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作物学报 ›› 2022, Vol. 48 ›› Issue (3): 580-589.doi: 10.3724/SP.J.1006.2022.11015

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

小麦矮秆突变体je0098的遗传分析与其矮秆基因定位

付美玉1,2(), 熊宏春2, 周春云2, 郭会君2, 谢永盾2, 赵林姝2, 古佳玉2, 赵世荣2, 丁玉萍2, 徐延浩1,*(), 刘录祥2,*()   

  1. 1长江大学农学院/主要粮食作物产业化湖北省协同创新中心, 湖北荆州 434025
    2中国农业科学院作物科学研究所 /农作物基因资源与基因改良国家重大科学工程/国家农作物航天诱变技术改良中心, 北京 100081
  • 收稿日期:2021-02-03 接受日期:2021-06-16 出版日期:2022-03-12 网络出版日期:2021-06-28
  • 通讯作者: 徐延浩,刘录祥
  • 作者简介:E-mail: 17862825179@163.com
  • 基金资助:
    国家自然科学基金项目(31801346);中国农业科学院基本科研业务费专项(Y2020YJ09);国家重点研发计划项目(2016YFD0102100)

Genetic analysis of wheat dwarf mutant je0098 and molecular mapping of dwarfing gene

FU Mei-Yu1,2(), XIONG Hong-Chun2, ZHOU Chun-Yun2, GUO Hui-Jun2, XIE Yong-Dun2, ZHAO Lin-Shu2, GU Jia-Yu2, ZHAO Shi-Rong2, DING Yu-Ping2, XU Yan-Hao1,*(), LIU Lu-Xiang2,*()   

  1. 1Hubei Collaborative Innovation Center for Grain Industry/College of Agriculture, Yangtze University, Jingzhou 434025, Hubei, China
    2Institute of Crop Sciences, Chinese Academy of Agricultural Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement/National Center of Space Mutagenesis for Crop Improvement, Beijing 100081, China
  • Received:2021-02-03 Accepted:2021-06-16 Published:2022-03-12 Published online:2021-06-28
  • Contact: XU Yan-Hao,LIU Lu-Xiang
  • Supported by:
    National Natural Science Foundation of China(31801346);Chinese Academy of Agricultural Sciences Basal Research Fund(Y2020YJ09);National Key Research and Development Program of China(2016YFD0102100)

摘要:

倒伏易引发小麦严重减产, 发掘和利用优异矮秆基因是培育高产抗倒伏小麦新品种的关键。本研究以京411 (WT)及其经EMS诱变获得的产量相关性状优良的矮秆突变体je0098为试验材料, 对其株高进行遗传分析, 结合外显子捕获测序和遗传连锁分析定位矮秆基因。3年田间株高数据统计分析表明, je0098与WT相比株高降低15 cm, 组织细胞学观察结果显示, je0098与WT相比节间细胞长度缩短18%, 暗示je0098的矮化是由于节间细胞长度变短所致; 赤霉素敏感性分析表明, je0098为赤霉素敏感型矮秆突变体。利用WT和je0098杂交构建的由344个单株组成的F2分离群体, 结合F2:3家系表型数据, 选取矮秆纯合和高秆单株构建混池, 对两亲本和子代混池分别进行外显子捕获测序, 在2D染色体上定位到一个具有降秆效应的数量性状位点(QTL)。结合全基因组重测序所得SNP位点, 在2D染色体开发了6个KASP分子标记, 对F2单株进行基因分型。利用QTL IciMapping作图软件构建遗传连锁图谱, 结合3年田间表型数据, 将矮秆基因定位在20.77~28.84 Mb区间内, 遗传距离为11.48 cM。本研究结果为突变体je0098矮秆基因的功能研究以及育种利用奠定了基础。

关键词: 小麦, 株高, 矮秆基因, BSA, 分子标记

Abstract:

Lodging easily causes severe decrease in wheat yields. Identification and utilization of favorable dwarfing genes is the key to develop new varieties with high yield and lodging resistance. In this study, a dwarf mutant je0098 as material was induced by EMS mutagenesis from Jing 411 (WT) and had fine characteristics in yield components. We mapped the dwarfing gene through genetic analysis of plant height, and combining with exon capture sequencing and genetic linkage analysis. Statistical analyses of plant height in three-year field experiment suggested that plant height of je0098 was 15 cm lower than that of WT. Histocytological analysis of je0098 and WT indicated that the internode cell length of je0098 was about 18% shorter than that of WT, suggesting that the shorter internode cell length caused the dwarfism of je0098. Gibberellic acid treatment showed that je0098 was a gibberellic acid-sensitive dwarf mutant. An F2 segregation population consisting of 344 individuals was constructed by crossing WT and je0098. Combining with the phenotypic data of F2:3 families, dwarf homozygous and tall individuals were selected to construct progeny pools. Exon capture sequencing was performed on the two parents and progeny pools, respectively. A quantitative trait locus (QTL) with effects on reduced height was identified on chromosome 2D. Based on SNPs detected by genome-wide sequencing, six KASP markers were developed on chromosome 2D to genotype F2 individuals. Genetic linkage map was constructed using QTL IciMapping. Combining with phenotype data of three-year field experiment, the dwarfing gene was mapped in the range of 20.77-28.84 Mb with genetic distance of 11.48 cM. These results will lay the foundation for further functional research of je0098 and its application in wheat breeding.

Key words: wheat, plant height, dwarf gene, BSA, molecular marker

图1

野生型(WT)与突变体je0098的表型比较 *表示野生型与突变体之间在P < 0.05水平差异显著, **表示野生型与突变体之间在P < 0.01水平差异显著。"

图2

野生型(WT)与je0098的节间细胞长度和赤霉素敏感性比较 *表示在P < 0.05水平差异显著, **表示在P < 0.01水平差异显著。"

表1

两年F2:3家系株高统计"

年份
Year
表型
Phenotype
观察值
Observed count (O)
期望值
Expected count (E)
(O-E)2/E χ2 P (df=1)
2019 矮秆表型 Dwarf phenotype 68 83.5 2.877 3.836 0.050
非矮秆表型 Non-dwarf phenotype 266 250.5 0.959
合计 Total 334 334.0 3.836
2020 矮秆表型 Dwarf phenotype 69 82.5 2.209 2.945 0.086
非矮秆表型 Non-dwarf phenotype 261 247.5 0.736
合计 Total 330 330.0 2.945

图3

欧氏距离关联分析拟合图 红色虚线表示拟合值的99百分位数。"

图4

2D染色体上QTL连锁图谱与相应物理图谱 红线表示以2018年J411/je0098 F2群体单株株高为表型检测到的LOD曲线, 绿线和蓝线分别表示以2019年和2020年J411/je0098 F2:3家系株高平均值为表型检测到的LOD曲线。"

图5

附图1 2015年WT和je0098的农艺性状比较 WT和je0098在中圃场和昌平试验点的千粒重和穗长: (A)千粒重; (B)穗长。**表示在P < 0.01水平差异显著。"

表2

候选区间SNP位点功能预测"

基因
Gene
突变位点
Position (bp)
野生型
Wild type
突变型
Mutated type
突变类型
Mutation type
基因功能
Gene function
TraesCS2D01G059800 24,925,674 C T 错义突变
Missense mutant
RNA结合家族蛋白
RNA-binding family protein
TraesCS2D01G062900 26,616,391 C T 错义突变
Missense mutant
PHD锌指蛋白
PHD finger protein
TraesCS2D01G052400LC 22,812,351 C T UTR突变
UTR mutant
MBOAT家族蛋白
MBOAT family protein
TraesCS2D01G053600LC 23,479,083 C T 内含子突变
Intron mutant
类烯醇化酶
Enolase-like
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