Journal of Nuclear Agricultural Sciences ›› 2021, Vol. 35 ›› Issue (9): 1953-1963.DOI: 10.11869/j.issn.100-8551.2021.09.1953

• Induced Mutations for Plant Breeding·Agricultural Biotechnology •     Next Articles

Fine Mapping of Node Numbers on the Main Stem QTLs in Soybean Based on BAS and SLAF-Seq

YANG Yuhua1(), BAI Zhiyuan1, WEI Baoguo1, LEI Yang2, ZHANG Ruijun1,*()   

  1. 1 Center for Agricultural Genetic Resources Research, Shanxi Agricultural University/Institute of Crop Germplasm Resources, Shanxi Academy of Agricultural Sciences, Key Laboratory of Crop Gene Resources and Germplasm Enhancement on Loess Plateau, Ministry of Agriculture, Shanxi Key Laboratory of Genetic Resources and Genetic Improvement of Minor Crops, Taiyuan, Shanxi 030031
    2 College of Horticulture, Shanxi Agricultural University, Taiyuan, Shanxi 030031
  • Received:2020-08-20 Accepted:2020-10-04 Online:2021-09-10 Published:2021-07-22
  • Contact: ZHANG Ruijun

基于BSA和SLAF-Seq技术对大豆主茎节数QTL精细定位

杨玉花1(), 白志元1, 卫保国1, 雷阳2, 张瑞军1,*()   

  1. 1 山西农业大学农业基因资源研究中心/农业部黄土高原作物基因资源与种质创制重点实验室/杂粮种质资源发掘与遗传改良山西省重点实验室,山西 太原 030031
    2 山西农业大学园艺学院,山西 太原 030031
  • 通讯作者: 张瑞军
  • 作者简介:杨玉花,女,助理研究员,主要从事大豆种质资源研究。E-mail: yhms0325@163.com
  • 基金资助:
    国家重点研发计划项目(2016YFD0101500);国家重点研发计划项目(2016YFD0101504);山西省青年基金项目(201901D211563);山西省农业科学院博士后基金(YCX2020BH4)

Abstract:

To provide technical supports for marker assisted breeding and accelerate the cloning and functional verification of candidate genes of node numbers on the main stem in soybean, high throughput sequencing was used to detect the association region (QTL), fine mapping of the QTL were then conducted with the InDel markers developed based on the re-sequencing of parents. In this study, 102 recombinant inbred lines (RIL) derived from the cross between C025 with few node numbers on the main stem and Zhong119 with many node numbers on the main stem were used as experimental materials. Two mixed pools were constructed from 30 inbred lines with extremely few or extremely many node numbers on the main stem respectively. Five quantitative trait locus (QTL) related to the node number on the main stem of soybean were detected on chromosome 4 by the Bulked segregant analysis (BSA) and specific-locus amplified fragment sequencing (SLAF-Seq) high throughput sequencing method. In order to further narrow down the association regions, insertion-deletion (InDel) information between the association regions were obtained according to the re-sequencing of parental lines and the InDel markers were developed. The genotype of F 2 population was firstly analyzed by the InDel markers, and the major locus was mapped in the third associated region. Then eight co-dominant InDel markers were developed in the major region and all RIL lines were genotyped, nine individuals were obtained and the major region was divided into six exchange types. Combined with phenotypic analysis, the node number on the main stem was mapped between InDel markers Chr04-38 and Chr04-46, a region with only 171.9 kb, including six candidate genes. Therefore, the major locus of the node number on the main stem of soybean main stem was fine mapped. In short, the combination of BSA and high-throughput sequencing can effectively and rapidly detect the association regions of the node number on the main stem of soybean, furtherly combined the re-sequencing of parents to develop the tightly linked InDel markers can realize fine mapping of the major region. The developed InDel Markers Chr04-38 and Chr04-46 are tightly linked with the node numbers on the main stem of soybean, which can be used in the molecular marker assisted breeding for node numbers on the main stem in soybean.

Key words: soybean, node numbers on the main stem, BSA, SLAF-Seq, fine mapping, InDel markers

摘要:

为了加速大豆主茎节数候选基因的的克隆和功能验证,并为大豆主茎节数分子标记辅助育种提供分子基础,本研究通过高通量测序检测与大豆主茎节数相关数量性状区间(QTL),结合双亲重测序信息开发QTL区间InDel分子标记,实现了大豆主茎节数相关主效QTL区间的精细定位。本研究以少主茎节数C025材料为母本,多主茎节数中119为父本杂交衍生的重组自交系(RIL)102个株系为试验材料,取自交系中极端少主茎节数30株和极端多主茎节数30株,构建两个极端混池,利用传统分群分析法(BSA)和全基因组特异性位点扩增片段测序手段(SLAF-Seq)相结合的方法在4号染色体检测到与大豆主茎节数相关的5个QTL。为了进一步缩小QTL区间,依据双亲材料的高通量重测序信息,获取QTL区间的插入缺失位点(InDel)信息,并开发InDel标记。首先利用InDel标记在F2群体进行基因型分析,结果主效位点落在第3个QTL区间。其次在主效区间开发8个共显性InDel标记,结合RIL群体全部株系进行表型鉴定,最终获得9个交换单株,将主效区间分为6种交换类型,结合表型分析最终将大豆主茎节数位点精细定位到InDel标记Chr04-38和Chr04-46之间,其区间只有171.9 kb,包含候选基因6个,实现了大豆主茎节数的精细定位。本研究通过高通量测序与极端混池相结合的方法可以高效快速地检测与大豆主茎节数相关区间,并结合双亲重测序信息开发关联区间InDel分子标记,精细定位大豆主茎节数。本研究开发的Indel标记Chr04-38和Chr04-46与大豆主茎节数紧密连锁,有利于后期大豆主茎节数分子标记辅助育种。

关键词: 大豆, 主茎节数, BSA, SLAF-Seq, 精细定位, InDel标记