Metabolomic approach to investigate Dactylis glomerata L. from the VIR collection.

The perennial grass cocksfoot (Dactylis glomerata L.) is a valuable early highly nutritious crop used as green fodder in agricultural production. The species is widespread across the Eurasian continent; it is characterized by plasticity and high ecological and geographical variability. The article considers the metabolic profiles of 15 accessions of the cocksfoot from the collection of the N.I. Vavilov Institute of Plant Genetic Resources (VIR). The material is represented by varieties and wild forms of various origin: the European part of the Russian Federation, Norway and Finland. The study was carried out using gas-liquid chromatography coupled with mass spectrometry. The study and comparison of groups of metabolites of cocksfoot accessions of various ecological and geographical origin was carried out. Statistical processing included the calculation of the main parameters of variability, factor analysis of the correlation system (Q- and R-technique), cluster analysis by Ward's method and discriminant analysis. The variability of the quantitative and qualitative composition of the substances identified was revealed. Based on statistical processing of the results obtained, five groups of cocksfoot accessions were identified, differing in the profile of metabolites. One of the groups with a similar composition of metabolites consisted of accessions from one ecological and geographical region; another, of accessions of different origin. Significant differences were noted in the metabolomic profiles of a late-maturing wild cocksfoot accession from the Republic of Karelia at the booting stage from early- and mid-maturing accessions at the heading stage; it contained the largest number of free amino acids and the smallest number of identified primary and secondary metabolites. Wild-growing accession k-44020 from Norway surpassed other wild-growing accessions in the content of free amino acids, sugars and phosphates at the heading stage. Wild-growing accessions differed from breeding varieties with a high content of proline and threonine, indicators of high resistance to lack of moisture and high air temperature.

Alkaloid content variability in the seeds of narrow-leafed lupine accessions from the VIR collection under the conditions of the Russian Northwest.

Alkaloid content was assessed in the seeds of 59 narrow-leafed lupine (Lupinus angustifolius L.) accessions from the VIR collection in the environments of Leningrad Province. The selected set included accessions of different statuses (wild forms, landraces, and advanced cultivars) and different years of introduction to the collection. Alkaloids were analyzed using gas-liquid chromatography coupled with mass spectrometry. Concentrations of main alkaloids: lupanine, 13-hydroxylupanine, sparteine, angustifoline and isolupanine, and their total content were measured. The total alkaloid content variability identified in the seeds of the studied set of accessions was 0.0015 to 2.017 %. In most cases, the value of the character corresponded to the accession's status: modern improved cultivars, with the exception of green manure ones, entered the group with the range of 0.0015-0.052 %, while landraces and wild forms showed values from 0.057 to 2.17 %. It is meaningful that the second group mainly included accessions that came to the collection before the 1950s, i. e., before the times when low-alkaloid cultivars were intensively developed. Strong variability of the character across the years was observed in the accessions grown under the same soil and climate conditions in both years. In 2019, the average content of alkaloids in the sampled set was 1.9 times higher than in 2020. An analysis of weather conditions suggested that the decrease in alkaloid content occurred due to a significant increase in total rainfall in 2020. Searching for links between the content of alkaloids and the type of pod (spontaneously non-dehiscent, or cultivated, spontaneously dehiscent, or wild, and intermediate) showed a tendency towards higher (approximately twofold in both years of research) total alkaloid content in the accessions with the wild pod type and the nearest intermediate one compared to those with the pod non-dehiscent without threshing. The correlation between the average total alkaloid content and seed color, reduced to three categories (dark, or wild, light, or cultivated, and intermediate), was significantly stronger in the group with dark seeds (5.2 times in 2019, and 3.7 times in 2020). There were no significant differences in the percentage of individual alkaloids within the total amount either between the years of research or among the groups with different pod types or the groups with different seed coat colors.

Metabolomic prof iles of Ribes nigrum L. and Lonicera caerulea L. from the collection of the N.I. Vavilov Institute in the setting of Northwest Russia.

Recently, the trend of using fruit and berry crops as ingredients for functional and dietary nutrition, the development and implementation of f lavors, pigments, new medicines and dietary supplements has been actualized. Because the direction of use depends on the biochemical properties of fruits, which are determined not only by species and varietal characteristics, but also by reproduction conditions, the study of the biochemical composition of fruits grown in various regions of the world continues to be relevant. In this regard, the collection of N.I. Vavilov Institute (VIR), which has a wide diversity of fruit and berry crops, is of great interest for study. Ribes nigrum fruits have a balanced set of sugars, organic acids, essential oils, microelements, a high content of vitamins, anthocyanins, pectins. Lonicera caerulea fruits are characterized by high values of phenolic substances: biof lavonoids, hydroxycinnamic acids, f lavonols, polyphenols, anthocyanins, as well as vitamins, carotenoids, iridoid glycosides and other natural antioxidants. The investigation of L. caerulea and R. nigrum fruit's accessions from the VIR collection using gas-liquid chromatography with mass spectrometry allows us to obtain new information about the biochemical characteristics of fruits, to identify L. caerulea and R. nigrum varieties with optimal economically valuable characteristics, to determine the specif icity of L. caerulea and R. nigrum metabolomic spectra in the setting of Northwest Russia. As a result of the analysis, typical compounds of the metabolomic prof ile of each culture were identif ied. Organic acids, phenol-containing compounds and polyols prevailed in L. caerulea, while mono- and oligosaccharides, in R. nigrum. The qualitative composition of the black currant varieties 'Malen'kii Printz', 'Dobriyi Dzhinn', 'Tisel', 'Orlovskii Val's', and blue honeysuckle 'S 322-4', 'Malvina', 'Leningradsky Velikan' was optimal for food consumption; the varieties of blue honeysuckle 'Bazhovskaya' and black currant 'Aleander' had a good representation of biologically active compounds, which makes samples attractive as raw materials for the production of biologically active additives, including with the use of microorganisms' cultures.

Comparative analysis of wild and cultivated Lathyrus L. species to assess their content of sugars, polyols, free fatty acids, and phytosterols.

Under the condition of climate change, the need for crops resistant to abiotic and biotic stresses is increasing. Lathyrus spp. are characterized by a high nutritional value of their green biomass. The grass pea is one of the most resistant to drought, waterlogging, cold, salinity, diseases and pests among cultivated legumes, and it is grown at minimal cost. The creation of new Lathyrus L. sorts with an improved nutrient composition of nutrients will allow to obtain high-quality feed in areas with extremely unstable weather conditions. In this connection, we studied the patterns of variability in the parameters of the carbohydrate complex (sugars, their lactone and methyl forms), polyols (including phenol-containing alcohols), phytosterols, free fatty acids (FFA) and acylglycerols in the green mass of 32 samples of Lathyrus sativus L., L. tuberosus L., L. sylvestris L., L. vernus (L.) Bernh., L. latifolius L., L. linifolius (Reichard) Bassler. from the VIR collection, reproduced in the Leningrad region in contrasting conditions 2012, 2013.The content of identified compounds varied depending on the genotype, species, and weather conditions. High temperatures and high level of precipitation in 2013 contributed to the accumulation of monosaccharides, in more colder and drier conditions in 2012 - oligosaccharides, most of polyols and FFA. The cultivated species (L. sativus) was distinguished by its high sugar content, and the wild species as follows: L. latifolius by FFA; L. linifolius by ononitol, myo-inositol, and glycerol 3-phosphate; L. vernus by MAG and methylpentofuranoside. The species cultivated in culture (L. sativus) was distinguished by a high sugar content, wild species: L. latifolius - by FFA, L. linifolius - ononitol, myo-inositol and glycerol-3-phosphate, L. vernus - MAG and methylpentofuranoside. According to our results, the studied samples are promising for the selection of Lathyrus varieties with high nutrition quality and stress-resistant.

Metabolomic approach to search for fungal resistant forms of Aegilops tauschii Coss. from the VIR collection.

Broadening of the genetic diversity of donors of resistance to biotic environmental factors is a challenging problem concerning Triticum L., which can be solved by using wild relatives of wheat, in particular, Aegilops tauschii Coss., in breeding programs. This species, believed to be the donor of D genome of common wheat (T. aestivum L.), is a source of some traits important for breeding. This greatly facilitates the possibility of crossing Ae. tauschii with common wheat. Aegilops L. species are donors of effective genes for resistance to fungal diseases in wheat. For instance, genes that determine resistance to rust agents in common wheat were successfully introgressed from Ae. tauschii into the genome of T. aestivum L. The aim of our study was to identify differences in metabolomic profiles of Ae. tauschii forms (genotypes), resistant or susceptible to such fungal pathogens as Puccinia triticina f. sp. tritici and Erysiphe graminis f. sp. tritici. These indicators may be used as biochemical markers of resistance. A comparative analysis of groups of Ae. tauschii accessions showed that metabolomic profiles of the forms with or without resistance to fungal pathogens differed significantly in the contents of nonproteinogenic amino acids, polyols, phytosterols, acylglycerols, mono- and oligosaccharides, glycosides, phenolic compounds (hydroquinone, kempferol), etc. This fact was consistent with the previously obtained data on the relationship between Fusarium resistance in oats (Avena sativa L.) and certain components of the metabolomic profile, such as acylglycerols, nonproteinogenic amino acids, galactinol, etc. Thus, our studies once again confirmed the possibility and effectiveness of the use of metabolomic analysis for screening the genetic diversity of accessions in the VIR collection, of Ae. tauschii in particular, in order to identify forms with a set of compounds in their metabolomic profile, which characterize them as resistant. Ae. tauschii accessions with a high content of pipecolic acids, acylglycerols, galactinol, stigmasterol, glycerol, azelaic and pyrogallic acids, campesterol, hydroquinone, etc., can be used for creating wheat and triticale cultivars with high resistance to fungal pathogens causing powdery mildew, brown rust, and yellow rust.

Alkaloids of narrow-leaved lupine as a factor determining alternative ways of the crop's utilization and breeding.

Narrow-leaved lupine (Lupinus angustifolius L.), a valuable leguminous crop adapted to a wide range of climatic conditions, has a very short history of domestication. For many centuries it was used mainly as a green manure, since the success and prospects of the multi-purpose use of the species depend on its breeding improvement, in particular, on a particular concentration of alkaloids in seeds and green mass. The first varieties of scientific breeding were created only in the 1930s after the appearance of low-alkaloid mutants. Despite wide prospects for use in various areas of the national economy, unstable productivity and susceptibility to diseases hinder the production of this crop. Obviously, breeders deal only with a small part of the gene pool of the species and limited genetic resources, using mainly low-alkaloid (sweet) genotypes to create new varieties. The genetic potential of the species can be used more efficiently. At the same time, it is rational to create highly alkaloid (bitter) varieties for green manure, while food and feed varieties should not lose their adaptive potential, in particular, resistance to pathogens, due to the elimination of alkaloids. In this regard, it seems to be a productive idea to create 'bitter/sweet' varieties combining a high content of alkaloids in the vegetative organs and low in seeds, which can be achieved by regulating the synthesis/transport of alkaloids in the plant. The paper discusses the current state of use of the species as a green manure, fodder, food plant. Information is given on the quantity and qualitative composition of narrow-leaved lupine alkaloids, their applied value, in particular, fungicidal, antibacterial, insecticidal, the use of lupine alkaloids as active principles of drugs. Along with promising breeding considerations, the possibility of using technologies for processing raw high-alkaloid materials with the accompanying extraction of valuable ingredients for pharmaceuticals is discussed. Information is briefly presented about the genomic resources of the species and the prospects for their use in marker-assistant selection and genome editing.