Selective influence of mannitol on culture of callus tissues of camelina sativa
DOI:
https://doi.org/10.31548/dopovidi2022.01.009Keywords:
camelina sativa, osmotic stress, nutrient medium, mannitol, callus, in vitroAbstract
Camelina sativa is able to provide stable yields in unfavorable natural conditions with low technological and material costs. The specific composition of camelina sativa’s oil makes it possible to use it for various purposes, in particular, food, medical, technical, energy, etc. Despite the prospects of camelina sativa, its sown areas in Ukraine remain insignificant. The main factor holding back the increase in the area under this crop is the lack of highly productive varieties adapted to adverse growing conditions.
Drought is one of the most common adverse abiotic environmental factors that plants face. Under conditions of osmotic stress, plants undergo significant physiological and biochemical changes, which reduces the viability and productivity of the organism. Adaptation of plants to the stress factor occurs at the molecular, cellular and organismal levels due to physiological-biochemical and anatomical-morphological adaptations. Involvement of biotechnological methods in the general scheme of selection process allows to increase efficiency of creation of genotypes with desirable signs. Various osmotically active substances that reduce the external water potential of the culture medium are used as a selective factor for the selection of in vitro drought-resistant samples of agricultural plants. The concentration of the selective agent in the nutrient medium, which is selected experimentally for each species, is crucial.
The aim of our research was to analyze the effect of mannitol on callus culture of camelina sativa to determine the optimal concentration of selective factor for the selection of in vitro drought-resistant forms.
The research was conducted in the educational and scientific laboratory «Biotechnology» of Uman National University of Horticulture. As a selective factor, mannitol was used, which was added in various concentrations (2, 4, 6, 8, 10, 12 %) to the modified nutrient medium according to the Murashige-Skuga recipe. Morphogenic microcalls obtained from explants of camelina sativa varieties Stepovy 1, Klondike, Peremoha and Yevro 12 were planted on the nutrient substrate. At the end of the passage, the viability of microcalls, growth intensity and morphogenic characteristics of induced biomaterials were determined.
Significant differences in the resistance of genotypes to osmotic stress were observed at 6 % of mannitol concentration. The least stable was the biomaterial obtained from Klondike explants with a tissue preservation of 10.5 %. For other genotypes, the survival of biomaterials ranged from 36.0 to 48.2 %.
For ranking the cellular structures of camelina sativa according to the level of resistance to osmotic stress, the concentration of mannitol in the nutrient medium over 8% is optimal. The highest survival rate of biomaterial was observed in the callus of the Peremoha variety (38.3 %). For Stepovy 1, Yevro 12 and Klondike varieties, the share of viable materials was 26.0, 11.6 and 3.4 %, respectively.
At 10 % mannitol concentration, only callus tissues induced from explants of Stepovy 1 and Peremoha varieties remained viable, the share of stable callus lines was 5.6 and 17.0 %, respectively. Further increase in the concentration of the selective factor was lethal to the callus of all genotypes.
The level of resistance of the biomaterial to the selective factor is indicated by the difference between the growth of biomass under stress and optimal cultivation conditions. The studied genotypes of camelina sativa differed in terms of callus tissue proliferation both in the control variant and in the presence of mannitol in the nutrient medium.
According to the results of a comparative analysis of the influence of the studied factors on the preservation and intensity of callus tissue proliferation of camelina sativa under conditions of osmotic stress, it was found that the greatest impact was the concentration of mannitol in the nutrient medium (79 %). The share of the genotype of the donor plant explant and the combined interaction of both factors was 11 and 10 %, respectively.
Thus, in the process of research revealed the stressful effect of mannitol on the callus tissue of camelina sativa. It was found that, depending on the genotype, the concentration of mannitol at the level of 8–10 % is optimal for the selection of in vitro drought-resistant culture forms. High resistance to osmotic stress is characterized by biomaterials obtained from the varieties Stepovy 1 and Peremoha. Isolated camelina sativa cell lines with high resistance to osmotic stress have been identified, which can be used by drought resistance gene donors in breeding research.
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