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作物学报 ›› 2025, Vol. 51 ›› Issue (7): 1827-1837.doi: 10.3724/SP.J.1006.2025.43028

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

转录因子ZmMYB153通过ABA信号调节气孔运动增强玉米苗期抗旱性

张建鹏1(), 王国瑞3, 别海2, 叶飞宇3, 马晨晨3, 梁小菡3, 鲁晓民3, 尚霄丽4,*(), 曹丽茹3,*()   

  1. 1濮阳职业技术学院, 河南濮阳 457000
    2郑州市农业科技研究院, 河南郑州 450002
    3河南省农业科学院粮食作物研究所, 河南郑州 450002
    4河南农业大学, 河南郑州 450002
  • 收稿日期:2024-07-03 接受日期:2024-12-12 出版日期:2025-07-12 网络出版日期:2025-01-10
  • 通讯作者: *曹丽茹, E-mail: caoliru008@126.com; 尚霄丽, E-mail: xiaoli820218@163.com
  • 作者简介:E-mail: jianpeng0218@163.com
  • 基金资助:
    国家自然科学基金项目(32201708);河南省重大科技专项(221100110300);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-02-08)

Transcription factor ZmMYB153 enhances drought tolerance in maize seedlings by regulating stomatal movement through ABA signaling

ZHANG Jian-Peng1(), WANG Guo-Rui3, BIE Hai2, YE Fei-Yu3, MA Chen-Chen3, LIANG Xiao-Han3, LU Xiao-Min3, SHANG Xiao-Li4,*(), CAO Li-Ru3,*()   

  1. 1Puyang Vocational and Technical College, Puyang 457000, Henan, China
    2Zhengzhou Agricultural Science and Technology Research Institute, Zhengzhou 450002, Henan, China
    3Grain Crop Research Institute, Henan Academy of Agricultural Sciences, Zhengzhou 450002, Henan, China
    4Henan Agricultural University, Zhengzhou 450002, Henan, China
  • Received:2024-07-03 Accepted:2024-12-12 Published:2025-07-12 Published online:2025-01-10
  • Contact: *E-mail: caoliru008@126.com; E-mail: xiaoli820218@163.com
  • Supported by:
    National Natural Science Foundation of China(32201708);Major Science and Technology Project of Henan Province(221100110300);China Agriculture Research System of MOF and MARA(CARS-02-08)

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摘要：

干旱影响玉米的生长与发育, 最终导致产量损失。为挖掘玉米抗旱关键基因并解析其抗旱分子机制, 从玉米干旱-复水转录组中鉴定到1个响应干旱胁迫基因GRMZM2G050550, 命名为ZmMYB153。基因进化树及亚细胞定位分析发现, ZmMYB153蛋白与其他物种中的同源蛋白具有高度的相似性, ZmMYB153定位在细胞核中。组织表达模式分析显示, 该基因在叶片中的表达量最高。PEG模拟干旱条件下, ZmMYB153基因的表达水平显著上升。脱落酸(abscisic acid, ABA)处理条件下, 该基因的表达呈现出先升后降的趋势。通过分析干旱胁迫下不同抗旱强度自交系中ZmMYB153基因的表达量, 发现该基因在强抗旱玉米自交系郑6722 (Z6722)中的表达量显著高于其在中抗旱玉米自交系B73中的表达量。为进一步验证ZmMYB153基因的生物学功能, 利用转基因技术获得该基因的过表达株系。干旱胁迫后, 与野生型(wild type, WT)相比, 过表达ZmMYB153植株(overexpression line-1, OE-1; overexpression line-2, OE-2)具有较高的叶片相对含水量(relative water content, RWC), 超氧化物歧化酶(superoxide dismutase, SOD)、过氧化物酶(peroxidase, POD)和过氧化氢酶(catalase, CAT)等氧化酶酶活性, 具有较低的离子渗透率以及丙二醛(malondialdehyde, MDA)含量。通过观察干旱条件下的叶片气孔状态, 发现过表达ZmMYB153株系的气孔闭合程度显著高于WT, 失水率显著低于WT。这些结果表明, ZmMYB153基因具有正向调节玉米响应干旱胁迫的功能。干旱胁迫下的ZmNYB153过表达株系中ABA信号通路相关基因ZmABI1和ZmPYL10的表达水平被显著上调或者下调, 气孔运动相关基因ZmSLAC1的表达量显著高于WT。综上所述, ZmMYB153基因可能通过参与ABA信号途径, 调节叶片气孔活动, 从而影响植物的干旱耐受性。

关键词: 玉米, MYB, 干旱, 气孔, ABA

Abstract:

Drought significantly impacts corn growth and development, ultimately causing yield losses. To identify key drought-resistant genes in maize and elucidate their molecular mechanisms, we analyzed the maize drought-rehydration transcriptome and identified a drought-responsive gene, GRMZM2G050550, named ZmMYB153. Phylogenetic analysis and subcellular localization revealed that the ZmMYB153 protein shares a high degree of similarity with homologous proteins in other species and is localized in the nucleus. Tissue-specific expression analysis showed that ZmMYB153 is most highly expressed in leaves. Under polyethylene glycol (PEG)-simulated drought conditions, the expression of ZmMYB153 was significantly upregulated. Similarly, under abscisic acid (ABA) treatment, its expression followed a dynamic pattern, initially increasing and then decreasing over time. Expression analysis of ZmMYB153 in different drought-resistant maize inbred lines under drought stress revealed that its expression level in the highly drought-resistant inbred line Zheng 6722 (Z6722) was significantly higher than in the moderately drought-resistant inbred line B73. To further investigate the biological function of ZmMYB153, transgenic maize lines overexpressing the gene were generated. Under drought stress, compared to wild-type (WT) plants, ZmMYB153 overexpression lines (OE-1 and OE-2) exhibited higher leaf relative water content (RWC), enhanced activities of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), and lower ion leakage and malondialdehyde (MDA) content. Observations of leaf stomatal status under drought conditions revealed that stomatal closure was significantly greater in ZmMYB153 overexpression lines than in WT plants, leading to a significantly reduced water loss rate. These findings suggest that ZmMYB153 plays a positive regulatory role in maize responses to drought stress. Furthermore, under drought stress, the expression levels of ABA signaling pathway genes ZmABI1 and ZmPAYL10 were significantly altered in ZmMYB153 overexpression lines, while the expression of the stomatal movement-related gene ZmSLAC1 was significantly higher than in WT plants. In summary, ZmMYB153 likely regulates leaf stomatal activity through its involvement in the ABA signaling pathway, thereby enhancing drought tolerance in maize.

Key words: maize, MYB, drought, stoma, abscisic acid

表1

试验所用的引物序列"

基因名称
Gene ID		 引物名称
Primer name		 引物序列
Primer sequence (5'-3')		 目的
Purpose
GRMZM2G050550 ZmMYB153-ORF-F ATGGCGTCGTCTTCTCCGTCC ZmMYB153基因的获得
Acquisition of ZmMYB153 gene
ZmMYB153-ORF-R CTACTCGATCTTGGCCATACCC
GRMZM2G050550 ZmMYB153-OE-AscⅠ-F TTGGCGCGCCATGGCGTCGTCTTCTCCGTCC 过表达载体的构建
ZmMYB153-OE-BamHⅠ-R CCGGATCCCTACTCGATCTTGGCCATACCC Construction of overexpression vectors
GRMZM2G050550 ZmMYB153-GFP-SpeI-F GCACTAGTGCGTCGTCTTCTCCGTCC 亚细胞定位载体构建Construction of subcellular localization vectors
ZmMYB153-GFP-BamHI-R GCGGATCCCTCGATCTTGGCCATACCC
GRMZM2G050550 Q-ZmMYB153-F CCGACAACGCCATCAAGAAC 荧光定量PCR Real-time PCR
Q-ZmMYB153-R TGGTTGCTAGAGTCGCTGAG
GRMZM2G300125 Q-ZmABI1-F CACATCGTGGTCGCCAACT 荧光定量PCR Real-time PCR
Q-ZmABI1-R CCACCTGCTGATTCTACCCTTT
GRMZM2G063882 Q-ZmPYL10-F GGCAACACCAAGGACGAGA 荧光定量PCR Real-time PCR
Q-ZmPYL10-R CAGTAGCAGTAGCAGTCCCTCAAA
GRMZM2G106921 Q-ZmSLAC1-F CGGTGAAGCCGAATGACA 荧光定量PCR Real-time PCR
Q-ZmSLAC1-R TGATTGAGGACACGAGGGAG
18sRNA Zm-18S-F CCATCCCTCCGTAGTTAGCTTCT 荧光定量内参
Real-time PCR internal reference
Zm-18S-R CCTGTCGGCCAAGGCTATATAC

图1

ZmMYB153进化树分析和蛋白序列比对 A: ZmMYB153与不同物种中同源蛋白的进化树分析; B: ZmMYB153与不同物种中同源蛋白的序列比对。"

图2

ZmMYB153亚细胞定位分析 Merged: 叠加图; GFP: 绿色荧光蛋白图; Bright field: 明场; 35S::GFP为转入空载体的原生质体; 35S::ZmMYB153-GFP为转入目的基因载体的原生质体。标尺为10 μm。"

图3

ZmMYB153在不同组织及处理下的表达情况 A: ZmMYB153在20%PEG处理下的表达情况; B: ZmMYB153在ABA处理下的表达情况; C: ZmMYB153在不同组织或部位的表达情况。 不同小写字母表示二者之间具有显著性差异(P < 0.05)。"

图4

ZmMYB153在抗旱性不同材料中的表达情况 **表示在0.01概率水平差异显著。Z6722: 郑6722。"

图5

ZmMYB153的过表达提高了玉米的抗旱性 A: ZmMYB153的过表达株系在正常、干旱及复水下的表型鉴定, 标尺为5 cm; B: 离子渗透率测定; C: RWC测定。BT表示处理之前; D5表示处理干旱5d; R3表示复水3 d。RWC: 相对含水量。**表示在0.01概率水平差异显著。WT: 野生型; OE-1、OE-2: 过表达ZmMYBL53植株。"

图6

ZmMYB153的过表达株系在正常、干旱和复水下的生理指标测定 缩写同图5。**表示在0.01概率水平差异显著。"

图7

ZmMYB153过表达株系在正常, 干旱及复水下的气孔开闭情况和离体失水率测定 A: ZmMYB153过表达株系在正常、干旱及复水下的气孔开闭情况照片, 标尺为5 μm; B: ZmMYB153过表达株系在正常、干旱及复水下的气孔开闭情况量化统计; C: 叶片离体失水率。FO表示气孔完全开放, PO表示气孔部分开放, FC表示气孔完全闭合。缩写同图5。"

图8

在不同处理下ZmMYB153的过表达株系中抗旱相关基因的表达情况 缩写同图5。*表示在0.05概率水平上差异显著, **表示在0.01概率水平差异显著。"

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