[Effect of Triton X-100 on genetic segregation and associated monocotyledonous and dicotyledonous traits in sugarbeet (Beta vulgaris L.)].

The effect of Triton X-100 (TX-100) on the ratio of phenotypic classes and the expression of morphological traits in the progeny of sugar beet hybrids (N12 and N2) was investigated. It was shown that the TX-100 exposition on the unopened flower buds of sugar beets has different effects on hybrid progenies. In agamospermic progeny of hybrid plant No 12km-4, a significant decrease in the heteroallelic (heterozygous) phenotypic classes of alcohol dehydrogenase (ADH1) fraction was determined in the nonagamospermic progeny of hybrid plant No 2km-2 appearance of sugar beet seedlings with one cotyledon leaf was detected. The obtained results indicate the high efficiency of the epimutagenic effect of TX-100 on the early stages of plant ontogenesis.

[Role of polyteny and chromosome-membrane interactions in plant genetic processes].

Existing data in the literature on polyteny in plants and in eukaryotes in general, data on the structure of interphase chromosomes and their association with the nuclear membrane are analyzed in the paper. In light of these data, the results of our studies on the ratios of phenotypic classes of marker enzymes in agamospermy (apomictic) progeny of sugar beet are considered. The given data allow to state that chromosome polyteny and their association with nuclear membrane provide not only hereditary information, but also determine the ratio of genotypes and phenotypes in progeny.

[Violation of the law of uniformity of the first generation of hybrids].

Variability was revealed in the isozyme spectra in the seed progeny from crosses of two beet plants homozygous for contrasting alleles of marker enzyme loci.

[Autosegregation of enzyme loci in agamospermous progenies of triploid plants of sugar beet (Beta vulgaris L.)].

The ratios of the phenotypic classes of glucosephosphate isomerase (GP12) and malate dehydrogenase (MDH1 and MDH2) were studied in agamospermous progenies of triploid sugar beet plants. The ratio of the phenotypic classes of these enzymes corresponds to the calculations based on the assumption of polyteny of chromosomes carrying alleles of the enzyme loci accompanied by the loss of extra copies of the alleles in the first division of a cell entering embryogenesis. An increase in the gene dosage due to polyteny leads to the appearance in the progeny with a definite frequency of alleles that were absent in the original parental plant. The notions of meiotic autosegregation and mitotic autosegregation characteristic of meiotic and mitotic agamospermy are introduced, as well as the term locus polygenotype characterizing not only the allelic composition and the number of chromosomes, but also the number of chromatids carrying alleles of the marker locus in the cell before its entry into embryogenesis.

[Effect of colchicine and Triton X-100 on expression of the enzyme-encoding genes in nongerminating seeds of sugarbeet (Beta vulgaris L.)].

The expression of the enzyme-coding genes, controlling glucose-phosphate isomerase (GPI), malate dehydrogenase (MDH), and alcohol dehydrogenase (ADH), was examined in nongerminating seeds of sugarbeet after Triton X-100 (TX-100) and colchicine treatment. Two types of changes revealed included modification of the enzymatic loci expression (change of the isozyme electrophoretic mobility) and inactivation of standard profiles. In the MDH and GPI systems, these processes were found to be associated. Complete isozyme modification was accompanied with the disappearance of standard profiles. In the ADH system, the treatment with TX-100 and colchicine gave rise to two independent processes, including silencing of the Adh1 locus and the appearance of the ADH isozymes with abnormal electrophoretic mobility, which were probably the products of the Adh2 locus. It was suggested that the effect of TX-100 and colchicine on the expression of the enzyme-encoding genes examined depended on the intracellular localization of the encoded enzymes.

[Triton X-100 induces heritable changes of morphological characters in Triticum aestivum L].

The effect of the nonionic detergent polyethylene glycol octylphenyl ester (Triton X-100, TX-100) on the spring common wheat cultivar Alem was studied under laboratory and field conditions. Treatment of seeds and vegetating plants with 0.1 or 0.01% TX-100 (aqueous solution) changed the spike morphology in all plants of the first posttreatment generation. The changes were inherited by the second generation without additional treatment with TX-100. Square-headed dense spikes with doubled spikelets of the duospiculum type (an additional spikelet at the top of the main one), elongate dense and lax spikes, mid-dense spikes, and fusiform spikes were observed. An epigenetic nature was assumed for the observed changes.

[Marker characters of variation induced in Triticum aestivum L. by nicotinic acid].

A search for markers of epigenetic variability in spring common wheat form Genotroph-1, obtained after the treatment of seeds and growing plants of spring common wheat cultivar Kazakhstanskaya-126 with nicotinic acid of innate origin (0.1-0.01% water solutions), and in a winter wheat line Gostianum-88 along with a common wheat cultivar Alem obtained on the basis of Genotroph-1 was conducted under laboratory conditions. The possibility of using isoenzymes as markers for studying variation induced by natural nicotinic acid with high frequency was demonstrated. Enzyme glutamate oxaloacetate transaminase-2, as a marker by which modified plants and lines obtained on the basis of these plants differ from the original cultivar, was identified.

[Polymorphism of PCR profiles and expression of alleles at the locus Adh1 in agamospermous progeny of beet root Beta vulgaris L].

A modification of the ISSR amplification method based on using a combination of microsatellite and specific unique primer is proposed and tested. This modification simplifies the detected PCR profiles and allows the examination of DNA regions containing definite genes. Combinations of microsatellite primer Mic2 (5'-gacag-acaga-cagac-a-3') and one of the primers specific to the Adh1 locus, which controls alcohol dehydrogenase (ADH1) in sugar beet, were employed in this work. The microsatellite primer was used in combination with the following specific primers: Adh1f (5'-agagt-gttgg-agagg-gtgtg-ac-3') containing the binding site at the fourth exon of gene Adh1, or Adh1r (5'-act(ct)a-cagca-ag(ct)cc-(ct)ac(ct)g-ctcc-3') that binds to the fifth exon of the same gene. In the agamospermous progeny of individual heterozygous diploid plants of sugar beet with the Adh1-F/Adh1-S genotype, polymorphism of PCR profiles obtained in plants of each of three phenotypic classes (FF, FS, and SS) was detected. Among plants of the progeny from an individual plant that represents the heterozygous phenotypic class FS, differences were revealed not only between the PCR profiles but also in the relative activity of allele isozymes of ADH1.

[Subcellular localization of isozymes of NAD-dependent malate dehydrogenase in sugar beet Beta vulgaris L].

Subcellular localization of isozymes of NAD-dependent malate dehydrogenase (MDH) in sugar beet was studied. Isozymes ss and 11 controlled by loci Mdh2 and Mdh3, respectively, were shown to locate in mitochondria, whereas isozyme pp controlled by locus Mdh1, in microbodies. All examined samples lack hybrid MDH isozymes, which could testify to the interaction between products of nonallelic Mdh genes. This can be explained by the localization of nonallelic isozymes in various compartments of the cell and organelles.

[Analysis of natural samples for studying the variation in sugar beat (Beta vulgaris L.) agamospermous seed progeny].

The existence of natural genetic and natural mixed samples of seeds was theoretically grounded and experimentally demonstrated. A natural genetic sample is detected when analyzing the seed agamospermous progeny in the sequence coinciding with the sequence of seed setting, i.e., in the order they are located on a branch. The method of natural genetic samples has demonstrated the presence of individual regions with nonrandom distribution of phenotypic classes of agamospermous seeds on the branches of sugar beet plants. This phenotype distribution reflects the state of somatic cells on extended branch regions and the manifestation of this state in seed-bud cells. In turn, the genotype of each seed reflects the state of the seed-bud cell (apozygote) that gave rise to embryo development. We also demonstrated heterogeneity in the ratios of enzyme phenotypes of the seeds set on individual branches, regarding it as a result of the changes arising in somatic tissues at the moment of branching. Analysis of the ratios of seed phenotypes on individual branches of the same plant was named the method of natural mixed samples. The combination of natural genetic and natural mixed samples provides for identifying and analyzing various modes of seed setting (gamospermy and agamospermy) and studying the factors that influence the variation in these processes. We postulated that the nonrandom distribution and ratio of the seed phenotypes on plant branches resulted from a nonrandom endoreduplication of the homologous regions in homologous chromosomes carrying the marker locus alleles.

[Molecular differences in Adh1-F and Adh1-S alleles in the sugar beet Beta vulgaris L]. / Molekuliarnye razlichiia allelei Adh1-F i Adh1-S u sakharnoi svekly Beta vulgaris L.

We compared nucleotide sequences of exon 4 and part of exon 5 of alleles F and S of the Adh1 locus controlling alcohol dehydrogenase in sugar beet. The Adh1-F and Adh1-S sequences of the examined fragment were shown to differ by two nucleotides. Adenine (A) and cytosine (C) of Adh1-F were substituted by respectively thymine (T) and adenine (A) in Adh1-S. Consequently, glutamine and asparagine from the F subunit of ADH1 are replaced by valine and lysine, respectively. Because of differences in the amino acid content, the F subunit is by two elementary charges more negatively charged electrically than the S subunit, which correlates with differences in their electrophoretic mobility. Comparison of the examined Adh1 fragment of sugar beet with its counterparts in other plants showed that the sites bearing substitutions in the former species are classed as variable.