Background: Investigations that are focused on arbuscular mycorrhizal fungus (AMF) biodiversity is still limited. The analysis of the AMF taxa in the North Caucasus, a temperate biodiversity hotspot, used to be limited to the genus level. This study aimed to define the AMF biodiversity at the species level in the North Caucasus biotopes. Methods: The molecular genetic identification of fungi was carried out with ITS1 and ITS2 regions as barcodes via sequencing using Illumina MiSeq, the analysis of phylogenetic trees for individual genera, and searches for operational taxonomic units (OTUs) with identification at the species level. Sequences from MaarjAM and NCBI GenBank were used as references. Results: We analyzed >10 million reads in soil samples for three biotopes to estimate fungal biodiversity. Briefly, 50 AMF species belonging to 20 genera were registered. The total number of the AM fungus OTUs for the “Subalpine Meadow” biotope was 171/131, that for “Forest” was 117/60, and that for “River Valley” was 296/221 based on ITS1/ITS2 data. The total number of the AM fungus species (except for virtual taxa) for the “Subalpine Meadow” biotope was 24/19, that for “Forest” was 22/13, and that for “River Valley” was 28/24 based on ITS1/ITS2 data. Greater AMF diversity, as well as number of OTUs and species, in comparison with that of forest biotopes, characterized valley biotopes (disturbed ecosystems; grasslands). The correlation coefficient between “Percentage of annual plants” and “Glomeromycota total reads” r = 0.76 and 0.81 for ITS1 and ITS2, respectively, and the correlation coefficient between “Percentage of annual plants” and “OTUs number (for total species)” was r = 0.67 and 0.77 for ITS1 and ITS2, respectively. Conclusion: High AMF biodiversity for the river valley can be associated with a higher percentage of annual plants in these biotopes and the active development of restorative successional processes.
Plants known as obligate aerobes developed different mechanisms to overcome the damage incurred under oxygen limitation. One of the survival strategies to have commonly appeared in hydrophytic plants is the escape strategy, which accelerates plant axial organs’ growth in order to escape hypoxic conditions as soon as possible. The present study aimed to distinguish the alterations in coleoptile elongation, viability and metabolic profiles in coleoptiles of slow- and fast-growing rice varieties. All the parameters were tested at 3, 5 and 7 days after sowing, to highlight changes during seedling development in normal and submerged conditions. The obtained results indicated that coleoptile elongation correlated with higher resistance to oxygen deprivation. GS-MS-based metabolic profiling indicated that coleoptiles of the fast-growing cultivar accumulated higher amounts of sugar phosphates, disaccharides, fatty acid derivatives and sterols, which are important for maintaining growth, membrane stability and viability. The slow-growing variety was characterized by a greater abundance of carboxylates, including lactate and phosphoric acid, indicating an energy crisis and cytosol acidification, leading to cell damage and low tolerance. Therefore, a metabolomics approach could be used for phenotyping (chemotyping) in the large-scale screening of newly developed varieties with higher tolerance to oxygen deprivation.
The study is aimed at revealing the effects of Rhizophagus irregularis inoculation on the transcriptome of Medicago lupulina leaves at the early (second leaf formation) and later (flowering) stages of plant development. A pot experiment was conducted under conditions of low phosphorus (P) level in the substrate. M. lupulina plants were characterized by high mycorrhizal growth response and mycorrhization parameters. Library sequencing was performed on the Illumina HiseqXTen platform. Significant changes in the expression of 4863 (padj < 0.01) genes from 34049 functionally annotated genes were shown by Massive Analysis of cDNA Ends (MACE-Seq). GO enrichment analysis using the Kolmogorov–Smirnov test was performed, and 244 functional GO groups were identified, including genes contributing to the development of effective AM symbiosis. The Mercator online tool was used to assign functional classes of differentially expressed genes (DEGs). The early stage was characterized by the presence of six functional classes that included only upregulated GO groups, such as genes of carbohydrate metabolism, cellular respiration, nutrient uptake, photosynthesis, protein biosynthesis, and solute transport. At the later stage (flowering), the number of stimulated GO groups was reduced to photosynthesis and protein biosynthesis. All DEGs of the GO:0016036 group were downregulated because AM plants had higher resistance to phosphate starvation. For the first time, the upregulation of genes encoding thioredoxin in AM plant leaves was shown. It was supposed to reduce ROS level and thus, consequently, enhance the mechanisms of antioxidant protection in M. lupulina plants under conditions of low phosphorus level. Taken together, the obtained results indicate genes that are the most important for the effective symbiosis with M. lupulina and might be engaged in other plant species.
The arbuscular mycorrhizal fungi (AMF) effect on the plant metabolome is an actual question of plant biology. Its alteration during host plant development and at different phosphorus supplies is of special interest. The aim of this study was to evaluate the effect of Rhizophagus irregularis (Błaszk., Wubet, Renker & Buscot) C. Walker & A. Schüßler inoculation and/or phosphorus treatment on the root metabolome of Medicago lupulina L. subsp. vulgaris Koch at the first true leaf, second leaf, third leaf development stages, the lateral branching initiation, the flowering and the mature fruit stages. The assessment of metabolic profiles was performed using GC-MS. In total, 327 metabolites were annotated: among them 20 carboxylic acids, 26 amino acids, 14 fatty acids and 58 sugars. The efficient AM was characterized by the upregulation of the metabolism of proteins, carbohydrates and lipids, as well as an increase in the content of phosphates. The tricarboxylic acid abundance was generally lower during mycorrhization. Fourteen metabolic markers of the efficient AM symbiosis were identified. The lateral branching initiation stage was shown to have key importance. Long-lasting metabolomic profiling indicated variances in mycorrhization and Pi supply effects at different key stages of host plant development.
Background: Number of investigations, which are focused on arbuscular mycorrhiza fungi (AMF) biodiversity is still limited. The analysis of AMF taxa in the North Caucasus, a temperate biodiversity hotspot, used to be limited by the genera level. The aim of this study was to define AMF biodiversity at the species level in the North Caucasus biotopes. Methods: Molecular genetic identification of fungi was carried out with ITS1 and ITS2 regions as barcodes, sequencing by Illumina MiSeq, analysis of phylogenetic trees for individual genera, searching the operational taxonomic units (OTUs) with identification at the species level. Sequences from MaarjAM and NCBI GenBank were used as references. Results: We analyzed >10 million reads in soil samples for 3 biotopes to estimate fungal biodiversity. 50 AMF species from 20 genera were found. The valley biotopes (disturbed ecosystems, grasslands) were characterized by a greater AMF diversity, as well as the number of OTUs and species, in comparison with the forest biotopes. Conclusion: Our hypothesis is that high AMF biodiversity for the river valley can be associated with a higher percentage of annual plants in these biotopes, as well as with the active development of restorative successional processes.
Tobacco BY-2 cell culture is one of the most widely used models in plant biology. The main advantage of BY-2 suspension cultures is the synchronization of cell development and the appearance of polar elongation. In batch culture, BY-2 cells passed through the lag, proliferation, elongation, and stationary phases. During this process, the composition of the growth medium changed dramatically. Sucrose was rapidly eliminated; hexose first accumulated and then depleted. The medium’s pH initially decreased and then rose with aging. As a result of the crosstalk between the internal and external stimuli, cells pass through complicated systemic rearrangements, which cause metabolomic alterations. The early stages were characterized by high levels of amino acids and sterols, which could be interpreted as the result of synthetic activity. The most intense rearrangements occurred between the proliferation and active elongation stages, including repression of amino acid accumulation and up-regulation of sugar metabolism. Later stages were distinguished by higher levels of secondary metabolites, which may be a non-specific response to deteriorating conditions. Senescence was followed by some increase in fatty acids and sterols as well as amino acids, and probably led to self-destructive processes. A correlation analysis revealed relationships between metabolites’ covariation, their biochemical ratio, and the growth phase.