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A novel R3H protein, OsDIP1, confers ABA-mediated adaptation to drought and salinity stress in rice

Abstract

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.

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Fig. 1
Fig. 2
Fig. 3

source of the apoplastic ROS production) inhibitor diphenylene iodonium (DPI), H2O2 scavenger (dimethylthiourea, DMTU) and catalase (CAT) treated with 50 μM ABA treatment. 10 μM DPI, 100 mM DMTU, and 100 U CAT were used. (C) Time course of changes in the expression of OsDIP1 under PEG NaCl treatments. 10% PEG6000 and 100 mM NaCl were used to treat 10-d old rice plants. (D) The expression pattern of OsDIP1 exposed to 80 μM fluridon (an ABA inhibitor) under PEG or NaCl stress. Data are mean ± SD (n = 8)

Fig. 4
Fig. 5
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Fig. 7
Fig. 8

Abbreviations

ABA:

Abscisic acid

APX:

Ascorbate peroxidase

CAT:

Catalase

DAPI:

4',6-Diamidino-2-phenylindole, dihydrochloride

DMTU:

Dimethylthiourea

DPI:

Diphenylene iodonium

E. coli :

Escherichia coli

H2DCF-DA:

2′,7′-Dichlorofluorescein diacetate

H2O2 :

Hydrogen peroxidase

NCED:

9-Cis-epoxycarotenoid dioxygenase

LUC/REN:

Firefly luciferase/renilla

MDA:

Malondialdehyde

NaCl:

Sodium chloride

PEG:

Polyethylene glycol

Rbohs:

Respiratory burst oxidase homologs

SOD:

Superoxide dismutase

YFP:

Yellow fluorescence protein

ZFP:

Zinc finger protein

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Acknowledgements

We are grateful to Prof Wenhua Zhang from Nanjing Agricultural University for supplying pSUPER1300 and pCAMBIA1301 vectors, and Prof. Hongxuan Lin from Shanghai Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences for providing rice protoplast transformation vectors to study the transactivation of TFs.

Funding

This work was supported by the National Natural Science Foundation of China (grants 31901202, 31672228), National Distinguished Expert Project (WQ20174400441), Project for High-level Talents of Foshan University (gg07102, gg05003/071), and by the Australian Research Council (DP150101663).

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Huang, L., Fu, W., Ji, E. et al. A novel R3H protein, OsDIP1, confers ABA-mediated adaptation to drought and salinity stress in rice. Plant Soil (2022). https://doi.org/10.1007/s11104-022-05428-y

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Keywords

  • Abscisic acid
  • Antioxidant defense
  • Water stress
  • OsDIP1
  • ABA