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ZmDnaJ-ZmNCED6 module positively regulates drought tolerance via modulating stomatal closure in maize
Thursday, 2024/12/05 | 08:18:31

Anyi Dong, Nan Wang, Tinashe Zenda, Xiuzhen Zhai, Yuan Zhong, Qian Yang, Yue Xing,

 Huijun Duan, Xiaocui Yan

Plant Physiology and Biochemistry; Volume 218, January 2025, 109286

Abstract

Heat Shock Protein plays a vital role in maintaining protein homeostasis and protecting cells from stress stimulation. As one of the HSP40 proteins, DnaJ is a stress response protein widely existing in plant cells. The function and regulatory mechanism of ZmDnaJ, a novel chloroplast-localized type-III HSP40, in maize drought tolerance were characterized. Tissue-specific expression analysis showed that ZmDnaJ is highly expressed in the leaves, and is strongly drought-induced in maize seedlings. Overexpression of ZmDnaJ improved maize drought tolerance by enhancing stomatal closure and increasing ABA content to mediate photosynthesis. In contrast, the CRISPR-Cas9 knockout zmdnaj mutant showed lower relative water content and high sensitivity to drought stress. Moreover, Y2H, BiFC and Co-IP analyses revealed that ZmDnaJ interacts with an ABA synthesis-related protein ZmNCED6 to regulate drought tolerance. Similarly, ZmNCED6 overexpressed lines showed stronger oxidation resistance, enhanced photosynthetic rate, stomatal closure and ABA content, whilst the CRISPR-Cas9 knockout mutant showed sensitive to drought stress. More importantly, ZmDnaJ could regulate key drought tolerance genes (ZmPYL10ZmPP2C44ZmEREB65ZmNCED4ZmNCED6 and ZmABI5), involved in ABA signal transduction pathways. Taken together, our findings suggest that ZmDnaJ-ZmNCED6 module improves drought tolerance in maize.

 

See https://www.sciencedirect.com/science/article/pii/S0981942824wxXH4qGPPw5_aNJfnqyy6rxBBskZM

 

Fig. 1. Molecular identification of ZmDnaJ gene and its response to drought stress. (A) the relative expression level of ZmDnaJ gene in PEG simulated drought conditions within 24 h. The lowest expression level detected in transgenic plants served as the reference and was set to ‘1’. ZmActin was used as the internal control. (B) subcellular localization of ZmDnaJ protein in tobacco leaf. Empty GFP was used as a control. (C) target sequence alignment of ZmDnaJ gene knockout plants compared with the wild type. (D) phenotypes of ZmDnaJ gene under normal and drought stress conditions. Scale bars = 8 cm. (E) relative expression level of ZmDnaJ. Relative expression level was calculated as the fold change of the expression value of ZmDnaJ in transgenic leaves to that of wild type and was normalized by using ZmActin as reference. (F) soil water content. (G) survival rate. Data were obtained through 14 days of drought and three days of rehydration. Note: The values shown are means ± SD. Statistical analysis using Student's t-test revealed the significant differences (∗, P < 0.05; ∗∗, P < 0.01).

 

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