析, 结果显示，大猿叶虫4个地理种群间的遗传距离大小与其相对地理距离的远近不相符, 其遗传变异程度与其滞育的特征也不相符。据此认为线粒体COⅠ基因的PCR-RFLP分析可以用于大猿叶虫不同地理种群的识别。

Identification and genetic diversity of four geographic populations of Colaphellus bowringi Baly (Coleoptera: Chrysomelidae) in China by PCRRFLP

To clarify the levels of genetic diversity and search population rapid identification method of the cabbage beetle Colaphellus bowringi Baly,seven restriction endonucleases were used to analyze the amplified products of mtDNA CO Ⅰ gene in four populations of Colaphellus bowringi,i.e.,Longnan population of Jiangxi,Xiushui population of Jiangxi,Tai'an population of Shandong and Harbin population of Heilongjiang in China.The results showed that Ase Ⅰ,Mbo Ⅰ,Nla Ⅲ and Rsa Ⅰ did not produce restriction fragment-length polymorphism among C.bowringi populations.The polymorphism was detected among the four populations with Alu Ⅰ and Dra Ⅰ.However,the restriction fragment-length polymorphism was found either among four populations or within Tai'an population of Shandong with Hae Ⅲ. Four haplotypes were found in all individuals.Obviously,the populations could be distinguished with unique haplotypes.According to the proportion of restriction fragments shared,the genetic distances among the four populations were calculated with POPGEN3.2,and a cladogram was produced with MEGA3.1.The results indicated that the genetic distance of C.bowringi among four geographic populations did not match with their geographic distance,neither their genetic variance with their diapause variance.We so conclude that PCR-RFLP analysis of mtDNA CO Ⅰ gene can be used to identify different geographic populations of C.bowringi.