One of the directions of apple breeding in the Middle Urals is the development of varieties with a long-term storability. The ability of apples to maintain their consumer qualities for a long period is one of the most important indicators of the variety. A significant role in the storage of apples is played by the amount of ethylene produced in them. The paper presents the results of identification of genes involved in the control of ethylene biosynthesis in apple varieties selected by the Sverdlovsk horticultural breeding station. A total of 21 apple varieties were analyzed. The main objective of the study was to detect Md-ACO1-1 and Md-ACS1-2 alleles in the homozygous state. The combination of these alleles in one genotype reduces the production of ethylene in fruits, which contributes to their long-term storability. The analysis showed the availability of polymorphism in the two studied genes. The Md-ACO1 gene is characterized by the availability of two alleles in most varieties. The homozygous Md-ACO1-1 allele was identified in the Isetskoe pozdnee variety. Analysis of the Md-ACS1 gene revealed the predominance of the Md-ACS1-1 allele form. The Md-ACS1-2 allelic form was observed only in heterozygous samples. No combination of Md-ACO1-1 and Md-ACS1-2 alleles was found in the homozygous state. However, heterozygous forms are also of interest for breeding. They can serve as a source of a character of reduced ethylene biosynthesis when creating varieties with a long-term storability. Such varieties are Sokol yasnyy, Aksena, Rozovatoe zimnee, Sverdlovchanin, Isetskoe pozdnee, Blagaya vest'. The comparison of fruit storage periods and the genotype of the variety has been made. Allelesassociated with a reduced level of ethylene biosynthesis are typical both for the varieties with low and high storability.

Background. Clonal apple rootstocks are one of the main components of intensive gardening. The degree of rootstock damage by fire blight affects the resistance of the variety–rootstock combination. The paper presents a study on marking quantitative trait loci (QTL) of resistance to fire blight Erwinia amylovora in clonal apple rootstock.Materials and methods. A collection of 20 rootstock forms was analyzed. For the study, SCAR markers GE-8019, AE10-375 and microsatellite marker CH-F7-FB1 were used.Results. Polymorphism was observed for all three markers, and their various combinations in one genotype were revealed. It was previously noted that genotypes that carry all three markers were more resistant than those that lack them. The presence of all three markers was observed only in forms 62-396 (В10), 16-1 and 2-9-102. The other genotypes did not have the GE8019 marker. The AE10-375 marker was identified in eight clonal rootstocks. Microsatellite marker CH-F7-FB1 was present in all tested rootstocks. However, polymorphism was detected there. Most genotypes had a 174 bp fragment, but a 210 bp fragment was identified in two of the 20 forms. Clonal rootstock 70-20-21 proved heterozygous for this marker. The analyzed collection also included samples that had only the microsatellite marker: G16, Malysh Budagovskogo, Paradizka Budagovskogo (B9), 54-118 (В118), 57-491, 70-20-20 (В119), 70-20-21, 71-7-22, 76-3-6, 83-1-15, 87-7-12, and 2-12-10. The study of rootstock forms on the basis of resistance to metabolites of the fire blight pathogen was carried out under laboratory conditions using the E. amylovora culture filtrate in vitro on leaf explants. Most of the studied genotypes had different combinations of markers. However, the experiments showed that forms 62-396 and 14-1 with two out of three markers (AE10-375 and CH-F7-FB1) phenotypically manifested the trait of resistance to metabolites of E. amylovora.

At Pushkin and Pavlovsk Laboratories of the N.I. Vavilov All-Russian Institute of Plant Genetic Resources (VIR) a diverse collec tion of local apple cultivars is maintained. Some of the cultivars are widely used in breeding programs for their ecological plasticity, increased adaptation to abiotic stress and disease resistance, still there have been no large-scale studies of these local cultivars for fruit storage ability. Fruit softening during storage is an important problem for apple production. Retention of desirable firmness after prolonged storage is one of the key requirements for new apple cultivars. Expansin and ethy lene biosynthesis related genes are known to be involved in control of fruit softening in apple, and gene specific molecular markers have been reported. In this study the polymorphism and allelic configuration of ethylene and expansin biosynthesis related genes Md-ACS1, Md-ACO1 and Md-Exp7 involved in control of fruit softening in 87 local apple cultivars from VIR Collection of Plant Genetic Resources were analyzed. PCR markers Md-ACS1, Md-ACO1 and SSR-marker Md-Exp7 were used in the study. The allele frequencies in the collection generally coincided with the data from previous studies. Md-ACS1 allele 2 associated with reduced ethylene production was found only in three local cultivars, while all the studied local cultivars were heterozygous for the Md-ACO1 locus, as well as most modern Russian apple cultivars. Half of the studied local cultivars were heterozygous for Md-Exp7 (198 : 202). Thirteen local cultivars with rare Md-Exp7 alleles (206, 210 and 212) were detected. No association was found between the Md-Exp7 genotype and the cultivars’ maturation time. The obtained results can be used for additional evaluation of the cultivars’ potential for breeding.

АНАЛИЗ ГЕНЕТИЧЕСКОЙ КОЛЛЕКЦИИ СОРТОВ И ГИБРИДНЫХ ФОРМ ТОМАТА ПО УСТОЙЧИВОСТИ К КЛАДОСПОРИОЗУ С ИСПОЛЬЗОВАНИЕМ ДНК-МАРКЕРОВ

The Michurinsk State Agrarian University (Michurinsk SAU) is one of the leading centers for breeding apple clonal rootstocks. A diverse collection of apple rootstocks, founded in 1930s by V.I. Budagovsky, is maintained at the Michurinsk SAU. In the present study, 87 rootstocks from this collection were analyzed using 18 SSR markers to assess their genetic diversity and relatedness. The detected polymorphism level was rather high compared to the previous estimates of apple rootstock genetic variability. A total of 199 alleles were detected with an average of 11.1 alleles per locus. Among the detected alleles, 67 (33.67%) were rare and 43 (21.61%) were unique. The average PIC value was 0.73, and the expected and observed heterozygosity averaged 0.76 and 0.69, respectively. All the studied accessions except two could be identified with the used marker set. Cluster analysis revealed several groups according to the rootstocks’ pedigrees and genetic origin. Furthermore, Structure analysis revealed two main groups of the studied rootstock accessions. No significant differentiation of the studied sample according to dwarfing ability was detected, while weak differentiation was detected according to leaf color. SSR genotyping data can be used for rootstock fingerprinting and pedigree verification and will facilitate collection management. In addition, data on the genetic diversity and structure of the studied collection may be useful for further development of the Michurinsk SAU rootstock breeding program.

Apple clonal rootstocks are the basis of modern intensive horticulture, providing a rapid increase in yield and convenience of fruit trees cultivation. Production of clonal rootstocks under high humidity often causes powdery mildew infection caused by the pathogenic fungus Podosphaera leucotricha Salm., which significantly reduces the productivity of stoolbed. Growing powdery mildew resistant genotypes is the most appropriate way to combat this disease and allows reducing the use of fungicides. To accelerate the search for resistant forms, molecular markers associated with resistance genes have been developed. However, these markers have not been used to study clonal rootstocks. The aims of the work were the field assessment of powdery mildew resistance of apple clonal rootstocks from the collection of the Michurinsk State Agrarian University and the screening of the collection for Pl-1, Pl-2, Pl-w and Pl-d resistance genes. The results of a three-year field evaluation of powdery mildew resistance of 80 rootstocks allowed us to distinguish five main groups ranging from very low to highly resistant. A group of 57 accessions was classified as powdery mildew resistant. The search for resistance genes was performed using the AT20 SCAR (Pl-1 gene), OPU02 SCAR (Pl- 2 gene), EM DM01 (Pl-d gene), and EM M02 (Pl-w gene) markers. The Pl-d and Pl-1 genes identified in 33 (41.25 %) and 31 (38.75 %) accessions, respectively, were the most common in the collection. The Pl-w gene was detected only in two accessions. Identification of the Pl-2 gene with the OPU02 SCAR marker did not reveal a fragment of the expected size. Thirty accessions with different powdery mildew resistance scores had two genes, Pl-1 and Pl-d, and highly resistant forms G16 and 14-1 had a combination of the Pl-d and Pl-w genes. These accessions can be used as donors of powdery mildew resistance for breeding new apple clonal rootstocks.