应用RAPD 标记和细胞质基因组PCR-RFLP技术研究大花蕙兰的遗传多样性

园艺学报 ›› 2006, Vol. 33 ›› Issue (2): 349-355.

应用RAPD 标记和细胞质基因组PCR-RFLP技术研究大花蕙兰的遗传多样性

甘　娜^{1, 2};谭向红¹;陈其兵¹;魏育明²;郑有良^2*

(¹四川农业大学林学园艺学院, 雅安625014; ²四川农业大学小麦研究所, 成都611830; ³四川师范大学地理与资源科学学院, 成都610066)

收稿日期:2005-06-16 修回日期:2005-10-24 出版日期:2006-04-25 发布日期:2006-04-25

Genetic Diversity in Cymbidium Based on RAPD Markers and PCR-RFLP Analyses of Organellar DNAs

Gan Na^{1, 2};Tan Xianghong1;Chen Qibing¹;Wei Yuming²;Zheng Youliang^2*

(¹College of Forestry and Horticulture, Sichuan Agricultural University, Ya'an 625014, China; ² Triticeae Research Institute,Sichuan Agricultural University, Chengdu 611830, China; 3 Faculty of Geography and Resources Science, Sichuan Normal University, Chengdu 610066, China)

Received:2005-06-16 Revised:2005-10-24 Online:2006-04-25 Published:2006-04-25

摘要/Abstract

摘要： 利用RAPD、叶绿体和线粒体基因组PCR-RFLP标记系统评价了大花蕙兰20个品种的遗传多
样性。在50个RAPD引物分析中, 有36个引物(72.0% ) 能揭示材料间的多态性, 材料间遗传相似系数为0.503～0.765, 平均0.598, 根据遗传相似系数进行聚类分析表明, RAPD标记能将所有材料区分开。在7个叶绿体基因组( cpDNA) 的PCR-RFLP分析中, 6个标记(87.5% ) 可扩增出1至多条清晰的谱带; 扩增产物经7种限制性内切酶消化后, 6个标记的19种引物/酶组合共检测到53条DNA片段, 其中多态性片段有37条, 占69.8%; 材料间遗传相似系数变化范围为0.571～0.949, 平均值为0.766。在8个线粒体基因组(mtDNA) 的PCR-RFLP标记分析中, 只有3个(37.5%) 标记能得到1条清晰的谱带; 利用7种限制性内切酶对3个标记的扩增产物消化后, 在10种标记/酶组合中, 共检测到33条酶切片段, 其中21条(63.6%) 具有多态性; 遗传相似系数为0.634～1.000, 平均0.829。这些结果表明, RAPD标记揭示的大花蕙兰遗传多样性最高, 其次为cpDNA PCR-RFLP标记, 而mtDNA PCR-RFLP标记揭示的遗传多样性最低。

关键词: 大花蕙兰, RAPD, PCR-RFLP, cpDNA, mtDNA

Abstract: The genetic diversity among 20 Cymbidium accessionswas investigated by RAPD markers and PCR-RFLP analyses of organellar DNAs. In RAPD analyses, the amplified roducts of 36 primers (72.0% ) were polymorphic. The RAPD-based genetic similarity (GS) among 20 Cymbidium accessions ranged from
01503 to 01765, with the mean of 0.598. Based on genetic similarity matrix resulting from RAPD makers, the genetic relationships among 20 Cymbidium accessions were estimated by UPGMA ( unweighted pair group method with arithmetic means) clustering analysis. It was found that all 20 Cymbidium accessions could be distinguished by RAPD markers. In cpDNA PCR-RFLP analyses, 6 out of 7 markers (87.5% ) could produce one or more than one distinct bands by direct electrophoresis in 2% agrose gels. After the amplified products were digested by 7 restriction enzymes, a total of 53 bandswere detected in 19 cpDNA PCR-RFLP marker / enzyme combinations, among which 37 bands (69.8% ) were polymorphic. The cpDNA PCR-RFLP-based genetic similarity (GS) among 20 Cymbidium accessions ranged from 0.571 to 0.949, with the mean of 0.766. Of the 8 mtDNA PCR-RFLP markers, 3 markers (37.5% ) could p roduce one distinct band with no polymorphism detected by direct electrophoresis in 2% agrose gels. After the amplified productswere digested by 7 restriction enzymes, a total of 33 bands were detected in 10 mtDNA PCR-RFLP marker/enzyme combinations,among which 21 bands (63.6% ) were polymorphic. The mtDNA PCR2RFLP2based genetic similarity (GS) among 20 Cymbidium accessions ranged from 0.634 to 1.000, with the mean of 0.829. These results suggested that relatively higher level of genetic polymorphism in Cym bidium could be detected by RAPD markers, whereas relatively lower level genetic polymorphism could be estimated by mtDNA PCR-RFLP markers.

Key words: Cymbidium, RAPD, PCR-RFLP, cpDNA, mtDNA

甘　娜;;谭向红;陈其兵;魏育明;郑有良. 应用RAPD 标记和细胞质基因组PCR-RFLP技术研究大花蕙兰的遗传多样性[J]. 园艺学报, 2006, 33(2): 349-355.

Gan Na;;Tan Xianghong;Chen Qibing;Wei Yuming;Zheng Youliang. Genetic Diversity in Cymbidium Based on RAPD Markers and PCR-RFLP Analyses of Organellar DNAs[J]. ACTA HORTICULTURAE SINICA, 2006, 33(2): 349-355.

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