ROS-dependent induction of oxidative damage can be used as a trigger initiating genetically determined non-specific protection in plant cells and tissues. Plants are potentially able to withstand various specific (toxic, osmotic) factors of abiotic effects, but do not have sufficient or specific sensitivity to form an adequate effective response. In this work, we demonstrate one of the possible approaches for successful cold acclimation through the formation of effective protection of photosynthetic structures due to the insertion of the heterologous FeSOD gene into the tobacco genome under the control of the constitutive promoter and equipped with a signal sequence targeting the protein to plastid. The increased enzymatic activity of superoxide dismutase in the plastid compartment of transgenic tobacco plants enables them to tolerate the oxidative factor of environmental stresses scavenging ROS. On the other hand, the cost of such resistance is quite high and, when grown under normal conditions, disturbs the arrangement of the intrachloroplastic subdomains leading to the modification of stromal thylakoids, probably significantly affecting the photosynthesis processes that regulate the efficiency of photosystem II. This is partially compensated for by the fact that, at the same time, under normal conditions, the production of peroxide induces the activation of ROS detoxification enzymes. However, a violation of a number of processes, such as the metabolism of accumulation, and utilization and transportation of sugars and starch, is significantly altered, which leads to a shift in metabolic chains. The expected step for further improvement of the applied technology could be both the use of inducible promoters in the expression cassette, and the addition of other genes encoding for hydrogen peroxide-scavenging enzymes in the genetic construct that are downstream in the metabolic chain.
In the common chickweed Stellaria media, two antimicrobial peptides (AMPs), SmAMP1.1a and SmAMP1.2a, have been shown to be proteolytically released as products of the expression of a single gene, proSmAMP1. In this study, the gene proSmAMP1 was introduced into two potato varieties, Zhukovsky ranny and Udacha. These early-maturing varieties were shown to be susceptible to early blight caused by Alternaria spp. Most transgenic lines of either variety having strong expression of the target gene demonstrated high levels of resistance to Alternaria spp. during three years of cultivation, but did not otherwise differ from the initial varieties. Disease severity index (DSI) was introduced as a complex measure of plant susceptibility to early blight, taking into account the diameter of lesions caused by the Alternaria spp., the fungus sporulation intensity and its incubation period duration. Across all transgenic lines, the DSI inversely correlated both with the target gene expression and the copy number in the plant genome. Our results are promising for improving the resistance of potato and other crops to early blight by expression of AMPs from wild plants.
Ex situ collections of algae, cyanobacteria, and plant materials (cell cultures, hairy and adventitious root cultures, shoots, etc.) maintained in vitro or in liquid nitrogen (−196 °C, LN) are valuable sources of strains with unique ecological and biotechnological traits. Such collections play a vital role in bioresource conservation, science, and industry development but are rarely covered in publications. Here, we provide an overview of five genetic collections maintained at the Institute of Plant Physiology of the Russian Academy of Sciences (IPPRAS) since the 1950–1970s using in vitro and cryopreservation approaches. These collections represent different levels of plant organization, from individual cells (cell culture collection) to organs (hairy and adventitious root cultures, shoot apices) to in vitro plants. The total collection holdings comprise more than 430 strains of algae and cyanobacteria, over 200 potato clones, 117 cell cultures, and 50 strains of hairy and adventitious root cultures of medicinal and model plant species. The IPPRAS plant cryobank preserves in LN over 1000 specimens of in vitro cultures and seeds of wild and cultivated plants belonging to 457 species and 74 families. Several algae and plant cell culture strains have been adapted for cultivation in bioreactors from laboratory (5–20-L) to pilot (75-L) to semi-industrial (150–630-L) scale for the production of biomass with high nutritive or pharmacological value. Some of the strains with proven biological activities are currently used to produce cosmetics and food supplements. Here, we provide an overview of the current collections’ composition and major activities, their use in research, biotechnology, and commercial application. We also highlight the most interesting studies performed with collection strains and discuss strategies for the collections’ future development and exploitation in view of current trends in biotechnology and genetic resources conservation.
1 April 2018 in Moscow University Biological Sciences Bulletin
An express assay for screening of potato transformants by their GFP fluorescence intensities is developed. In comparison to the widely used methods of transgenic plant screening by PCR, Real-Time...
Late blight is one of the most economically important diseases affecting potato and causing a significant loss in yield. The development of transgenic potato plants with enhanced resistance to infection by Phytophthora infestans may represent a possible approach to solving this issue. A comparative study of the leaf response in control potato plants (S.tuberosum L. cultivar Skoroplodnyi), control transgenic plants expressing the reporter gene of thermostable lichenase (transgenic licBM3 line) and transgenic plants expressing cyanobacterial hybrid genes ∆9-acyl-lipid desaturase (transgenic desC lines) and ∆12-acyl-lipid desaturase (transgenic desA lines) to infection with P. infestans has been performed. The expression of desaturase genes in potato plants enhanced their tolerance to potato late blight agents as compared with the control. The lipid peroxidation level raised in the leaves of the control and transgenic desA plants on third day after inoculation with P. infestans zoospores and remained the same in the transgenic desC plants. The number of total phenolic compounds was increased as early as on the second day after infection in all studied variants and continued to remain the same, except for transgenic desC plants. Accumulation of flavonoids, the main components of the potato leaf phenolic complex, raised on the second day in all studied variants, remained unchanged on the third day in the control plants and decreased in most transgenic plants expressing desaturase genes. The results obtained in our study demonstrate that the expression of genes of Δ9- and Δ12-acyl-lipid desaturases in potato plants enhanced their resistance to P. infestans as compared with the control non-transgenic plants due to concomitant accumulation of phenolic compounds, including flavonoids, in the leaves. All these changes were more pronounced in transgenic desC plants, which indicates that the Δ9-acyllipid desaturase gene appears to be a potential inducer of the production of biological antioxidants in plant cells.
A comprehensive study was conducted to develop an effective protocol for the clonal micropropagation and subsequent acclimatization of three mint (Mentha) cultivars: ‘Pamyati Kirichenko’, ‘Botanicheskaya’, and ‘Serebryanoe Chudo’. This research investigated the influence of nutrient Murashige–Skoog medium composition with the addition of growth regulators (BAP/NAA) on in vitro morphogenesis, as well as the impact of two acclimatization methods to in vivo conditions (direct field planting and a two-stage acclimatization approach) on field performance, yield, and essential oil composition in three mint cultivars. The results revealed a distinct cultivar-specific response. The ‘Pamyati Kirichenko’ and ‘Botanicheskaya’ cultivars showed high efficiency on the BAP/NAA medium and good survival rates with both acclimatization methods. In contrast, the ‘Serebryanoe Chudo’ cultivar exhibited pronounced sensitivity to the medium with growth regulators and a low survival rate upon direct planting (40.5%). The two-stage acclimatization method proved critically important for this cultivar, enabling an increase in survival to 68%, as well as improvements in yield, essential oil output, and the restoration of its cultivar-specific linalool profile (59.5% linalool). It was established that, for cost-effective production, an individual selection of technological parameters for each genotype is necessary at all stages of microclonal propagation.