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 technology has increasingly become more and more important over last years. They can be used to improve the breeding of agricultural plants. Grapevine is one of the oldest and most important cultured plants. This article presents a review of the world's main achievements in grapevine genetics and markerassisted selection after the Vitis genome sequence. The process of creating new forms of grapes, as well as all crops, is based on the use of existing genetic diversity. For this reason, the problem of a detailed study of the gene pool of the genus Vitis, wild popu lations and varieties created by men during the long cultivation history of this crop becomes more and more important every year. The genome of Vitis vinifera L. is the fourth sequenced nuclear genome of higher plants. A number of genes and quantitative trait loci were identified and mapped by various research groups, and so were sets of DNA markers for the genes of economically valuable traits. The genes for resistance to the most harmful fungal pathogens Plasmopara vitikola, Erysiphe necator are the most studied. Advancements in genetic mapping and the use of DNA markers in traditional breeding made it possible to refine the genetic mechanism of seedlessness, an important trait in the breeding of table grapes. The study of the genetic control of the content of substances that determine the organoleptic properties of wines has also progressed. Marker-assisted selection (MAS) is used in practice increasingly. Markers associated with disease resistance genes currently used for elimination of susceptible seedlings at the initial stage of large-scale breeding programs carried out in Germany, Italy and the United States. Thus, advances in molecular biology of grapevines creates a conducive situation for active use of DNA-marker technology in this culture.Key words: grapevine genome; DNA markers; identification of genes; molecular marking; markerassisted selection.В последнее время все более важную роль приобретают ДНК-тех-нологии, которые эффективно могут быть использованы для опти-мизации селекции. Виноград Vitis vinifera L. -одно из древнейших и ценнейших культурных растений. В статье приведен обзор основных мировых достижений в области генетики и селекции винограда с периода секвенирования генома виноградного рас -тения. Процесс создания новых форм винограда, как и всех куль-турных растений, базируется на использовании существующего генетического разнообразия. По этой причине проблема деталь-ного изучения генофонда рода Vitis: дикорастущих популяций и сортов, созданных человеком на протяжении длительной исто-рии возделывания данной культуры, -с каждым годом стано вится все актуальней. Геном винограда -четвертый из секвенированных ядерных геномов высших растений. Работами различных исследо-вательских коллективов был определен и картирован ряд генов и локусов количественных признаков виноградного растения, а также наборы ДНК-маркеров генов хозяйственно ценных при-знаков. Основные успехи достигнуты в изучении генов, опреде-ля ющих устойчивость к наиболе...
DNA markers are widely used in grapevine breeding to create forms with combined resistance genes. Downy mildew is one of the most common fungal diseases of the vine in the world. Growing grapevines with increased resistance allows to reduce the number of chemical treatments. The decrease in the use of pesticides is especially significant for viticulture of table varieties, since berries are directly consumed by humans for food. Currently, more than 20 resistance genes have been identified by molecular methods, and DNA markers for many genes have been developed. The genes Rpv3 (inherited from North American grape species) and Rpv12 (derived from V. amurensis) are among the most effective and have an additive effect. The study of 14 table grape varieties for the presence of the Rpv3 gene and 8 varieties for the presence of the Rpv12 gene was performed by using DNA-marker analysis. The analysis included varieties that could inherit these genes from the parent forms, according to their ancestry. The study was conducted using an automatic genetic analyzer ABI Prism 3130 and special software GeneMapper and PeakScanner, DNA-markers were taken from literature sources. According to the results of DNA-marker analysis, 9 varieties were identified, including 2 seedless varieties, with the Rpv3299-279 allele in the genotypes, which determines resistance to downy mildew, and 3 table varieties with the Rpv12 gene in the genotypes. One table grape genotype was identified with Rpv3 and Rpv12.
Recently, the importance of studying the biodiversity of the Vitaceae Juss family. increases markedly, while new data appear on the gene pool of cultivated and wild-growing grapes Vitis vinifera L. Modern methods of analyzing the polymorphism of populations of grape plants, both wild-growing and cultural, are actively used. According to the literature, a number of studies have been conducted in the Russian Federation to study native varieties and wild forms of grapes in the Crimea, Dagestan, along the banks of the Don and other regions, but there is very little information about Kuban wild and ancient forms of grapes. The article presents the results of a study of wild forms of the genus Vitis L., growing in places of ancient human settlements on the territory of the Utrish State Nature Reserve of the Krasnodar Territory. For the first time, ampelographically and genetically samples of isolated populations of wild grapes growing on the territory of the Utrish State Nature Reserve have been studied and described. Several habitats of wild grapes, presumably belonging to the Vitis vinifera ssp species, have been studied. Silvestris Gmel. or its variants var. Tipica Negr. - typically wild forest grapes. A detailed description of phytocenoses in which the studied plant forms grow is carried out. Ecological and geographical characteristics of their habitats are given. Samples were taken for further genetic analysis. DNA-marker analysis of plant genotypes by microsatellite loci VVMD27, VVMD5, VS2, VrZAG79 recommended for genotyping of grapes was performed. It was shown that some of the samples are polymorphic to each other, while others are genetically close. When analyzing genotypes, rare alleles were detected for the analyzed SSR loci.
Table grapes are a valuable dietary product. Seedless grapes are in high demand among consumers. For this reason, the breeding of seedless varieties is one of the popular trends in modern viticulture, along with the production of environmentally friendly products. Downy mildew (Plasmopara viticola (Berk. & M.A. Curtis) Berl. & De Toni) is one of the most common fungal diseases of the grapevine. Most downy mildew resistant grape accessions belong to North American species like Vitis aestivalis Michx., V. berlandieri Planch., V. cinerea (Engelm. ex A. Gray) Engelm. ex Millard, V. riparia Michx., V. rupestris Scheele, etc. The search for donors of resistance genes is an urgent task. Rpv3 is one of the most significant resistance genes from a number of North American grape varieties. The aim of this work is to identify the downy mildew resistance gene Rpv3 in seedless grape varieties by means of DNA-marker analysis. The grape varieties with rudimentary development of seed in berries and with North American species in the pedigree were chosen as the object of the study. The varieties “Dunavski lazur” and “Seyve Villard 12-375” with reference alleles were used as the positive control, while V. vinifera L. was used as the negative control. UDV305 and UDV737 DNA-markers were used in this study to identify the allelic type of the Rpv3 gene. The work was performed using the polymerase chain reaction. The reaction products were separated by capillary electrophoresis using the ABI Prism 3130 automatic genetic analyzer. Evaluation of the results was done using the GeneMapper and PeakScanner software. Functional alleles of the downy mildew resistance gene Rpv3 were revealed in grape varieties “Kishmish zaporozhskiy”, “Lady Patricia”, “Remaily seedless”, “Pamyati Smirnova” and “Shayan”. Rpv3299-279, one of the seven known haplotypes, was identified in all the varieties. The pedigree analysis of the studied varieties indicated that the parental forms – “Seyve Villard” and “Seibel” hybrids – are presumably the donors of the gene. Grape accessions with the identified Rpv3 gene can be used in seedless varieties breeding as donors of resistance to downy mildew.
Работа выполнена при финансовой поддержке Российского фонда фундаментальных исследований и Администрации Краснодарского края (грант № 19-416-230025 р_а).
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