陕西省主要保存家蚕品种亲缘关系的AFLP分析
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摘要
家蚕（Bombyx mori）作为鳞翅目类的模式生物和重要的经济昆虫，其起源进化问题一直受到关注，这也是家蚕遗传育种的理论基础。家蚕存在多种生态类型和系统类群，如中国系统、日本系统、热带系统和欧洲系统等。家蚕品种资源间的亲缘关系和遗传距离，对家蚕起源进化研究，种质资源鉴定、保存和利用，家蚕实用品种的亲本选配等，都有重要的理论和现实意义。
为了更好地开发西部蚕区家蚕品种资源，以及在家蚕新品种选育中的利用提供分子水平的理论依据。本研究选择陕西省保存的具有代表性的53份家蚕品种资源，利用AFLP分子标记技术分析其亲缘关系和遗传距离。
1适于家蚕滞育期蚕卵DNA提取方法的建立
DNA提取是家蚕分子遗传学研究的基本技术，也是品种鉴定和纯度检测研究中较为常见的问题。本实验中使用3种DNA抽提方法进行对比，建立适合滞育期蚕卵的基因组DNA提取方法，达到快速高效满足分子实验要求的目标。
2家蚕遗传多样性分析
通过筛选出的14对引物组合对53个品种基因组DNA样品的AFLP扩增共获得535条带，其中多态性条带数472条，每对引物扩增的多态性比率为88.22%，显示出不同家蚕品种间存在丰富的遗传多样性。
3家蚕遗传相似性分析与聚类分析
使用NTSYS-pc2.1软件计算各家蚕品种间遗传距离。53个家蚕品种间遗传距离在0.0728~0.6019间。
基于UPGMA法及NJ法聚类分析得到53个家蚕品种聚类分析图，以及各个系统的聚类分析图。中系品种单独聚为一类，欧系品种和日系品种聚为一大类。其中基于UPGMA法聚类一化性三眠的西北地方品种明显区别于其它蚕品种聚为不同的类群。
Silkworm(Bombyx mori) is an important model and economic Lepidoptera insert, hasalways been concerned about the origin and evolution. It is the theoretical basis of thesilkworm genetics and breeding. Silkworm have a variety of ecological types and systemsgroups, such as the Chinese system, Japanese system, Tropical system and European system.Phylogenetic relationships among silkworm breeds and genetic distance of silkworm origin,evolution, germplasm identification, preservation and use of varieties of silkworm parentselection practice, has important theoretical significance and practical. The genetic diversityof53silkworm (Bombyx mori.) varieties conserved in Shaanxi province was analyzed byAFLP（amplified fragment length polymorphism）method in order to utilize germplasmresources in silkworm breeding and provides evidences for researches on the selectiveimproved variety of silkworm from Northwest area in China.
1Methods of DNA extraction from diapause of silkworm eggs
Extraction of DNA from silkworm is the basic work of molecular genetic researchtechnology, and also is the most common problems of identification and purity testing. In thispaper, by comparing the result of three methods on DNA extraction, we found a better wayand DNA extracted from a single eggs could meet the requirement for basic molecularresearch.
2Genetic diversity of silkworm
Total535bands were amplified by14pairs of primers combinations, and472(88.22%)of which showed polymorphisms.
3Genetic Distance and Cluster Analysis
Calculate the genetic distance between silkworm varieties by NTSYS-pc2.1. the genetic distances of53varieties are between0.1934-0.3491. The cluster analysis of53silkwormvarieties and the each system by UPGMA and NJ. Study subjects were divided into threegroups, the Japanese line and European line have the closet genetic distance than Chinese line,four monovoltine trimolter varisties origined from Northwest area in China belonged to aseparate group by Cluster Analysis using UPGMA.
为了更好地开发西部蚕区家蚕品种资源，以及在家蚕新品种选育中的利用提供分子水平的理论依据。本研究选择陕西省保存的具有代表性的53份家蚕品种资源，利用AFLP分子标记技术分析其亲缘关系和遗传距离。
1适于家蚕滞育期蚕卵DNA提取方法的建立
DNA提取是家蚕分子遗传学研究的基本技术，也是品种鉴定和纯度检测研究中较为常见的问题。本实验中使用3种DNA抽提方法进行对比，建立适合滞育期蚕卵的基因组DNA提取方法，达到快速高效满足分子实验要求的目标。
2家蚕遗传多样性分析
通过筛选出的14对引物组合对53个品种基因组DNA样品的AFLP扩增共获得535条带，其中多态性条带数472条，每对引物扩增的多态性比率为88.22%，显示出不同家蚕品种间存在丰富的遗传多样性。
3家蚕遗传相似性分析与聚类分析
使用NTSYS-pc2.1软件计算各家蚕品种间遗传距离。53个家蚕品种间遗传距离在0.0728~0.6019间。
基于UPGMA法及NJ法聚类分析得到53个家蚕品种聚类分析图，以及各个系统的聚类分析图。中系品种单独聚为一类，欧系品种和日系品种聚为一大类。其中基于UPGMA法聚类一化性三眠的西北地方品种明显区别于其它蚕品种聚为不同的类群。
Silkworm(Bombyx mori) is an important model and economic Lepidoptera insert, hasalways been concerned about the origin and evolution. It is the theoretical basis of thesilkworm genetics and breeding. Silkworm have a variety of ecological types and systemsgroups, such as the Chinese system, Japanese system, Tropical system and European system.Phylogenetic relationships among silkworm breeds and genetic distance of silkworm origin,evolution, germplasm identification, preservation and use of varieties of silkworm parentselection practice, has important theoretical significance and practical. The genetic diversityof53silkworm (Bombyx mori.) varieties conserved in Shaanxi province was analyzed byAFLP（amplified fragment length polymorphism）method in order to utilize germplasmresources in silkworm breeding and provides evidences for researches on the selectiveimproved variety of silkworm from Northwest area in China.
1Methods of DNA extraction from diapause of silkworm eggs
Extraction of DNA from silkworm is the basic work of molecular genetic researchtechnology, and also is the most common problems of identification and purity testing. In thispaper, by comparing the result of three methods on DNA extraction, we found a better wayand DNA extracted from a single eggs could meet the requirement for basic molecularresearch.
2Genetic diversity of silkworm
Total535bands were amplified by14pairs of primers combinations, and472(88.22%)of which showed polymorphisms.
3Genetic Distance and Cluster Analysis
Calculate the genetic distance between silkworm varieties by NTSYS-pc2.1. the genetic distances of53varieties are between0.1934-0.3491. The cluster analysis of53silkwormvarieties and the each system by UPGMA and NJ. Study subjects were divided into threegroups, the Japanese line and European line have the closet genetic distance than Chinese line,four monovoltine trimolter varisties origined from Northwest area in China belonged to aseparate group by Cluster Analysis using UPGMA.
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