The E3 ligase TaGW2 mediates transcription factor TaARR12 degradation to promote drought resistance in wheat

Shumin Li, Yifang Zhang, Yuling Liu, Peiyin Zhang, Xuemin Wang, Bin Chen, Li Ding, Yingxiong Nie, Fangfang Li, Zhenbing Ma, Zhensheng Kang, Hude Mao
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Abstract

Drought stress limits crop yield, but the molecular modulators and their mechanisms underlying the trade-off between drought resistance and crop growth and development remain elusive. Here, a Grain width and weight2 (GW2)-like RING finger E3 ligase, TaGW2, was identified as a pivotal regulator of both kernel development and drought responses in wheat (Triticum aestivum). TaGW2 overexpression enhances drought resistance but leads to yield drag under full irrigation conditions. In contrast, TaGW2 knockdown or knockout attenuates drought resistance but remarkably increases kernel size and weight. Furthermore, TaGW2 directly interacts with and ubiquitinates the type-B Arabidopsis response regulator TaARR12, promoting its degradation via the 26S proteasome. Analysis of TaARR12 overexpression and knockdown lines indicated that TaARR12 represses the drought response but does not influence grain yield in wheat. Further DNA affinity purification sequencing combined with transcriptome analysis revealed that TaARR12 down-regulates stress-responsive genes, especially group-A basic leucine zipper (bZIP) genes, resulting in impaired drought resistance. Notably, TaARR12 knockdown in the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated nuclease 9 (Cas9)-mediated tagw2 knockout mutant leads to significantly higher drought resistance and grain yield compared to wild-type plants. Collectively, these findings show that the TaGW2–TaARR12 regulatory module is essential for drought responses, providing a strategy for improving stress resistance in high-yield wheat varieties.
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E3 连接酶 TaGW2 介导转录因子 TaARR12 降解,促进小麦的抗旱性
干旱胁迫限制了作物产量,但抗旱性与作物生长发育之间权衡的分子调控因子及其机制仍未确定。在这里,一种类似谷粒宽度和重量2(GW2)的RING指E3连接酶TaGW2被鉴定为小麦(Triticum aestivum)籽粒发育和干旱响应的关键调控因子。TaGW2 过表达可增强抗旱性,但在充分灌溉条件下会导致产量下降。相反,TaGW2 基因敲除或基因敲除会削弱抗旱性,但会显著增加籽粒的大小和重量。此外,TaGW2 与拟南芥 B 型响应调节因子 TaARR12 直接相互作用并泛素化,促进其通过 26S 蛋白酶体降解。对 TaARR12 过表达和基因敲除株系的分析表明,TaARR12 可抑制干旱响应,但不会影响小麦的籽粒产量。进一步的 DNA 亲和纯化测序结合转录组分析表明,TaARR12 下调了胁迫响应基因,尤其是 A 组碱性亮氨酸拉链(bZIP)基因,导致抗旱性受损。值得注意的是,与野生型植株相比,在聚类规则间隔短回文重复(CRISPR)/CRISPR相关核酸酶9(Cas9)介导的tagw2基因敲除突变体中敲除TaARR12可显著提高抗旱性和谷物产量。总之,这些研究结果表明,TaGW2-TaARR12调控模块对干旱响应至关重要,为提高高产小麦品种的抗逆性提供了一种策略。
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