BnaTZF1A/1C在甘蓝型油菜干旱胁迫反应中的功能及作用机制研究

甘蓝型油菜是我国重要的油料作物，随着栽培地区的北移与西扩，干旱成为制约种植规模、影响产量与品质的关键因素之一。RR-TZF是植物特有的串联锌指蛋白，通过诱导mRNA的降解参与转录后调控。我们从甘蓝型油菜中鉴定出两个与TZF1高度同源的基因BnaTZF1A/1C，在拟南芥中的过表达能显著提高抗旱能力，因此推测BnaTZF1A/1C可能与甘蓝型油菜的抗旱性密切相关，也具有与RR-TZF蛋白类似的作用机制。本项目拟采用转基因、RIP-seq和EMSA等技术手段系统研究BnaTZF1A/1C对甘蓝型油菜抗旱性及生长发育等的影响，进一步鉴定该锌指蛋白作用的靶mRNA以及互作方式，并开展靶基因的功能分析，深入解析该基因的分子调控机制。项目研究结果将有助于揭示TZF1蛋白在甘蓝型油菜干旱胁迫反应中的作用机理，挖掘新的功能基因，为调控和改良甘蓝型油菜耐旱性提供理论基础和基因资源。

Brassica napus is one of the most important oil crop in China. Drought has become one of the vital factors that restricting the scale of cultivation and affecting yield and quality, with the northward migration and westward expansion of cultivation areas. It is one of the key problems to study the mechanism of drought resistance and improve the drought resistance of B. napus. RR-TZFs (Arginine-rich motif-tandem CCCH zinc finger) characterized by two tandem-arrayed CCCH-type zinc fingers, are the plant specific CCCH proteins, which control the individual development and defense response of plants. RR-TZFs are involved in plant resistance regulation by inducing mRNA degradation, but the target mRNAs and regulation pathways are not yet clear. Our previous studies showed that overexpression of AtTZF1 significantly enhanced the drought resistance of plants, and found that there were 2 homologous genes BnaTZF1A/1C in B. napus. The results of simulation experiments showed that BnaTZF1A/1C was closely related to drought stress. It is speculated that BnaTZF1A/1C participates in drought stress response by binding unknown target mRNAs and regulating their stability. This project aims to complete the following main tasks: study of the effects of BnaTZF1A/1C on plant growth and development under drought stress in details, screening potential target mRNAs and verification by sequencing of transgenic rapeseed, RNA-binding protein immunoprecipitation sequence (RIP-Seq) and other techniques, and studying the combine mechanism of BnaTZF1A/1C and target mRNAs and the regulation ways by prokaryotic expression, protoplast expression, EMSA, qRT-PCR and others. The results of the project will help to reveal the mechanism of Arginine-rich motif-tandem CCCH zinc finger TZF1 in the response to drought stress in B. napus, and provide a theoretical basis and genetic resources for regulating and improving the drought tolerance of B. napus.