线辣椒SSR-PCR反应体系优化及多态性标记筛选  

Optimization of SSR-PCR Reaction System and Selection for Polymorphic Primers in Chili Pepper(Capsicum annuum var.longunt)

作　　者：李全辉[1] 李江[1] 邵登魁[1] 王亚艺[1] 侯全刚[1] 钟启文[1] 

机构地区：[1]青海大学农林科学院,西宁810016

出　　处：《西北农业学报》2017年第7期1047-1053,共7页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：青海省农林科学院中青年基金(2014-NKY-04);青海省遗传与生理重点实验室项目~~

摘　　要：以循化线辣椒果实黄色突变体H0809和高代自交系0599-1为试材,采用L_9(3~4)正交试验设计,研究线辣椒SSR-PCR反应体系的主要成分对扩增结果的影响,并对420对SSR分子标记进行筛选。结果表明,最优的反应体系为:总体积20μL,基因组DNA 20 ng,dNTP 100μmol·L^(-1),引物0.6μmol·L^(-1),Mg^(2+)2.5 mmol·L^(-1),Taq DNA聚合酶1.5μmol;通过筛选,获得72对具有多态性的SSR标记,多态性率为17.1%,平均每个标记扩增出2.67个等位基因。本试验建立的SSR-PCR反应体系和筛选的多态性标记可用于线辣椒种质资源分析、分子标记辅助育种、分子遗传图谱构建及基因克隆等研究。In the study,the factors affecting the SSR-PCR results of pepper were studied with L9（3^4） orthogonal design,and the polymorphism of 420 primer pairs were screened by yellow pepper fruit mutant HJ 2002 and high generation inbred 0599-1.The results showed that optimal reaction system was the total 20 μL reaction system containing 20 ng of DNA template,100 μmol·L^-1 dNTP,0.6 μmol·L^-1 primer,2.5 mmol·L^-1 Mg^2＋ and 1.5 μmol of Taq DNA polymerase enzyme.All the 420 primer pairs were selected at random for yielding amplification products,of which 72 primer pairs showed polymorphism between two different pepper varieties.The polymorphism rate was 17.1%while the average alleles per locus was 2.67.This optimized SSR— PCR system and polymorphism primer could be applied for further research on germplasm analysis,molecular marker — assisted breeding,construction of molecular genetic linkage maps and gene cloning in chili pepper.

关 键 词：线辣椒 SSR 体系优化 引物筛选 

分 类 号：S641.3[农业科学—蔬菜学]