Journal of Integrative Agriculture ›› 2024, Vol. 23 ›› Issue (7): 2164-2177.DOI: 10.1016/j.jia.2023.07.023

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普通小麦P型质膜H+-ATP酶的鉴定以及与种子休眠和萌发相关的TaHA7的功能描述

  

  • 收稿日期:2023-05-03 接受日期:2023-07-04 出版日期:2024-07-20 发布日期:2024-07-01

Identification of P-type plasma membrane H+-ATPases in common wheat and characterization of TaHA7 associated with seed dormancy and germination

Bingli Jiang*, Wei Gao*, Yating Jiang, Shengnan Yan, Jiajia Cao, Litian Zhang, Yue Zhang, Jie Lu, Chuanxi Ma, Cheng Chang#, Haiping Zhang#   

  1. Key Laboratory of Wheat Biology and Genetic Improvement on Southern Yellow & Huai River Valley, Ministry of Agriculture and Rural Affairs/College of Agronomy, Anhui Agricultural University, Hefei 230036, China
  • Received:2023-05-03 Accepted:2023-07-04 Online:2024-07-20 Published:2024-07-01
  • About author:Bingli Jiang, E-mail: 3104764960@qq.com; Wei Gao, E-mail: 1226575882@qq.com; #Correspondence Haiping Zhang, E-mail: zhhp20@163.com; Cheng Chang, E-mail: changtgw@126.com * These authors contributed equally to this study.
  • Supported by:
    This work was supported by grants from the University Synergy Innovation Program of Anhui Province, China (GXXT-2021-058), the National Natural Science Foundation of China (U20A2033), the Natural Science Foundation of Anhui Province, China (2108085MC98), the Key Scientific and Technological Breakthroughs of Anhui Province, China (2021d06050003), the Anhui Province Education Department Sciences Research Project, China (YJS20210212), the Scientific Research Project of Higher Education in Anhui Province, China (2022AH050924 and 2022AH050885), and the Jiangsu Collaborative Innovation Center for Modern Crop Production, China (JCIC-MCP).

摘要:

穗发芽严重降低小麦产量、品质以及种子活力。种子休眠性弱或缺乏是穗发芽发生的根本原因,但相关分子机制尚不清楚。 P型质膜(PM)H+-ATP酶(HAs)是植物生长、发育和抵御逆境的核心因子。尽管HAs在多种植物中的功能已被验证,但其在小麦种子萌发和休眠中的作用尚不清楚。本研究利用生物信息学方法在小麦中共鉴定了28个P型PM H+-ATP酶家族成员(命名为TaHA1-28),并将其分为5个亚族。综合转录组和基因公共表达数据库分析结果,共筛选出6个候选基因(TaHA7/-12/-14/-16/-18/-20)可能参与调控小麦种子休眠和萌发。其中,TaHA7的表达量随着种子萌发的进程而增加,推测TaHA7可能参与了小麦种子休眠和萌发的调控。随后,在强休眠和弱休眠小麦品种中对TaHA7进行全长克隆、序列变异分析、功能标记开发,并利用2个自然群体共同验证了该基因序列变异与种子休眠和萌发表型显著相关。同时,利用济麦22和京411背景的EMS突变体以及转基因拟南芥株系进一步验证了TaHA7的2种等位变异与种子休眠和萌发表型显著相关。此外,我们测定了济麦22和京411背景的EMS突变体和转基因拟南芥种子中P型PM H+-ATP酶的活性以及TaHA7拟南芥同源基因突变体和回补株系中GA和ABA信号通路中关键基因的相对表达量,推测TaHA7很可能通过影响H+-ATP酶的活性以及GA和ABA的合成、代谢以及信号转导途径,最终调控小麦种子休眠和萌发。本文结果为深入研究TaHAs基因调控小麦种子休眠和萌发的分子机制奠定了重要的理论基础。

Abstract:

The P-type plasma membrane (PM) H+-ATPases (HAs) are crucial for plant development, growth, and defense.  The HAs have been thoroughly characterized in many different plants.  However, despite their importance, the functions of HAs in germination and seed dormancy (SD) have not been validated in wheat.  Here, we identified 28 TaHA genes (TaHA1-28) in common wheat, which were divided into five subfamilies.  An examination of gene expression in strong- and weak-SD wheat varieties led to the discovery of six candidate genes (TaHA7/-12/-14/-16/-18/-20).  Based on a single nucleotide polymorphism (SNP) mutation (C/T) in the TaHA7 coding region, a CAPS marker (HA7) was developed and validated in 168 wheat varieties and 171 Chinese mini-core collections that exhibit diverse germination and SD phenotypes.  We further verified the roles of the two allelic variations of TaHA7 in germination and SD using wheat mutants mutagenized with ethyl methane sulphonate (EMS) in ‘Jimai 22’ and ‘Jing 411’ backgrounds, and in transgenic Arabidopsis lines.  TaHA7 appears to regulate germination and SD by mediating gibberellic acid (GA) and abscisic acid (ABA) signaling, metabolism, and biosynthesis.  The results presented here will enable future research regarding the TaHAs in wheat.


Key words: wheat ,  P-type plasma membrane H+-ATPase ,  seed dormancy ,  abscisic acid ,  gibberellin