冬小麦TaMYBS1-like基因的克隆及功能分析

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

摘要/Abstract

摘要：

目的东农冬麦1号（Dn1）是首个能在高寒地带大面积种植、返青率超过85%的强抗寒冬小麦（Triticum aestivum）品种，探索冬小麦MYB类转录因子TaMYBS1-like基因的功能，以及低温和干旱胁迫对该基因表达的影响。方法利用RT-PCR法从冬小麦Dn1中克隆了TaMYBS1-like基因的cDNA序列，并对其进行生物信息学分析。于4、0、-10、-25 ℃处理后，对冬小麦叶片和分蘖节进行取样，待冬小麦（Dn1）长至三叶期时用20% PEG-6000处理，在0、3、6、12、24 h时对冬小麦叶片和分蘖节进行取样。利用RT-qPCR技术分析该基因在低温和干旱胁迫下冬小麦叶片和分蘖节中的表达模式。构建表达载体，利用亚细胞定位确定其蛋白位置，通过农杆菌介导法将TaMYBS1-like基因转入拟南芥中，并进行低温和干旱胁迫处理，分析验证TaMYBS1-like基因功能。结果 TaMYBS1-like基因全长903 bp，共编码300个氨基酸，编码蛋白为不稳定亲水性蛋白，主要定位于细胞核；表达模式分析发现，低温胁迫时，叶片和分蘖节中TaMYBS1-like基因的表达量与4 ℃时相比均有显著提高。干旱胁迫3、6、12、24 h时叶片TaMYBS1-like基因的表达量均显著高于对照组，过表达TaMYBS1-like基因提高了拟南芥植株的基础生理代谢、细胞活性和抗氧化酶系统活性，减轻了细胞膜的损伤。结论冬小麦TaMYBS1-like基因的异源过表达提高了拟南芥的抗寒和抗旱能力。

关键词: 冬小麦, TaMYBS1-like基因, 抗低温, 抗旱性

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

王欣悦, 邵庆一, 陈宇姝, 胡昳凡, 赵楠, 王嫱, 王敏丽, 王雪松, 金忠民, 刘丽杰. 冬小麦TaMYBS1-like基因的克隆及功能分析[J]. 生物技术通报, 2026, 42(6): 218-226.

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.

图/表 7

图1 TaMYBS1-like蛋白的二级结构及曲线趋势（A）和三级结构（B）

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

图2 不同低温和干旱胁迫下冬小麦叶片和分蘖节中TaMYBS1-like基因表达量的变化不同字母表示在0.05水平上差异显著（P0.05），下同

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

图3 TaMYBS1-like在烟草表皮细胞中的亚细胞定位

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

图4 TaMYBS1-like T3代转基因拟南芥低温胁迫下的表型观察

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

图5 WT、TaMYBS1-like T3代转基因拟南芥低温胁迫下的生理指标

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

图6 TaMYBS1-like T3代转基因拟南芥干旱胁迫下的表型观察

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

图7 WT、TaMYBS1-like T3代转基因拟南芥干旱胁迫下的生理指标

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

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