Cloning and Functional Analysis of the TaMYBS1-like Gene in Winter Wheat

doi:10.13560/j.cnki.biotech.bull.1985.2026-0028

Abstract

Abstract:

Objective Dongnong Winter Wheat 1 (Dn1) is the first highly cold-tolerant winter wheat (Triticum aestivum) variety suitable for large-scale cultivation in high-latitude cold regions, with an overwintering survival rate exceeding 85% after winter. This study was conducted to investigate the function of the TaMYBS1-like gene in winter wheat and its expression responses to low-temperature and drought stress. Methods The cDNA sequence of the TaMYBS1-like gene from winter wheat (Dn1) was cloned using RT-PCR and subjected to bioinformatic analysis. Samples of leaves and tiller nodes were collected under low-temperature treatments at 4, 0, -10, and -25 ℃. At the three-leaf stage, seedlings were treated with 20% PEG-6000, and leaf and tiller node samples were taken at 0, 3, 6, 12, and 24 h. RT-qPCR was used to analyze the gene expression patterns in leaves and tiller nodes under different low-temperature and drought treatments. An expression vector was constructed, and subcellular localization was performed to determine the protein localization. The TaMYBS1-like gene was transferred into Arabidopsis thaliana via Agrobacterium-mediated transformation for low-temperature and drought stress assays to validate its function. Results TaMYBS1-like belongs to the MYB transcription factor family. The full-length cDNA of TaMYBS1-like is 903 bp, encoding 300 amino acids. The encoded protein is an unstable hydrophilic protein primarily localized in the nucleus. Expression pattern analysis revealed that under low-temperature stress, TaMYBS1-like expression levels at 0, -10, and -25 ℃ were significantly higher compared to those at 4 ℃. In leaves, TaMYBS1-like expression was significantly higher at 3, 6, 12, and 24 h of drought stress compared to the control. Overexpression of TaMYBS1-like enhanced basic physiological metabolism, cell activity, and antioxidant enzyme system activity, while reducing cell membrane damage. Conclusion Overexpression of the winter wheat TaMYBS1-like gene improves plant cold and drought tolerance.

Key words: winter wheat, TaMYBS1-like gene, cold tolerance, drought tolerance

WANG Xin-yue, SHAO Qing-yi, CHEN Yu-shu, HU Yi-fan, ZHAO Nan, WANG Qiang, WANG Min-li, WANG Xue-song, JIN Zhong-min, LIU Li-jie. Cloning and Functional Analysis of the TaMYBS1-like Gene in Winter Wheat[J]. Biotechnology Bulletin, 2026, 42(6): 218-226.

Figures/Tables 7

Fig. 1 Secondary structure prediction curve (A) and tertiary structure (B) of the TaMYBS1-like protein

Fig. 2 Changes in TaMYBS1-like gene expression in leaves and tiller nodes of winter wheat under different low-temperature and drought stressesDifferent letters indicate significant differences at the 0.05 level (P0.05). The same below

Fig. 3 Subcellular localization of TaMYBS1-like in tobacco epidermal cells

Fig. 4 Phenotypic observation of TaMYBS1-like T3 transgenic Arabidopsis thaliana under low temperature stress

Fig. 5 Physiological indicators of WT and TaMYBS1-like T3 transgenic Arabidopsis under cold stress

Fig. 6 Phenotypic observation of TaMYBS1-like T3 transgenic Arabidopsis thaliana under drought stress

Fig. 7 Physiological indicators of WT and TaMYBS1-like T3 transgenic Arabidopsis thaliana under drought stress

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