异鳞石山棕ISSR-PCR反应体系的建立及其亲缘关系分析  

Establishment of ISSR-PCR Reaction System and Analysis on Genetic Relationships of Guihaia heterosquama

作　　者：彭慧 胡焕 何溪 李名扬[1] 李志能[1] 李先源[1] Peng Hui;Hu Huan;He Xi;Li Mingyang;Li Zhineng;Li Xianyuan(College of Horticulture and Landscape Architecture,Southwest University,Chongqing,400715)

机构地区：[1]西南大学园艺园林学院,重庆400715

出　　处：《分子植物育种》2021年第3期905-913,共9页Molecular Plant Breeding

基　　金：中央高校基本科研业务费专项资金(XDJK2014C178)资助。

摘　　要：为进一步明确异鳞石山棕(Guihaia heterosquama)的分类地位以及异鳞石山棕的遗传多样性本研究以异鳞石山棕及其近缘种共10种40份植物为材料,利用L_(16)(4^(5))正交设计实验建立异鳞石山棕的最佳ISSR-PCR反应体系(总体积20μL,包括2μL 10×Buffer,2.5μL Mg^2+(25 mmol/L),0.7μL dNTPs(10 mmol/L),0.5μL引物(10μmol/L),模板DNA 50 ng,Taq DNA聚合酶1.25 U),并通过筛选的ISSR引物对40份植物进行亲缘关系分析。结果表明筛选出的14条引物共扩增出209个位点多态性位点200个,平均多态性位点百分率为95.7%。异鳞石山棕与两广石山棕(Guihaia grossifibrosa)遗传相似系数最大(0.718),亲缘关系最近,与多裂棕竹(Rhapis multifidi)遗传相似系数最小(0.565),亲缘关系相较于其他种最远。10种植物共聚为3大类,异鳞石山棕和石山棕(Guihaia argyrata)、两广石山棕聚到一起。研究结果支持将异鳞石山棕划分至石山棕属(Guihaia)的观点,为异鳞石山棕的分类提供分子水平上的证据,同时也为后续异鳞石山棕的遗传多样性、种质资源的开发利用提供参考依据。In order to further clarify the taxonomic status and genetic diversity of Guihaia heterosquama.In this study,ISSR-PCR optimal reaction system of G.heterosquana(total volume 20μL,including 2μL 10×Buffer,2.5μL Mg^2+(25 mmol/L),0.7μL dNTPs(10 mmol/L),0.5μL primer(10μmol/L),DNA template 50 ng,Taq DNA polymerase 1.25 U)was established with L_(16)(4^(5))orthogonal design,and genetic relationship of 10 species,40 plants of G.heterosquama and its related species were analyzed by using ISSR molecular marker.The results showed that a total of 209 sites were amplified by 14 primers selected,among which 200 were polymorphic points.The average polymorphism percentage reached 95.7%.Between G.heterosquama and G.grossifibrosa,the genetic similarity coefficients was the largest(0.718),the genetic relationship was the closest.Between G.heterosquama and Rhapis multifidi,the genetic similarity coefficients was the smallest(0.565),the genetic relationship was the farthest compared with other species.10 species were divided into three categories,and G.heterosquama,G.argyrata and G.grossifibrosa were clustered together.The research indicate of supporting the idea of classifying G.heterosquama into Guihaia,providing a molecular evidence for the taxonomic status of G.heterosquama,and also provided a reference for the genetic diversity,the development and utilization of germplasm resources of G.heterosquama.

关 键 词：异鳞石山棕(Guihaia heterosquama) ISSR-PCR 反应体系 亲缘关系 

分 类 号：S687[农业科学—观赏园艺]