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Acta Agronomica Sinica ›› 2025, Vol. 51 ›› Issue (7): 1827-1837.doi: 10.3724/SP.J.1006.2025.43028

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• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

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 Online:2025-07-12 Published: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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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

Table 1

Primer sequences used in the study"

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

Fig. 1

Evolution tree analysis and protein sequence alignment analysis of ZmMYB153 A: evolutionary tree analysis of homologous proteins of ZmMYB153 in different species; B: protein sequence alignment of ZmMYB153 homologs in different species."

Fig. 2

Subcellular localization results of ZmMYB153 protein Merged: overlay diagram; GFP: green fluorescent protein map; Bright field: bright field map; 335S::GFP is a protoplast transferred into an empty vector; 35S::ZmMYB153-GFP is the protoplast transferred into the target gene vector. Bar: 10 μm."

Fig. 3

Expression of ZmMYB153 in different tissues and treatments A: expression of ZmMYB153 at 20% PEG; B: expression of ZmMYB153 under ABA treatment; C: expression of ZmMYB153 in different tissue sites. Different lowercase letters indicate significant differences between the two (P < 0.05)."

Fig. 4

Expression of ZmMYB153 in materials differing in drought resistance ** indicates significant difference at the 0.01 probability level. Z6722: Zheng 6722."

Fig. 5

Overexpression of ZmMYB153 improved the drought resistance of maize A: phenotypic identification of overexpression strains of ZmMYB153 under normal, drought, and rehydration conditions, bar: 5 cm; B: measurement of ion permeability; C: RWC determination. BT represents before treatment; D5 represents the treatment of drought for five days; R3 represents rehydration for three days. RWC: relative water content. ** indicates significant difference at the 0.01 probability level. WT: wild type; OE-1, OE-2: overexpression line-1 and line-2."

Fig. 6

Physiological indicators of overexpressed ZmMYB153 strain under normal, drought and rehydration conditions Abbreviations are the same as those given in Fig.5. ** indicates significant difference at the 0.01 probability level."

Fig. 7

Measurement of stomatal opening and closing and in vitro water loss rate of ZmMYB153 overexpression strain under normal, drought, and rehydration conditions A: Photos of stomatal opening and closing of ZmMYB153 overexpressing strains under normal, drought and rehydration conditions, bar: 5 μm; B: quantitative statistics of stomatal opening and closing of ZmMYB153 overexpressing strains under normal, drought and rehydration conditions; C: leaf dehydration rate in vitro. FO indicates that the pores are completely open, PO indicates pores are partial opening, and FC indicates pores are complete closure, respectively. Abbreviations are the same as those given in Fig. 5."

Fig. 8

Expression of drought resistance related genes in overexpression strains of ZmMYB153 under different treatments Abbreviations are the same as those given in Fig. 5. * indicates a significant difference at the 0.05 probability level, and ** indicates a significant difference at the 0.01 probability level."

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