猕猴桃属植物的分子系统学研究
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摘要
植物分子系统学已成为植物系统学研究的主要组成部分，广泛地应用于植物系统学研究的各级分类单元上，并体现出了巨大的应用价值和理论意义，猕猴桃属(Actinidia Lindl．)隶属于猕猴桃科(Actinidiaceae)，为多年生藤本植物，雌雄异株，是一种原产我国的重要果树资源。但猕猴桃属植物的分类依据一直以形态为主，许多物种界定模糊，分类地位未能最终确定。而且该属是目前已知被子植物中唯一的叶绿体严格父系遗传而线粒体严格母系遗传的植物类群。本研究采用分子标记和测序技术从猕猴桃属核-质三基因组开展分子系统发育研究，主要结果如下：
1．利用随机扩增多态性DNA(RAPD)技术对猕猴桃属植物31种、9变种、13个品种及2种近缘属植物进行了分析。从144个引物筛选出的22个随机引物扩增了220个DNA多态性片段，其中猕猴桃属内多态性片段188个，该属具有丰富的遗传多样性。运用NTSYS-pc软件进行遗传相似性分析，计算遗传距离，构建UPGMA聚类表征图，并运用Phylip软件包的相关软件进行Wagner简约分支分析，构建系统分支树，其结果表明：(1)净果组为单系类群且相对原始，其它三组均不构成单系类群；支持净果组作为单一组的划分，不支持现有其它组、系的划分。(2)该属物种按地理分布聚类，分支分化与地理分布区域呈现出一定的相关性，表明地理环境在猕猴桃属分化及系统发育过程中起重要作用。本文还讨论了猕猴桃属植物遗传多样性、属下分类体系及其部分种(变种)间的相互关系，该属需要适当修订。RAPD适用于该属遗传多样性和分类学研究。
2．本研究利用扩增片段长度多态性(AFLP)标记技术对国家猕猴桃种质资源圃所收集保存猕猴桃属植物38种、17变种及2个可疑分类群和1近缘属物种共96份材料行了遗传多样性分析和系统发育分析。运用4个选择性引物组合共扩增出105个可靠的AFLP遗传标记，其中多态性标记103个，多态性标记百分比为98％，分类群内平均多态性标记百分比为27.01％。运用NTSYS-pc软件进行遗传相似性分析，计算遗传距离，构建UPGMA聚类表征图，并运用PHYLIP软件包的相关软件进行Wagner简约分支分析，构建系统分支树，其结果表明：(1)聚类分析反映出了各分类群间的亲缘关系，与传统依据形态分类提及的近缘关系大体一致。(2)分支分析支持净果组为单系类群且相对原始，其它三组均不构成单系类群；支持净果组作为单一组的划分，不支持现有其它组系的划分。(3)该属物种按地理分布聚类，分支分化与地理分布区域呈现出一定的相关性，表明地理环境在猕猴桃属分化及系统发育过程中起重要作用，该属植物的遗传分化还在强烈地进行。(4)猕猴桃属现有种的划分过细，可以
Molecular systematics has been invited into the phylogenetic study of plants, and widely used on all taxonomic categories with great application potential and theoretical significance. The genus Actinidia belonging to the Actinidiaceae family, is dioecious, perennial liane, and has become an important fruit resource native to China. While the present taxonomy of the genus Actinidia was mainly based on morphological characteristics, and caused some classification difficulties for certain species, which taxonomic positions were not decided yet. So far, the genus Actinidia has been found as the only angiosperm with strictly paternal inheritance of chloroplast, but maternal inheritance for mitochondria. In this study, the phylogenetic relationships of Actinidia were reviewed using molecular markers and sequencing techniques form three genomes included the nucleus, chloroplast and mitochondrion. The main achievements are as follow:
1. Forty taxa of Actinidia and two closely related genera were analyzed by random amplified polymorphic DNA (RAPD) to determine the genetic diversity and phylogenetic relationships within genus Actinidia, including 31 species, 9 varieties, and 13 cultivars of commercially important species A. chinensis and A. deliciosa. Twenty-two highly efficient primers prescreened from 144 oligonucleotide 10-base primers amplified total 220 polymorphic DNA bands and 180 polymorphic DNA bands within genus Actinidia, respectively . The RAPD diversity is high within the genus and more than 75% of the diversity resided among the taxa. Based on the RAPD data, cluster and cladistic analyses were performed using UPGMA clustering method with the NTSYS-pc 1.80c and Wagner, Mixed Parsimony Method with
1．利用随机扩增多态性DNA(RAPD)技术对猕猴桃属植物31种、9变种、13个品种及2种近缘属植物进行了分析。从144个引物筛选出的22个随机引物扩增了220个DNA多态性片段，其中猕猴桃属内多态性片段188个，该属具有丰富的遗传多样性。运用NTSYS-pc软件进行遗传相似性分析，计算遗传距离，构建UPGMA聚类表征图，并运用Phylip软件包的相关软件进行Wagner简约分支分析，构建系统分支树，其结果表明：(1)净果组为单系类群且相对原始，其它三组均不构成单系类群；支持净果组作为单一组的划分，不支持现有其它组、系的划分。(2)该属物种按地理分布聚类，分支分化与地理分布区域呈现出一定的相关性，表明地理环境在猕猴桃属分化及系统发育过程中起重要作用。本文还讨论了猕猴桃属植物遗传多样性、属下分类体系及其部分种(变种)间的相互关系，该属需要适当修订。RAPD适用于该属遗传多样性和分类学研究。
2．本研究利用扩增片段长度多态性(AFLP)标记技术对国家猕猴桃种质资源圃所收集保存猕猴桃属植物38种、17变种及2个可疑分类群和1近缘属物种共96份材料行了遗传多样性分析和系统发育分析。运用4个选择性引物组合共扩增出105个可靠的AFLP遗传标记，其中多态性标记103个，多态性标记百分比为98％，分类群内平均多态性标记百分比为27.01％。运用NTSYS-pc软件进行遗传相似性分析，计算遗传距离，构建UPGMA聚类表征图，并运用PHYLIP软件包的相关软件进行Wagner简约分支分析，构建系统分支树，其结果表明：(1)聚类分析反映出了各分类群间的亲缘关系，与传统依据形态分类提及的近缘关系大体一致。(2)分支分析支持净果组为单系类群且相对原始，其它三组均不构成单系类群；支持净果组作为单一组的划分，不支持现有其它组系的划分。(3)该属物种按地理分布聚类，分支分化与地理分布区域呈现出一定的相关性，表明地理环境在猕猴桃属分化及系统发育过程中起重要作用，该属植物的遗传分化还在强烈地进行。(4)猕猴桃属现有种的划分过细，可以
Molecular systematics has been invited into the phylogenetic study of plants, and widely used on all taxonomic categories with great application potential and theoretical significance. The genus Actinidia belonging to the Actinidiaceae family, is dioecious, perennial liane, and has become an important fruit resource native to China. While the present taxonomy of the genus Actinidia was mainly based on morphological characteristics, and caused some classification difficulties for certain species, which taxonomic positions were not decided yet. So far, the genus Actinidia has been found as the only angiosperm with strictly paternal inheritance of chloroplast, but maternal inheritance for mitochondria. In this study, the phylogenetic relationships of Actinidia were reviewed using molecular markers and sequencing techniques form three genomes included the nucleus, chloroplast and mitochondrion. The main achievements are as follow:
1. Forty taxa of Actinidia and two closely related genera were analyzed by random amplified polymorphic DNA (RAPD) to determine the genetic diversity and phylogenetic relationships within genus Actinidia, including 31 species, 9 varieties, and 13 cultivars of commercially important species A. chinensis and A. deliciosa. Twenty-two highly efficient primers prescreened from 144 oligonucleotide 10-base primers amplified total 220 polymorphic DNA bands and 180 polymorphic DNA bands within genus Actinidia, respectively . The RAPD diversity is high within the genus and more than 75% of the diversity resided among the taxa. Based on the RAPD data, cluster and cladistic analyses were performed using UPGMA clustering method with the NTSYS-pc 1.80c and Wagner, Mixed Parsimony Method with
引文
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