论文标题:兰属和石斛属植物的遗传多样性研究
Genetic Diversity Studies of Cymbidium and Dendrobium Based on RAPD and AFLP
论文作者 王玉东
论文导师 周晓云；王慧中,论文学位 硕士,论文专业 生物化工
论文单位 浙江工业大学,点击次数 112,论文页数 69页File Size5308k
2004-04-01论文网 http://www.lw23.com/lunwen_54034097/ RAPD,AFLP,Cymbidium,Dendrobium,遗传多样性,银染
RAPD,AFLP,Cymbidium,Dendrobium,genetic variation,silver staining
本文采用RAPD技术和AFLP技术对兰属植物14种材料以及采用AFLP技术对石斛属13种材料进行了遗传多样性检测。 从100个随机引物中筛选出12个随机引物，对兰属植物14种材料进行随机扩增，共得到254条带，多态性带有243条，多态性百分率为95.7％。扩增出的DNA带的分子大小在300～2200bp之间。聚类分析结果表明，在相似系数0.46的水平上可将14种兰属材料分为Ⅰ类和Ⅱ类。Ⅰ类包括邱北冬蕙兰、春剑、春兰、峨眉春蕙、寒兰、珍珠矮、墨兰、建兰。Ⅱ类包括兔耳兰、大雪兰、独占春、文山红柱兰、虎头兰、黄蝉兰。 用3对选择性引物对E+ACT／M+CAA、E+ACT／M+CAC、E+ACT／M+CAG分别对兰属植物14种材料进行扩增，在100～300bp之间共得到279条带，多态性带223条，多态性百分率为79.9％。聚类分析结果表明，在相似系数0.72的水平上可将14种兰属材料分为两类Ⅰ类和Ⅱ类。Ⅰ类包括兔耳兰、邱北冬蕙兰、春剑、春兰、峨眉春蕙、寒兰、珍珠矮、墨兰、建兰。Ⅱ类包括大雪兰、独占春、文山红柱兰、虎头兰、黄蝉兰。 运用RAPD和AFLP技术对兰属植物14种材料所得的聚类分析结果基本一致，也与Du Puy & Cribb分类系统接近。两种结果都得出建兰与墨兰，寒兰与峨眉春蕙，虎头兰和黄蝉兰以及大雪兰与独占春之间的亲缘关系最近。 选用3对选择性引物对E+ACT/M+CAC、E+从C/M+CAC、E+ACA/M+CAC分别对石解属植物13种材料进行扩增，得到非常丰富的条带，在100~3O0bp之间共得到346条带，多态性带342条，多态性百分率为98.8%。聚类分析结果表明，在相似系数0.56处，可将13种材料分为I、H、m、W类。I类包括串珠石解、铁皮石解、广东石解、尖刀唇石解和晶帽石解。H类包括细叶石解、滇桂石解、报春石解、玫瑰石解、球花石解。m类包括鼓褪石解。W类包括美花石解。这与形态分类有差别，表明石解属植物的遗传多样性和分类有待进一步研究。
In this study, the genetic variations of 14 species of Cymbidium were revealed by using RAPD and AFLP markers and the genetic variations of 13 species of Dendrobium were revealed by using AFLP markers.Twelve primers selected from 100 random primers amplified 254 frangments in 14 species of Cymbidium, in which 243 segments are polymorphic. The rate of polymorphism was 95.7%. And all the segments were ranging from 200bp to 2200bp. The cluster analysis with UPGMA suggested that the tested materials were clustered into two groups on coefficient level 0.46. Group I included C.qiubeiense, C.goeringii var. longibracteatum, C.goeringii, C.faberi var. omeiense, C.kanra,n C.nanulum, C.sinense,C.ensifolium. Group II included C. lancifolium, C.mastersii, C.eburneum, C.wenshanense, C.hookerianum, C.iridioides.Three selective primer combinations were used to amplify in 14 species of Cymbidium, which were E+ACT/M+CAA, E+ACT/M+CAC, E+ACT/M+CAG. Ranging from 100bp to 300bp, a total of 279 AFLP markers were produced, among which 223 polymorphic bands werepolymorphic. The polymorphism rate was 79.9%. The cluster analysis with UPGMA indicated that the 14 species were also clustered into two groups on coefficient level 0.72. Group I included C. lancifolium, C. qiubeiense, C. goeringii var. longibracteatum, C. goeringii, C. faberi var. omeiense, C. kanran, C. nanulum, C. sinense, C. ensifolium. Group II included C. mastersii, C. eburneum, C. wenshanense, C. hookerianum, C. iridioides.The results of UPGMA cluster analysis obtained from RAPD data and AFLP data were in essence consistent, and also very closed to Du Puy & Cribb morphological classification. Two results showed that the genetic distances are closest between C. ensifolium and C. sinense, C. kanran andC. faberi var. omeiense, C. mastersii and C. eburneum.Three selective primer combinations were used to detect the genetic variations in 13 species of Dendrobium , which were E+ACT/M+CAC,E +AAC/M+CAC,E+ACA/M+CAC.Abundant bands were gotten. Between lOObp and 300bp, a total of 346 DNA bands were amplified, 342 of which were polymorphic. The rate of polymorphism was 98.8%. The cluster analysis with UPGMA indicated that the 13 species were clustered into four groups on coefficient level 0.56. Group I included D. falconeri, D. officinale, D. wilsonii, D. heterocarpum, D. crystallinum. Group II includedD. hancockii, D. guangxiense, D. primulinum, D. crepidatum, D. thyrsiflorum. Group III included D. chrysotoxum. Group IV included D.loddigesii. The results were different from that of morphological study, indicating that classification of Dendrobium remain to study further.
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