Page 111 - Zbornik_Konf_2019

https://doi.org/10.15414/2019.9788055220703

4 International Scientific Conference Abstracts Book
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CULTIVATION OF TOBACCO PLANTS EXPRESSING desC AND desA GENES OF
DESATURASE CYANOBACTERIA IN CONDITIONS OF OSMOTIC STRESSES
Tetiana Kyrpa‐Nesmiian, Mykola Kuchuk

Institute of Cell Biology and Genetic Engineering NAS of Ukraine, Kyiv, Ukraine;
E-mail.: t-kirpa@ukr.net
The study of the adaptation of plants to abiotic stress is one of the most promising
directions today. One of the mechanisms to increase the resistance of plants to the influence of
temperatures (high, low, frost), lack of moisture, the influence of wind, salinity of soils is an
increase in the proportion of unsaturated fatty acids in the lipids of membrane. With the
increase in the proportion of unsaturated fatty acids in the membrane lipids, the viscosity of
the membranes increases, and resistance to abiotic stresses increases.
Desauturases are enzymes that promote the formation of double bonds in fatty acids
and thus convert them from saturated to unsaturated. Cyanobacteria are among the ancient
organisms that have not been mutated for the last two million years. Which points to high
adaptability. The desC gene encoding the Δ9-acyl-lipid desaturase of cyanobacteria
Synechococcus vulcanus and the desA gene encoding Δ12-acyl-lipid desaturase of
cyanobacteria Synechocystis sp. PCC 6803 was used in the work.
By genetic Agrobacterium tumefaciens-mediated transformation, Nicotiana tabacum
tobacco plants that simultaneously express the desC and desA genes were obtained. Plants
were planted on a medium of MS with the addition of mannitol (concentration 100 mM
200 mm) for further work. The parameters of malondialdehyde, the superoxide dismutase
enzyme, and the level of gene expression on the activity of the reporter protein were checked.
Wild-type tobacco plants and transgenic plants that have in their genome expressing the
gfp::licBM3 gene are used as controls.
Malondialdehyde is a marker of oxidative degradation of unsaturated fatty acids. The
mechanism of action of SOD consists in the successive restoration and oxidation of superoxide
anionic radicals of the metal of the active center of the enzyme. The activity of SOD is carefully
studied since this enzyme plays an important role in protecting cells and tissues from
oxidative degradation. Since the desaturase genes are in the frame of reading from the
genome of the reporter protein of the thermostable lichenase, the activity of the thermostable
lichenase after the cultivation of the plants on MS medium and MS with the addition of
mannitol is checked.
It was found that the highest level of activity of thermostable lichenase was in plants
with desaturase genes, indicating increased expression of desaturase genes. Most damages
were detected in plants with the gen gfp::licBM3. Less degenerative changes have been found
in plants containing in their genome and expressing genes of desaturase of cyanobacteria.
Keywords: abiotic stress, desaturase.

4 International Scientific Conference Agrobiodiversity Nutrition, Health and Quality of Human and Bees Life |111
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September 11–13, 2019