基于TRAP分子标记的铁皮石斛野生居群的分析与鉴别  

Molecular identification of wild populations of Dendrobium officinale based on target region amplification polymorphism

作　　者：刘玲[1] 吴睿[1] 牛志韬 薛庆云[1] 刘薇[1] 丁小余[1] 

机构地区：[1]南京师范大学生命科学学院,江苏南京210023

出　　处：《药学学报》2016年第12期1926-1933,共8页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(31170300);江苏省科技厅产学研合作项目(BY2014001-01)

摘　　要：本文根据铁皮石斛多糖合成途径中磷酸烯醇式丙酮酸羧化酶基因(PEPC)、尿苷二磷酸葡萄糖焦磷酸化酶基因(UGP)和生物碱合成过程中苯丙氨酸解氨酶基因(PAL)构建目标区域扩增多态性(TRAP)分子标记,对铁皮石斛野生居群及其杂交后代进行分子鉴别。结果表明,在54对TRAP引物中,筛选出的7对引物可将铁皮石斛野生居群鉴别开。同时,在杂交后代中均能检测到父母本特征条带和新产生条带,能够有效地将亲本及其杂交后代鉴别开来。聚类分析表明,当以云南广南铁皮石斛为亲本时,正反杂交后代均先与其聚为一支,再与另一亲本聚为一支,体现了子代与云南广南铁皮石斛的亲缘关系较近。采用TRAP分子标记对铁皮石斛野生居群及其杂交后代进行分子鉴别,有利于选择优质居群进行人工培育,实现了杂交后代的早期鉴别,提高种质质量的控制,为改善铁皮石斛品质提供理论依据。In this study, TRAP molecular markers were used in identification of wild populations and hybrids of Dendrobium officinale, based on the sequences of genes encoding phosphoenolpyruvate carboxylase（PEPC）, UDP-glucose pyrophosphorylase（UGP） and phenylalanine ammonia-lyase（PAL）. Seven polymorphic target region amplification polymorphism（TRAP） primers were selected from 54 primer combinations and used in the identification of wild populations. Moreover, hybrids had female polymorphic bands, male polymorphic bands and heterozygous bands, which suggest that seven TRAP markers are able to identify the hybrids from their parents. Furthermore, the UPGMA dendrogram revealed that when sample from Guangnan in Yunnan province was used as one parent, reciprocal hybrids grouped with them in first, and then grouped with the other parent. The results indicated that the hybrids were closer to D. officinale from Guangnan population. This study identified the wild populations and hybrids of D. officinale by TRAP molecular markers, which is useful in selection of good varieties for artificial cultivation and early identification of hybrids. The study provides a method in the control of stability of germplasm and quality of D. officinale.

关 键 词：铁皮石斛 TRAP分子标记 野生居群 杂交后代 鉴别 

分 类 号：R931[医药卫生—生药学]