中国烟草学报2018，Vol.24Issue(3)：37-45,9.DOI:10.16472/j.chinatobacco.2017.289
利用HRM技术快速筛选低镉烟草突变体
Rapid screening of low-cadmium tobacco mutants by HRM technology
摘要
Abstract
Low-cadmium tobacco germplasm resources are essential for tobacco breeding and production safety.In this study,we treated Guiyan No.1 with EMS,and planted 2020 M2 lines including 10 individuals,then extracted bulk DNA from leaves of three individuals.49 materials with putative mutation in NtHMA4 were screened using high-resolution melting(HRM)curve analysis.Among them,21 mutants with SNP mutation were confirmed by DNA sequencing,of which 15 were non-synonymous mutations caused amino acid changed and the rest were synonymous mutations.Subsequently,cadmium content in the leaves of eight mutants decreased significantly from 16.19％to 42.08％compared to wild type.Ten M3progeny with non-synonymous mutations,which can germinate normally,were detected by HRM.Results showed that HRM technology could not only quickly screen mutants in tobacco EMS mutagenesis,but could also be used for rapid and accurate genotyping of heterozygous mutant offspring combined with DNA sequencing.关键词
镉转运基因/诱变/烟草/低镉/HRMKey words
cadmium transporter gene/mutagenesis/tobacco/low-cadmium/HRM引用本文复制引用
张超,张吉顺,王仁刚,曾吉凡,王自力,任学良,舒庆尧,武立权,赵海军..利用HRM技术快速筛选低镉烟草突变体[J].中国烟草学报,2018,24(3):37-45,9.基金项目
烟草行业重点实验室项目:利用分子标记辅助筛选低镉和低降烟碱烟草种质研究(合同号:110201403019) （合同号:110201403019）
中国烟草总公司贵州省公司科技项目《贵州烤烟种质创制及新品种选育研究》(中烟黔科[2016]6号) （中烟黔科[2016]6号）
