Abstract
The cuticle plays an important role in plant interactions with pathogens and with their surroundings. The cuticle acts as both a physical barrier against physical stresses and pathogens and a chemical deterrent and activator of the plant defense response. Cuticle production in tomato plants is regulated by several transcription factors, including SlSHINE3, an ortholog of the Arabidopsis WIN/SHN3. Here we used a SlSHINE3-overexpressing (SlSHN3-OE) and silenced (Slshn3-RNAi) lines and a mutant in SlCYP86A69 (Slcyp86A69)—a direct target of SlSHN3—to analyze the roles of the leaf cuticle and cutin content and composition in the tomato plant’s defense response to the necrotrophic foliar pathogen Botrytis cinerea and the biotrophic bacterial pathogen Xanthomonas campestris pv. vesicatoria. We showed that SlSHN3, which is predominantly expressed in tomato fruit epidermis, also affects tomato leaf cuticle, as morphological alterations in the SlSHN3-OE leaf tissue resulted in shiny, stunted and permeable leaves. SlSHN3-OE leaves accumulated 38 % more cutin monomers than wild-type leaves, while Slshn3-RNAi and Slcyp86A69 plants showed a 40 and 70 % decrease in leaf cutin monomers, respectively. Overexpression of SlSHN3 resulted in resistance to B. cinerea infection and to X. campestris pv. vesicatoria, correlated with cuticle permeability and elevated expression of pathogenesis-related genes PR1a and AOS. Further analysis revealed that B. cinerea-infected Slshn3-RNAi plants are more sensitive to B. cinerea and produce more hydrogen peroxide than wild-type plants. Cutin monomer content and composition differed between SlSHN3-OE, Slcyp86A69, Slshn3-RNAi and wild-type plants, and cutin monomer extracted from SlSHN3-OE plants altered the expression of pathogenesis-related genes in wild-type plants.
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Acknowledgments
Work in M.L.’s laboratory was partially supported by BARD. A.A. is the incumbent of the Adolpho and Evelyn Blum Career Development Chair of Cancer Research. Work in A.A.’s laboratory was supported by the Israel Science Foundation (ISF) and the European Research Council (ERC) SAMIT project. Work in S.B.'s laboratory was supported by the ISF.
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Supplementary material 2 Bacterial disease symptoms on SlSHN3-OE (SHN3-OE) and wild-type (WT) plants. Bacterial infection-induced necrotic spots can be clearly seen on shoot internodes of infected WT, but not SHN3-OE plants (TIFF 2347 kb)
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Supplementary material 3 GC/MS analysis of cutin composition in SlSHN3-OE (SHN3-OE), Slcyp86A69 and Slshn3-RNAi (RNAi) plants. Fold change of individual cutin monomers in SlSHN3-OE, Slcyp86A69 (cyp86A69) and Slshn3-RNAi as compared with wild-type (WT) plants (TIFF 324 kb)
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Supplementary material 4 Effect of cutin monomers on in vitro B. cinerea spore germination. Percentage of B. cinerea spore germination (a) and colony diameter (b) on PDA plates supplemented with SlSHN3-OE (SHN3-OE), Slcyp86A69 (cyp86A69), SlSHN3-RNAi (RNAi) or wild-type (WT) cutin monomers (TIFF 273 kb)
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Supplementary material 5 Effect of cutin monomers on X. campestris pv. vesicatoria—disease symptoms. Wild-type (WT) leaves were inoculated with X. campestris pv. vesicatoria supplemented with 0.01 μg/μl cutin monomers extracted from either SlSHN3-OE or WT plants, and WT plants inoculated with X. campestris pv. vesicatoria only served as the control. (a) Quantitative analysis of symptoms as reflected by spot number on different shoot internodes. Values are presented as mean ± SE (n = 10). (b) Mean ± SE (n = 6) of log CFU per gram tissue (TIFF 271 kb)
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Buxdorf, K., Rubinsky, G., Barda, O. et al. The transcription factor SlSHINE3 modulates defense responses in tomato plants. Plant Mol Biol 84, 37–47 (2014). https://doi.org/10.1007/s11103-013-0117-1
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DOI: https://doi.org/10.1007/s11103-013-0117-1