MdHIR4 transcription and translation levels associated with disease in apple are regulated by MdWRKY31
Here we describe that the regulation of MdWRKY31 on MdHIR4 in transcription and translation levels associated with disease in apple.
The phytohormone salicylic acid (SA) is a main factor in apple (Malus domestica) production due to its function in disease resistance. WRKY transcription factors play a vital role in response to stress. An RNA-seq analysis was conducted with ‘Royal Gala’ seedlings treated with SA to identify the WRKY regulatory mechanism of disease resistance in apple. The analysis indicated that MdWRKY31 was induced. A quantitative real-time polymerase chain reaction (qPCR) analysis demonstrated that the expression of MdWRKY31 was induced by SA and flg22. Ectopic expression of MdWRKY31 in Arabidopsis and Nicotiana benthamiana increased the resistance to flg22 and Pseudomonas syringae tomato (Pst DC3000). A yeast two-hybrid screen was conducted to further analyze the function of MdWRKY31. As a result, hypersensitive-induced reaction (HIR) protein MdHIR4 interacted with MdWRKY31. Biomolecular fluorescence complementation, yeast two-hybrid, and pull-down assays demonstrated the interaction. In our previous study, MdHIR4 conferred decreased resistance to Botryosphaeria dothidea (B. dothidea). A viral vector-based transformation assay indicated that MdWRKY31 evaluated the transcription of SA-related genes, including MdPR1, MdPR5, and MdNPR1 in an MdHIR4-dependent way. A GUS analysis demonstrated that the w-box, particularly w-box2, of the MdHIR4 promoter played a major role in the responses to SA and B. dothidea. Electrophoretic mobility shift assays, yeast one-hybrid assay, and chromatin immunoprecipitation-qPCR demonstrated that MdWRKY31 directly bound to the w-box2 motif in the MdHIR4 promoter. GUS staining activity and a protein intensity analysis further showed that MdWRKY31 repressed MdHIR4 expression. Taken together, our findings reveal that MdWRKY31 regulated plant resistance to B. dothidea through the SA signaling pathway by interacting with MdHIR4.
KeywordsMalus domestica MdWRKY31 SA Disease resistance Botryosphaeria dothidea
Y-JH, Y-YL, and X-YZ conceived and designed the experiments. X-YZ performed the research. C-HQ, X-YZ, HJ, M-SZ and C-XY analyzed the data. X-YZ and Y-YL wrote this paper.
This work was supported by grants from the National Key Research and Development Program (2018YFD1000200), the National Natural Science Foundation of China (31772275, U1706202), the Natural Science Fund for Excellent Young Scholars of Shandong Province (ZR2018JL014).
Compliance with ethical standards
Conflict of interest
All authors have no conflicts of interest to declare.
- Jung HW, Hwang BK (2007) The leucine-rich repeat (LRR) protein, CaLRR1, interacts with the hypersensitive induced reaction (HIR) protein, CaHIR1, and suppresses cell death induced by the CaHIR1 protein. Mol Plant Pathol 8:503–514. https://doi.org/10.1111/j.1364-3703.2007.00410.x CrossRefGoogle Scholar
- Lewis LA, Polanski K, de Torres-Zabala M, Jayaraman S, Bowden L, Moore J, Kulasekaran S (2015) Transcriptional dynamics driving MAMP-triggered immunity and pathogen effector-mediated immunosuppression in Arabidopsis leaves following infection with Pseudomonas syringae pv tomato DC3000. Plant Cell 27:3038–3064. https://doi.org/10.1105/tpc.15.00471 CrossRefGoogle Scholar
- Marsberg A, Kemler M, Jami F, Nagel JH, Postma-Smidt A, Naidoo S, Wingfield MJ, Crous PW, Spatafora JW, Hesse CN, Robbertse B, Slippers B (2017) Botryosphaeria dothidea: a latent pathogen of global importance to woody plant health. Mol Plant Pathol 18:477–488. https://doi.org/10.1111/mpp.12495 CrossRefGoogle Scholar
- Pieterse CMJ, Van der Does D, Zamioudis C, Leon-Reyes A, Van Wees SCM (2012) Hormonal modulation of plant immunity. Annu Rev Cell Dev Biol 28:489–521. https://doi.org/10.1146/annurev-cellbio-092910-154055 CrossRefGoogle Scholar
- Qi Y, Tsuda K, Nguyen LV, Wang X, Lin J, Murphy AS, Glazebrook J, Thordal-Christensen H, Katagiri F (2011) Physical association of Arabidopsis hypersensitive induced reaction proteins (HIRs) with the immune receptor RPS2. J Biol Chem 286:31297–31307. https://doi.org/10.1074/jbc.M110.211615 CrossRefGoogle Scholar
- Rostoks N, Schmierer D, Kudrna D, Kleinhofs A (2003) Barley putative hypersensitive induced reaction genes: genetic mapping, sequence analyses and differential expression in disease lesion mimic mutants. Theor Appl Genet 107:1094–1101. https://doi.org/10.1007/s00122-003-1351-8 CrossRefGoogle Scholar
- Sarowar S, Kim YJ, Kim EN, Kim KD, Hwang BK, Islam R, Shin JS (2005) Overexpression of a pepper basic pathogenesis-related protein 1 gene in tobacco plants enhances resistance to heavy metal and pathogen stresses. Plant Cell Rep 24:216–224. https://doi.org/10.1007/s00299-005-0928-x CrossRefGoogle Scholar
- Sarris PF, Duxbury Z, Huh SU, Ma Y, Segonzac C, Sklenar J, Derbyshire P, Cevik V, Rallapalli G, Saucet SB, Wirthmueller L, Menke FLH, Sohn KH, Jones JDG (2015) A plant immune receptor detects pathogen effectors that target WRKY transcription factors. Cell 161:1089–1100. https://doi.org/10.1016/j.cell.2015.04.024 CrossRefGoogle Scholar
- Van der Does D, Leon-Reyes A, Koornneef A, Van Verk MC, Rodenburg N, Pauwels L, Van Wees SC (2013) Salicylic acid suppresses jasmonic acid signaling downstream of SCFCOI1-JAZ by targeting GCC promoter motifs via transcription factor ORA59. Plant Cell 25:744–761. https://doi.org/10.1105/tpc.112.108548 CrossRefGoogle Scholar
- Vlot AC, Dempsey DMA, Klessig DF (2009) Salicylic acid, a multifaceted hormone to combat disease. Annu Rev Phytopathol 47:177–206. https://doi.org/10.1146/annurev.phyto.050908.135202 CrossRefGoogle Scholar
- Wang B, Liang X, Gleason ML, Zhang R, Sun G (2018) Comparative genomics of Botryosphaeria dothidea and B. kuwatsukai, causal agents of apple ring rot, reveals both species expansion of pathogenicity-related genes and variations in virulence gene content during speciation. IMA Fungus 9:243. https://doi.org/10.5598/imafungus.2018.09.02.02 CrossRefGoogle Scholar
- Xue H, Cao S, Li H, Zhang J, Niu J, Chen L, Zhang F, Zhao D (2017) De novo transcriptome assembly and quantification reveal differentially expressed genes between soft-seed and hard-seed pomegranate (Punica granatum L.). PLoS ONE 12:e0178809. https://doi.org/10.1371/journal.pone.0178809 CrossRefGoogle Scholar
- Zhang G, Dong YL, Zhang Y, Li YM, Wang XJ, Han QM, Guo J, Huang LL, Kang ZS (2009) Cloning and characterization of a novel hypersensitive-induced reaction gene from wheat infected by stripe rust pathogen. J Phytopathol 157:722–728. https://doi.org/10.1111/j.1439-0434.2009.01556.x CrossRefGoogle Scholar
- Zhang Q, Yong D, Zhang Y, Shi X, Li B, Li G, Liang W, Wang C (2016) Streptomyces rochei A-1 induces resistance and defense-related responses against Botryosphaeria dothidea in apple fruit during storage. Postharvest Biol Technol 115:30–37. https://doi.org/10.1016/j.postharvbio.2015.12.013 CrossRefGoogle Scholar
- Zhang W, Dong C, Zhang Y, Zhu J, Dai H, Bai S (2018) An apple cyclic nucleotide-gated ion channel gene highly responsive to Botryosphaeria dothidea infection enhances the susceptibility of Nicotiana benthamiana to bacterial and fungal pathogens. Plant Sci 269:94–105. https://doi.org/10.1016/j.plantsci.2018.01.009 CrossRefGoogle Scholar
- Zhou L, Cheung MY, Zhang Q, Lei CL, Zhang SH, Sun SSM, Lam HM (2009) A novel simple extracellular leucine-rich repeat (eLRR) domain protein from rice (OsLRR1) enters the endosomal pathway and interacts with the hypersensitive-induced reaction protein 1 (OsHIR1). Plant Cell Environ 32:1804–1820. https://doi.org/10.1111/j.1365-3040.2009.02039.x CrossRefGoogle Scholar