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      The cytosolic iron–sulphur cluster assembly mechanism in grapevine is one target of a virulent Crinkler effector from Plasmopara viticola

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          Abstract

          Grapevine downy mildew is one of the most devastating diseases in grape production worldwide, but its pathogenesis remains largely unknown. A thorough understanding of the interaction between grapevine and the causal agent, Plasmopara viticola, is helpful to develop alternative disease control measures. Effector proteins that could be secreted to the interaction interface by pathogens are responsible for the susceptibility of host plants. In this study, a Crinkler effector, named PvCRN17, which is from P. viticola and showed virulent effects towards Nicotiana benthamiana previously, was further investigated. Consistently, PvCRN17 showed a virulent effect on grapevine plants. Protein–protein interaction experiments identified grapevine VAE7L1 ( Vitis protein ASYMMETRIC LEAVES 1/2 ENHANCER 7‐Like 1) as one target of PvCRN17. VAE7L1 was found to interact with VvCIA1 and VvAE7, thus it may function in the cytosolic iron–sulphur cluster assembly (CIA) pathway. Transient expression of VAE7L1 in Vitis riparia and N. benthamiana leaves enhanced the host resistance to oomycete pathogens. Downstream of the CIA pathway in grapevine, three iron–sulphur (Fe‐S) proteins showed an enhancing effect on the disease resistance of N. benthamiana. Competitive co‐immunoprecipitation assay showed PvCRN17 could compete with VvCIA1 to bind with VAE7L1 and VvAE7. Moreover, PvCRN17 and VAE7L1 were colocalized at the plasma membrane of the plant cell. To conclude, after intruding into the grapevine cell, PvCRN17 would compete with VCIA1 to bind with VAE7L1 and VAE7, demolishing the CIA Fe‐S cluster transfer complex, interrupting the maturation of Fe‐S proteins, to suppress Fe‐S proteins‐mediated defence responses.

          Abstract

          Two indispensable components of the cytosolic iron–sulphur cluster assembly pathway in grapevine are abducted to the plasma membrane by a virulent CRN effector from Plasmopara viticola to impede grapevine defence responses.

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          The plant immune system.

          Many plant-associated microbes are pathogens that impair plant growth and reproduction. Plants respond to infection using a two-branched innate immune system. The first branch recognizes and responds to molecules common to many classes of microbes, including non-pathogens. The second responds to pathogen virulence factors, either directly or through their effects on host targets. These plant immune systems, and the pathogen molecules to which they respond, provide extraordinary insights into molecular recognition, cell biology and evolution across biological kingdoms. A detailed understanding of plant immune function will underpin crop improvement for food, fibre and biofuels production.
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            Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method.

            Collective efforts of several laboratories in the past two decades have resulted in the development of various methods for Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana. Among these, the floral dip method is the most facile protocol and widely used for producing transgenic Arabidopsis plants. In this method, transformation of female gametes is accomplished by simply dipping developing Arabidopsis inflorescences for a few seconds into a 5% sucrose solution containing 0.01-0.05% (vol/vol) Silwet L-77 and resuspended Agrobacterium cells carrying the genes to be transferred. Treated plants are allowed to set seed which are then plated on a selective medium to screen for transformants. A transformation frequency of at least 1% can be routinely obtained and a minimum of several hundred independent transgenic lines generated from just two pots of infiltrated plants (20-30 plants per pot) within 2-3 months. Here, we describe the protocol routinely used in our laboratory for the floral dip method for Arabidopsis transformation. Transgenic Arabidopsis plants can be obtained in approximately 3 months.
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              Pivoting the plant immune system from dissection to deployment.

              Diverse and rapidly evolving pathogens cause plant diseases and epidemics that threaten crop yield and food security around the world. Research over the last 25 years has led to an increasingly clear conceptual understanding of the molecular components of the plant immune system. Combined with ever-cheaper DNA-sequencing technology and the rich diversity of germ plasm manipulated for over a century by plant breeders, we now have the means to begin development of durable (long-lasting) disease resistance beyond the limits imposed by conventional breeding and in a manner that will replace costly and unsustainable chemical controls.
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                Author and article information

                Contributors
                yan.xu@nwafu.edu.cn
                Journal
                Mol Plant Pathol
                Mol Plant Pathol
                10.1111/(ISSN)1364-3703
                MPP
                Molecular Plant Pathology
                John Wiley and Sons Inc. (Hoboken )
                1464-6722
                1364-3703
                07 September 2022
                December 2022
                : 23
                : 12 ( doiID: 10.1111/mpp.v23.12 )
                : 1792-1806
                Affiliations
                [ 1 ] State Key Laboratory of Crop Stress Biology in Arid Areas College of Horticulture, Northwest A & F University Yangling China
                [ 2 ] Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China Ministry of Agriculture, College of Horticulture, Northwest A & F University Yangling China
                [ 3 ] College of Horticulture Northwest A & F University Yangling China
                Author notes
                [*] [* ] Correspondence

                Yan Xu, State Key Laboratory of Crop Stress Biology in Arid Areas, College of Horticulture, Northwest A & F University, Yangling, China.

                Email: yan.xu@ 123456nwafu.edu.cn

                Author information
                https://orcid.org/0000-0002-1935-7822
                https://orcid.org/0000-0003-3304-3579
                Article
                MPP13266 MPP-OA-22-117.R1
                10.1111/mpp.13266
                9644279
                36071584
                6966a973-eda3-4d00-9699-a3f755b47139
                © 2022 The Authors. Molecular Plant Pathology published by British Society for Plant Pathology and John Wiley & Sons Ltd.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.

                History
                : 25 July 2022
                : 18 April 2022
                : 17 August 2022
                Page count
                Figures: 9, Tables: 1, Pages: 15, Words: 8736
                Funding
                Funded by: National Natural Science Foundation of China , doi 10.13039/501100001809;
                Award ID: 31672115
                Award ID: 31872054
                Categories
                Original Article
                Original Articles
                Custom metadata
                2.0
                December 2022
                Converter:WILEY_ML3GV2_TO_JATSPMC version:6.2.0 mode:remove_FC converted:09.11.2022

                Plant science & Botany
                cia pathway,crn effector,fe‐s proteins,grapevine,plasmopara viticola,virulence

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