木犀科苦丁茶种质资源的RAPD分析  

RAPD Analysis of Germplasm Resources of Kudingcha in Oleaceae

作　　者：郑道君[1,2] 梁远发[3] 刘国民[1,2] 鄢东海[3] 令狐昌弟[3] 田永辉[3] 

机构地区：[1]海南大学热带生物资源教育部重点实验室,海口570228 [2]海南大学苦丁茶研究所,海口570228 [3]贵州省茶叶科学研究所,贵州湄潭564100

出　　处：《中国农业科学》2008年第12期4164-4172,共9页Scientia Agricultura Sinica

基　　金：国家自然科学基金资助项目(39860048);贵州省自然科学基金资助项目(黔科合J字[2005]2033号)

摘　　要：【目的】从分子水平上揭示木犀科苦丁茶植物物种间的遗传差异、亲缘关系,为木犀科苦丁茶的物种分类、鉴定和遗传育种提供依据。【方法】利用RAPD分子标记技术对木犀科苦丁茶8个物种,包括粗壮女贞[Ligustrum robustum(Roxb.)Blume]、丽叶女贞(L.henryi Hemsl.)、日本女贞(L.japonicum Thunb.)、日本毛女贞(L.japonicum Thunb.var.pubscens Koidz)、女贞(L.lucidum Ait.)、序梗女贞(L.pedunculare Rehd)、牛矢果(Osmanthus masumuranus Hayata)和川滇蜡树(L.delavayanm Hariot),共21份种质材料进行亲缘关系和遗传多样性分析,计算不同种质材料间的Jaccard遗传相似性系数,并采用UPGMA方法进行聚类分析。【结果】20个引物共获得427条谱带,多态性百分率(PPB)为97.7%,个体间遗传相似系数在0.1522～0.8322,平均遗传相似系数为0.5466,木犀科苦丁茶各物种间存在着较大的遗传差异;基于RAPD遗传相似系数的UPGMA聚类分析图可把供试材料完全分开并表明它们之间的亲缘关系。【结论】RAPD分子标记技术可有效地应用于木犀科苦丁茶种质资源的遗传多样性分析、亲缘关系及分类地位研究;日本毛女贞与序梗女贞的亲缘关系在所有的供试物种中最近,而与日本女贞的关系较远。作者据此认为,日本毛女贞是否应该划分为日本女贞种下一变种或是可以独立划分为一个种的问题尚需进一步研究;RAPD的分析结果支持紫茎女贞(L.pururascens Y.C.Yang)归并为粗壮女贞。【Objective】The genetic difference and relationship among germplasm resources of Kudingcha in Oleaceae were revealed at molecular levels on this study,which could be considered as the significant reference bases for the classification and identification and breeding of Kudingcha species in Oleaceae.【Method】The random amplified polymorphic DNA marker（RAPD） was applied to detect the genetic relationships and diversity among 21 germplasm materials of Kudingcha in Oleaceae,which involved in 8 species,i.e.Ligustrum robustum（Roxb.） Blume,L.henryi Hemsl.,L.japonicum Thunb.,L.japonicum Thunb.var.pubscens Koidz,L.lucidum Ait.,L.pedunculare Rehd,Osmanthus masumuranus Hayata,L.delavayanm Hariot.The Jaccard similarity coefficient of 21 germplasm materials was calculated,and the UPGMA was used for cluster analysis.【Result】20 RAPD primers selected were applied for the amplification on the 21 germplasm materials mentioned above.427 bands were obtained,and the percentage of polymorphic bands（PPB） was 97.7%.The genetic similarity coefficients（GS） ranged from 0.1522 to 0.8322 with an average of 0.5466.There were significant genetic difference among germplasm materials of Kudingcha species in Oleaceae,and UPGMA cluster based on GS of RAPD could distinguish all test germplasm materials clearly and indicated relationship of 8 species mentioned above.【Conclusion】RAPD markers could be used for the studies of genetic diversity and relationship and classification of germplasm resources of Kudingcha in Oleaceae.Analysis results of RAPD showed that L.japonicum Thunb.var.pubscens Koidz has closer genetic relationship with L.pedunculare Rehd and further genetic relationship with L.japonicum Thunb.among all tested species.The authors suggested that the further research be needed to study whether L.japonicum Thunb.var.pubscens Koidz should be classified into a variata of L.japonicum Thunb.,or considered as an independent species.The analysis results supported that L.pururascens Y.C.Yang should be combined into L.

关 键 词：木犀科 苦丁茶 种质资源 亲缘关系 遗传多样性 RAPD 

分 类 号：S571.1[农业科学—茶叶生产加工]