College of Life Science, Sichuan Agricultural University, No. 46, Xinkang Road, Ya'an, 625014, Sichuan, China.
Xichang College, 615013, Xichang, Sichuan, China.
Plant Physiol Biochem. 2020 Feb;147:43-53. doi: 10.1016/j.plaphy.2019.12.004. Epub 2019 Dec 4.
The WRKY transcription factor family includes plant-specific transcription factors that are widely involved in plant biotic and abiotic stress responses, growth and development. Tartary buckwheat is a type of small grain with strong resistance to adverse growing conditions. No systematic exploration of the WRKY family in Tartary buckwheat has yet been reported. In this paper, we report the FtWRKY46 gene from Tartary buckwheat and study its role in salt tolerance. FtWRKY46 has transcriptional activation activity in yeast, and FtWRKY46 fused to yellow fluorescent protein localizes to the nucleus. Further studies have found that its transcriptional activation region is located at the N-terminus. A yeast one-hybrid assay indicated that FtWRKY46 could bind to a W-box and activate reporter gene expression. Similarly, transient cotransfection showed that FtWRKY46 could specifically bind to W-box regions and activate reporter gene expression in plants. Furthermore, ectopic expression of FtWRKY46 could enhance Arabidopsis tolerance to salt stress. More specifically, the seed germination rate, root length, chlorophyll content and proline content were significantly higher in transgenic plants ectopically expressing FtWRKY46 than in WT plants after salt stress (P < 0.05), while MDA levels were significantly lower than in WT plants (P < 0.05). Additionally, salt treatment increased the expression of stress-related genes. To summarize, our results suggest that ectopic expression of FtWRKY46 enhance the stress tolerance of transgenic plants by modulating ROS clearance and stress-related gene expression.
WRKY 转录因子家族包括植物特异性转录因子,广泛参与植物生物和非生物胁迫反应、生长和发育。苦荞是一种对不良生长条件具有较强抗性的小粒谷物。目前尚未有关于苦荞 WRKY 家族的系统研究。本文从苦荞中报告了 FtWRKY46 基因,并研究了其在耐盐性中的作用。FtWRKY46 在酵母中具有转录激活活性,并且 FtWRKY46 与黄色荧光蛋白融合后定位于细胞核。进一步的研究发现,其转录激活区位于 N 端。酵母单杂交试验表明,FtWRKY46 可以与 W-box 结合并激活报告基因的表达。同样,瞬时共转染表明,FtWRKY46 可以在植物中特异性结合 W-box 区域并激活报告基因的表达。此外,异位表达 FtWRKY46 可以增强拟南芥对盐胁迫的耐受性。更具体地说,在盐胁迫后,异位表达 FtWRKY46 的转基因植物的种子发芽率、根长、叶绿素含量和脯氨酸含量均显著高于 WT 植物(P < 0.05),而 MDA 水平显著低于 WT 植物(P < 0.05)。此外,盐处理增加了与胁迫相关的基因的表达。综上所述,我们的结果表明,异位表达 FtWRKY46 通过调节 ROS 清除和与胁迫相关的基因表达来增强转基因植物的胁迫耐受性。