生物技术通报 ›› 2023, Vol. 39 ›› Issue (10): 136-147.doi: 10.13560/j.cnki.biotech.bull.1985.2023-0339

• 研究报告 • 上一篇    下一篇

低磷胁迫下番茄转录因子WRKY6功能分析

陈浩婷1(), 张玉静1, 刘洁1, 代泽敏1, 刘伟1, 石玉1, 张毅1(), 李天来1,2()   

  1. 1.山西农业大学园艺学院,太谷 030801
    2.沈阳农业大学园艺学院,沈阳 110866
  • 收稿日期:2023-04-11 出版日期:2023-10-26 发布日期:2023-11-28
  • 通讯作者: 张毅,男,博士,教授,研究方向:设施蔬菜栽培与生理;E-mail: harmony1228@163.com
    李天来,男,博士,教授,研究方向:设施蔬菜栽培与生理;E-mail: ltl@syau.edu.cn
  • 作者简介:陈浩婷,女,博士研究生,研究方向:设施蔬菜栽培与生理;E-mail: 746852776@qq.com
  • 基金资助:
    山西省研究生教育创新项目(2021Y325);山西省基础研究计划(202203021211263)

Functional Analysis of WRKY6 Gene in Tomato Under Low-phosphorus Stress

CHEN Hao-ting1(), ZHANG Yu-jing1, LIU Jie1, DAI Ze-min1, LIU Wei1, SHI Yu1, ZHANG Yi1(), LI Tian-lai1,2()   

  1. 1. College of Horticulture, Shanxi Agricultural University, Taigu 030801
    2. College of Horticulture, Shenyang Agricultural University, Shenyang 110866
  • Received:2023-04-11 Published:2023-10-26 Online:2023-11-28

摘要:

本研究旨在分析番茄转录因子WRKY6在低磷胁迫应答中的作用,为研究番茄耐低磷分子机制及挖掘提高番茄低磷耐受性和磷素利用率的基因资源奠定理论基础。试验以野生型番茄Ailsa Craig为材料,以其cDNA为模板克隆基因SlWRKY6,利用根癌农杆菌介导转化法构建RNAi-SlWRKY6和OE-SlWRKY6转基因番茄株系,并对野生型、RNAi-SlWRKY6和OE-SlWRKY63种不同基因型的番茄植株进行低磷胁迫处理,在处理第18天对叶片与根系进行表型观察和生理生化指标测定。表型观察结果显示,与RNAi-SlWRKY6转基因番茄相比,OE-SlWRKY6转基因番茄植株较矮壮,叶片数量较多,受低磷胁迫影响较小,表现出较强的耐低磷性。生理生化指标分析结果表明,与野生型相比,OE-SlWRKY6转基因番茄根系与叶片的有机磷和总磷含量显著升高,根系酸性磷酸酶活性和部分有机酸含量显著增加,磷转运体表达量显著降低。而RNAi-SlWRKY6转基因番茄受到低磷胁迫后,这些指标的变化趋势与过表达株系相反。低磷胁迫后,LePT1的相对表达量在OE-SlWRKY6植株根系中呈下降趋势,与RNAi-SlWRKY6植株中变化相反。在野生型、RNAi-SlWRKY6和OE-SlWRKY6植株根系中LePT2的相对表达量均呈整体上升趋势,而LePT3则呈整体下降趋势。由上述可知,SlWRKY6能够响应低磷胁迫,其表达量高低与番茄植株的低磷耐受性呈正相关,且可能影响磷转运体基因对低磷的响应。该研究结果可为进一步揭示番茄WRKY转录因子家族的功能及其对低磷胁迫的响应机制提供理论依据。

关键词: 番茄, 转录因子, SlWRKY6, 低磷胁迫, 功能分析, 磷含量, 酸性磷酸酶

Abstract:

The purpose of this study is to analyze the role of tomato transcription factor SlWRKY6 in response to low-phosphorus stress, and to lay a theoretical foundation for studying the molecular mechanism of low-phosphorus tolerance in tomato and exploring tomato germplasm resources of improving low-phosphorus tolerance and phosphorus utilization. Wild-type tomato Ailsa craig was used as the material and its cDNA was used as the template to clone SlWRKY6, and the RNAi-SlWRKY6 and OE-SlWRKY6 transgenic tomato plant lines were constructed by agrobacterium tumefacien-mediated transformation. Tomato plants with three different genotypes of wild type, RNAi-SlWRKY6 and OE-SlWRKY6, were subjected to low phosphorus. On the day 18 of low-phosphorus treatment, the results of phenotype and physiological and biochemical indexes in the leaves and roots were determined. Phenotypic observation showed that OE-SlWRKY6 transgenic tomato plants were of thicker and shorter after low phosphorus treatment while compared with RNAi-SlWRKY6 transgenic tomato plants, they had a higher number of leaves, and were less affected by low-phosphorus stress, and showed stronger low-phosphorus tolerance. The analysis results of physiological and biochemical indexes showed that the organic phosphorus and total phosphorus content in the roots and leaves of transgenic tomato plant OE-SlWRKY6 significantly increased compared with wild type, while the acid phosphatase activity and some organic acid contents in the roots significantly increased. The expressions of phosphorus transporters also significantly reduced. However, the trend of changes in these indicators in the transgenic tomato plant RNAi-SlWRKY6 under low-phosphorus stress was opposite to that in transgenic tomato plant OE-SlWRKY6. After low-phosphorus stress, the relative expressions of LePT1 decreased in the roots of OE-SlWRKY6 plants, which was opposite to that of RNAi-SlWRKY6 plants. The relative expressions of LePT2 in the roots of wild type, RNAi-SlWRKY6 and OE-SlWRKY6 plants showed an overall increasing trend, while the relative expressions of LePT3 showed an overall decreasing trend. According to the above results, SlWRKY6 responded to low-phosphorus stress and its expression was positively correlated with the low-phosphorus tolerance of tomato plants, and might affect the response of phosphorus transporter genes to low phosphorus. The results of this study may provide certain theoretical basis for further revealing the function of tomato WRKY transcription factor family and its response mechanism to low-phosphorus stress.

Key words: tomato, transcription factors, SlWRKY6, low-phosphorus stress, functional analysis, phosphorus content, acid phosphatase