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Cloning and functional analysis of ZmGRAS13 gene in maize

SHE Meng1,2,ZHENG Deng-Yu2,KE Zhao2,WU Zhong-Yi2,ZOU Hua-Wen1,*,ZHANG Zhong-Bao2,*   

  1. 1 College of Agriculture, Yangtze University, Jingzhou 434025, Hubei, China; 2 Institute of Biotechnology, Beijing Academy of Agriculture and Forestry Sciences, Beijing Key Laboratory of Agricultural Gene Resources and Biotechnology, Beijing 100097, China
  • Received:2023-10-18 Revised:2024-01-12 Accepted:2024-01-12 Published:2024-02-19
  • Supported by:
    This study was supported by the Beijing Natural Science Foundation (6222009), the Beijing Academy of Agricultural and Forestry Sciences (KJCX20230203), and the Beijing Academy of Agricultural and Forestry Sciences Biotechnology Sharing Platform in 2023.

Abstract:

GRAS family is a plant-specific transcription factor, which plays an important role in regulating plant growth and development and responding to stresses. Exploring the function of GRAS family genes in maize (Zea mays L.) provides the important genetic resources for the creation of new maize germplasm. In this study, ZmGRAS13 gene (Zm00001eb401210) was cloned, and its basic characteristics, tissue expression characteristics, and the relative expression patterns under stresses were analyzed by bioinformatics and qRT-PCR. Bioinformatics showed that the full-length coding sequence of this gene was 1638 bp, encoding 545 amino acids. ZmGRAS13 protein had no transmembrane structure, and the molecular weight of 60.79 kD, the theoretical isoelectric point of 5.86, and had a conserved domain unique to the GRAS family. The analysis of 2 kb sequence upstream of the gene promoter indicated that the sequence contained cis-acting elements related to stresses, hormone response, and light response. The qRT-PCR analysis showed that ZmGRAS13 gene was expressed in different tissues of maize, and the relative expression level in stem was the highest. At the same time, the gene has different degrees of induced expression under different abiotic stress treatment conditions. The transient expression experiment of maize protoplasts demonstrated that ZmGRAS13 protein was localized in the nucleus. On 1/2 MS solid medium containing different concentrations of NaCl, mannitol, abscisic acid (ABA), jasmonic acid (JA), and salicylic acid (SA), respectively, the root length of ZmGRAS13 transgenic Arabidopsis lines was significantly longer than the control. In the soil, transgenic Arabidopsis lines grew better than the control under high salt and drought treatments, and the green leaf rate was higher than the control. Compared with the wild type, the content of stress resistance physiological index MDA decreased, the chlorophyll content increased, and the activities of POD and CAT increased in the transgenic ZmGRAS13 Arabidopsis thaliana, and the difference was significant difference. In conclusion, ZmGRAS13 gene may be involved in the regulation of maize growth and development, response to stresses and hormone signal transduction pathway. This study provides an important reference for the further analysis of the biological function of ZmGRAS13 in maize.

Key words: maize, ZmGRAS13, transcription factors, osmotic stress, salt stress, heterologous expression

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