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为研究苹果小吉丁虫Agrilus mali Matsumura的种群遗传结构,采用磁珠富集法以生物素标记探针(AC)_(12)和(AG)_(12)构建苹果小吉丁虫微卫星文库,根据阳性克隆测序获得的微卫星侧翼序列设计引物,筛选和开发苹果小吉丁虫多态性微卫星标记,并利用开发的微卫星标记对其不同地理种群的遗传结构进行分析。结果显示,从微卫星文库中的300个阳性克隆中筛选得到248个含有微卫星片段的克隆子,阳性克隆率为82.7%,其中完美型微卫星131个,占52.8%;不完美型90个,占36.3%;混合型27个,占10.9%,表明磁珠富集法开发新微卫星标记的效率较高。从获得的248个苹果小吉丁虫微卫星中筛选获得7个多态性微卫星位点,这7个位点在苹果小吉丁虫8个地理种群样本中的等位基因数为10~23个,有效等位基因数为1.674~12.218个,多态信息含量为0.304~0.796。8个苹果小吉丁虫种群的观测杂合度为0.095~0.676,期望杂合度为0.469~0.755,Shannon指数为0.791~1.621,遗传距离为0.071~0.788。采自新疆维吾尔自治区的7个苹果小吉丁虫种群之间遗传相似度较高,但其与辽宁省种群的遗传相似度较低。
In order to study the population genetic structure of Agrilus mali Matsumura, the microsatellite DNA library of Agrimonia sibirica was constructed by magnetic bead enrichment method with biotinylated probe (AC) _ (12) and (AG) _ (12) Primers were designed based on the microsatellite flanking sequences obtained from positive cloning and sequencing, and the microsatellite markers of apple jilin were screened and developed. The genetic structure of different geographical populations was analyzed by using the developed microsatellite markers. The results showed that 248 clones containing microsatellite fragments were screened out from 300 positive clones in microsatellite libraries. The positive clonality rate was 82.7%, of which 131 were perfect microsatellites (52.8%), and 90 , Accounting for 36.3%; 27 hybrids, accounting for 10.9%, indicating that the magnetic bead enrichment method is more efficient in developing new microsatellite markers. Seven microsatellite loci were obtained from 248 Apis cerana cepacia microsatellites. The number of alleles in these 7 loci was between 10 and 23 , The effective number of alleles was 1.674 ~ 12.218, and the polymorphic information content was 0.304 ~ 0.796. The observed heterozygosity of Populus euchinensis population was 0.095 ~ 0.676, the expected heterozygosity was 0.469 ~ 0.755, Shannon index was 0.791 ~ 1.621, the genetic distance is 0.071 ~ 0.788. The genetic similarity among seven apple jacinthi populations collected from Xinjiang Uygur Autonomous Region was high, but their genetic similarity with Liaoning Province population was low.