skip to main content
US FlagAn official website of the United States government
dot gov icon
Official websites use .gov
A .gov website belongs to an official government organization in the United States.
https lock icon
Secure .gov websites use HTTPS
A lock ( lock ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

Explore Research Products in the PAR
It may take a few hours for recently added research products to appear in PAR search results.


Title: Transcription factors VviWRKY10 and VviWRKY30 co-regulate powdery mildew resistance in grapevine
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.  more » « less
Award ID(s):
1901566
PAR ID:
10563269
Author(s) / Creator(s):
; ; ; ; ; ;
Publisher / Repository:
Oxford University Press on behalf of American Society of Plant Biologists.
Date Published:
Journal Name:
Plant Physiology
Volume:
195
Issue:
1
ISSN:
0032-0889
Page Range / eLocation ID:
446 to 461
Subject(s) / Keyword(s):
Grapevine, Powdery mildew, WRKY, immunity
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
More Like this
  1. ; ; ; ; ; ; ; ; ; ; et al (, New Phytologist)
    Summary Calcium‐dependent protein kinases (CDPKs) play vital roles in metabolic regulations and stimuli responses in plants. However, little is known about their function in grapevine.Here, we report thatVpCDPK9andVpCDPK13, two paralogousCDPKsfromVitis pseudoreticulataaccession Baihe‐35‐1, appear to positively regulate powdery mildew resistance. The transcription of them in leaves of ‘Baihe‐35‐1’ were differentially induced upon powdery mildew infection. Overexpression ofVpCDPK9‐YFPorVpCDPK13‐YFPin theV. viniferasusceptible cultivar Thompson Seedless resulted in enhanced resistance to powdery mildew (YFP, yellow fluorescent protein). This might be due to elevation of SA and ethylene production, and excess accumulation of H2O2and callose in penetrated epidermal cells and/or the mesophyll cells underneath.Ectopic expression ofVpCDPK9‐YFPin Arabidopsis resulted in varied degrees of reduced stature, pre‐mature senescence and enhanced powdery mildew resistance. However, these phenotypes were abolished inVpCDPK9‐YFPtransgenic lines impaired in SA signaling (pad4sid2) or ethylene signaling (ein2). Moreover, both of VpCDPK9 and VpCDPK13 were found to interact with and potentially phosphorylate VpMAPK3, VpMAPK6, VpACS1 and VpACS2in vivo(ACS, 1‐aminocyclopropane‐1‐carboxylic acid (ACC) synthase; MAPK, mitogen‐activated protein kinase).These results suggest thatVpCDPK9andVpCDPK13contribute to powdery mildew resistance via positively regulating SA and ethylene signaling in grapevine. 
    more » « less
  2. ; ; ; ; ; ; ; ; (, Journal of Integrative Plant Biology)
    Abstract Study on the regulation of broad‐spectrum resistance is an active area in plant biology.RESISTANCE TO POWDERY MILDEW 8.1(RPW8.1) is one of a few broad‐spectrum resistance genes triggering the hypersensitive response (HR) to restrict multiple pathogenic infections. To address the question how RPW8.1 signaling is regulated, we performed a genetic screen and tried to identify mutations enhancing RPW8.1‐mediated HR. Here, we provided evidence to connect an annexin protein with RPW8.1‐mediated resistance inArabidopsisagainst powdery mildew. We isolated and characterizedArabidopsis b7‐6mutant. A point mutation inb7‐6at theAt5g12380locus resulted in an amino acid substitution in ANNEXIN 8 (AtANN8). Loss‐of‐function or RNA‐silencing ofAtANN8led to enhanced expression ofRPW8.1, RPW8.1‐dependent necrotic lesions in leaves, and defense against powdery mildew. Conversely, over‐expression ofAtANN8compromised RPW8.1‐mediated disease resistance and cell death. Interestingly, the mutation in AtANN8 enhanced RPW8.1‐triggered H2O2. In addition, mutation in AtANN8 led to hypersensitivity to salt stress. Together, our data indicate that AtANN8 is involved in multiple stress signaling pathways and negatively regulates RPW8.1‐mediated resistance against powdery mildew and cell death, thus linking ANNEXIN's function with plant immunity. 
    more » « less
  3. ; ; ; ; ; ; ; ; ; ; et al (, Journal of Experimental Botany)
    Abstract Powdery mildew fungi are obligate biotrophic pathogens that only invade plant epidermal cells. There are two epidermal surfaces in every plant leaf: the adaxial (upper) side and the abaxial (lower) side. While both leaf surfaces can be susceptible to adapted powdery mildew fungi in many plant species, there have been observations of leaf abaxial immunity in some plant species including Arabidopsis. The genetic basis of such leaf abaxial immunity remains unknown. In this study, we tested a series of Arabidopsis mutants defective in one or more known defense pathways with the adapted powdery mildew isolate Golovinomyces cichoracearum UCSC1. We found that leaf abaxial immunity was significantly compromised in mutants impaired for both the EDS1/PAD4- and PEN2/PEN3-dependent defenses. Consistently, expression of EDS1–yellow fluorescent protein and PEN2–green fluorescent protein fusions from their respective native promoters in the respective eds1-2 and pen2-1 mutant backgrounds was higher in the abaxial epidermal cells than in the adaxial epidermal cells. Altogether, our results indicate that leaf abaxial immunity against powdery mildew in Arabidopsis is at least partially due to enhanced EDS1/PAD4- and PEN2/PEN3-dependent defenses. Such transcriptionally pre-programmed defense mechanisms may underlie leaf abaxial immunity in other plant species such as hemp and may be exploited for engineering adaxial immunity against powdery mildew fungi in crop plants. 
    more » « less
  4. ; ; (, Frontiers in Plant Science)
    An integral part of plant immunity is transcription reprogramming by concerted action of specific transcription factors that activate or repress genes through recruitment or release of RNA polymerase II (Pol II). Pol II is assembled into Pol II holoenzyme at the promoters through association with a group of general transcription factors including transcription factor IIB (TFIIB) to activate transcription. Unlike other eukaryotic organisms, plants have a large family of TFIIB-related proteins with 15 members in Arabidopsis including several plant-specific TFIIB-related proteins (BRPs). Molecular genetic analysis has revealed important roles of some BRPs in plant reproductive processes. In this study, we report that Arabidopsis knockout mutants for BRP1, the founding member of the BRP protein family, were normal in growth and development, but were hypersusceptible to the bacterial pathogenPsuedomonas syringae. The enhanced susceptibility of thebrp1mutants was associated with reduced expression of salicylic acid (SA) biosynthetic geneISOCHORISMATE SYNTHASE 1(ICS1) and SA-responsivePATHOGENESIS-RELATED(PR) genes. Pathogen-induced SA accumulation was reduced in thebrp1mutants and exogenous SA rescued thebrp1mutants for resistance to the bacterial pathogen. In uninfected plants, BRP1 was primarily associated with the plastids but pathogen infection induced its accumulation in the nucleus. BRP1 acted as a transcription activator in plant cells and binded to the promoter ofICS1. These results collectively indicate that BRP1 is a functionally specialized transcription factor that increasingly accumulates in the nucleus in response to pathogen infection to promote defense gene expression. 
    more » « less
  5. ; ; ; ; ; ; ; ; ; ; et al (, New Phytologist)
    Summary Biotrophic pathogens are believed to strategically manipulate sugar transport in host cells to enhance their access to carbohydrates. However, mechanisms of sugar translocation from host cells to biotrophic fungi such as powdery mildew across the plant–haustorium interface remain poorly understood.To investigate this question, systematic subcellular localisation analysis was performed for all the 14 members of the monosaccharide sugar transporter protein (STP) family inArabidopsis thaliana. The best candidate AtSTP8 was further characterised for its transport properties inSaccharomyces cerevisiaeand potential role in powdery mildew infection by gene ablation and overexpression in Arabidopsis.Our results showed that AtSTP8 was mainly localised to the endoplasmic reticulum (ER) and appeared to be recruited to the host‐derived extrahaustorial membrane (EHM) induced by powdery mildew. Functional complementation assays inS. cerevisiaesuggested that AtSTP8 can transport a broad spectrum of hexose substrates. Moreover, transgenic Arabidopsis plants overexpressingAtSTP8showed increased hexose concentration in leaf tissues and enhanced susceptibility to powdery mildew.Our data suggested that the ER‐localised sugar transporter AtSTP8 may be recruited to the EHM where it may be involved in sugar acquisition by haustoria of powdery mildew from host cells in Arabidopsis. 
    more » « less
  • Citation Formats
  • MLA
  • APA
  • Chicago
  • BibTeX
  • Save / Share this Record
  • Send to Email