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作物学报 ›› 2019, Vol. 45 ›› Issue (2): 196-203.doi: 10.3724/SP.J.1006.2019.84100

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

基于重测序的陆地棉InDel标记开发与评价

吴迷,汪念,沈超,黄聪,温天旺,林忠旭()   

  1. 华中农业大学植物科学技术学院 / 作物遗传改良国家重点实验室, 湖北武汉 430070
  • 收稿日期:2018-07-19 接受日期:2018-10-08 出版日期:2019-02-12 网络出版日期:2018-11-16
  • 通讯作者: 林忠旭
  • 基金资助:
    本研究由湖北省技术创新专项资助(2018ABA082)

Development and evaluation of InDel markers in cotton based on whole-genome re-sequencing data

Mi WU,Nian WANG,Chao SHEN,Cong HUANG,Tian-Wang WEN,Zhong-Xu LIN()   

  1. National Key Laboratory of Crop Genetic Improvement / College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei, China
  • Received:2018-07-19 Accepted:2018-10-08 Published:2019-02-12 Published online:2018-11-16
  • Contact: Zhong-Xu LIN
  • Supported by:
    This study was supported by the Technology Innovation Program of Hubei Province(2018ABA082)

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摘要：

碱基插入/缺失(InDel)是基因组中丰富的遗传变异形式。InDel以其密度高、易于基因型分型等优点成为分子标记开发的理想来源。本研究利用262份陆地棉品系重测序数据鉴定的InDel位点, 在全基因组范围内设计了3206个InDel标记并挑选均匀分布的320个标记进行验证。320个标记筛选出87个多态性标记, 多态性率为26.88%。利用多态性标记对不同地理来源的262份陆地棉种质资源进行基因分型, 共检测到160个等位位点; 多态性信息含量(PIC)为0.0836~0.3750, 平均值为0.3073; 基因多样性指数变异范围为0.0874~0.5000, 平均值为0.3876, 表明我国陆地棉遗传基础相对狭窄。群体结构分析将262份陆地棉品系大致划分为2个亚群, 聚类分析和主成分分析的结果与之基本一致。采用混合线性模型(Mixed linear model)对6个纤维品质性状的关联分析检测到65个关联位点(P < 0.01), 各位点对表型变异贡献率为2.57%~8.12%。本研究旨在利用重测序数据开发全基因组范围的可用于凝胶检测的InDel标记, 为棉花种质资源研究和分子标记辅助选择育种提供便捷工具。

关键词: 陆地棉, InDel标记, 遗传多样性, 群体结构, 关联分析

Abstract:

Insertion and deletion (InDel) are abundant forms of genetic variation in the genome. InDel has been recognized as an ideal source for marker development due to its high-density distribution and genotyping efficiency. In this study, the whole genome re-sequencing data of 262 upland cotton accessions were applied to identify 3206 InDel markers, and 320 markers with uniform distribution across the genome were selected to be evaluated. Eighty-seven polymorphic markers were identified, accounting for 26.88% of screened markers. A total of 160 allelic loci were detected using the 87 polymorphic markers in the 262 upland cotton accessions with an average polymorphic information content (PIC) of 0.3073 (ranging from 0.0836 to 0.3750) and an average genetic diversity of 0.3876 (ranging from 0.0874 to 0.5000), indicating a relatively low genetic diversity. Population structure analysis revealed extensive admixture and identified two subgroups, clustering analysis and principal component analysis supported the subgroups identified by STRUCTURE. Association analysis were performed by MLM (Mixed linear model), and 65 marker loci were associated with fiber quality traits (P < 0.01), explaining 2.57%-8.12% of the phenotypic variation. Genome-wide and gel based InDel markers developed based on re-sequencing data in this study provide a facile tool for cotton germplasm resources research and molecular marker assisted selection breeding.

Key words: Upland cotton, InDel marker, genetic diversity, population structure, association analysis

图1

标记HAU_ID_D11-01在部分品系中扩增结果"

图2

K值与ln P(D)、ΔK值折线图 A: ln P(D)与K值变化折线图; B: ΔK值随K值变化折线图。"

图3

基于InDel标记的262份陆地棉品系群体结构图"

图4

基于InDel标记的262份陆地棉品系的PCA图 NIR: 中国西北内陆棉区; NSEMR: 中国北方特早熟棉区; SCR: 中国南方棉区; SU: 前苏联; USA: 美国; YRR: 中国黄河流域棉区; YtRR: 中国长江流域棉区。"

图5

基于遗传距离的系统发生树 红色线条: G1, 包含182份种质材料; 绿色线条: G2, 包含80份种质材料。"

表1

多效应标记位点"

标记位点
Marker locus		 染色体
Chromosome		 位置
Position (bp)		 性状
Trait
HAU_ID_A01-15 A01 93250996 FUHML, FU, SF
HAU_ID_A03-09 A03 57007522 FE, FU, SF
HAU_ID_A08-06 A08 39475602 FE, MV
HAU_ID_A08-07 A08 47285210 FE, MV
HAU_ID_A08-14 A08 98729870 FU, SF
HAU_ID_A09-04 A09 23151235 FUHML, FS
HAU_ID_A10-01 A10 1109653 MV, SF
HAU_ID_A10-09 A10 57152725 FS, FU, MV
HAU_ID_D01-10 D01 43635771 FS, FU
HAU_ID_D02-01 D02 1069504 FUHML, FE
HAU_ID_D02-08 D02 45682280 MV, SF
HAU_ID_D04-07 D04 47451029 FE, MV
HAU_ID_D06-06 D06 35163294 FUHML, FU, SF
HAU_ID_D06-07 D06 39113925 FUHML, FU, SF
HAU_ID_D07-04 D07 23469098 FU, SF
HAU_ID_D07-09 D07 47924875 FUHML, FS, FE, FU, SF
HAU_ID_D09-10 D09 49075327 FUHML, SF
HAU_ID_D11-13 D11 65414930 FE, SF
HAU_ID_D12-10 D12 51191458 FUHML, FS, FU, SF
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