木奶果种质资源的遗传多样性分析及DNA指纹图谱构建

Genetic diversity analysis and DNA fingerprinting construction with Baccaurea ramiflora Lour. germplasm resources

  • 摘要: 【目的】对来自广西、云南、海南和广东的63份木奶果种质资源进行遗传多样性分析,并构建对应的DNA指纹图谱,为木奶果种质资源的分类鉴定、保护和利用提供理论参考。【方法】选取3份地理位置相差较远的种质,从75条SCoT引物中筛选出条带清晰、重复性好和多态性丰富的引物,对63份供试种质进行多态性扩增,利用Popgene 1.32和NTsys 2.10e进行遗传多样性和聚类分析,并依据扩增结果构建DNA指纹图谱。【结果】筛选出的6条SCoT引物从63份木奶果种质资源中共扩增出192条带,其中多态性条带188条,多态性比率为97.86%;每条引物扩增的条带数为26~36条,平均为32条,多态性条带数为26~36条,平均每条引物可扩增出31.33条多态性条带,多态性比率为90.00%~100.00%;多态性信息含量值(PIC)为0.9052~0.9527,平均0.9319,Shannon’s信息指数(I)在0.2459~0.3878,Nei’s基因多样性(H')为0.1476~0.2412,等位基因数(Na)为1.9000~2.0000,有效等位基因数(Ne)为1.2324~1.3700。6条SCoT引物扩增出的多态性条带存在明显差异,单条引物可区分的种质份数为49~63份,平均为58份。供试材料间的遗传相似系数(GS)为0.5781~0.9115,平均为0.7496,在遗传相似性系数为0.7100处被分为5个组,其中A组为2份广西的种质、B组为1份广西的种质;C组为4份云南的种质;D组为海南和广东的全部种质;E组为剩余的50份种质。将进一步筛选出的5条SCoT引物(SCoT32、SCoT66、SCoT68、SCoT73和SCoT74)用于初步构建63份木奶果种质资源的DNA指纹图谱,每份种质均有唯一对应的图谱。【结论】供试的63份木奶果种质资源具有丰富的遗传多样性,种质间的亲缘关系与地域有一定的相关性。所筛选的6条SCoT引物具有较强的多态性,适用于木奶果种质资源的评价鉴定、遗传多样性分析及指纹图谱构建等研究。

     

    Abstract: 【Objective】To conduct genetic diversity analysis of 63 Baccaurea ramiflora Lour. germplasm resources from Guangxi,Yunnan,Hainan and Guangdong and to establish a preliminary DNA fingerprint,so as to provide a reference for varieties classification,identification,protection,and utilization of B. ramiflora Lour.【Method】Three germplasm resources whose geographical locations were highly different were selected,and the SCoT primers with best quality,repeatability and polymorphism were screened from 75 SCoT primers to perform amplification and electrophoresis on 63 germplasm resources. Genetic diversity and clustering analysis were performed by Popgene 1.32 and NTsys 2.10e and the results were used to construct the fingerprint profiles.【Result】The 6 SCoT primers selected from 63 germplasm resources were used to amplify 192 bands,including 188 polymorphic bands with a polymorphism ratio of 97.86%. The number of bands amplified by each primer ranged from 26 to 36,with an average of 32,and the number of polymorphic bands ranged from 26 to 36,with an average of 31.33 polymorphic bands amplified by each primer,and the polymorphic rate ranged from 90.00% to 100.00%. The polymorphic information content values(PIC)ranged from 0.9052 to 0.9527, with a mean of 0.9319. The Shannon’s information index(I)ranged from 0.2459 to 0.3878,the Nei’s gene diversity(H') ranged from 0.1476 to 0.2412,the observed number of alleles(Na)ranged from 1.9000 to 2.0000,the number of effective alleles(Ne)ranged from 1.2324 to 1.3700. There were significant differences in polymorphic bands amplified by 6 SCoT primers. The number of germplasms that could be distinguished by single primer was 49-63,with an average of 58. According to the results of cluster analysis,the genetic similarity coefficients(GS)of 63 germplasm resources ranged from 0.5781 to 0.9115,with a mean of 0.7496. At the similarity coefficient of 0.7100,the tested materials were divided into five groups:group A included two germplasm resources from Guangxi;group B included a germplasm resource from Guangxi;group C inclu-ded 4 germplasm resources from Yunnan;group D included all germplasm resources from Hainan and Guangdong;group E included the remaining 50 germplasm resources. 5 SCoT primers(SCoT32,SCoT66, SCoT68,SCoT73,SCoT74)were further screened out to construct the DNA fingerprinting of 63 germplasm resources and each resource had specific DNA fingerprint code in the database.【Conclusion】The 63 germplasm resources are rich in genetic diversity,and there is a certain correlation between the kinship of the germplasm and the geographical area;the six SCoT primers screened are of high polymorphism,which can provide reference for the future studies on evaluation,genetic diversity analysis and fingerprinting on B. ramiflora Lour. germplasm resources.

     

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