Development and characterization of a line with substitution of chromosome 4B of wheat Triticum aestivum L. on chromosome 4Hmar of wild barley Hordeum marinum ssp. gussoneanum (4x).

Introgressive hybridization is the main method of broadening the genetic diversity of bread wheat. Wild barley Hordeum marinum ssp. gussoneanum Hudson (2n = 4x = 28) has useful agronomical traits, such as high resistance to stress factors, that could be a potential source of new genes for bread wheat improvement. This study aimed to evaluate the possibility of introgression of H. marinum chromosomes into the genome of bread wheat using an incomplete amphiploid H. marinum ssp. gussoneanum (4x)-T. aestivum (Pyrotrix 28) (2n = 54) carrying the cytoplasm of wild barley. For this purpose, we crossed the line of bread wheat variety Pyrotrix 28 with an incomplete amphiploid, and then selected cytogenetically stable 42-chromosome plants with a high level of fertility in hybrid progeny. Genomic in situ hybridization (GISH) revealed a pair of H. marinum chromosomes in the genome of these plants. C- banding analysis confirmed that bread wheat chromosome 4B was replaced by wild barley chromosome 4Hmar. SSR markers Xgwm368 and Xgwm6 confirmed the absence of chromosome 4B, and EST markers BAWU808 and BAW112 identified chromosome 4Hmar in the genome of the isolated disomic wheat-barley substitution line. The study of this line showed that the substitution of chromosome 4B with chromosome 4Hmar resulted in a change of some morphological traits. It included intense anthocyanin coleoptile coloration, specific for H. marinum, as well as a lack of purple coloration of the ears in the leaf sheath, specific for Pyrotrix 28. Line 4Hmar(4B) showed increased performance for several traits, including plant height, number of spikes and tillers per plant, spikelet and grain number in the main spike, grain number per plant, but it had decreased values of 1000-grain weight compared to wheat. Cytogenetic stability and fertility of line 4Hmar(4B) indicated a high compensation ability of barley 4Hmar for wheat chromosome 4B and confirmed their homeology.

Erratum to: "Gas chromatography-mass spectrometry in the taxonomy of Miscanthus".

Vavilovskii Zhurnal Genetiki i Selektsii = Vavilov Journal of Genetics and Breeding. 2019;23(8):1076-1081 (in Russian) Page 1081, in Acknowledgements instead of This work was supported by State Budgeted Project АААА-А17-117092070032-4. should read This work was supported by State Budgeted Project 0259-2019-0011. The original article can be found under DOI 10.18699/VJ19.583.

[The Inheritance of Endosperm Storage Proteins by the Line of the Saratovskaya 29 Cultivar of Common Wheat from its Parental Forms].

We ran a comparative analysis of storage proteins (gliadins, high- (HMW) and low-molecular-weight (LMW) glutenins, puroindolines, and exogenous α-amylase pest inhibitors) in the Saratovskaya 29 cultivar line from the collection of a genetic engineering laboratory, its parental forms (Albidum 24 and Lyutescens 55/11), and distant ancestors (Poltavka, Selivanovskiy Rusak, Sarroza, and tetraploid Beloturka). It was confirmed that the allelic states of storage proteins in the Gli-1, Gli-2 and Glu-1 loci originate from ancestral forms from the collection of the Vavilov Institute of Plant Industry. Moreover, new alleles were found in Lyutescense 55/11 (Glu-Ala) and Selivanovskiy Rusak (Glu-B1b) cultivars from the collection of the Institute of Cytology and Genetics. A new allelic state, Ha, was observed in the loci of the Poltavka cultivar as a soft-grain cultivar, and the ha allele was found in the hard-grain Albidum 24 and Sarroza cultivars. It was found that the expression rate of exogenous α-amylase inhibitors of pests in the Saratovskaya 29 cultivar line is lower than that of ancestral cultivars (Albidum 24, Sarroza, Poltavka, and Beloturka). Such inhibitors are absent in the paternal form Lyutescense 55/11. A high expression rate of protein pest inhibitors for exogenous α-amylases and puroindolines was observed in the Poltavka cultivar. The allelic composition of Glu-1 loci was newly studied in the Sarroza cultivar, which has some promising features. The Saratovskaya 29 cultivar line, on the basis of which a wide range of diverse lines were created in the Institute of Cytology and Genetics, is isogenic for all of the studied traits.

[Prospects for application of breakthrough technologies in breeding: The CRISPR/Cas9 system for plant genome editing].

Integration of the methods of contemporary genetics and biotechnology into the breeding process is assessed, and the potential role and efficacy of genome editing as a novel approach is discussed. Use of molecular (DNA) markers for breeding was proposed more than 30 years ago. Nowadays, they are widely used as an accessory tool in order to select plants by mono- and olygogenic traits. Presently, the genomic approaches are actively introduced into the breeding processes owing to automatization of DNA polymorphism analyses and development of comparatively cheap methods of DNA sequencing. These approaches provide effective selection by complex quantitative traits, and are based on the full-genome genotyping of the breeding material. Moreover, biotechnological tools, such as doubled haploids production, which provides fast obtainment of homozygotes, are widely used in plant breeding. Use of genomic and biotechnological approaches makes the development of varieties less time consuming. It also decreases the cultivated areas and financial expenditures required for accomplishment of the breeding process. However, the capacities of modern breeding are not limited to only these advantages. Experiments carried out on plants about 10 years ago provided the first data on genome editing. In the last two years, we have observed a sharp increase in the number of publications that report about successful experiments aimed at plant genome editing owing to the use of the relatively simple and convenient CRISPR/Cas9 system. The goal of some of these experiments was to modify agriculturally valuable genes of cultivated plants, such as potato, cabbage, tomato, maize, rice, wheat, barley, soybean and sorghum. These studies show that it is possible to obtain nontransgenic plants carrying stably inherited, specifically determined mutations using the CRISPR/Cas9 system. This possibility offers the challenge to obtain varieties with predetermined mono- and olygogenic traits.

[Induced expression of Serratia marcescens ribonuclease III gene in transgenic Nicotiana tabacum L. cv. SR1 tobacco plants].

Transgenic Nicotiana tabacum L. cv. SR1 plants, characterized by an increase in the level of dsRNA-specific hydrolytic activity after induction by wounding, were obtained. The Solanum lycopersicum anionic peroxidase gene promoter (new for plant genetic engineering) was for the first time used for the induced expression of the target Serratia marcescens RNase III gene. Upon infection with the tobacco mosaic virus (TMV), the transgenic plants of the obtained lines did not differ significantly from the control group in the level of TMV capsid protein accumulation. In general, no delay in the development of the infection symptoms was observed in transgenic plants as compared with the control group. The obtained transgenic plants represent a new model for the study of the biological role of endoribonucleases from the RNase III family, including in molecular mechanisms of resistance to pathogens.

[Transgenic Expression of Serratia marcescens Native and Mutant Nucleases Modulates Tobacco Mosaic Virus Resistance in Nicotiana tabacum L].

Extracellular Serratia marcescens nuclease is an extremely active enzyme which non-specifically degrades RNA and DNA. Its antiviral activity was previously shown both in animals and in plants when applied exogenously. Transgenic tobacco plants (Nicotiana tabacum L cv. SR1) expressing S. marcescens chimeric, mutant, and intracellular mutant nuclease gene variants were regenerated and challenged with tobacco mosaic virus. The transgenic plants exhibited a higher level of resistance to the virus infection than the control non-transgenic plants. The resistance was evidenced by the delay of the appearance of mosaic symptoms and the retarded accumulation of viral antigen. Thus, these results reveal that modulations of both extracellular nuclease activity and intracellular RNA/DNA binding can protect plants against viral diseases.

[Virus-Induced Silencing as a Method for Studying Gene Functions in Higher Plants].

The method of virus-induced gene silencing (VIGS) based on posttranscriptional gene silencing (PTGS) is a promising new method for the study of plant gene functions. In the current review we analyzed works on the development and improvement of this method, including the creation of new viral constructions for different plant species, the search for new reporter genes for the control of VIGS efficiency, and the development of new efficient methods of infection.

[Development and study of spring bread wheat variety Pamyati Maystrenko with introgression of genetic material from synthetic hexaploid Triticum timopheevii zhuk. x Aegilops tauschii Coss].

Synthetic hexaploids are bridges for transferring new genes that determine resistance to stress factors from wild-type species to bread wheat. In the present work, the method of developing the spring bread wheat variety Pamyati Maystrenko and the results of its study are described. This variety was obtained using one of the immune lines produced earlier via the hybridization of the spring bread wheat variety Saratovskaya 29 with the synthetic hexaploid T. timopheevii Zhuk. x Ae. tauschii Coss. The C-staining of chromosomes in the Pamyati Maystrenko variety revealed substitutions of 2B and 6B chromosomes by the homeologous chromosomes of the G genome of T. timopheevii and the substitution of chromosome 1D by an orthologous chromosome ofAe. tauschii. It was found that this variety is characterized by resistance to leaf and stem rust, powdery mildew, and loose smut as well as by high grain and bread-making qualities. The role of the alien genetic material introgressed into the bread-wheat genome in the expression of adaptive and economically valuable traits in the Pamyati Maystrenko variety is discussed.

[Comparative Characteristic of Triticum aestivum/Triticum durum and Triticum aestivum/Triticum dicoccum hybrid lines by genomic composition and resistance to fungal diseases under different environmental conditions].

The genetic diversity of common wheat hybrid lines Triticum aestivum/Triticum durum and Triticum aestivum/Triticum dicoccum (2n = 42, F(6-7)) using chromosome-specific microsatellite (SSR) markers and C-staining of chromosomes was studied. Cluster analysis of data obtained by 42 SSR markers indicated that the hybrid lines can be broken into three groups according to their origin. There were two cases of complete genetic similarity between lines 183(2)-2/184(1)-6 and-208-3/213-1, which were obtained using common wheat as the parental plants. In cross combinations, when the stabilization of the nuclear genome of hexaploid lines occurred against a background of the cytoplasmic genome of tetraploid wheats, there was a high level of divergence between sister lines, in some cases exceeding 50%. The evaluation of the degree of susceptibility of the lines to powdery mildew, leaf and stem rust, and septoria leafblotch was performed under different environmental conditions. It was shown that resistance to powdery mildew and leaf rust significantly depended on the region where assays were conducted. An evaluation of the field data showed that he lines 195-3, 196-1, and 221-1 with T. durum genetic material displayed complex resistance to fungal pathogens in Western Siberia and the Republic of Belarus. For lines 195-3 and 196-1, one shows a possible contribution of chromosomes 4B and 5B in the formation of complex resistance to diseases. Hybrid lines with complex resistance can be used to expand the genetic diversity of modern common wheat cultivars for genes of immunity.

[Physiological and agrochemical properties of different symbiotic genotypes of pea (Pisum sativum L.)].

Physiological characters of symbiotic mutants of pea were studied: nodulation, activities of nitrogenase and nitrate reductase, chlorophyll content in leaves and their water-holding capacity, biomass accumulation, and nitrogen forms. The parameters reflecting the genotype state of the macrosymbiont under soil conditions considerably varied. Supernodulation mutants stood out against symbiotic pea genotypes by high contents of chlorophyll and nonprotein nitrogen compounds, high nitrogenase activity, and low nitrate reductase activity. The efficiency of the legume-rhizobium symbiosis was largely mediated by the macrosymbiont genotype. The highest atmospheric nitrogen fixation (50-80%) was observed in the parental pea varieties. Despite the highest nitrogenase activity in the nodules, the supernodulation mutants were inferior to the parental varieties by the nitrogen fixation capacity (40-60%), which was due to their low productivity.

Development of genetic research in the USSR.

Two periods of the development of genetic research in the USSR with reference to its current trends of plant and animal genetics, cytogenetics, and molecular genetics are reviewed. A short list of priority areas is established: the maintenance and use of unique gene pools of plants and animals; the domestication of animals and cultivation of new plants; the development of programmes for mathematical treatment of genetic data banks. It is suggested to consider them within the framework of international projects. The idea is to promote the collaborative efforts of scientists on an international scale.

The occurrence of Ds-like sequences in cereal genomes.

The occurrence of DNA sequences similar to the Ds-element of sh-m5933 maize (Ds-like sequences) was studied in other representatives of the Gramineae. The approximate number of copies of such sequences found under gentle and stringent conditions of washing was determined by dot-hybridization. It was shown that in the maize genome the number of copies of Ds-like sequences exceeds about ten-fold the content of such sequences found in wheat, rye and barley genomes. Quantitative differences in Ds-like sequences between wheat species with various genomes and ploidies (when estimated per genome) as well as between different H. vulgare varieties was not determined. The various melting points (Tm) of DNA-duplexes formed when the Ds-element is hybridized with wheat, rye and barley DNA respectively do not show significant differences and are essentially lower than the Tm of the Ds-element (by 8°-9°C). Thus, these duplexes have 9-11% of nucleotide substitutions in comparison to Ds sh-m5933. The data obtained permit one to suppose the presence of a series of Ds-like sequences heterogenous for the length and degree of homology to the Ds-element isolated from the shrunken locus (sh-m5933) of maize DNA.