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荷花遗传多样性的ISSR标记分析
刘艺平1, 李 创2, 李 娜1
1.河南农业大学 林学院;2.新郑市南龙湖管委会 市政建设环保局
摘要:
【目的】探讨荷花(Nelumbo nucifera Gaertn.)资源的遗传多样性和亲缘关系,为荷花资源的保护利用、品种鉴定及育种效率提高提供参考。【方法】利用ISSR分子标记技术,对6份野生半野生荷花品系和27份荷花栽培品种进行遗传多态性分析,从40条引物中筛选扩增效果较好的5条引物用于试验。采用POPGEN32软件计算供试材料的等位基因数(Na)、有效等位基因数(Ne)、Nei’s基因多样度(He)和Shannon多态性信息指数(I),利用UPGMA方法进行聚类分析。【结果】用筛选出的5条引物(UBC807、UBC811、UBC836、UBC840、UBC880)分别对33份荷花材料基因组DNA进行扩增,共扩增出54条清晰条带,其中多态性条带25条,多态性条带比例为46.3%。应用引物UBC811、UBC840和UBC880的15个特异性位点绘制了33份荷花的DNA指纹图谱,这些位点组合后可以将33份荷花材料逐一区分开。33份荷花资源的Ne为1.276 6,He为0.192 0,I为0.323 7,且野生半野生品系的各项参数值均低于栽培品种。利用UPGMA法将供试荷花资源分为4个类群,6个野生半野生荷花品系均在第Ⅰ类群,3个中、美杂种莲种系和13个中国莲种系的品种聚在第Ⅱ类群,藕莲和花莲分属于第Ⅰ类群和第Ⅱ类群,第Ⅲ和第Ⅳ类群分别仅包括1个和2个品种。【结论】供试33份荷花资源的遗传多样性水平较低。聚类结果显示,荷花资源间的亲缘关系与地理分布距离相关,中、美杂种莲种系品种与中国莲种系品种的亲缘关系较近。加强对全国各地荷花资源的调查与保护,是荷花遗传多样性保护的重要工作。
关键词:  荷花  ISSR  遗传多样性
DOI:
分类号:
基金项目:河南省重大科技专项(072102150001)
ISSR markers based on genetic diversity analysis of Nelumbo nucifera Gaertn.
Abstract:
【Objective】The research was conducted to explore the genetic diversity and phylogenetic relationship of Nelumbo nucifera Gaertn.and improve protection and utilization of the lotus accessions,cultivar identification and breeding efficiency.【Method】The DNA polymorphism of 6 wild and semi-wild lotus stains and 27 lotus cultivars were analyzed using inter-simple sequence repeat (ISSR) markers.5 out of the 40 primers subjected to screening were used.By POPGEN32,the average values of alleles,effective number of alleles,the average Nei’s gene diversity and the average Shannon’s information index were analyzed.The cluster analysis was conducted by UPGMA (unweighted pair group method with arithmetic average).【Result】Using 5 selected primers (UBC807,UBC811,UBC836,UBC840,and UBC880),54 DNA bands were amplified,of which 46.3% were polymorphic.Based on the 15 polymorphic loci from the primers UBC811,UBC840,and UBC880,the ISSR fingerprints were constructed and the 33 lotus accessions were separated.The average value of effective number of alleles,Nei’s gene diversity and Shannon’s information index were 1.276 6,0.192 0 and 0.323 7,respectively.The indexes of the 6 lotus strains were lower than those of cultivars.The 33 lotus accessions were divided into four groups by UPGMA.The 6 lotus strains belonged to the first group and the second group included the 3 cultivars of Sino-American hybrids species constitution and the 13 cultivars of Chinese lotus species constitution.The rhizome cultivars and the flower cultivars belonged to the first and second group,respectively.【Conclusion】The tested 33 lotus accessions exhibited a low level of genetic diversity,which may result from their asexual mode of reproduction,survival environment and the studied accessions.The genetic relationship of lotus accessions is related to their geographic distribution.Sino-American hybrids species constitution and Chinese lotus species constitution are genetically close.It is the important work of lotus genetic diversity protection to strengthen investigation and protection of the lotus resources around China.
Key words:  Nelumbo nuciferaGaertn.  ISSR  genetic diversity