仲彬草属物种细胞质基因组PCR-RFLP分析  

PCR-RFLP analysis of plasmon in Kengyilia populations

作　　者：张利[1] 周永红[1] 郑有良[1] 刘世贵[2] 

机构地区：[1]四川农业大学小麦研究所 [2]四川大学生命科学学院,四川成都610042

出　　处：《草业学报》2006年第3期115-122,共8页Acta Prataculturae Sinica

基　　金：国家自然科学基金(30270099和30470135);长江学者和创新团队发展计划(IRT0453);四川省教育厅;科技厅重点项目资助

摘　　要：应用PCR-RFLP技术对32份仲彬草属和2份外类群共34份材料细胞质基因组DNA进行遗传变异分析。选用的16个引物中4个叶绿体引物和5个线粒体引物可扩增出稳定而明显的PCR产物,其扩增产物用15种限制性内切酶进行酶切。结果发现,所有的135种引物/酶组合中,得到清晰稳定的83种引物/酶组合,其中在32份仲彬草属材料中有40种引物/酶组合(占48.2%)能检测到多态性,但多态性水平较低。32份仲彬草属物种间的遗传相似系数在0.833以上,平均值为0.903。聚类分析表明,细胞质基因组PCR-RFLP标记能将全部供试材料区分开,32份仲彬草属材料聚为4类。同种不同居群细胞质间的遗传变异很小,分别聚在一起;种间的细胞质遗传差异大于种内不同居群间的遗传差异;而形态相似、地理分布一致的物种有聚类在一起的倾向。因此,利用细胞质基因组PCR-RFLP标记不仅可检测到仲彬草属种间多态性,也可检测到种内多态性。细胞质基因组PCR-RFLP标记能为仲彬草属物种系统学研究提供有价值的资料。Plasmon genetic variation of 32 Kengyilia accessions and 2 outgroups was investigated using PCR-RFLP technology. Four cpDNA and 5 mtDNA primers which produced steady and clear bands were selected from 16 plasmon primers. The PCR products were digested with 15 restriction endonucleases and 83 of 135 marker/enzyme combinations generated bands. Forty out of 83 （48.2 %） marker/enzyme combinations revealed polymorphisms, but the level of polymorphisms was low. Genetic similarity coefficient （GS） values were above 0. 833, with an average of 0. 903. Cluster analysis showed that 32 Kengyilia accessions could be differentiated by plasmon PCR--RFLP technology and they were classified into four groups. The different accessions of a species were clustered together and showed little variation. The interspecific genetic variation was greater than the intraspecific genetic variations. Species with similar morphological characters and species from the same areas or neighboring geographical regions were clustered together. The results are basically comparable with those obtained from our studies using RAPD, RAMP and ISSR. Plasmon PCR--RFLP technology can therefore be used to detect both interspecific and intraspecific genetic variations in Kengyilia. It could also be used as an effective method for the systematic study of Kengyilia.

关 键 词：仲彬草属 PCR—RFLP标记 细胞质基因组 聚类分析 亲缘关系 

分 类 号：S543.9[农业科学—作物学]