Progress in germplasm identification and genotyping methods in the study of three table grape varieties

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Kuban State Agrarian University, Krasnodar, Russia

Progress in germplasm identification and genotyping methods in the study of three table grape varieties Cost action FA1003: East-West Collaboration for Grapevine Diversity Exploration and Mobilization of Adaptive Traits for Breeding.

Troshin Leonid Petrovich

Dr.Sci.Biol., professor

Milovanov Alexander Valerievich

training master

Mahovickiy Bogdan Alexandrovich

postgraduate student

Derived by combinative breeding three new high-yielding table grape varieties Preobragenie, Victor and Jubilej Novocherkassk with elevated resistance to Phylloxera and fungal diseases and suitable for own-rooted cultivation became a basis for solutions a two hundred year problem in grape growing [1-12, 16-17, 19-30].

It plays an important role a market demand for spreading of these and similar "optimal" hybrid forms of spectacular appearance as a consumer of breeding material, it is difficult to visually distinguishable by morphometry of leaves and therefore needed to recognize when attracting an interactive ampelography [4, 13-15, 18].

Material and Methods

Three table grape sibling varieties were taken for studies, bred by amateur wine-grower Victor Nikolaevich Kraynov from city Novocherkassk under supervising of a professional Ivan Aleksandrovich Kostrikin. The breeding scheme was the following: Talisman [Frumoasa Alba (Guzal kara x Saves Villard 20-473) x Vostorg (Dawn North x Dolores) x Russian early] x Kishmish Luchistyi (Cardinal x Kishmish rozovyi, pink) [2, 6-7, 9, 19, 22-23] (Fig. 1 and 2).

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Fig. 1. Cluster of Talisman [25] Fig. 2. Cluster of Kishmish Luchistyi [25]

While these three (Preobragenie, Victor and Jubilej Novocherkassk) varieties have quite similar phenotypes (Fig. 3-17) its become necessary to conduct comparative morphometric study of leaves [13-15].

Fig. 3-6. Open bud, shoot tip, matured leaf and cluster of Preobragenie

Preobragenie. The variety has very early ripening period - 110-115 days. The clusters are large, in range from 0,7 to 1,5 kg, conic, without berry shot. The berries are very large, 12-18 g, oval-oblong, pink, with harmonious taste. The flesh is fleshy-juicy, the skin is edible. The productivity is high. It can be cultivated as own-rooted plant having medium-power growth. The shoot ripening is good, rooting ability of cuttings is easily, the compatibility with rootstocks is good. The frost resistance is -21°C. Resistance to fungal diseases is of 3,0-3,5 points. The transportability is good with very high market ability. Damaging by wasps is medium. This variety has similar morphological characters as Victor, but vigor of growth is higher respect of it. The variety has ability to give an additional crop on stepson [19].

Fig. 7-10. Open bud, shoot tip, matured leaf and cluster of Victor

Victor. Variety is very early ripening period 110 days. Clusters are of medium loose, from 0,7 to 1 kg, tapered, are not exposed breakage, very interesting. Berries are large, 12-18 g, elongated, dark pink, harmonious taste. Sredneplotnaya pulp, peel eaten. 1-2 seeds per berry pieces. Can be cultivated in own-rooted culture and medium-power growth. Shoots ripen well, cuttings take root easily, compatibility with the stocks is good, freezeproof to -23 °C. To fungal diseases is relatively stable - 3 points. Portability is good, very high marketability. Wasps damaged in the medium. Morphologically similar to Preobragenie, but has a more moderate force growth. Able to give a second crop on stepson [19].

Fig. 11-14. Open bud, shoot tip, matured leaf and cluster of Jubilee Novocherkassk

Jubilee Novocherkassk. Variety is very early ripening period: 110-115 days. The clusters are large, 0,8 to 1,6 kg, tapered, are not exposed breakage. Berries are very large, 12-18 g, oblong-oval, white and pink, harmonious taste. Flesh fleshy-juicy, the skin is eaten. Can be cultivated in own-rooted culture and medium-power growth. Shoots ripen well, cuttings root easily, compatibility with the stocks is good. Average frost resistance, can withstand temperatures of -23 °C. Resistance to fungal diseases 3 points. Transportability and high marketability. Wasps damaged in the medium. Morphologically similar to the sort Victor and Preobragenie, but differs from them in a more subtle berry. Currently proliferate growers Russia and Ukraine [19, 25].

The task of research: a comparison of morphological features descriptor leaves of three grape varieties growing in the testing plots of OAO "Juzhnaja", Temryuk district of the Krasnodar Territory, and to find out their phenotypic identity and DNA- analysis of this siblings.

To determine the morphological features of leaves of compared three varieties of siblings: Preobragenie, Victor and Jubilee Novocherkassk first made their scan and detailed measurement on every sheet of 22 quantitative traits (Fig. 15-30, Table 1).

Fig. 15-16. The leaves of bushes number 4 and number 18 varieties Preobragenie

Рис. 17-18. Double-sided sheet of bush number 9 varieties Preobragenie

Fig. 19-20. Double-sided sheet number 14 of the bush varieties Preobragenie

Fig. 21-22. The leaves of bushes number 2 and number 4 varieties Victor

Fig. 23-24. Double-sided sheet number 5 of the bush varieties Victor

Fig. 25-26. The leaves of bushes number 15 and number 17 varieties Victor

Fig. 27-28. The leaves of the bush number 4 varieties Jubilee Novocherkassk

Fig. 29-30. The leaves of bushes number 23 and 24 varieties Jubilee Novocherkassk

Description scanned 426 raised, that is, adult leaves was conducted using SIAMS Photolab. Information about OIV codes are automatically stored in Excel-table in the following order: 065-1 or 651 in our work, - the length of the leaf blade, 065-2, or 652, - width of the leaf blade, 092 - the length of the petiole, 601 - length of midrib; 602 - length of the upper lateral veins; 603 - length of the lower lateral veins; 604 - length from the intersection of the lower lateral veins to its lower edge 605 - distance to top bay, 606 - distance to lower bay, 607 - alpha angle, 608 - beta angle, 609 - angle zeta, 610 - angle gamma, 611 - the width of the intersection of the lower lateral veins of the lower edge, 612 - the length of the far right tooth, 613 - the width of the far right tooth, 614 - length of the right lower tooth; 615 - the width of the right lower tooth; 616 - the number of teeth the upper side of the blade, 617 - the width of the upper side of the blade, 618 - the distance between the lower lobes; 619 - length from the point of attachment of the petiole to the leaf blade to the end point of the lower teeth [13-15].

Technical work of measurement leafs was performed by A. Milovanov and B. Mahovitskii (Table 1). Provision of selected natural leaf samples carried out by agronomists of OAO "Juzhnaja", Temryuk district of the Krasnodar Territory O.B. Belizovskii and L.A. Lashenko, for which we express our sincere gratitude to them.

Table 1. Data of interactive measurement adult leaves of three grape varieties and 22 original settings

For comparative studies, leaves of varieties were also evaluated 18 additional index features [15].

Results of biometric data analysis of interactive three measurements of 426 leaves of grapes are shown in Table 2 and 3.

Table 2. Variation analysis of the data leaves of the three grape varieties on 22 ampelographic featured

where: ? x - arithmetic mean, sx - error of the mean, lim - the interval of variation, min - minimum value, max - the maximum value, CV - coefficient of variation, %.

According to the data presented in Table 3 and below the Table 4, you can make the following conclusion. The coefficients of variation CV directly measured 22 ampelographic characteristics of leaves, without their biometric transformation from Preobragenie vary within wide ranges: 10,1 - 43,5 under his average of 19,7% in CV Victor almost in the same range: 10,7 - 40,6 at 19,9%, and the cultivar Jubilee Novocherkassk in a smaller amplitude: 14,6 - 35,9 at 19,4%.

Table 4. Variation analysis of these calculations 18 index traits of leaves in three grape varieties

If we analyze the coefficients of variation of 18 index traits of leaves in the same three grape varieties (Table 4), they are reflected in the following figures: 9,3 - 29,2 at 18,5%, 9,3 - 30,3 at 18,9% and 8,7 - 31,7 at 17,5%. The overall mean CV of all index attributes accounted for more than 18% - almost 2% less than the original, that is, direct measurement of the 22 signs.

When combined with 40 CV source and index the basis of their average values ??in the cultivars were at the same level: 19,5, 19,8 and 18,9.

Carried out by the Student's evaluation of differences expressivity 40 ampelographic characteristics in the studied grape varieties has resulted in: between varieties Preobragenie and Victor significant arithmetic differences were found (t < 0,92 for P > 0.05) - the leaves they were twins phenotypic and grade Jubilee of Novocherkassk in comparison with his two above-named siblings metrically different in 21 and 22 cases out of 40 (t > 2,57 at P < 0.01).

For a contemporary analysis of the differences of genotypes was used molecular genetic method. When DNA isolation and PCR reactions were involved in the standard of our research methods and parameters previously highlighted in the literature [30].

DNA extraction was performed upgraded CTAB-method [30]. The paper studied the following protoclones grapes: Victor 29-5 (1), Victor 29-4 (2), Preobragenie 24-6 (3), Preobragenie 24-1 (4), Jubilee Novocherkassk 7-3 (5), Jubilee Novocherkassk 7-1 (6). For a description was used one microsatellite primer: VrZag79.

As a result, imaging and separation of products of the amplification on the acrylamide gel were obtained the following results.

m.w. 1 2 3 4 5 6 m.w.

* m.w. - molecular weight marker

Table 5. The molecular weight of the identified alleles

When comparing it can be concluded that, despite the close relationship, the individuals have different alleles on the number of nucleotides.

Recent evidence table grapes Preobragenie, Victor and Jubilee Novocherkassk an indisputable scientific and production value, and so they create a real basis for profitable production. Ampelometric information of leaf varieties of Preobragenie and Victor is very similar, which is phenotypically geminate and varieties Jubilee Novocherkassk - markedly different from the previous two siblings.

Molecular-genetic specifically PCR-method revealed that varieties Victor, the Preobragenie and the Jubilee Novocherkassk are different genotypes.

Along with these three table grape siblings-"heroes" in OAO "Juzhnaja " and OOO "Fanagoria-Agro" on the Anapa-Taman zone of Krasnodar region notable are varieties Anthonii Velikii, Anjuta, Bogotyanovskii, Gelios (Arcadia rozovaya), Gurman rannii, Dolgozhdannyi, Kishmish Vengerskii, Kubattik, Livia-k, Nizina, Roschfor-k, Super-Extra:

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