Productivity and stress-tolerance of transgenic tobacco plants with constitutive expression of rapeseed glutathione synthetase gene BnGSH

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Summary: Glutathione is the most important part of plant antioxidant defense system. Biosynthesis of glutathione in the cells is performed by two enzymes: glutamylcysteine ligase and glutathione synthetase, the latter catalyzing the attachment of glycine to a dipeptide glutamylcysteine. In literature there is information on the improvement of heavy metal-tolerance of transgenic plants due to the increase in the expression level of glutathione synthetase genes. However there is not enough data on the tolerance of these plants to other types of abiotic stress. Therefore the aim of our research was to make transgenic tobacco plants with constitutive expression of rapeseed glutathione synthetase gene BnGSH and to estimate their growth parameters in normal conditions and under salt, drough and cold stress. Using agrobacterial transformation method, we generated 17 lines of transgenic plants containing rapeseed BnGSH gene under control of 35S promoter. The presence of transgenes was confirmed by PCR method and histochemical analysis of the activity of GUS reporter gene. 12 lines with the highest expression of BnGSH gene were chosen on the basis of the results of RT-PCR. We performed morphological analysis, including measurements of stem hight, leaf area, flower length, fresh and dry weight of shoots and root length. Some transgenic plants demonstrated increased productivity in normal conditions as well as under NaCl stress. However, no change in drought and cold tolerance was observed in transgenic plants.

About the authors

Bulat R Kuluev

Institute of Biochemistry and Genetics, Ufa Sci. Center of RAS

Author for correspondence.
Email: Kuluev@bk.ru

Senior Researcher. Laboratory of molecular biology and nanobiotechnology

Russian Federation, Ufa, Russia

Zoya A Berezhneva

Institute of Biochemistry and Genetics, Ufa Sci. Center of RAS

Email: berezhneva-z@yandex.ru

Postgraduate. Laboratory of molecular biology and nanobiotechnology

Russian Federation, Ufa, Russia

Elena V Mikhaylova

Institute of Biochemistry and Genetics, Ufa Sci. Center of RAS

Email: mikhele@list.ru

Senior laboratory assistant. Laboratory of molecular biology and nanobiotechnology

Russian Federation, Ufa, Russia

Bogdan N Postrigan

Institute of Biochemistry and Genetics, Ufa Sci. Center of RAS

Email: postrigan@bk.ru

Researcher. Laboratory of molecular biology and nanobiotechnology

Russian Federation, Ufa, Russia

Aleksey V Knyazev

Institute of Biochemistry and Genetics, Ufa Sci. Center of RAS

Email: knyazev@anrb.ru

Senior Researcher. Laboratory of molecular biology and nanobiotechnology

Russian Federation, Ufa, Russia

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2. Fig. 1. Morphometric parameters of transgenic tobacco plants overexpressing the BnGSH gene under normal growth conditions: a – stem height; b – leaf area; c – fresh weight of shoot; d – dry weight of shoot; ДТ – wild type, 1–14 – lines of transgenic plants. n = 5. * p < 0,01

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3. Fig. 2. Morphometric parameters of transgenic tobacco plants overexpressing the BnGSH gene under NaCl salinity: a – stem height; b – fresh weight of shoot; c – dry weight of shoot; d – root growth in length when grown under normal conditions; e – root growth in length when grown under action of 50 мМ NaCl; f – root growth in length when grown under action of 100 мМ NaCl. ДТ – wild type, 1–14 – lines of transgenic plants. n = 5. * p < 0,01. In the histograms a-c with a light-gray column indicate morphometric parameters under normal conditions and a dark gray column – morphometric parameters under the action of NaCl

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4. Fig. 3. Morphometric parameters of transgenic tobacco plants overexpressing the BnGSH gene under low positive temperatures and drought: a – fresh weight of shoot under action of +12 °C; b – dry weight of shoot under action of +12 °C; c – root growth in length when grown under action of +12 °C; d – fresh weight of shoot under action of drought; e – dry weight of shoot under action of drought; f – relative water content (RWC). ДТ – wild type, 1-14 – lines of transgenic plants. n = 5. * p < 0,01

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Copyright (c) 2017 Kuluev B.R., Berezhneva Z.A., Mikhaylova E.V., Postrigan B.N., Knyazev A.V.

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This work is licensed under a Creative Commons Attribution 4.0 International License.
 


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