The perspectives of metabolomics studies of potato plants

According to FAO (Food and Agricultural Organization of the United Nation), potato is the fourth crop in terms of food production after rice, wheat and maize, and the first among the tubers and roots. The importance of potato is difficult to overestimate; it is a valuable source of carbohydrates, antioxidants and vitamins. A huge number of investigations are focused on the study of metabolic processes occurring in the potato plant in order to elucidate the mechanisms responsible for productivity and accumulation of compounds that determine taste and nutritional quality, keeping quality of tubers, plant resistance, etc. The sum of metabolites, which is produced as a result of metabolic network activity, is defined as metabolome. Complex studies of metabolic diversity with the use of modern state-of-the-art chromatography approaches and highly precise detection of individual compounds revealed specificity of metabolic spectra from subcellular to organism levels and its amazing plasticity under the influence of a variety of internal and external stimuli. Metabolomic approaches are already in use for phenotyping available species, lines and varieties as well as for evaluation of potato plant resistance to environmental challenges and for detection of changes in tubers during storage. Metabolome profiling is widely employed to study differences between genetically modified forms of potatoes from untransformed relatives. A limited number of systemic studies on potatoes combines metabolome investigation with genome, transcriptome and proteome analysis and suggests an important role of the genome in the determination of metabolic rates. It is obvious that the search for biochemical markers depends on standartization of cultivation techniques, sample preparation and subsequent analysis similar to what has been developed for progress in genomic and transcriptomic studies. In the future, potato metabolome studies might complete classical and molecular approaches to develop new lines and varieties

SWEET transporters of Medicago lupulina in the arbuscular-mycorrhizal system in the presence of medium level of available phosphorus

Arbuscular mycorrhiza (AM) fungi receive photosynthetic products and sugars from plants in exchange for contributing to the uptake of minerals, especially phosphorus, from the soil. The identification of genes controlling AM symbiotic efficiency may have practical application in the creation of highly productive plant-microbe systems. The aim of our work was to evaluate the expression levels of SWEET sugar transporter genes, the only family in which sugar transporters specific to AM symbiosis can be detected. We have selected a unique “host plant–AM fungus” model system with high response to mycorrhization under medium phosphorus level. This includes a plant line which is highly responsive to inoculation by AM fungi, an ecologically obligate mycotrophic line MlS-1 from black medick (Medicago lupulina) and the AM fungus Rhizophagus irregularis strain RCAM00320, which has a high efficiency in a number of plant species. Using the selected model system, differences in the expression levels of 11 genes encoding SWEET transporters in the roots of the host plant were evaluated during the development of or in the absence of symbiosis of M. lupulina with R. irregularis at various stages of the host plant development in the presence of medium level of phosphorus available for plant nutrition in the substrate. At most stages of host plant development, mycorrhizal plants had higher expression levels of MlSWEET1b, MlSWEET3c, MlSWEET12 and MlSWEET13 compared to AM-less controls. Also, increased expression relative to control during mycorrhization was observed for MlSWEET11 at 2nd and 3rd leaf development stages, for MlSWEET15c at stemming (stooling) stage, for MlSWEET1a at 2nd leaf development, stemming and lateral branching stages. The MlSWEET1b gene can be confidently considered a good marker with specific expression for effective development of AM symbiosis between M. lupulina and R. irregularis in the presence of medium level of phosphorus available to plants in the substrate

Sugar transporters of the SWEET family and their role in arbuscular mycorrhiza

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

Structural Elucidation of Cisoid and Transoid Cyclization Pathways of a Sesquiterpene Synthase Using 2-Fluorofarnesyl Diphosphates

Sesquiterpene skeletal complexity in nature originates from the enzyme-catalyzed ionization of (trans,trans)-farnesyl diphosphate (FPP) (1a) and subsequent cyclization along either 2,3-transoid or 2,3-cisoid farnesyl cation pathways. Tobacco 5-epi-aristolochene synthase (TEAS), a transoid synthase, produces cisoid products as a component of its minor product spectrum. To investigate the cryptic cisoid cyclization pathway in TEAS, we employed (cis,trans)-FPP (1b) as an alternative substrate. Strikingly, TEAS was catalytically robust in the enzymatic conversion of (cis,trans)-FPP (1b) to exclusively (≥99.5%) cisoid products. Further, crystallographic characterization of wild-type TEAS and a catalytically promiscuous mutant (M4 TEAS) with 2-fluoro analogues of both all-trans FPP (1a) and (cis,trans)-FPP (1b) revealed binding modes consistent with preorganization of the farnesyl chain. These results provide a structural glimpse into both cisoid and transoid cyclization pathways efficiently templated by a single enzyme active site, consistent with the recently elucidated stereochemistry of the cisoid products. Further, computational studies using density functional theory calculations reveal concerted, highly asynchronous cyclization pathways leading to the major cisoid cyclization products. The implications of these discoveries for expanded sesquiterpene diversity in nature are discussed

Fusicoccin Counteracts the Toxic Effect of Cadmium on the Growth of Maize Coleoptile Segments

The effects of cadmium (Cd; 0.1–1000 μM) and fusicoccin (FC) on growth, Cd2+ content, and membrane potential (Em) in maize coleoptile segments were studied. In addition, the Em changes and accumulation of Cd and calcium (Ca) in coleoptile segments treated with Cd2+ combined with 1 μM FC or 30 mM tetraethylammonium (TEA) chloride (K+-channel blocker) were also determined. In this study, the effects of Ca2+-channel blockers [lanthanum (La) and verapamil (Ver)] on growth and content of Cd2+ and Ca2+ in coleoptile segments were also investigated. It was found that Cd at high concentrations (100 and 1000 μM) significantly inhibited endogenous growth of coleoptile segments and simultaneously measured proton extrusion. FC combined with Cd2+ counteracted the toxic effect of Cd2+ on endogenous growth and significantly decreased Cd2+ content (not the case for Cd2+ at the highest concentration) in coleoptile segments. Addition of Cd to the control medium caused depolarization of Em, the extent of which was dependent on Cd concentration and time of treatment with Cd2+. Hyperpolarization of Em induced by FC was suppressed in the presence of Cd2+ at 1000 μM but not Cd2+ at 100 μM. It was also found that treatment of maize coleoptile segments with 30 mM TEA chloride caused hyperpolarization of Em and decreased Cd2+ content in coleoptile segments, suggesting that, in the same way as for FC, accumulation of Cd2+ was dependent on plasma membrane (PM) hyperpolarization. Similar to FC, TEA chloride also decreased Ca2+ content in coleoptile segments. La and Ver combined with Cd2+ (100 μM) significantly decreased Cd content in maize coleoptile segments, but only La completely abolished the toxic effect of Cd2+ on endogenous growth and growth in the presence of FC. Taken together, these results suggest that the mechanism by which FC counteracts the toxic effect of Cd2+ (except at 1000 μM Cd2+) on the growth of maize coleoptile segments involves both stimulation of PM H+-ATPase activity by FC as well as Cd2+-permeable, voltage-dependent Ca channels, which are blocked by FC and TEA chloride-induced PM hyperpolarization

Racial differences in human platelet PAR4 reactivity reflect expression of PCTP and miR-376c.

Racial differences in the pathophysiology of atherothrombosis are poorly understood. We explored the function and transcriptome of platelets in healthy black (n = 70) and white (n = 84) subjects. Platelet aggregation and calcium mobilization induced by the PAR4 thrombin receptor were significantly greater in black subjects. Numerous differentially expressed RNAs were associated with both race and PAR4 reactivity, including PCTP (encoding phosphatidylcholine transfer protein), and platelets from black subjects expressed higher levels of PC-TP protein. PC-TP inhibition or depletion blocked PAR4- but not PAR1-mediated activation of platelets and megakaryocytic cell lines. miR-376c levels were differentially expressed by race and PAR4 reactivity and were inversely correlated with PCTP mRNA levels, PC-TP protein levels and PAR4 reactivity. miR-376c regulated the expression of PC-TP in human megakaryocytes. A disproportionately high number of microRNAs that were differentially expressed by race and PAR4 reactivity, including miR-376c, are encoded in the DLK1-DIO3 locus and were expressed at lower levels in platelets from black subjects. These results suggest that PC-TP contributes to the racial difference in PAR4-mediated platelet activation, indicate a genomic contribution to platelet function that differs by race and emphasize a need to consider the effects of race when developing anti-thrombotic drugs

Molecular-Genetic and Epidemiologic Examination of Personnel Subjected to Occupational Irradiation

It is given an evaluation of the genetic polymorphisms (Hp, Tf, Gc; 6-PGD, EsD, ACP, PGM1, microsatellite loci CSF1PO и F13AO1, detoxicating genes GSTT1, GSTM1 and GSTP1), individual radiosensitivity (by the criterion of ribosome gene fragments) and DNA-damage rate (Comet-assay) in two cohorts comprised by VNIIEF personnel subjected chronically to gamma-neutron ionizing radiation and to β-radiation of Tritium in comparison with the effects in non-irradiated cohort. There are discussed data on the influence of the occupational irradiation, age and genotype on the rate of structural genome damage, as well as on the activity of the human repair system activity and health status