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A novel R3H protein, OsDIP1, confers ABA-mediated adaptation to drought and salinity stress in rice
Plant and Soil ( IF 4.9 ) Pub Date : 2022-04-25 , DOI: 10.1007/s11104-022-05428-y
Liping Huang , WenXuan Fu , E. Ji , Mohsin Tanveer , Sergey Shabala , Min Yu , Mingyi Jiang

Aims

Dehydration responsive element-binding factors (DBFs) have recently emerged as essential components of stress tolerance mechanisms in plants. In this work, we investigated the role of OsDIP1 that operate upstream of DBFs as a regulator of drought and salinity tolerance in rice.

Methods

Experiments were conducted on rice plants (Oryza sativa) and included combination of physiological (plant phenotyping) and molecular methods. The latter involved real-time PCR (RT-qPCR); yeast two-hybrid, BiFC assay, GST pull-down, CoIP and firefly luciferase (LUC)/ renilla (REN) assays.

Results

OsDIP1 expression was induced by hydrogen peroxide (H2O2), ABA (abscisic acid), drought (polyethylene glycol treatment), and salt stress. Overexpression of OsDIP1 in rice enhanced drought and salinity tolerance while knocking out OsDIP1 by CRISPR-Cas9 editing resulted in drought and salt sensitive phenotype. The activity and gene expression of antioxidant defense enzymes, superoxide dismutase (SOD), catalase (CAT), was increased in OsDIP1-overexpressed transgenic rice plants, while the content of malondialdehyde (MDA) was decreased. In contrast, the content and gene expression of SODCc2 and CatB, decreased, and the content of MDA was increased in knockout of OsDIP1 rice plants, suggesting that overexpression of OsDIP1 enhances the antioxidant capacity of rice plants. The yeast two-hybrid screening test revealed that OsDIP1 interacted with ZFP36, a key zinc finger transcription factor involved in ABA-induced antioxidant defense. Moreover, OsDIP1 could modulate some key ABA-responsive genes via interacting with ZFP36.

Conclusions

In this work, we show that DIP1 plays a central role in modulating drought and salinity stress tolerance in rice.



中文翻译:

一种新的 R3H 蛋白 OsDIP1 赋予 ABA 介导的水稻对干旱和盐分胁迫的适应能力

目标

脱水反应元件结合因子 (DBF) 最近已成为植物胁迫耐受机制的重要组成部分。在这项工作中,我们研究了在 DBF 上游运行的 OsDIP1 作为水稻耐旱和耐盐性的调节因子的作用。

方法

实验在水稻植物(Oryza sativa)上进行,包括生理学(植物表型)和分子方法的结合。后者涉及实时 PCR (RT-qPCR);酵母双杂交、BiFC 测定、GST 下拉、CoIP 和萤火虫荧光素酶 (LUC)/海肾 (REN) 测定。

结果

OsDIP1表达由过氧化氢(H 2 O 2)、ABA(脱落酸)、干旱(聚乙二醇处理)和盐胁迫诱导。OsDIP1在水稻中的过表达增强了干旱和耐盐性,而通过 CRISPR-Cas9 编辑敲除OsDIP1导致干旱和盐敏感表型。OsDIP1过表达的转基因水稻中抗氧化防御酶超氧化物歧化酶(SOD)、过氧化氢酶(CAT)的活性和基因表达增加,而丙二醛(MDA)的含量降低。相比之下,SODCc2CatB的含量和基因表达, OsDIP1基因敲除后, MDA 含量增加, 表明OsDIP1的过表达增强了水稻的抗氧化能力。酵母双杂交筛选试验表明,OsDIP1 与 ZFP36 相互作用,ZFP36 是参与 ABA 诱导的抗氧化防御的关键锌指转录因子。此外,OsDIP1 可以通过与 ZFP36 相互作用来调节一些关键的 ABA 反应基因。

结论

在这项工作中,我们表明 DIP1 在调节水稻的干旱和盐胁迫耐受性中起核心作用。

更新日期:2022-04-26
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