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The OsWRKY6 transcriptional cascade functions in basal defense and Xa1-mediated defense of rice against Xanthomonas oryzae pv. oryzae

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Abstract

Main conclusion

The rice protein OsWRKY6 directly activates OsWRKY45 and OsWRKY47 expression, and also activates OsPR1a and OsPR1b through the two OsWRKYs, and this transcriptional module participates in Xa1-mediated defense against the pathogen Xanthomonas oryzae pv. oryzae.

Abstract

Biotic stress, the pathogen Xanthomonas oryzae pv. oryzae (Xoo) in particular, negatively impacts worldwide productivity and yield in the staple crop rice (Oryza sativa). OsWRKY transcription factors are involved in various biotic stress responses in rice, and OsWRKY6 specifically acts as an important defense regulator against Xoo. However, the relationship between OsWRKY6 and other OsWRKYs, as well as its role in resistance (R) gene-mediated defense, have yet to be studied in depth. Here, we characterized a transcriptional cascade triggered by OsWRKY6 that regulated defense against Xoo infection mediated by the NBS-LRR protein Xa1. OsWRKY45 and OsWRKY47 were identified as direct transcriptional targets of OsWRKY6, and their two gene products reciprocally activated their two genes. Furthermore, OsWRKY6 activated OsPR1a and OsPR1b via the OsWRKY45 and OsWRKY47. Two OsWRKY6 RNAi knockdown lines showed significantly reduced defense even against an incompatible Xoo infection, and the expression of OsWRKY6 was not regulated by OsWRKY51 and OsWRKY88. This study reveals that a novel downstream transcriptional pathway activated by OsWRKY6 is involved in Xa1-mediated defense against Xoo.

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Abbreviations

BLB:

Bacterial leaf blight

GFP:

Green fluorescent protein

rLUC:

Renilla luciferase

RMP:

Rice mesocotyl protoplast

Xoo :

Xanthomonas oryzae Pv. oryzae

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Acknowledgements

This study was carried out with the support of “Research Program for Agricultural Science & Technology Development (Project No. PJ01246303)”, National Institute of Agricultural Sciences, Rural Development Administration, Republic of Korea.

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Author notes

  1. Duk-Ju Hwang

    Present address: Mediprogen Inc.1447, Pyungchang-gun, 25354, Republic of Korea

  2. Jong Hee Im

    Present address: Department of Horticulture, Michigan State University, East Lansing, MI, 48824, USA

  3. Changhyun Choi

    Present address: National Institute of Crop Science, Rural Development Administration, 180 Hyeoksin-ro, Wanju-gun, 55365, Republic of Korea

Authors and Affiliations

  1. National Institute of Agricultural Science, Rural Development Administration, Jeonju, 54874, Republic of Korea

    Jong Hee Im, Changhyun Choi, Sang Ryeol Park & Duk-Ju Hwang

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  1. Jong Hee Im
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Correspondence to Duk-Ju Hwang.

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Supplementary Information

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425_2022_3830_MOESM1_ESM.pptx

Supplementary file1 Fig. S1 The expression analysis of defense related genes in OsWRKY6OX and OsWRKY6KD. a Gene expression of 11 defense related OsWRKYs in OsWRKY6OX. b Gene expression of OsWRKY11, 24, 62, 77 and 104 in two OsWRKY6KD lines. Total RNA was isolated from three-week-old WT and OsWRKY6OX or OsWRKY6KD seedlings and cDNA synthesis was carried out. qPCR was performed with gene-specific primers. OsActin was used as a gene expression control. Fig. S2 Distribution of W-box and W-box-like element 1 (WLE1) motifs in OsWRKY45, 47 and 76 promoters. a OsWRKY45 promoter. b OsWRKY47 promoter. c OsWRKY76 promoter. Closed triangle: W-box; open triangle: WLE1. The letters a, b, c, and d marked on each promoter indicate the regions amplified by ChIP-qPCR. Anti-hemagglutinin (HA) antibody was used as a negative control. Fig. S3 Gene expressions of defense marker genes in OsWRKY6 and 45 and 47 overexpressing lines. Gene expression of OsPR1a, OsPR1b, BETV1 and Chitianse were analyzed in OsWRKY6OX (a), OsWRKY45OX (b) and OsWRKY47OX (c). Total RNA was isolated from the WT and designated transgenics and cDNA was synthesized with it. The gene expressions were determined with qPCR with gene-specific primers. OsActin was used as an expression control. Values are means of triplicate-independent biological repeats with SEs. ** P < 0.001, * P < 0.01, Fig. S4 Chromatin immunoprecipitation (ChIP)-qPCR of OsPR1a and OsPR1b promoters by OsWRKY6. a and b The schematic draw of OsPR1a (a) and OsPR1b (b) promoters. Closed triangle: W-box; open triangle: WLE1. The letters a and b marked on each promoter indicate the regions amplified by ChIP-qPCR. c ChIP-qPCR result of OsPR1a. d ChIP-qPCR result of OsPR1b. Anti-hemagglutinin (HA) antibody was used as a negative control. Fig. S5 Gene expression analysis of OsWRKY6 in OsWRKY45OX and OsWRKY47OX. Total RNA was isolated from three-week-old WT and two transgenic lines of OsWRKY45 and OsWRKY47. cDNA was synthesized with the RNA and OsWRKY6 expressions were determined with specific primers. OsActin was used as an expression control. Table S1 List of primers in this study (PPTX 173 KB)

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Im, J.H., Choi, C., Park, S.R. et al. The OsWRKY6 transcriptional cascade functions in basal defense and Xa1-mediated defense of rice against Xanthomonas oryzae pv. oryzae. Planta 255, 47 (2022). https://doi.org/10.1007/s00425-022-03830-5

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