V A Pukhal'skiĭ

Vavilov Institute of General Genetics, Moskva, Moscow, Russia

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Publications (22)7.81 Total impact

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    ABSTRACT: Antimutagenic activity on human RD cells was studied for beta-purothionin Tk-AMP-BP isolated from seeds of wheat Triticum kiharae, which has a higher stress resistance. Cadmium chloride at 5 x 10(-6) M was used as a mutagen. The numbers of DNA breaks in mutagen-treated and intact cells were inferred from the single-stranded to double-stranded DNA ratio and expressed as protection coefficients. The protective effect was simultaneously assayed for aqueous plant extracts known to possess antioxidant properties. Wheat thionin was the most active among all of the antimutagens examined; its protection coefficient reached 85-88% at micromolar peptide concentrations (8-32 microg/ml). Thus, wheat beta-purothionin was for the first time demonstrated to be highly efficient in protecting human cell DNA from the damaging effect of cadmium chloride.
    Genetika 09/2011; 47(9):1267-70. · 0.37 Impact Factor
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    ABSTRACT: Using C-banding method and in situ hybridization with the 45S and 5S rRNA gene probes, six hexaploid species of the genus Avena L. with the ACD genome constitution were studied to reveal evolutionary karyotypic changes. Similarity in the C-banding patterns of chromosomal and in the patterns of distribution of the rRNA gene families suggests a common origin of all hexaploid species. Avena fatua is characterized by the broadest intraspecific variation of the karyotype; this species displays chromosomal variants typical of other hexaploid species of Avena. For instance, a translocation with the involvement of chromosome 5C marking A. occidentalis was discovered in many A. fatua accessions, whereas in other representatives of this species this chromosome is highly similar to the chromosome of A. sterilis. Only A. fatua and A. sativa show slight changes in the morphology and in the C-banding pattern of chromosome 2C. These results can be explained either by a hybrid origin of A. fatua or by the fact that this species is an intermediate evolutionary form of hexaploid oats. The 7C-17 translocation was identified in all studied accessions of wild and weedy species (A. sterilis, A. fatua, A. ludoviciana, and A. occidentalis) and in most A. sativa cultivars, but it was absent in A. byzantina and in two accessions of A. sativa. The origin and evolution of the Avena hexaploid species are discussed in context of the results.
    Genetika 06/2011; 47(6):783-95. · 0.37 Impact Factor
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    ABSTRACT: A novel antifungal peptide, LAMP-Ia, was isolated from sand-elymus (Leymus arenarius) seeds. Expression of a synthetic gene encoding this peptide in Escherichia coli cells was obtained. The target peptide was expressed as a fusion with thioredoxin. Identity of the recombinant peptide to native LAMP-Ia was confirmed by chromatography, mass spectrometry, and amino acid sequencing. LAMP-Ia displayed a high inhibitory activity in respect of a number of phytopathogenic fungi in in vitro assays, which opens up possibilities for the gene encoding it to be used for genetic transformation of plants and for engineering pathogen-resistant crops.
    Genetika 12/2010; 46(12):1645-51. · 0.37 Impact Factor
  • V A Pukhal'skiĭ, S P Martynov, E N Bilinskaia
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    ABSTRACT: Study of necrosis genotypes of 72 Russian cultivars of winter common wheat has confirmed a tendency towards "washing off" of genotypes with the Ne1 gene. Fifty-six percent of cultivars have the genotype ne1ne1Ne2Ne2, and 44% have the genotype ne1ne1ne2ne2; i.e., they are free of hybrid necrosis genes. The results of the study indicate that the diversity of the original ancestors in the groups of cultivars with the ne1ne 1Ne2Ne2 and ne1ne1ne2ne2 genotypes is almost the same. This determines the instability of the tendency towards a higher prevalence of the nel ne 1Ne2Ne2 genotype in recent years. The changes in the diversity of the original ancestors with time have shown an increase in the diversity index. These processes may somewhat decrease the rate of genetic erosion caused by the fact that the Ne1Ne1ne2ne2 falls out of breeding. The routes of transmission of necrosis gene alleles from ancestors to descendants have been traced using extended pedigrees, and this information has been used to identify the probable donors and sources of hybrid necrosis gene alleles. In most cases, the cultivars Mironovskaya 808 and Krasnodarskaya 39 are the putative sources of the Ne2allele (60.6 and 27.3% of all cases, respectively). The old cultivar Gostianum 237 from Saratov oblast is the putative source of the Ne2 allele in the cultivar Krasnodarskaya 39. The cultivars Bezostaya 1 and Odesskaya 51 (whose pedigree also includes Bezostaya 1) are the donors of the recessive genotype ne1nelne2ne2 in 93.5% of cases. The old Ukrainian cultivar Ukrainka is the most frequent source of recessive alleles. The strength of the Ne2 allele has been estimated in 36 cultivars. The results indicate that modifier genes affect the expression of tumor necrosis genes.
    Genetika 11/2010; 46(11):1516-24. · 0.37 Impact Factor
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    ABSTRACT: The primary structure of the coat protein (CP) gene was examined for pathogenic strain MS-1 and vaccine strain VIROG-43M of the cucumber green mottle mosaic virus (CGMMV). In CP amino acid composition, strains MS-1 and VIROG-43M are typical representatives of CGMMV: their CPs have 98-100% homology to CPs of other tobamoviruses of the group. The CP gene has the same nucleotide composition in pathogenic MS-1 and vaccine VIROG-43M, indicating that strain attenuation is not determined by this gene. The CP amino acid sequences of the two Russian strains are fully identical to the CP sequences of two Greek strains, GR-3 and GR-5. However, the nucleotide sequences of their genes differ in 13 bp, testifying to the difference between the Russian and Greek strains.
    Genetika 12/2007; 43(11):1461-7. · 0.37 Impact Factor
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    ABSTRACT: The dynamics of genetic transformations at gliadin-coding loci in the winter common wheat cultivars produced in Serbia and Italy over 40 years of scientific breeding was studied. It was demonstrated that a number of alleles unique for the wheat cultivars of each country were substituted with the alleles of a limited number of donor cultivars, in particular, cultivar Bezostaya 1 and its derivatives. On the background of preserved heterogeneity values during various time periods of breeding in each country, the genetic diversity in the total region decreased, as demonstrated by similarity in the sets of alleles of gliadin loci and their frequencies in the modem cultivars of these two Southern European countries. This decrease in the genetic diversity is an erosion of genetic resources within the region, which results in a loss of unique coadapted gene complexes.
    Genetika 12/2007; 43(11):1478-85. · 0.37 Impact Factor
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    ABSTRACT: Ninety-four lines of Triticum dicoccum isolated from 86 wheat accessions from Vavilov All-Russia Research Institute of Plant Industry (VIR, Russia) and INRA (Clermont-Ferrand, France) germ-plasm collections were studied using C-banding technique. Visual comparison of karyotypes of different accessions was performed to establish genetic relationships and evaluate features inherent for ecological--geographical groups. The level of C-banding polymorphism in the whole sample of tetraploid emmer proved to be relatively low. The diversity within groups was mostly higher than the differences between them. The material studied contained 39 lines carrying 16 different types of chromosomal rearrangements including single and multiple translocations and inversions. The level of translocation polymorphism was comparable with that detected earlier for polyploid wheat species. The frequencies of individual translocation types varied from 18 (T7A:5B) to 1 (nine types). Analysis of the distribution of the most frequent translocations & A:5B suggested that it has significant adaptive value on the territory of Europe. Similarity of the C-banding patterns of European emmer and the accessions with the same translocation of the Asian origin points to their possible common origin. The occurrence of the same translocation in several T. dicoccoides accessions from Syria and Lebanon may indicate that such forms of wild emmer could have taken part in the origin of cultivate emmer from Western Europe. Similarity of the C-banding patterns of some chromosomes of European emmer and spelt could serve as an indirect evidence of their close genetic relationships.
    Genetika 11/2007; 43(11):1517-33. · 0.37 Impact Factor
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    ABSTRACT: Differential C-banding of chromosomes and in situ hybridization with the probes pTa71 and pTa794 were used for a comparative cytogenetic study of the three tetraploid oat species with the A and C genomes: Avena insularis, A. magna, and A. murphyi. These species were similar in the structure and patterns of C-banding of several chromosomes as well as in the location of the loci 5S rRNA genes and main NOR sites; however, they differed in the number and localization of minor 45S rDNA loci as well as in the morphology and distribution of heterochromatin in some chromosomes. According to the data obtained, A. insularis is closer to A. magna, whereas A. murphyi is somewhat separated from these two species. Presumably, all the three studied species originated from the same tetraploid ancestor, and their divergence is connected with various species-specific chromosome rearrangements. The evolution of A. murphyi is likely to have occurred independently of the other two species.
    Genetika 07/2007; 43(6):747-61. · 0.37 Impact Factor
  • S P Martynov, T V Dobrotvorskaia, V A Pukhal'skiĭ
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    ABSTRACT: Genealogical analysis was used to study the dynamics of genetic diversity in Russian cultivars of winter common wheat from 1929 to 2005. The Shannon diversity index of the total set of released cultivars remained almost unchanged, although the number of original ancestors (landraces and genetic lines) increased almost tenfold in the period under study. This was explained in terms of the dependence of the modified Shannon diversity index on two parameters, the number of original ancestors and the mean coefficient of parentage. Significant direct effects were revealed: a positive effect of the former parameter and a negative of the latter. As a result, the increase in the number of original ancestors was compensated by the increase in relatedness of cultivars. Genetic erosion of realized diversity was observed, as a half of Russian landraces were lost. Although the mean coefficient of parentage did not reach its critical value (R = 0.25), cultivars of some regions (Central and Volga-Vyatka) proved to be closely related. A favorable gradual decrease in the mean coefficient of parentage was observed in the past 15 years. A set of modem winter wheat cultivars, which were introduced in the Russian State Catalog from 2002 to 2005, displayed a cluster structure. The overwhelming majority of cultivars formed two clusters originating from Bezostaya 1 (67% of cultivars) and Mironovskaya 808 (31%).
    Genetika 11/2006; 42(10):1359-70. · 0.37 Impact Factor
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    ABSTRACT: Two catalogs of alleles of gliadin-coding loci, controlling synthesis of a storage protein of wheat caryopsis, gliadin, were compared. One catalogue comprises the alleles detected according to the electrophoretic patterns in starch gels; the other, in polyacrylamide gels. Determination of the allelic state of gliadin-coding loci in 31 previously not studied cultivars of winter common wheat allowed us to construct a matching system for the alleles compiled in the two catalogs, which gives the possibility to compare the results of wheat cultivar analyses performed at different scientific institutions.
    Genetika 09/2006; 42(8):1107-16. · 0.37 Impact Factor
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    ABSTRACT: Cytogenetic examination showed that three diploid oat species containing the As genome are highly similar in karyotype structure and chromosome C-banding patterns. Avena strigosa is more similar to A. wiestii, while A. hirtula is to an extent separated from the two species, differing in the C-banding pattern of chromosome 6. The karyotypes of all three species harbor a small acrocentric chromosome, which is absent from diploid oat species containing other variants of the A genome. The results made it possible to assume genome specificity of the rearrangement resulting in this chromosome.
    Genetika 01/2006; 41(12):1718-24. · 0.37 Impact Factor
  • S P Martynov, T V Dobrotvorskaia, V A Pukhal'skiĭ
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    ABSTRACT: Based on genealogical analysis, the genetic diversity of 78 spring durum wheat cultivars released in Russia in 1929-2004 have been examined. The temporal trends of change in diversity were studied using series of n x m matrices (where n is the number of the cultivars and m is the number of original ancestors) and calculating coefficients of parentage in sets of cultivars released in particular years. The pool of original ancestors of spring durum wheat cultivars includes 90 landraces and old varieties, more than a half (57%) of which originate from European countries, including Russia and Ukraine (45%). The original ancestors strongly differ in the frequency of presence in the cultivar pedigrees. Landraces Beloturka, Sivouska, Kubanka (T. durum Desf.), Transbaikalian emmer, Yaroslav emmer (T. dicoccum Schuebl.), Poltavka (T. aestivum L.), and the original ancestors of cultivars Kharkov 46, Narodnaya, and Melanopus 1932 enter in the pedigrees of more than half of cultivars created within the framework of various breeding programs. At that, their distribution by cultivars from different breeding centers strongly varies. Analysis of temporal dynamics of genetic diversity, based on genetic profiles and coefficients of parentage, has shown that the genetic diversity of Russian durum wheats increased during the period examined. Nevertheless, genetic erosion of the local material-a loss of approximately 20% of the pool of Russian original ancestors-has been found. The contribution of the original ancestors to the pedigrees of different cultivars, constructed in different breeding centers and recommended for cultivation in different regions, has been estimated. The variation of the released cultivars was highest in the Lower Volga region and lowest in the Ural region. In all, the lower threshold of genetic diversity in all regions does not reach the critical level, corresponding to the similarity of half-sibs. The set of modern cultivars included in the Russian Official List 2004 has a cluster structure.
    Genetika 11/2005; 41(10):1358-68. · 0.37 Impact Factor
  • T V Dobrotvorskaia, S P Martynov, V A Pukhal'skiĭ
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    ABSTRACT: Using genealogy analysis, we studied genetic diversity of 340 cultivars of spring bread wheat that were released on the territory of Russia in 1929-2003. Trends in the temporal change of genetic diversity were inferred from analysis of a set of n x m matrices, where n is the number of the released cultivars and m is the number of original ancestors. The pool of original ancestors of the spring bread wheat cultivars for the total period of study included 255 landraces, of which 88 were from the former USSR and modern Russia. The original ancestors showed great differences in their presence in the cultivar sets examined and, consequently, in their importance for the gene pool of Russian spring wheats. The distributions of contributions of dominant original ancestors to cultivar diversity were significantly different in different regions, indicating that the ancestors were specific for the cultivation conditions. During the last 75 years, the genetic diversity of the spring bread wheat cultivars has been increasing owing to the wide use of foreign material in Russian breeding programs. However, our analysis showed that about 60 landraces, including the Russian ones, were lost during the studied time period. The lost part makes up 35% of the gene pool of the Russian original ancestors. It is reasonable to assume that the lost landraces carried a gene complex f or adaptation to specific Russian environments. Specificity of the contributions of the original ancestors in the sets of cultivars produced in different breeding centers was established. A comparative analysis of genetic similarity of cultivars was carried out using coefficients of parentage. Significant differences in this parameter between breeding institutes and regions of cultivation were revealed.
    Genetika 12/2004; 40(11):1509-22. · 0.37 Impact Factor
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    ABSTRACT: Intraspecific divergence of hexaploid wheat Triticum spelta was studied by chromosome C-banding in 41 accessions of different geographic origins. The spelt accessions did not differ in karyotype structure or heterochromatin distribution from common wheat, but showed greater intraspecific polymorphism for chromosome rearrangements (translocations, inversions) and banding patterns. On evidence of C-banding patterns, spelt was assumed to occupy an intermediate position between tetraploid and hexaploid wheat species. Accessions of the Asian spelt subspecies had more diverse banding patterns than European accessions. A relatively high frequency of chromosome rearrangements was observed in Iranian accessions. Visual analysis revealed high uniformity of chromosome banding patterns in T. spelta populations of Afghanistan, Spain, and Germany (Bavarian group), suggesting a significant role of the founder effect in their evolution.
    Genetika 11/2004; 40(10):1352-69. · 0.37 Impact Factor
  • S V Goriunova, E Z Kochieva, N N Chikida, V A Pukhal'skiĭ
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    ABSTRACT: RAPD analysis was carried out to study the genetic variation and phylogenetic relationships of polyploid Aegilops species, which contain the D genome as a component of the alloploid genome, and diploid Aegilops tauschii, which is a putative donor of the D genome for common wheat. In total, 74 accessions of six D-genome Aegilops species were examined. The highest intraspecific variation (0.03-0.21) was observed for Ae. tauschii. Intraspecific distances between accessions ranged 0.007-0.067 in Ae. cylindrica, 0.017-0.047 in Ae. vavilovii, and 0.00-0.053 in Ae. juvenalis. Likewise, Ae. ventricosa and Ae. crassa showed low intraspecific polymorphism. The among-accession difference in alloploid Ae. ventricosa (genome DvNv) was similar to that of one parental species, Ae. uniaristata (N), and substantially lower than in the other parent, Ae. tauschii (D). The among-accession difference in Ae. cylindrica (CcDc) was considerably lower than in either parent, Ae. tauschii (D) or Ae. caudata (C). With the exception of Ae. cylindrica, all D-genome species--Ae. tauschii (D), Ae. ventricosa (DvNv), Ae. crassa (XcrDcrl and XcrDcrlDcr2), Ae. juvenalis (XjDjUj), and Ae. vavilovii (XvaDvaSva)--formed a single polymorphic cluster, which was distinct from clusters of other species. The only exception, Ae. cylindrica, did not group with the other D-genome species, but clustered with Ae. caudata (C), a donor of the C genome. The cluster of these two species was clearly distinct from the cluster of the other D-genome species and close to a cluster of Ae. umbellulata (genome U) and Ae. ovata (genome UgMg). Thus, RAPD analysis for the first time was used to estimate and to compare the interpopulation polymorphism and to establish the phylogenetic relationships of all diploid and alloploid D-genome Aegilops species.
    Genetika 06/2004; 40(5):642-51. · 0.37 Impact Factor
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    ABSTRACT: Generation of transgenic tobacco plants, producing double-stranded RNA with no homology to tobacco genome sequences is reported. The RNA synthesis is mediated by a construct containing an inverted repeat of the pBR322 tetracycline-resistance gene fragment under control of the 35S CaMV promoter. Analysis of the resistance of transgenic plants to the tobacco mosaic virus revealed the changes in the protein spectra of the infected plants. The 25- and 30-kDa proteins found were not detected in the extracts of normal plants. Amino acid sequencing of the 30-kDa peptide with subsequent computer database search revealed the homology of this protein to the hydrolases belonging to the group of plant beta-glucanases. The role of the novel polypeptides in an increase of the resistance of transgenic plants to TMV, and also the possibility of the regulation of their expression by nonhomologous dsRNA are discussed.
    Genetika 05/2004; 40(4):531-7. · 0.37 Impact Factor
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    ABSTRACT: Based on analysis of gliadin patterns in common wheat cultivars developed at the Research Institute of Agriculture of the Southeast, profile dynamics in gliadin loci has been surveyed for the period of over eight decades. It was shown that long-term breeding of the wheat cultivars involved gradual replacement of alleles characteristic of ancient cultivars for those widely spread in the world, which are probably linked with alleles that currently confer advantage to their carriers. The process of reduction of inter-population genetic diversity in wheat (with special reference to the allele frequency dynamics at gliadin loci) is discussed. This process is responsible for genetic erosion of the species.
    Genetika 11/2003; 39(10):1338-46. · 0.37 Impact Factor
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    ABSTRACT: A comparison of similarity indices between 64 durum wheat cultivars calculated using pedigree analysis and RAPD method showed a correspondence between these two approaches to estimation of genetic diversity. The associations between the results of RAPD clustering and coefficients of parentage (chi 2 test) and the coefficient of correlation between similarity matrices were statistically significant. However, the correlation was rather weak while pedigree analysis and RAPD method did not yield completely identical estimates of genetic diversity in the set of cultivars studied.
    Genetika 10/2003; 39(9):1237-46. · 0.37 Impact Factor
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    ABSTRACT: RAPD genome analysis of 53 species and cultivars of the genus Lycopersicon (Tourn.) Mill. revealed their high genetic polymorphism (Tourn.) Mill., based on which their phylogenetic relationships were inferred. In total, 248 polymorphic DNA fragments were amplified. Intraspecific polymorphism was maximum (79%) in L. peruvianum and minimum (9%) in L. parviflorum. In general, genome divergence among cross-pollinating tomato species was substantially higher than in self-pollinating species. An UPGMA dendrogram constructed from the RAPD patterns was consisted with the Lycopersicon phylogeny inferred from the molecular data of RFLP, ISSR, and microsatellite analyses and with a classification based on morphological characters. The relationships of taxa within the genus Lycopersicon are discussed.
    Genetika 10/2002; 38(9):1298-303. · 0.37 Impact Factor
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    ABSTRACT: Inter-simple sequence repeat (ISSR) analysis was for the first time used to study the genetic diversity and phylogenetic relationships in 54 wild accessions and cultivars of the genus Lycopersicon. Analysis involved 14 ISSR primers homologous to microsatellite repeats and containing additional selective anchor nucleotides. In total, 318 ISSR fragments were amplified for the wild and cultivated tomato genomes. The interspecific polymorphism revealed with the ISSR primers was 95.6%. Species-specific ISSR fragments were detected for each tomato species. The highest number (more than 20) of species-specific fragments were obtained for L. esculentum sensu lato, although the intraspecific variation of ISSR patterns was low. UPGMA cluster analysis was used to construct a dendrogram and to estimate the genetic distances between the species of the genus Lycopersicon; between populations of L. peruvianum, L. pimpinellifolium, and L. esculentum; and between tomato cultivars. The ISSR-based phylogeny was generally consistent with Lycopersicon taxonomy based on morphological and molecular evidence, suggesting the applicability of ISSR analysis for genotyping and phylogenetic studies in tomato.
    Genetika 09/2002; 38(8):1133-42. · 0.37 Impact Factor

Publication Stats

51 Citations
7.81 Total Impact Points

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Institutions

  • 2002–2010
    • Vavilov Institute of General Genetics
      Moskva, Moscow, Russia
  • 2007
    • Институт молекулярной биологии им. В.А. Энгельгардта Российской академии наук
      Moskva, Moscow, Russia
  • 2001
    • Russian Academy of Sciences
      • Engelhardt Institute of Molecular Biology
      Moscow, Moscow, Russia