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作物学报 ›› 2024, Vol. 50 ›› Issue (8): 1896-1906.doi: 10.3724/SP.J.1006.2024.43007

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

基于大刍草渗入系的玉米抗旱优异等位基因挖掘

刘爽(), 李珅, 王东梅, 沙小茜, 何冠华, 张登峰, 李永祥, 刘旭洋, 王天宇, 黎裕, 李春辉()   

  1. 作物基因资源与育种全国重点实验室 / 中国农业科学院作物科学研究所, 北京 100081
  • 收稿日期:2024-01-24 接受日期:2024-04-01 出版日期:2024-08-12 网络出版日期:2024-04-22
  • 通讯作者: * 李春辉, E-mail: lichunhui@caas.cn
  • 作者简介:E-mail: 1971556590@qq.com
  • 基金资助:
    财政部和农业农村部国家现代农业产业技术体系建设专项(CARS-02-03);中国农业科学院科技创新工程项目(CAAS-ZDRW202004)

Superior allele genes mining for drought tolerance in maize based on introgression line from a cross between maize and teosinte

LIU Shuang(), LI Shen, WANG Dong-Mei, SHA Xiao-Qian, HE Guan-Hua, ZHANG Deng-Feng, LI Yong-Xiang, LIU Xu-Yang, WANG Tian-Yu, LI Yu, LI Chun-Hui()   

  1. State Key Laboratory of Crop Gene Resources and Breeding / Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2024-01-24 Accepted:2024-04-01 Published:2024-08-12 Published online:2024-04-22
  • Contact: * E-mail: lichunhui@caas.cn
  • Supported by:
    China Agriculture Research System of MOF and MARA(CARS-02-03);Innovation Program of Chinese Academy of Agricultural Sciences(CAAS-ZDRW202004)

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摘要：

干旱是影响玉米生产的主要非生物胁迫之一。为了挖掘玉米优异抗旱基因, 本研究基于墨西哥大刍草和玉米自交系PH4CV构建的BC2F6群体, 通过苗期抗旱性初步鉴定, 筛选出抗旱性强的渗入系TP180。干旱胁迫后发现渗入系TP180较轮回亲本PH4CV萎蔫程度更小, 并且复水后存活率显著高于PH4CV。全基因组基因型鉴定发现, 渗入系TP180含有0.6%的墨西哥大刍草导入。通过开展TP180和PH4CV不同水分条件下的转录组测序, 在渗入系TP180和PH4CV之间共鉴定了 2307个差异表达基因, 不同胁迫重叠差异表达基因共122个, 这些基因涉及生长素途径, 茉莉酸途径等, 包含多个转录因子。整合差异表达基因和含大刍草导入区段分析, 鉴定出2个抗旱候选基因Zm00001d033050和Zm00001d002025, 并进一步开展了RT-PCR分析。本研究为挖掘大刍草中抗旱基因资源提供了重要材料和信息基础。

关键词: 玉米, 大刍草, 渗入系, 抗旱, 转录组分析

Abstract:

Drought is one of the major abiotic stresses affecting maize production. In order to explore new genes for drought tolerance in maize, the BC2F6 population constructed on the basis of teosinte and PH4CV was screened for the drought-tolerant introgression lines TP180 through the preliminary identification of drought tolerance at seedling stage. After drought stress, the TP180 was less wilting than its recurrent parent PH4CV, and the survival rate of TP180 was significantly higher than PH4CV after rehydration. Genome-wide genotypic identification revealed that introgression line TP180 contained 0.6% of the teosinte genome. By transcriptomic analysis of TP180 and PH4CV under different water conditions, a total of 2307 differentially expressed genes were identified between TP180 and PH4CV, and 122 of the differentially expressed genes were identified under both two drought stresses conditions. These genes were related to the growth hormone pathway, jasmonic acid pathway, etc., and contained multiple transcription factors. Integrating the differentially expressed genes and the analysis of introgression region containing the teosinte genome, two drought-resistant candidate genes (Zm00001d033050 and Zm00001d002025) were identified and further validated by RT-PCR. This study provides an important germplasm and information for mining drought-resistant gene of the teosinte.

Key words: maize, teosinte, introgression line, drought tolerance, transcriptomic analysis

表1

本研究所使用的引物信息表"

引物名称Primer name 引物序列Primer sequence (5'-3')
GAPDH-F CCCTTCATCACCACGGACTAC
GAPDH-R AACCTTCTTGGCACCACCCT
ReTIFY9-F CAGCGGAACGTGCAGAAACA
ReTIFY9-R2 GCCTTGCGCTTCTCCATGAAC
ReEreb24-F CACCACTGCTACGCCAATG
ReEreb24-R ATTGAAAGCGACCGGGGTG

图1

TP180和PH4CV抗旱性分析与干旱处理存活率分析 A: 正常水分; B: 中度干旱胁迫; C: 重度干旱胁迫; D: 复水; E: 存活率。**表示在P < 0.01水平差异显著。"

图2

大刍草导入区段分析"

图3

不同处理下差异表达基因数目 A: Venn图描述所有差异表达基因的数量; B: 上调、下调的差异基因总数。WW: 正常水分; MD: 中度干旱胁迫; SD: 重度干旱胁迫。"

图4

不同处理下差异表达基因GO富集 A: 中度干旱胁迫上调表达基因; B: 中度干旱胁迫下调表达基因; C: 重度干旱胁迫上调表达基因; D: 重度干旱胁迫中度干旱胁迫下调表达基因。"

图5

差异表达中已知抗旱基因不同处理下表达量 FPKM代表差异显著基因的表达水平, WW代表正常水分(watered)条件下基因表达量, MD代表中度干旱胁迫(moderate-drought)条件下基因表达量, SD代表重度干旱胁迫(severe-drought)条件下基因表达量。*和**分别表示在P < 0.05和P < 0.01水平差异显著, ns表示无差异显著性。"

图6

候选抗旱基因不同处理下基因表达水平 A~B图为转录组数据, C~D图为RT-PCR实验结果, FPKM代表差异显著基因的表达量, Relative expression level为RT-PCR实验得到的基因表达量, WW代表正常水分(watered)条件下基因表达量, MD代表中度干旱胁迫(moderate-drought)条件下基因表达量, SD代表重度干旱胁迫(severe-drought)条件下基因表达量。*和**分别表示在P < 0.05和P < 0.01水平差异显著, ns表示无差异显著性。"

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