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A salicylic acid inducible mulberry WRKY transcription factor, MiWRKY53 is involved in plant defence response

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MiWRKY53 is expressed in response to various stresses and hormones. Although it is localized in the nucleus, it shows no transcriptional activation. Role of SA-mediated plant defence response is demonstrated.

Abstract

WRKY transcription factors are one the largest gene families in plants involved in almost every process in plants including development, physiological processes, and stress response. Salicylic acid (SA) is key regulator of biotic stress against various pathogens in plants acting via its multiple mechanisms to induce defence response. Herein, we have identified and functionally validated WRKY53 from mulberry (Morus indica var. K2). MiWRKY53 expressed differentially in response to different stress and hormonal treatments. MiWRKY53 belongs to group III of WKRY gene family, localized in nucleus, and lacks transcriptional activation activity in yeast. Hormone responsive behaviour of MiWRKY53 Arabidopsis overexpression (OE) transgenics preferentially was noted in root growth assay in response to Salicylic acid (SA). Arabidopsis overexpression plants also displayed alteration in leaf phenotype having wider leaves than the wild-type plants. PR-1 transcripts were higher in MiWRKY53 Arabidopsis OE plants and they displayed resistance towards biotrophic pathogen Pseudomonas syringae PstDC3000. MiWRKY53 Mulberry OE transgenics also depicted SA-responsive behaviour. Several hormones and stress-related cis-acting elements were also identified in the 1.2-Kb upstream regulatory region (URR) of MiWRKY53. Functional characterization of full-length promoter region revealed that it is induced by SA and further analysis of deletion constructs helped in the identification of minimal promoter responsible for its inducibility by SA. Altogether, the findings from this study point towards the SA preferential behaviour of MiWRKY53 and its function as regulator of plant defence response through SA-mediated mechanisms.

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Acknowledgements

NN acknowledges the Department of Science and Technology, Government of India for the INSPIRE fellowship, and PK is thankful to Department of Biotechnology, Government of India, New Delhi for financial support.

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Authors and Affiliations

  1. Department of Plant Molecular Biology, University of Delhi South Campus, Benito Juarez Road, New Delhi, 110021, India

    Nisha Negi & Paramjit Khurana

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  1. Nisha Negi
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  2. Paramjit Khurana
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NN and PK planned experiments. NN performed experiments. NN and PK analyzed data and PK reviewed the manuscript.

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Correspondence to Paramjit Khurana.

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Communicated by Amit Dhingra.

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Supplementary Information

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299_2021_2710_MOESM1_ESM.pdf

Confirmation of MiWRKY53 clone (1053 bp) by restriction digestion. b CDS sequence and protein sequence of MiWRKY53 (PDF 170 KB)

a

Pictorial representation of vector map of overexpression pMDC32::MiWRKY53 b PCR confirmation of Arabidopsis OE T4 lines using gene-specific primers c and hptII (hygromycin) primers. d Overexpression of MiWRKY53 as checked by real-time PCR using GSP (PDF 193 KB)

a

PCR amplification of MiWRKY53 promoter and its deletion constructs. b Arabidopsis transgenic confirmation of MiWRKY53 full length promoter and its deletion constructs. Arabidopsis T1 lines PCR confirmation with GSP. Arabidopsis T1 lines PCR confirmation with gene-specific primers and Kanamycin-specific PCR (PDF 389 KB)

299_2021_2710_MOESM4_ESM.pdf

PCR confirmation of M. indica var. K2 pBI121:MiWRKY53 transformants using a nptII, b gene-specific, c Relative transcript abundance in Morus indica cv. K2 overexpression lines harbouring MiWRKY53 (PDF 268 KB)

List of primers (PDF 159 KB)

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Negi, N., Khurana, P. A salicylic acid inducible mulberry WRKY transcription factor, MiWRKY53 is involved in plant defence response. Plant Cell Rep 40, 2151–2171 (2021). https://doi.org/10.1007/s00299-021-02710-8

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