DNA-marker based identification of the RPV3 gene determining downy mildew resistance in grapevines

Abstract

Downy mildew is one of the most common fungal diseases of the vine, caused by Plasmopara viticola. An effective way to control the spread of the pathogen is to cultivate resistant varieties. Cultivars of Vitis vinifera, being the basis of high-quality viticulture, practically do not possess genetic resistance to P. viticola, so screening for resistance donors is an important stage in breeding. One of the major resistance loci to downy mildew, the Rpv3 gene, was identified in the genotype of a complex interspecific hybrid of grapes Bianca. Later, it was found that this gene had seven haplotypes of resistance inherited from North American grape species, and that it was possible to identify the allelic status of the gene using DNA-markers UDV305, UDV737. However, only two haplotypes can be combined in one diploid form. To determine the Rpv3 gene in the grape gene pool we, using these markers, studied 35 different genotypes of grapevines, most of which are interspecies cultivars. Three varieties with known allelic status of the Rpv3 gene (Dunavski lazur, Noah, Seyve Villard 12-375) were included in the study as reference genotypes. The genotypes were studied through polymerase chain reaction with separation of amplification products by capillary electrophoresis in automatic genetic analyzer ABI Prism 3130. In the studied grape cultivars DNA marker analysis indentified the Rpv3 gene in sixteen genotypes of interspecific origin, including haplotype Rpv3299-279 found in twelve varieties, Rpv3321-312 – in three, and haplotype Rpv3null-271 – in one variety. Seyve Villard 12-375 turned out to be the donor of resistance gene in the most of the genotypes carrying Rpv3 in this study. The obtained data can be useful in selection of mildew resistant grape varieties and screening for hybridization pairs.

DNA-marker identification of Rpv3 and Rpv12 resistance loci in genotypes of table and seedless grape varieties

“…We have already started research for searching gene Rpv3 donors and some of them are published [13][14][15]. A large work in this direction was done by Di Gaspero and colleagues: DNA-marker analysis of the Rpv3 locus in 580 genotypes of various origins [10].…”

DNA marker identification of Rpv3 downy mildew resistance gene in seedless grape varieties

“…Образцы ДНК выделяли из молодых верхушек побегов растений, экстракцию ДНК осуществляли с помощью буфера на основе ЦТАБ (цетилтриметиламмоний бромида) (Rogers, Bendich, 1985). Полимеразную цепную реакцию (ПЦР) проводили в конечном объеме 25 мкл по ранее отработанному протоколу (Ilnitskaya et al, 2018). В исследовании использовали микросателлитные ДНК-маркеры UDV305 и UDV737, позволяющие идентифицировать аллельное состояние гена Rpv3.…”

In Vitro Collection for the Safe Storage of Grapevine Hybrids and Identification of the Presence of Plasmopara viticola Resistance Genes

“…The Rpv3 and Rpv12 loci, derived from North American, Eurasian, and Asian grapevines ( Vitis spp. ), respectively, are the most widely used Rpvs for resistance to P. viticola and are currently of relevance in viticultural breeding programs [ 34 , 35 , 36 , 37 , 38 ]. Resistance to downy mildew is thought to involve more than one gene [ 36 , 37 , 38 , 39 ].…”

“…), respectively, are the most widely used Rpvs for resistance to P. viticola and are currently of relevance in viticultural breeding programs [ 34 , 35 , 36 , 37 , 38 ]. Resistance to downy mildew is thought to involve more than one gene [ 36 , 37 , 38 , 39 ]. Therefore, to breed mildew-resistant grape varieties, it is highly desirable to combine as many resistance genes as possible, known as pyramiding, to provide not only effective but also durable resistance.…”