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.

Phenotypic traits differentiating the genetic resources of pea (Pisum sativum L.) by the type of use.

The paper presents an analysis of the data obtained for pea accessions from the VIR collection studied at the Adler Experiment Station in the setting of the Krasnodar Territory in 2017-2019. It was for the f irst time that these accessions were studied for a set of phenotypic traits. The object of the study was a sample of 494 pea accessions originated from 43 countries and 18 regions and territories of the Russian Federation. The work was carried out in compliance with the methodological guidelines developed at VIR. Statistica 13.3 software was employed for statistical data processing. An assessment of four qualitative, 10 quantitative and four phenological traits in the accessions made it possible to differentiate them by the type of use, that is, as dry, forage and garden peas. The varieties differing in the type of use signif icantly differed by the values of such traits as stem length, number of pods per plant, number of nodes to the f irst f lower, number of f lowers in the inf lorescence, the maximum number of seeds per pod, pod length, and a narrower pod of forage pea compared to that of dry and garden peas. The average values of these traits were recorded for the peas with different types of use. The maximum difference was noted between garden and forage pea varieties. Dry pea varieties occupied an intermediate position. The complex of phenotypic traits identif ied determines the differences between three types of pea use, which is important when selecting the initial material for breeding appropriate varieties.

Genetic resources of narrow-leaved lupine (Lupinus angustifolius L.) and their role in its domestication and breeding.

Narrow-leaved lupine (Lupinus angustifolius L.) is a cultivated multipurpose species with a very short history of domestication. It is used as a green manure, and for feed and food. This crop shows good prospects for use in pharmacology and as a source of f ish feeds in aquaculture. However, its genetic potential for the development of productive and adaptable cultivars is far from being realized. For crop species, the genetic base of the cultivated gene pool has repeatedly been shown as being much narrower than that of the wild gene pool. Therefore, eff icient utilization of a species' genetic resources is important for the crop's further improvement. Analyzing the information on the germplasm collections preserved in national gene banks can help perceive the worldwide diversity of L. angustifolius genetic resources and understand how they are studied and used. In this context, the data on the narrow-leaved lupine collection held by VIR are presented: its size and composition, the breeding status of accessions, methods of studying and disclosing intraspecif ic differentiation, the classif ications used, and the comparison of this information with available data on other collections. It appeared that VIR's collection of narrow-leaved lupine, ranking as the world's second largest, differed signif icantly from others by the prevalence of advanced cultivars and breeding material in it, while wild accessions prevailed in most collections. The importance of the wild gene pool for the narrow-leaved lupine breeding in Australia, the world leader in lupine production, is highlighted. The need to get an insight into the species' ecogeographic diversity in order to develop cultivars adaptable to certain cultivation conditions is shown. The data on the testing of VIR's collection for main crop characters valuable for breeders are presented. Special attention is paid to the study of accessions with limited branching as a promising gene pool for cultivation in relatively northern regions of Russia. They demonstrate lower but more stable productivity, and suitability for cultivation in planting patterns, which has a number of agronomic advantages. Analyzing the work with narrow-leaved lupine genetic resources in different national gene banks over the world helps shape the prospects of further activities with VIR's collection as the only source of promising material for domestic breeding.

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.

Selection of an optimal method for screening the collection of narrow-leaved lupine held by the Vavilov Institute for the qualitative and quantitative composition of seed alkaloids.

Narrow-leaved lupine (Lupinus аngustifolius L.) is a widely cultivated leguminous forage and green manure crop with a potential for human nutrition. However, the presence of secondary metabolites - alkaloids - in lupine seeds considerably affects the quality of raw produce, reducing its nutritive value; in addition, high concentrations of alkaloids are toxic to humans and animals. Therefore, plant breeders working with lupine need to gain knowledge about the variability of alkaloid content in seeds of different genotypes and search for the sources of their low concentrations in the crop's gene pool. The collection of narrow-leaved lupine genetic resources held by the N.I. Vavilov Institute of Plant Genetic Resources (VIR) offers wide opportunities for such search by means of mass screening. For its part, largescale gene pool screening requires the selection of an optimal technique to measure alkaloid content in seeds, so that it would be easily reproducible and as little labor-, time- and fund-consuming as possible. The results of the search for such method are presented. Qualitative and quantitative indices were compared when target compounds had been extracted with multicomponent mixtures and individual reagents (chloroform, methanol, etc.) and the extracts analyzed using gas chromatography-mass spectrometry. High-performance liquid chromatography combined with mass spectrometry was also employed. Five major alkaloids were found to be present in all types of extracts: lupanine, 13-hydroxylupanine (dominant ones), angustifoline, sparteine, and isolupanine. The fullest extraction of alkaloids was observed when the extractant with an added alkaline agent was used (425 mg/100 g). The lowest level of extraction was registered with chloroform (216 mg/100 g). The significance of the differences was confirmed statistically.