Background: Guar (Cyamopsis tetragonoloba (L.) Taub.), a short-day plant, is an economically valuable legume crop. Seeds of guar serve as a source of galactomannan polysaccharide, known as guar gum, which is in demand in the gas and oil industries. The rapid and complete maturation of guar seeds depends on the flowering time of a particular genotype. It is known that flowering in guar is controlled by several gene systems. However, no information about the process and mechanisms that trigger flowering in guar on the molecular and biochemical levels was previously reported. The aim of the study was to investigate the metabolic landscape underlying transition to the flowering in guar using GC-MS-metabolomic analysis. Results: 82 diverse guar genotypes (each in 8 replicates) from the VIR collection were grown under experimental conditions of high humidity and long photoperiod. In the stress environment some guar genotypes turned to flowering early (41 ± 1,8 days from the first true leaf appearance) while for others the serious delay of flowering (up to 95 ± 1,7 days) was observed. A total of 244 metabolites were detected by GC-MS analysis on the third true leaves stage of 82 guar genotypes. Among them some molecules were associated with the transition of the guar plants to flowering. Clear discrimination was observed in metabolomic profiles of two groups of «early flowering» and «delayed flowering» plants, with 65 metabolites having a significantly higher abundance in early flowering genotypes. Among them 7 key molecules were identified by S-plot, as potential biomarkers discriminating of «early flowering» and «delayed flowering» guar genotypes. Conclusions: The metabolomic landscape accompanying transition to flowering in guar was firstly described. The results obtained can be used in subsequent genomic research for identifying metabolite-gene associations and revealing genes responsible for the onset of flowering and photoperiod sensitivity of guar. In addition, the detected key metabolites associated with flowering of guar can be employed as biomarkers allowing rapid screening of breeding material for the potentially early flowering genotypes.

Guar (Cyamopsis tetragonoloba (L.) Taub.) is an annual legume crop native to India and Pakistan. Seeds of the plant serve as a source of galactomannan polysaccharide (guar gum) used in the food industry as a stabilizer (E412) and as a gelling agent in oil and gas fracturing fluids. There were several attempts to introduce this crop to countries of more northern latitudes. However, guar is a plant of a short photoperiod, therefore, its introduction, for example, to Russia is complicated by a long day length during the growing season. Breeding of new guar varieties insensitive to photoperiod slowed down due to the lack of information on functional molecular markers, which, in turn, requires information on guar genome. Modern breeding strategies, e.g., genomic predictions, benefit from integration of multi-omics approaches such as transcriptome, proteome and metabolome assays. Here we present an attempt to use transcriptome-metabolome integration to understand the genetic determination of flowering time variation among guar plants that differ in their photoperiod sensitivity. This study was performed on nine early- and six delayed-flowering guar varieties with the goal to find a connection between 63 metabolites and 1,067 differentially expressed transcripts using Shiny GAM approach. For the key biomarker of flowering in guar myo-inositol we also evaluated the KEGG biochemical pathway maps available for Arabidopsis thaliana. We found that the phosphatidylinositol signaling pathway is initiated in guar plants that are ready for flowering through the activation of the phospholipase C (PLC) gene, resulting in an exponential increase in the amount of myo-inositol in its free form observed on GC-MS chromatograms. The signaling pathway is performed by suppression of myo-inositol phosphate kinases (phosphorylation) and alternative overexpression of phosphatases (dephosphorylation). Our study suggests that metabolome and transcriptome information taken together, provide valuable information about biomarkers that can be used as a tool for marker-assisted breeding, metabolomics and functional genomics of this important legume crop.

Амплитуда изменчивости фотопериодической реакции генотипов гуара (Cyamopsis tetragonoloba (L.) Taub.) разного географического происхождения

Background.VIR’s potato collection is one of the oldest and richest; however, it is constantly exposed to viruses that negatively affect useful agronomic properties of tubers. Close monitoring of the phytosanitary state of potato accessions helps to select the most effective method of therapy for subsequent healing of infected plants and obtaining high-quality planting material.Materials and methods. The research was aimed at improving the health of 18 varieties ofSolanum tuberosumL. from the VIR collection. Testing for the presence of viruses was based on the ICA and RTPCR techniques, and the consequent healing was performed using the methods of meristem culture and cryotherapy.Results and conclusions. During the field test of potato plants, PVX, PVS and PVA were found to be the most common viruses. PSTVd was completely absent in all tested accessions. The effectiveness of in vitro healing of potato plants from viruses was assesses using meristem culture. The percentage of healed plants was 0% for PVS, 0% for PVX, 33.4% for PVA, 50% for PLRV, 72.3% for PVY, and 83.4% for PVM. Healing with meristem culture was shown to be the most effective against PVY and PVM. While assessing the effectiveness of post-cryogenic restoration of potato microplants, the level of post-cryogenic regeneration of the shoot tips in potato microplants was determined at 22.3% on average for a sample. The minimum was observed in k-16762 ‘Sagita N’ (5%), and the maximum in k-1378 ‘Marta’ (41.7%). Analysis of the effectiveness of potato recovery from viruses byin vitrocryotherapy showed that the percentage of recovered plants was 100% for PVY, 100% for PVA, 88.9% for PVM, 77.8% for PVS, 44.4% for PVX. Thus, the techniques of apical meristem culture and cryotherapy proved to be effective against PVY, PVA and PVM viruses. However, in the case of multiple infections, it is necessary to combine elements of different healing protocols to increase the effectiveness of the healing procedure.

Guar (Cyamopsis tetragonoloba (L.) Taub.) is an agricultural crop species new to Russia and is in demand by the gas, oil and food industries. Due to the progress of “omics” technologies and the marker-assisted selection, there is a huge interest in the studies that compare the metabolites of various guar varieties, employing metabolomics as a method of functional genomics. For a large-scale screening of guar germplasm from the VIR collection, it is important to choose an efficient method to extract metabolites from samples. The accuracy of the assessment of the content of metabolites in samples is crucial for distinguishing varieties within the crop, since the metabolome profiles of plants within the same species differ mainly in the quantitative ratio of metabolites, and not in their qualitative composition. In metabolome practice, two methods of extracting polar compounds are usually employed in the preparation of samples for GC-MS analysis. One of the widely used methods of sample preparation is the long-term extraction of metabolites from whole tissues with the aid of a methanol solvent. Another method of sample preparation is based on the short-term methanol extraction of metabolites from frozen and homogenized material. The advantages and disadvantages of these two methods revealed in the course of our work have prompted us to develop a new approach that avoids some difficulties in analyzing the metabolic profiles of leaves of various guar varieties. The method we suggested combines the advantages of the two above-mentioned approaches of sample preparation, namely eliminates the loss of metabolites due to centrifugation and ensures the complete destruction of all cell walls, ensuring the maximum extraction level of polar metabolites. The essence of the new method is that the leaf is rapidly frozen in liquid nitrogen with subsequent thawing in cold methanol. Thus, leaf tissues retain morphological integrity, and subsequent centrifugation, necessary for homogenization, is skipped. We have checked the effectiveness of this improved method by experiments with leaf samples of three guar genotypes. It has been shown that the amount of extracted metabolites increases more than 5-fold compared to extraction with methanol from fresh unfrozen leaf tissues and more than 2-fold compared to extraction with methanol after freezing and homogenization. Extraction of metabolites using the new method allows the GC-MS analysis of guar samples to be conducted with the least loss and high accuracy required to distinguish varieties.

Background. Guar ( Cyamopsis tetragonoloba (L.) Taub.) is a leguminous crop plant of tropical origin that has gained unprecedented popularity in recent years due to the presence of gum in its seeds. The use of guar gum in the oil and gas industry gives the crop strategic importance. This was the reason for its introduction to the Russian Federation (RF) at the beginning of the 21st century and for the active breeding of domestic cultivars. The demand for the guar collection has increased dramatically, serving as an impetus for its active study. Materials and methods. The materials of the study were guar accessions from the VIR collection: 50 accessions in 2023, and 30 most productive of them in 2024. The accessions were phenotyped for 13 traits important for breeding at Volgograd Experiment Station of VIR. The indicator of early maturity was assessed by indirect methods. Statistical processing of the research results was performed using the Statistica 13.3 software package. Results. Seed productivity of guar accessions was analyzed, its structure and interrelations among its defining characteristics. Differentiation of the guar gene pool for the studied traits was revealed. The most productive accessions were identified. Previously obtained data on the relationship of guar collection accessions to the photoperiod served as a basis for proposing a modified algorithm for determining the photoperiod sensitivity of accessions as an indirect indicator of their earliness. Conclusion. The data obtained for guar accessions from the VIR collection under the conditions of the Russian Federation will make it possible to use this germplasm effectively as source material for breeding domestic cultivars.