Journal of Stress Physiology & Biochemistry, Vol. 12 No. 1 2016, pp. 87-107  ISSN 1997-0838
Original Text Copyright (cc) 2015 by  Pradedova, Nimaeva, Putilina, Semenova, Sobenin and Salyaev



ORIGINAL ARTICLE
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Determination of Glutathione and Its Redox Status in Isolated Vacuoles of Red Beetroot Cells

E.V. Pradedova*, O.D. Nimaeva, T.E. Putilina, N.V. Semenova, A.M. Sobenin, R.K. Salyaev

  Siberian Institute of Plant Physiology and Biochemistry, SB RAS, Irkutsk, Russia

*E-Mail: praded@sifibr.irk.ru

Received January 20, 2016

The glutathione of the red beetroot vacuoles (Beta vulgaris L.) was measured using three well-known methods: the spectrofluorimetric method with orthophthalic aldehyde (OPT); the spectrophotometric method with 5.5'-dithiobis-2-nitrobenzoic acid (DTNB); the high-performance liquid chromatography (HPLC). The content of reduced (GSH) and oxidized glutathione (GSSG) differed depending on the research method. With OPT the concentration of glutathione was: GSH – 0.059 µmol /mg protein; GSSG – 0.019 µmol/mg protein and total glutathione (GSHtotal) – 0.097 µmol/mg protein. In the case of determining with DTNB the concentration of glutathione was: GSH – 0.091 µmol/mg protein; GSSG – 0.031 µmol/mg protein; GSHtotal – 0.153 µmol/mg protein. HPLC-defined concentration of glutathione was lower: GSH – 0.039 µmol/mg protein; GSSG – 0.007 µmol/mg protein; GSHtotal – 0.053 µmol/mg protein. Redox ratio of GSH/GSSG was also dependent on the method of determination: with OPT – 3.11; with DTNB – 2.96 and HPLC – 5.57. Redox ratio of glutathione in vacuoles was much lower than the tissue extracts of red beetroot, which, depending on the method of determination, was: 7.23, 7.16 and 9.22. The results showed the vacuoles of red beetroot parenchyma cells contain glutathione. Despite the low value of the redox ratio GSH/GSSG, in vacuoles the pool of reduced glutathione prevailed over the pool of oxidized glutathione.


Key words:   
Beta vulgaris L., vacuole, glutathione


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