基于转录组的小豆SSR分子标记开发及其应用  

Development and application of SSR molecular markers based on transcriptome sequencing of adzuki bean

作　　者：徐晓丹 冷淼 张明媛 柯希望[1] 殷丽华[1] 左豫虎[1] XU Xiaodan;LENG Miao;ZHANG Mingyuan;KE Xiwang;YIN Lihua;ZUO Yuhu(Heilongjiang Provincial Key Laboratory of Crop-Pest Interaction Biology and Ecological Control,National Coarse Cereals Engineering Research Center,Heilongjiang Bayi Agricultural University,Daqing,Heilongjiang 163319,China)

机构地区：[1]黑龙江省作物-有害生物互作生物学及生态防控重点实验室,国家杂粮工程技术研究中心,黑龙江八一农垦大学,黑龙江大庆163319

出　　处：《干旱地区农业研究》2023年第1期13-18,共6页Agricultural Research in the Arid Areas

基　　金：黑龙江省应用技术研究与开发计划(GA19B104);大庆市科学技术局指导项目(zd-2020-45);黑龙江八一农垦大学校启动计划(2031011068)。

摘　　要：为了开发小豆SSR(Simple Sequence Repeats)分子标记,2019—2020年,利用微卫星识别工具MISA和软件Primer 3搜索小豆资源QH1转录组测序得到的Unigene,获得3045个SSR分子标记,通过对标记的重复基序特点分析,发现标记的重复基序以单核苷酸重复、二核苷酸重复和三核苷酸重复为主,分别占标记总数的41.2%、26.4%和25.6%。利用可以锚定到小豆基因组并能成功设计引物的标记,构建了包括1505个SSR分子标记的物理图谱。从以上1505个标记中,随机选取分布于小豆11条染色体上的132个SSR标记进行有效性验证,发现有效扩增的标记为118个,有效率为89.4%。通过多态性标记筛选,获得6个多态性标记,利用这6个标记对36份小豆品种进行聚类分析,发现供试小豆材料可以分为4个类群。说明开发的SSR分子标记能够分析小豆的遗传背景差异,为小豆种质资源鉴定、遗传多样性分析以及优良性状基因定位研究提供可用的分子标记。To develop SSR(Simple Sequence Repeats)molecular markers for adzuki bean,MISA and Primer 3 were used to search Unigenes deriving from transcriptome sequencing of adzuki bean QH1 in 2019-2020,and 3045 SSR markers were obtained.By analyzing the characteristics of repeated motifs of markers,it was found that mononucleotide,dinucleotide and trinucleotide were the main motifs,accounting for 41.2%,26.4%and 25.6%of total markers,respectively.A physical map containing of 1505 SSR markers was constructed using markers that was anchored to the genome of adzuki bean and successfully designed with primers.From the above 1505 SSR markers,132 SSR markers distributed on 11 chromosomes of adzuki bean were randomly selected for validation.118 markers were effectively amplified with the efficiency of 89.4%.Six polymorphic markers were obtained through polymorphic marker screening,and the clustering analysis of 36 adzuki bean cultivars using these six markers revealed that the test materials were classified into four groups.This indicated that the developed SSR molecular markers were able to analyze the genetic background differences of adzuki bean and provide usable molecular markers for germplasm identification,genetic diversity analysis and gene mapping for good traits.

关 键 词：小豆 转录组 SSR 分子标记 

分 类 号：S521[农业科学—作物学] S388