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作物学报 ›› 2021, Vol. 47 ›› Issue (12): 2314-2323.doi: 10.3724/SP.J.1006.2021.04266

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

花生全基因组抗病基因鉴定及其对青枯菌侵染的响应分析

张欢**(), 罗怀勇**(), 李威涛, 郭建斌, 陈伟刚, 周小静, 黄莉, 刘念, 晏立英, 雷永, 廖伯寿, 姜慧芳*()   

  1. 中国农业科学院油料作物研究所 / 农业农村部油料作物生物学与遗传育种重点实验室, 湖北武汉 430062
  • 收稿日期:2020-12-08 接受日期:2021-04-14 出版日期:2021-12-12 网络出版日期:2021-06-15
  • 通讯作者: 姜慧芳
  • 作者简介:张欢, E-mail: 18198335427@163.com;
    罗怀勇, E-mail: huaiyongluo@caas.cn第一联系人：**同等贡献
  • 基金资助:
    国家自然科学基金项目(31761143005);国家自然科学基金项目(31971903);中央级科研院所基本业务费专项(1610172019008)

Genome-wide identification of peanut resistance genes and their response to Ralstonia solanacearum infection

ZHANG Huan**(), LUO Huai-Yong**(), LI Wei-Tao, GUO Jian-Bin, CHEN Wei-Gang, ZHOU Xiao-Jing, HUANG Li, LIU Nian, YAN Li-Ying, LEI Yong, LIAO Bo-Shou, JIANG Hui-Fang*()   

  1. Oil Crops Research Institute, Chinese Academy of Agricultural Sciences / Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture and Rural Affairs, Wuhan 430062, Hubei, China
  • Received:2020-12-08 Accepted:2021-04-14 Published:2021-12-12 Published online:2021-06-15
  • Contact: JIANG Hui-Fang
  • About author:First author contact:**Contributed equally to this work
  • Supported by:
    National Natural Science Foundation of China(31761143005);National Natural Science Foundation of China(31971903);Central Public-interest Scientific Institution Basal Research Fund(1610172019008)

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摘要：

花生是主要的油料作物之一, 在生产过程中受到多种病原微生物的危害。培育和选用抗病品种是防治病害最经济有效的途径之一, 而抗病基因是植物抵御病原微生物的重要基因。本文首次对花生抗病基因进行全基因组鉴定, 发掘抗病候选基因4156个, 其中RLK、RLP、NL、CNL、TNL这5种典型抗病基因分别有536、490、232、182和149个。抗病基因在染色体上分布不均匀, 多数抗病基因集中在B02染色体上。转录组测序发现, 抗病材料中特异表达的基因有111个, 感病材料中特异表达的基因有104个, 抗、感病材料均有表达的基因2216个、均不表达的有1725个。筛选出第1类响应青枯菌诱导的抗病基因5个, 第2类持续上调表达抗青枯病基因65个。qRT-PCR成功验证了1个抗病候选基因Arahy.5D95TJ。本文对花生抗病基因的鉴定分析, 为后续研究抗病基因功能与花生抗病的分子育种提供重要参考。

关键词: 花生, 抗病基因, 青枯病, 转录组

Abstract:

Peanut is one of the main oil crops, which is harmed by many pathogenic microorganisms during growth and development period. Breeding and selection of disease-resistant varieties is one of the most economical and effective ways to control disease, and disease resistance genes are important genes for plant resistance to pathogenic microorganisms. Here, the whole genome-wide identification of peanut disease resistance genes was carried out for the first time. A total of 4156 candidate disease resistance genes were identified. Among them, 536, 490, 232, 182, and 149 genes were RLK, RLP, NL, CNL, and TNL, respectively. The distribution of disease resistance genes was uneven on chromosomes, and most of them were concentrated on chromosome B02. Transcriptome profiling revealed that 111 genes were specifically expressed in resistant materials, 104 genes were specifically expressed in susceptible materials, 2216 genes were expressed in both resistant and susceptible materials, while 1725 genes were not expressed in both resistant and susceptible materials. Two kinds of differentiate expressed R genes were identified, including five genes in the first group responded to the infection of Ralstonia solanacearum at specific time and 65 genes in the second group which exhibited higher expressions in resistant cultivar than susceptible cultivar. A candidate gene Arahy.5D95TJ was successfully validated by qRT-PCR. In this study, the identification and analysis of peanut disease resistance genes provides the important reference for further research of their functions and molecular breeding of peanut disease resistance.

Key words: Arachis hypogaea, disease resistance gene, bacterial wilt, transcriptome profiling

表1

花生R基因的数量和分类"

类别
Type		 包含结构域
Domain contained		 基因数
Gene number		 占R基因比例
Proportion in R gene (%)
RLK TM, LRR, Kinase 536 12.90
RLP TM, LRR 490 11.79
CNL CC, TM, NBS, LRR 182 4.38
TNL TM, TIR, NBS, LRR 149 3.59
KIN TM, Kinase 1714 41.24
NL TM, NBS, LRR 232 5.58
CK CC, TM, Kinase 221 5.32
N TM, NBS 146 3.51
CN CC, TM, NBS 136 3.27
CTNL CC, TM, TIR, NBS, LRR 77 1.85
L LRR 76 1.83
T TM, TIR 65 1.57
TN TM, TIR, NBS 61 1.47
CL CC, TM, LRR 28 0.67
CLK CC, TM, LRR, Kinase 20 0.48
CNT CC, TM, NBS, TIR 15 0.36
CT CC, TM, TIR 6 0.14
TRAN TM 2 0.05
总计Total 4156

图1

花生R基因在染色体上的分布 绿色代表在抗病和感病材料中都表达; 红色是只在抗病材料中表达的抗病基因; 紫色是只在感病材料中表达的抗病基因; 黑色代表抗病基因在抗感材料中都不表达。"

图2

花生R基因FPKM值分布范围 RC: 抗病材料对照; RT: 抗病材料处理; SC: 感病材料对照; ST: 感病材料处理。"

图3

花生接种青枯菌后差异表达的2类R基因 RC: 抗病材料对照; RT: 抗病材料处理; SC: 感病材料对照; ST: 感病材料处理。RT12、RT24、RT48、RT72和RT96分别表示抗病材料接种青枯菌后12、24、48、72和96 h的样本; RC12、RC24、RC48、RC72和RC96分别表示抗病材料对照组(清水处理) 12、24、48、72和96 h的样本; ST12、ST24、ST48、ST72和ST96分别表示感病材料接种青枯菌后12、24、48、72和96 h的样本; SC12、SC24、SC48、SC72和SC96分别表示感病材料对照组(清水处理) 12、24、48、72和96 h的样本。"

图4

主效抗青枯病QTL区间差异表达R基因及验证 缩写同图3。"

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