应用RAPD技术研究不同种山羊草叶绿体DNA的遗传变异  

Phylogenetic Relations of Aegilops Species Based on RAPD Analysis of Chloroplast DNA

作　　者：王洁[1] 陈孝[1] 倪丕冲[1] 赵进东[2] 吴光耀[2] 

机构地区：[1]中国农业科学院作物育种栽培研究所,北京100081 [2]北京大学生命科学学院,北京100871

出　　处：《作物学报》1998年第5期520-528,共9页Acta Agronomica Sinica

基　　金：国家自然科学基金

摘　　要：采用随机扩增多态性DNA(RAPD)技术研究了山羊草属cpDNA的遗传变异性。用改良的“高盐低pH法”提取了15种山羊草材料和一种二粒小麦的叶绿体DNA,并建立了重复性较好的RAPD标准分析条件。在此标准条件下,检测了16个材料叶绿体DNA的多态性。经10个引物扩增,在得到的64个片段中,有6个片段是16个材料共有的,多态性达90％。聚类分析结果显示,关系较近的有小亚山羊草和欧山羊草,粘果山羊草和易变山羊草,高大山羊草和拟斯卑尔脱山羊草以及偏凸山羊草和粗山羊草。粗山羊草种内叶绿体DNA存在着一定程度的变异性。本研究将16个材料的cpDNA分成10个类型,与前人用限制性内切酶对cpDNA的分类结果基本吻合。不同的是本研究认为关系较密切的高大山羊草和拟斯卑尔脱山羊草叶绿体DNA类型不同,而小伞山羊草却与具有相同cpDNA类型的其它山羊草种关系较远。Good quality cpDNAs were extracted from 15 accessions of 13 Aegilop species and one accession of T. dicoccoides Korn by using 'high salt-low pH' method. Our primary study showed that the template concentration (from 25 ng to 100 ng)has little effect on RAPD patterns and non-specific bands were observed when DNA polymerase(Tag) with very high con centration was used. Reproducible amplification products were obtained with rigorously optimized reaction conditions. Sixty-four amplified fragments were generated by RAPD with 10 primers, among which 6 fragments were shared by all 16 accessions. The degree of polymorphism was quite high(90%). The cluster analysis using UPGMA indicated that Ae. columnaris Zhuk and Ae. biuncialis, Ae. kotschyi Boiss and Ae. variabilis Eig, Ae. longissma and Ae. speltoides Tausch, Ae. ventricosa Tausch and Ae. squrrosa L. are much more closely related. Intraspecific variation of cpDNAs was found in Ae. squrrosa. cpDNAs from the 16 accessions were classified into 10 types based on RAPDs. The relationships among the studied species are in good agree ment with most of those suggested by other researchers by using RFLPs. However, the present study found that the relationships between Ae. longissma and Ae. speltoides, Ae. umbellulata and other Aegilops species with 3 type plastome have slight difference from the previous studies. Finally, the phylogenetic relationships of Aegilops and some characteristics of RAPD were dis cussed.

关 键 词：山羊草属 叶绿体 DNA RAPD 遗传 变异 

分 类 号：S512.101[农业科学—作物学]