利用TRAP和SCAR标记评价铁皮石斛的遗传多样性与抗寒性  

Assessment of genetic diversity and cold resistance of Dendrobium officinale using TRAP and SCAR markers

作　　者：张君毅[1] 吴永辉 刘嘉 秦建立 张轶钧 ZHANG Junyi;WU Yonghui;LIU Jia;QIN Jianli;ZHANG Yijun(Department of Bioengineering and Biotechnology,Huaqiao University,Xiamen,Fujian 361021,China)

机构地区：[1]华侨大学生物工程与技术系,福建厦门361021

出　　处：《植物生理学报》2020年第4期743-751,共9页Plant Physiology Journal

基　　金：福建省科技计划项目(2015N0028)。

摘　　要：开发目标区域扩增多态性(TRAP)和特异序列扩增区域(SCAR)标记,对铁皮石斛(Dendrobium officinale)的3个生态类群46份种质进行遗传多样性评价、野生群体鉴别以及抗寒性筛选。以抗寒相关的CBF1/DREB1转录因子和钙依赖型蛋白激酶(CDPK1)基因设计TRAP固定引物,从57对TRAP引物中共筛选出21个引物组合,多态信息量(PIC)、分辨率(RP)和多态带百分比(PPB)平均分别为0.899、2.054和76.79%。Nei氏基因多样度(H)为0.217 7~0.291 7 (平均为0.258 1), Shannon信息指数(I)为0.325 1~0.442 1 (平均为0.389 2)。野生类群遗传多样性水平最高,仿野生类群和栽培类群基本一致。主成分分析(PCA)结果表明供试材料可分为野生和栽培两大类群。此外,开发了一个铁皮石斛转录因子(RAP2-4)的SCAR标记,并以该SCAR标记和TRAP固定引物共同筛选出了核心抗寒种质资源。本文研究策略拟为TRAP标记在育种中的应用提供参考。Target region amplified polymorphism(TRAP) and sequence-characterized amplified regions(SCAR) markers were developed to carry out the genetic diversity evaluation, wild population identification, and selection of cold-resistant Dendrobium officinale consisted of 46 germplasms from three ecological groups. TRAP-fixed primers were designed based on cold-resistance-related CBF1/DREB1 transcription factor genes and calcium-dependent protein kinase(CDPK1) genes. Totally 21 primer combinations were screened out from 57 TRAP primer pairs, and the average values of polymorphism information content(PIC), resolving power(RP), and the percentage of polymorphic band(PPB) were 0.899, 2.054, and 76.79%, respectively. Nei’s gene diversity(H) was 0.217 7–0.291 7(mean 0.258 1), and Shannon’s information index(I) ranged from 0.3251 to 0.4421(mean 0.389 2). The genetic diversity level of the wild group is the highest, and imitated wild group almost equals to cultivated group. Principal component analysis(PCA) showed that the tested samples were clustered into two groups, wild and cultivation. In addition, a SCAR marker linked to D. officinale transcription factor(RAP2-4) was developed, and core germplasm resources of cold-resistant D. officinale were selected out by the SCAR marker and TRAP-fixed primers. This research strategy would provide reference for the application of TRAP markers in breeding.

关 键 词：铁皮石斛 TRAP标记 抗寒性 遗传多样性 SCAR标记 

分 类 号：S567.239[农业科学—中草药栽培]