Search

US-12618079-B2 - Powdery mildew resistant grapevine plant

US12618079B2US 12618079 B2US12618079 B2US 12618079B2US-12618079-B2

Abstract

Provided herein are powdery mildew-resistant grapevine plants and methods for providing powdery mildew resistance to susceptible grapevine plants. Specifically, provided herein are grapevine plants including in their genome an impaired Erysiphe necator resistance-conferring gene, wherein the corresponding not impaired Erysiphe necator resistance conferring resistance-conferring gene designated VvMLO13 encodes a protein including the amino acid sequence of SEQ ID No. 1, or proteins having 95% sequence identity therewith. The impairment results in an absence of a protein comprising the amino acid sequence of SEQ ID No. 1, or proteins having 95% sequence identity therewith, in the grapevine plant and wherein the grapevine plant is resistant to powdery mildew.

Inventors

  • Lisa Giacomelli
  • Tieme Zeilmaker
  • Claudio Moser
  • Jeroen Nicolaas Albert Maria Rouppe Van Der Voort

Assignees

  • SCIENZA BIOTECHNOLOGIES 4 B.V.
  • FONDAZIONE EDMUND MACH

Dates

Publication Date
20260505
Application Date
20200618

Claims (7)

  1. 1 . A powdery mildew-resistant grapevine plant comprising in its genome a mutation in a gene having the nucleotide sequence of SEQ ID No. 2, wherein the mutation results in the absence of a protein comprising the amino acid sequence of SEQ ID No. 1, or a protein having 95% sequence identity therewith, and confers resistance to Erysiphe necator in said grapevine plant.
  2. 2 . The grapevine plant according to claim 1 , wherein the alleles of the gene with the mutation have the nucleotide sequences of SEQ ID No. 3, SEQ ID No. 4, and/or SEQ ID No. 5.
  3. 3 . The grapevine plant according to claim 2 , wherein the plant comprises in its genome two alleles selected from the group consisting of SEQ ID No. 3 and SEQ ID No. 4, SEQ ID No. 3 and SEQ ID No. 5, SEQ ID No. 4 and SEQ ID No. 5, SEQ ID No. 3 and SEQ ID No. 3, SEQ ID No. 4 and SEQ ID No. 4, or SEQ ID No. 5 and SEQ ID No. 5.
  4. 4 . The grapevine plant according to claim 1 , wherein the grapevine plant further comprises in its genome one or more Erysiphe necator resistance-conferring genes selected from the group consisting of VvMLO6, VvMLO7 and VvMLO11.
  5. 5 . A method for providing a powdery mildew-resistant grapevine plant, comprising mutating a gene having the nucleotide sequence of SEQ ID No. 2, wherein the mutation results in the absence of a protein comprising the amino acid sequence of SEQ ID No. 1, or a protein having 95% sequence identity therewith, in a powdery mildew-susceptible grapevine plant, thereby providing a grapevine plant that is resistant to powdery mildew.
  6. 6 . The method for providing a powdery mildew-resistant grapevine plant according to claim 5 , wherein the step of mutating the gene encoding a protein comprising the amino acid sequence of SEQ ID No. 1 comprises introducing a deletion, insertion, or substitution in an endogenous sequence comprising the nucleotide sequence of SEQ ID No. 2.
  7. 7 . A seed, fruit, or plant part of a grapevine plant according to claim 1 .

Description

CROSS REFERENCE TO RELATED APPLICATION This application is the United States national phase of International Patent Application No. PCT/EP2020/067007 filed Jun. 18, 2020, the disclosure of which is hereby incorporated by reference in its entirety. The Sequence Listing associated with this application is filed in electronic format via EFS-Web and is hereby incorporated by reference into the specification in its entirety. The name of the text file containing the Sequence Listing is 2208066_ST25.txt. The size of the text file is 29,578 bytes, and the text file was created on Dec. 6, 2022. BACKGROUND OF THE INVENTION Field of the Invention The present invention relates to powdery mildew resistant grapevine plants (Vitis spp.) and to methods and means for providing the present powdery mildew resistance to powdery mildew susceptible grapevine plants and to methods and means for providing the present powdery mildew resistant grapevine plants. Description of Related Art Erysiphe necator, also designated as Uncinula necator, is a fungus that causes powdery mildew disease symptoms in grapevine plants. The fungus is a common pathogen for Vitis species of which the most important species is Vitis vinifera. Grapevine plants require large amounts of pesticides, particularly fungicides, to prevent yield losses. Between 1992 and 2003, 73% of the fungicides sold in France, Italy, Spain and Germany, were used for grapevine protection, a crop that covers only 8% of the land used for agriculture in the considered countries (EUROSTAT, 2007). Grapevine powdery mildew (PM) caused by the fungus Erysiphe necator, is one of the most economically relevant diseases of grapevine worldwide. Erysiphe necator is an obligate biotroph that can infect all green tissues of grapevine and causes significant losses in yield and berry quality. PM symptoms include a white or grey powder covering of the upper and lower surfaces of the leaves. Fruit infections result in shriveling or cracking of the berries. The quality of the fruit is severely damaged, with increased acidity and decreased anthocyanin and sugar content. Powdery mildew is controlled with frequent applications of chemical fungicides. However, the intense application of chemical fungicides has several drawbacks. First of all, the effects on the environment of fungicides are well documented. Secondly, the costs of the chemicals and their applications can reach up to 20% of the total expenses for grape production in some areas. Thirdly, the development of resistant populations of the pathogen was already documented by Baudoin et al. (2008) and Dufour et al. (2011), strongly reducing the efficacy of chemical treatments. Therefore, there is increasing interest in the development of new alternative methods to chemical treatments. The generation of PM-resistant varieties is one of the best options to make sustainable grapevine cultivation a realistic possibility, preserving at the same time the incomes of the growers. A study carried out on “Chardonnay” production in California, showed that the use of a PM-resistant variety could save the growers around $720 per hectare, with a significant reduction of fungicide usage (Fuller et al., 2014). Most cultivars of the European grapevine (Vitis vinifera), which includes the world's finest and most widely planted wine and table grapevine cultivars, are highly susceptible to PM (Gadoury et al. 2003). In contrast, North American Vitis species co-evolved with E. necator and possess various levels of resistance to the pathogen (Fung et al., 2008). This resistance could be introgressed by crossing Vitis vinifera with one of the resistant American Vitis species, but breeding is a slow process in grapevine and the acceptance of resistant hybrids by producers and consumers has been limited in the past (Fuller et al., 2014). The use of technologies like genetic transformation or high-throughput marker-assisted selection can be used to obtain resistant grapevine cultivars with desirable grape properties for producers and consumers (Feechan et al., 2013a). The most common strategy to develop resistant plants is focused on the introgression of resistance genes (R-genes). R-genes encode proteins that recognize pathogen effectors and trigger a defense response, mediated by a signaling network in which plant hormones play a major role (Pavan et al., 2010). Resistance is manifested as a localized hypersensitive response at the site of infection (Bari and Jones, 2009). Resistance conferred by R-genes is scarcely durable, as mutations of pathogen effectors allow it to overcome resistance (Parlevliet et al., 1993). An alternative approach is based on the inactivation of susceptibility genes (S-genes), defined as genes whose loss-of-function results in recessively inherited resistance (Pavan et al., 2010). Some pathogens are able to suppress plant defense by activating plant proteins whose function is the negative regulation of plant immunity system. The genes encoding