Editorial
STRESS RESISTANCE IN PLANTS
Vitis rotundifolia Michx. is one of the species of the family Vitaceae, with resistance to both biotic and abiotic stresses. The present study reports new scientific knowledge about the inheritance of resistance to downy mildew, powdery mildew and frost by V. vinifera varieties from V. rotundifolia. Recombinant lines of three hybrid populations from the crossing of the maternal genotype ♀M. 31-77-10 with V. rotundifolia hybrids were used as the object of the study. As a result of laboratory screening, more than 40 % of recombinants of the ♀M. 31-77-10× ×[DRX-M5-734+DRX-M5-753+DRX-M5-790] population showed a high degree of frost resistance (–24 °C), while 6 % of transgressive recombinants were characterized by a very high degree of resistance (–27 °С). The maternal genotype ♀M. 31-77-10 does not carry alleles of resistance to powdery mildew at the Run1 locus and in the field suffers from powdery mildew much more than the paternal genotypes. The prevalence of powdery mildew on vegetative organs in the three recombinant populations over the years varies on average between 3.2–17.1, 0.3–17.7 and 0.6–5.2 %, respectively. As a result, almost all recombinant genotypes that received a resistant allele from the paternal genome are highly resistant to powdery mildew.
Wheatgrass Thinopyrum intermedium is a source of agronomically valuable traits for common wheat. Partial wheat–wheatgrass amphidiploids and lines with wheatgrass chromosome substitutions are extensively used as intermediates in breeding programs. Line Agis 1 (6Agi2/6D) is present in the cultivar Tulaykovskaya 10 pedigree. Wheatgrass chromosome 6Agi2 carries multiple resistance to fungal diseases in various ecogeographical zones. In this work, we studied the transfer of chromosome 6Agi2 in hybrid populations Saratovskaya 29×Tulaykovskaya 10 (S29×T10) and Tulaykovskaya 10×Saratovskaya 29 (T10×S29). Chromosome 6Agi2 was identified by PCR with chromosome-specific primers and by genomic in situ hybridization (GISH). According to molecular data, 6Agi2 was transmitted to nearly half of the plants tested in the F2 and F3 generations. A new breeding line 49-14 (2n = 42) with chromosome pair 6Agi2 was isolated and characterized in T10×S29 F5 by GISH. According to the results of our field experiment in 2020, the line had high productivity traits. The grain weights per plant (10.04±0.93 g) and the number of grains per plant (259.36±22.49) did not differ significantly from the parent varieties. The number of grains per spikelet in the main spike was significantly higher than in S29 (p ≤ 0.001) or T10 (p ≤ 0.05). Plants were characterized by the ability to set 3.77±0.1 grains per spikelet, and this trait varied among individuals from 2.93 to 4.62. The grain protein content was 17.91 %, and the gluten content, 40.55 %. According to the screening for fungal disease resistance carried out in the field in 2018 and 2020, chromosome 6Agi2 makes plants retain immunity to the West Siberian population of brown rust and to dominant races of stem rust. It also provides medium resistant and medium susceptible types of response to yellow rust. The possibility of using lines/varieties of bread wheat with wheatgrass chromosomes 6Agi2 in breeding in order to increase protein content in the grain, to confer resistance to leaf diseases on plants and to create multiflowered forms is discussed.
The Sr22, Sr35, and Sr25 genes attract the attention of bread wheat breeders with their effectiveness against Puccinia graminis f. sp. tritici race Ug99 and its biotypes. The effectiveness and impact of Sr22+Sr25 and Sr35+Sr25 gene combinations on agronomic traits have not yet been studied. In the present article, these traits were studied using the spring bread wheat lines L503/W3534//L503, L503/Sr35//L503/3/L503 carrying the Sr22+Sr25 and Sr35+Sr25 genes during 2016–2020. These lines were assessed for resistance to P. graminis f. sp. tritici under natural epiphytotics and to the Saratov, Lysogorsk and Omsk populations of the pathogen and to the PgtZ1 (TKSTF) and PgtF18.6 fungus isolates in laboratory conditions (TKSTF + Sr33). The presence of the studied Sr-genes was confirmed by using molecular markers. Prebreeding studies were conducted during 2018–2020 vegetation periods. Under the natural epiphytotics of the pathogen and in the laboratory conditions, the Sr22+Sr25 combination was highly effective, while Sr35+Sr25 was ineffective. For grain yield, the lines with the Sr22+Sr25 and Sr35+Sr25 genes were superior to the recipient cultivar L503 in one year (Sr22+Sr25 in 2019; Sr35+Sr25 in 2018), with a decrease in 2020, but in general there were no differences. For the period 2018–2020, both combinations showed a decrease in 1000 grains weight and an increase in the germination-earing period. The line with Sr22+Sr25 genes showed insignificant effects on gluten and dough tenacity, but the ratio of dough tenacity to extensibility was higher, and flour strength, porosity and bread volume were lower; in the line with Sr35+Sr25 genes, the gluten content was lower, but the strength, tenacity of the dough and the ratio of dough tenacity to extensibility were higher, flour strength and the porosity of the bread were at the recipient level, but the volume of bread was lower.
An original initial material of spring and winter bread wheat with group resistance to stem and leaf rust was developed using new donors of resistance to stem rust: winter soft wheat GT 96/90 (Bulgaria) and accession 119/4-06rw with genetic material of the species Triticum migushovae and (Aegilops speltoides and Secale cereale), respectively, a line of spring wheat 113/00i-4 obtained using the species Ae. triuncialis and T. kiharae, as well as spring accession 145/00i with genetic material of the species Ae. speltoides resistant to leaf rust. The transfer of effective Sr-genes to progeny was monitored using molecular markers. New lines underwent a field assessment of resistance to leaf and stem rust in the epiphytotic development of diseases in the Central Region of the Russian Federation, as well as in the North Caucasus and Western Siberia, and showed high resistance to these pathogens. Fourteen genotypes of spring wheat with group resistance to these diseases and parental forms that participated in the origin of the lines were evaluated for resistance to spot blotch (Cochliobolus sativus) and tan spot (Pyrenophora tritici-repentis) using isolates from Kazakhstan and Omsk in laboratory conditions. A highly resistant parental form of winter soft wheat from “Arsenal” collection 119/4-06rw (wheat-Ae. speltoides-rye hybrid 2n = 42) with group resistance to two spots, four medium-resistant genotypes to both isolates of tan spot from Kazakhstan and Omsk populations of the pathogen, as well as genotypes resistant to the Omsk isolate of P. triticirepentis (parental form 113/00i-4 and lines 1-16i, 6-16i, 9-16i) were isolated. Among the lines of winter wheat, four were identified with group resistance to spot blotch and tan spot. Additionally, the stress resistance of the lines to NaCl salinization and prolonged flooding of seeds with water was evaluated at the early stages of ontogenesis in laboratory conditions. Lines 33-16i, 37-16i, 32-16i and 9-16i showed a high ability to withstand excess moisture. Lines 33-16i, 37-16i, 32-16i and 3-16i were characterized by high salt tolerance, exceeding the average of 49.7 %. Among the winter genotypes, lines were identified with increased resistance to hypoxia (37-19w, 32-19w, 16-19w, 90-19w) and with increased salt tolerance (20-19w, 9-19w, 37-19w, 90-19w), significantly exceeding the standard cv. Moskovskaya 39. The listed lines are of interest as sources of resistance to anaerobic and salt stress, as well as donors of resistance to a group of fungal diseases: leaf and stem rust and tan spot. We attribute the increased level of resistance of the new initial material to the presence of alien translocations in the original parental forms involved in the origin of the lines.
Fusarium disease of oats reduces yield quality due to decreasing germination that is caused by then contamination of grain with mycotoxins produced by Fusarium fungi. The aim of this study was to characterize the resistance of naked breeding lines of oats to fungal grain infection and to contamination with T-2 and HT-2 toxins. Thirteen naked oat breeding lines and two naked varieties, Nemchinovsky 61 and Vyatskiy, as well as a husked variety Yakov, were grown under natural conditions in the Nemchinovka Federal Research Center in 2019–2020. The contamination of grain with fungi was determined by the mycological method and real-time PCR. The analysis of mycotoxins was carried out by ELISA. In oats, Alternaria (the grain infection was 15–90 %), Cochliobolus (1–33 %), Cladosporium (1–19 %), Epicoccum (0–11 %), and Fusarium (3–17 %) fungi prevailed in the grain mycobiota. The predominant Fusarium species were F. poae (its proportion among Fusarium fungi was 49–68 %) and F. langsethiae (29–28 %). The highest amounts of F. langsethiae DNA ((27.9–71.9)×10–4 pg/ng) and T-2/HT-2 toxins (790–1230 μg/kg) were found in the grain of husked oat Yakov. Among the analysed naked oat lines, the amount of F. langsethiae DNA varied in the range of (1.2–42.7)×10–4 pg/ng, and the content of T-2/HT-2 toxins was in the range of 5–229 μg/kg. Two oat breeding lines, 54h2476 and 66h2618, as well as a new variety, Azil (57h2396), can be characterized as highly resistant to infection with Fusarium fungi and contamination with mycotoxins compared to the control variety Vyatskiy.
Present-day wheat breeding for immunity exploits extensively closely related species from the family Triticeae as gene donors. The 2NS/2AS translocation has been introduced into the genome of the cultivated cereal Triticum aestivum from the wild relative T. ventricosum. It contains the Lr37, Yr17, and Sr38 genes, which support seedling resistance to the pathogens Puccinia triticina Eriks., P. striiformis West. f. sp. tritici, and P. graminis Pers. f. sp. tritici Eriks. & E. Henn, which cause brown, yellow, and stem rust of wheat, respectively. This translocation is present in the varieties Trident, Madsen, and Rendezvous grown worldwide and in the Russian varieties Morozko, Svarog, Graf, Marquis, and Homer bred in southern regions. However, the Sr38 gene has not yet been introduced into commercial varieties in West Siberia; thus, it remains of practical importance for breeding in areas where populations of P. graminis f. sp. tritici are represented by avirulent clones. The main goal of this work was to analyze the frequency of clones (a)virulent to the Sr38 gene in an extended West Siberian collection of stem rust agent isolates. In 2019–2020, 139 single pustule isolates of P. graminis f. sp. tritici were obtained on seedlings of the standard susceptible cultivar Khakasskaya in an environmentally controlled laboratory (Institute of Cytology and Genetics SB RAS) from samples of urediniospores collected on commercial and experimental bread wheat fields in the Novosibirsk, Omsk, Altai, and Krasnoyarsk regions. By inoculating test wheat genotypes carrying Sr38 (VPM1 and Trident), variations in the purity of (a)virulent clones were detected in geographical samples of P. graminis f. sp. tritici. In general, clones avirulent to Sr38 constitute 60 % of the West Siberian fungus population, whereas not a single virulent isolate was detected in the Krasnoyarsk collection. The Russian breeding material was screened for sources of the stem rust resistance gene by using molecular markers specific to the 2NS/2AS translocation. A collection of hybrid lines and varieties of bread spring wheat adapted to West Siberia (Omsk SAU) was analyzed to identify accessions promising for the region. The presence of the gene was postulated by genotyping with specific primers (VENTRIUP-LN2) and phytopathological tests with avirulent clones of the fungus. Dominant Sr38 alleles were identified in Lutescens 12-18, Lutescens 81-17, Lutescens 66-16, Erythrospermum 79/07, 9-31, and 8-26. On the grounds of the composition of the West Siberian P. graminis f. sp. tritici population, the Sr38 gene can be considered a candidate for pyramiding genotypes promising for the Novosibirsk, Altai, and Krasnoyarsk regions.
DEVELOPMENTAL GENETICS
CLE (CLV3/ESR) is one of the most important groups of peptide phytohormones: its members regulate the development of various plant organs and tissues, as well as interaction with some parasites and symbionts and response to environmental factors. In this regard, the identification and study of the CLE genes encoding the peptides of this group in cultivated plants are of great practical interest. Relatively little is known about the functions of CLE peptides in potato, since the CLE genes of the potato Solanum phureja Juz. et Buk. were characterized only in 2021. At the same time, potato includes plenty of tuberous species of the genus Solanum L., both wild and cultivated, and the diversity of its forms may depend on differences in the sequences of CLE genes. In this work, we performed a search for and analysis of the CLE gene sequences in three wild potato species (S. bukasovii Juz., S. verrucosum Schltdl., S. commersonii Dunal) and four cultivated species (S. chaucha Juz. et Buk., S. curtilobum Juz. et Buk., S. juzepczukii Juz. et Buk., S. ajanhuiri Juz. et Buk.). In total, we identified 332 CLE genes in the analyzed potato species: from 40 to 43 genes of this family for each potato species. All potato species taken for analysis had homologues of previously identified S. phureja CLE genes; at the same time, the CLE42 gene, which is absent from the S. phureja genome, is present in all other analyzed potato species. Polymorphism of CLE proteins of S. commersonii is significantly higher than that of other analyzed potato species, due to the fact that S. commersonii grows in places outside the growing areas of other potato species and this potato is probably not one of the ancestors of cultivated potato. We also found examples of polymorphism of domains of CLE proteins that carried different functions. Further study of potato CLE proteins will reveal their role in development, including regulation of productivity in this important agricultural crop.
Plant sugar transporters play an essential role in the organism’s productivity by carrying out carbohydrate transportation from source cells in the leaves to sink cells in the cortex. In addition, they aid in the regulation of a substantial part of the exchange of nutrients with microorganisms in the rhizosphere (bacteria and fungi), an activity essential to the formation of symbiotic relationships. This review pays special attention to carbohydrate nutrition during the development of arbuscular mycorrhiza (AM), a symbiosis of plants with fungi from the Glomeromycotina subdivision. This relationship results in the host plant receiving micronutrients from the mycosymbiont, mainly phosphorus, and the fungus receiving carbon assimilation products in return. While the efficient nutrient transport pathways in AM symbiosis are yet to be discovered, SWEET sugar transporters are one of the three key families of plant carbohydrate transporters. Specific AM symbiosis transporters can be identified among the SWEET proteins. The survey provides data on the study history, structure and localization, phylogeny and functions of the SWEET proteins. A high variability of both the SWEET proteins themselves and their functions is noted along with the fact that the same proteins may perform different functions in different plants. A special role is given to the SWEET transporters in AM development. SWEET transporters can also play a key role in abiotic stress tolerance, thus allowing plants to adapt to adverse environmental conditions. The development of knowledge about symbiotic systems will contribute to the creation of microbial preparations for use in agriculture in the Russian Federation.
Biotechnology in the postgenomic epoch BIOTECHNOLOGY IN THE POSTGENOMIC EPOCH
Soybean, Glycine max L., is one of the most important agricultural crops grown in a wide range of latitude. In this regard, in soybean breeding, it is necessary to pay attention to the set of genes that control the transition to the flowering stage, which will make it possible to adapt genotypes to local growing conditions as accurately as possible. The possibilities of soybean breeding for this trait have now significantly expanded due to identification of the main genes (E1–E4, GmFT2a, GmFT5a) that control the processes of flowering and maturation in soybean, depending on the day length. The aim of this work was to develop a panel of markers for these genes, which could be used for a rapid and efficient genotyping of domestic soybean cultivars and selection of plant material based on sensitivity to photoperiod and the duration of vegetation. Combinations of 10 primers, both previously developed and our own, were tested to identify different alleles of the E1–E4, GmFT2a, and GmFT5a genes using 10 soybean cultivars from different maturity groups. As a result, 5 combinations of dominant and recessive alleles for the E1–E4 genes were identified: (1) e1-nl(e1-as)/ e2-ns/e3-tr(e3-fs)/e4; (2) e1-as/e2-ns/e3-tr/E4; (3) e1-as/e2-ns/E3-Ha/e4; (4) E1/e2-ns/e3-tr/E4; (5) e1-nl/e2-ns/E3-Ha/E4. The studied cultivars contained the most common alleles of the GmFT2a and GmFT5a genes, with the exception of the ‘Cassidi’ cultivar having a rare dominant allele GmFT5a-H4. The degree of earliness of cultivars positively correlated with the number of recessive genes E1–E4, which is consistent with the data of foreign authors on different sets of cultivars from Japan and North China. Thus, the developed panel of markers can
The use of the gene pool of wild relatives, which have a significant reserve of genetic diversity, is of immediate interest for breeding common wheat. The creation and use of synthetic forms as “bridges” is an effective method of transferring valuable genetic material from wild relatives to cultivated wheat. For this purpose, genome addition, genome substitution and recombinant “secondary” synthetic forms have been created in the P.P. Lukyanenko National Center of Grain. The synthetic recombination form RS5 (BBAASDt ), in which the third genome consists of chromosomes of Aegilops speltoides (S) and Aegilops tauschii (Dt ), was obtained from crossing the synthetic forms Avrodes (BBAASS) and M.it./Ae. tauschii (BBAADt Dt ), in which the D genome from Ae. tauschii was added to the BBAA genomes of the durum wheat cultivar Mutico italicum. Introgression lines resistant to leaf rust, yellow rust and powdery mildew have been obtained from backcrosses with the susceptible common wheat cultivars Krasnodarskaya 99, Rostislav and Zhirovka. Twelve resistant lines that additionally have high technological characteristics of grain and flour have been selected. The cytological study (С-banding) has revealed chromosomal modifications in 6 of 8 lines under study. The rearrangements mainly affected the chromosomes of the D genome, 1D, 3D, 4D, 6D and 7D. It was found that in most cases the genetic material from the synthetic form RS5 in the studied lines was represented by substituted chromosomes from Ae. tauschii. In line 5791p17, the substitution of chromosomes 6D from Ae. tauschii and 7D from Ae. speltoides was revealed. Substitutions 4D(4Dt ), 6D(6Dt ) from Ae. tauschii and 7D(7S) from Ae. speltoides were obtained for the first time. Molecular analysis of 12 lines did not reveal effective leaf rust resistance genes, presumably present in synthetic forms of M.it./Ae. tauschii and Avrodes. It is assumed that the lines may carry previously unidentified genes for fungal disease resistance, in particular for resistance to leaf rust, from Ae. tauschii and Ae. speltoides.
The oxidative phosphorylation (OXPHOS) system of mitochondria supports all the vitally important energyconsuming processes in eukaryotic cells, providing them with energy in the form of ATP. OXPHOS enzymes (complexes I–V) are located in the inner mitochondrial membrane, mainly in the cristae subcompartment. At present, there is a large body of data evidencing that the respiratory complexes I, III2 and IV under in vivo conditions can physically interact with each other in diverse stoichiometry, thereby forming supercomplexes. Despite active accumulation of knowledge about the structure of the main supercomplexes of the OXPHOS system, its physical and functional organization in vivo remains unclear. Contemporary models of the OXPHOS system’s organization in the inner membrane of mitochondria are contradictory and presume the existence of either highly organized respiratory strings, or, by contrast, a set of randomly dispersed respiratory supercomplexes and complexes. Furthermore, it is assumed that ATP-synthase (complex V) does not form associations with respiratory enzymes and operates autonomously. Our latest data obtained on mitochondria of etiolated shoots of pea evidence the possibility of physical association between the respiratory supercomplexes and dimeric ATP-synthase. These data have allowed us to reconsider the contemporary concept of the phosphorylation system organization and propose a new subcompartmented oxphosomic model. According to this model, a substantial number of the OXPHOS complexes form oxphosomes, which in a definite stoichiometry include complexes I–V and are located predominantly in the cristae subcompartment of mitochondria in the form of highly organized strings or patches. These suprastructures represent “mini-factories” for ATP production. It is assumed that such an organization (1) contributes to increasing the efficiency of the OXPHOS system operation, (2) involves new levels of activity regulation, and (3) may determine the inner membrane morphology to some extent. The review discusses the proposed model in detail. For a better understanding of the matter, the history of development of concepts concerning the OXPHOS organization with the emphasis on recent contemporary models is briefly considered. The principal experimental data accumulated over the past 40 years, which confirm the validity of the oxphosomic hypothesis, are also provided.
This paper describes an attempt to analyze, with the aid of bioinformatics resources (programs and databases), the probable cause of the cross-interaction of antibodies against HPV16 L1 with antigenic protein HPV6 L1, which has been revealed in the investigation of the candidate vaccine obtained on the base of a plant expression system (tomato plants). In our opinion, the most likely reason for the cross-interaction of antibodies with antigens of different pathogenic HPV types is the similarity of their antigenic determinants. In this work, the amino acid sequences of HPV16 L1 and HPV6 L1 used for the development of a binary vaccine against cervical cancer and anogenital papillomatosis have been analyzed. For the analysis of antigenic determinants, the programs BepiPred-2.0: Sequential B-Cell Epitope Predictor, DiscoTope 2.0 Server and SYFPEITHI have been used. As a result of the analysis of probable B-cell linear determinants (epitopes), it has been found that in both types of HPV the proteins have approximately the same location and size of linear antigenic determinants; the difference is observed only in the form of small shifts in the size of several amino acid residues. However, there are some differences in the amino acid composition of epitopes; therefore, the possibility for cross-interaction of the antibodies with the antigens due to the similarity of linear antigenic determinants for B-cells is very small. The analysis of potential threedimensional epitopes for B-cells has shown that due to little difference between them the HPV16 L1 and HPV6 L1 proteins have no prerequisites for cross-interaction of the antibodies with the antigens belonging to the two different pathogenic HPV types. The analysis of probable linear epitopes for T-cells has revealed a common antigenic determinant in the two protein sequences. According to the rank made with the SYFPEITHI program, the amino acid sequence AQL(I)FNKPYWL is the second most likely antigenic determinant for T-cells. Meanwhile, the amino acid sequences of this determinant in HPV16 L1 and HPV6 L1 are virtually identical. There is a difference in only one position, but it is not critical due to the similarity of the physicochemical properties of amino acids, for which there is a replacement in the amino acid sequence of antigenic determinants. Consequently, some moderate cross-interaction of the antibodies to HPV16 L1 with the antigens of HPV6 L1 may be expected.