中国舞毒蛾六个地理种群的RAPD分析及SCAR标记构建

Analysis of RAPD markers and development of SCAR markers of six geographic populations of the gypsy moth,Lymantria dispar L.(Lepidoptera:Lymantriidae),from China

舞毒蛾Lymantria disparL.是世界性农林害虫,包含不同的亚种,其中亚洲舞毒蛾的雌蛾具有较强的飞行能力,已成为国际性的重要检疫性有害生物。然而,不同舞毒蛾亚种及种群间形态难辨,因此采用传统的手段鉴别舞毒蛾亚种种群是很困难的。本研究首先采用RAPD标记分析了中国舞毒蛾6个地理种群的遗传多态性。结果表明,所检测的舞毒蛾种群的遗传分化系数Gst为0.7571,由此推算出的平均有效迁移数(基因流参数)Nem为0.1604,说明不同舞毒蛾种群间的遗传分化程度较高,缺乏广泛的基因流动。本研究在RAPD遗传分析基础之上,筛选出了4个舞毒蛾种群的特异性遗传位点,然后对这些特异性位点进行了克隆测序、序列分析和位点特异性引物设计。结果表明,其中2个舞毒蛾种群的位点特异性引物可产生序列特征性扩增区域(SCAR)标记。经验证,这些标记可被用来鉴别特定的舞毒蛾地理种群,因此有助于对这些舞毒蛾地理种群的分布与扩散进行监测。

The gypsy moth,Lymantria dispar L.（Lepidoptera：Lymantriidae）,is one of the most important agricultural and forest insect pests.It is composed of several subspecies,among which the Asian gypsy moth female adults with stronger flight ability have become serious international quarantine pests.Unfortunately,it can be very difficult to distinguish these subspecies morphologically.Herein,the random amplified polymorphic DNA （RAPD） markers were used to analyze the genetic polymorphism of six gypsy moth populations collected from different provinces in China.The results showed that the genetic differentiation coefficient （Gst） among the gypsy moth populations was 0.7571 and the deduced effective number of migrants （Nem,a gene flow index） was 0.1604,suggesting that these populations are highly differentiated and the gene flow between them is very low.Based on genetic analyses,four population-specific loci were located,followed by cloning and sequencing,sequence analyses and locus-specific primer designing.Finally,the sequence characterized amplified region （SCAR） markers for two gypsy moth populations were developed.The results of primer testing showed that these SCAR markers could be used for rapid and accurate identification of the two gypsy moth populations,which is helpful for monitoring the distribution and dispersal of gypsy moth populations.