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Abstract Grapevine (Vitis vinifera) is an economically important fruit crop worldwide. The widely cultivated grapevine is susceptible to powdery mildew caused by Erysiphe necator. In this study, we used CRISPR-Cas9 to simultaneously knock out VviWRKY10 and VviWRKY30 encoding two transcription factors reported to be implicated in defense regulation. We generated 53 wrky10 single mutant transgenic plants and 15 wrky10 wrky30 double mutant transgenic plants. In a 2-yr field evaluation of powdery mildew resistance, the wrky10 mutants showed strong resistance, while the wrky10 wrky30 double mutants showed moderate resistance. Further analyses revealed that salicylic acid (SA) and reactive oxygen species contents in the leaves of wrky10 and wrky10 wrky30 were substantially increased, as was the ethylene (ET) content in the leaves of wrky10. The results from dual luciferase reporter assays, electrophoretic mobility shift assays and chromatin immunoprecipitation (ChIP) assays demonstrated that VviWRKY10 could directly bind to the W-boxes in the promoter of SA-related defense genes and inhibit their transcription, supporting its role as a negative regulator of SA-dependent defense. By contrast, VviWRKY30 could directly bind to the W-boxes in the promoter of ET-related defense genes and promote their transcription, playing a positive role in ET production and ET-dependent defense. Moreover, VviWRKY10 and VviWRKY30 can bind to each other's promoters and mutually inhibit each other's transcription. Taken together, our results reveal a complex mechanism of regulation by VviWRKY10 and VviWRKY30 for activation of measured and balanced defense responses against powdery mildew in grapevine.
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N -hydroxypipecolic acid and salicylic acid play key roles in autoimmunity induced by loss of the callose synthase PMR4
Abstract In Arabidopsis thaliana, the POWDERY MILDEW RESISTANT4 (PMR4)/GLUCAN SYNTHASE LIKE5 (GSL5) callose synthase is required for pathogen-induced callose deposition in cell wall defense. Paradoxically, pmr4/gsl5 mutants exhibit strong resistance to both powdery and downy mildew. The powdery mildew resistance of pmr4/gsl5 has been attributed to upregulated salicylic acid (SA) signaling based on its dependance on PHYTOALEXIN DEFICIENT4 (PAD4), which controls SA accumulation, and its abolishment by bacterial NahG salicylate hydroxylase. Our study revealed that disruption of PMR4/GSL5 also leads to early senescence. Suppressor analysis uncovered that PAD4 and N-hydroxypipecolic acid (NHP) biosynthetic genes ABERRANT GROWTH AND DEATH2-LIKE DEFENSE RESPONSE PROTEIN1 (ALD1) and FLAVIN-DEPENDENT MONOXYGENASE1 (FMO1) are required for early senescence of pmr4/gsl5 mutants. The critical role of NHP in the early senescence of pmr4/gsl5 was supported by greatly increased accumulation of pipecolic acid in pmr4/gsl5 mutants. In contrast, disruption of the SA biosynthetic gene ISOCHORISMATE SYNTHASE1/SA-INDUCTION DIFFICIENT 2 (ICS1/SID2), which greatly reduces SA accumulation, had little effect on impaired growth of pmr4/gsl5. Furthermore, while disruption of PAD4 completely abolished the powdery mildew resistance in pmr4/gsl5, mutations in ICS1/SID2, ALD1, or FMO1 had only a minor effect on the resistance of the mutant plants. However, disruption of both ICS1/SID2 and FMO1 abolished the enhanced immunity of the callose synthase mutants against the fungal pathogen. Therefore, while NHP plays a crucial role in the early senescence of pmr4/gsl5 mutants, both SA and NHP have important roles in the strong powdery mildew resistance induced by the loss of the callose synthase.
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- PAR ID:
- 10656163
- Publisher / Repository:
- Oxford University Press
- Date Published:
- Journal Name:
- Plant Physiology
- Volume:
- 198
- Issue:
- 1
- ISSN:
- 0032-0889
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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Accepted Manuscript
- Journal Article:
- https://doi.org/10.1093/plphys/kiaf163
