Olga A Shumakova

Publications (2)4.44 Total impact

• Article: Resveratrol content and expression of phenylalanine ammonia-lyase and stilbene synthase genes in cell cultures of Vitis amurensis treated with coumaric acid.

  Olga A Shumakova, Artem Y Manyakhin, Konstantin V Kiselev
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  ABSTRACT: The present study examines the effect of p-coumaric acid (CA), a precursor of stilbenes and isoflavonoids, on biosynthesis of resveratrol in cell cultures of Vitis amurensis. Earlier, we transformed V. amurensis with the rolB gene of Agrobacterium rhizogenes and showed increased level of resveratrol production in the rolB transgenic cell culture. We used control and the rolB-transgenic cell culture of V. amurensis as a model system in this study. CA was capable of increasing resveratrol production in the control and the rolB-transgenic cell cultures in 10.3 and 1.5 times, respectively. The CA-treated control and rolB transgenic calli produced up to 0.06% and 1.1% DW of resveratrol. Using quantitative real-time RT-PCR, we characterized the expression of phenylalanine ammonia-lyase (PAL) and stilbene synthase (STS) genes in the CA-treated control and rolB transgenic cell cultures. The expression of PAL genes remained essentially unchanged under 0.1 mM of CA, while expression of VaPAL1, VaPAL2, VaPAL3, and VaPAL5 genes was considerably decreased under 0.5 and 2 mM CA compared with the untreated cells. In the CA-treated calli, expression of VaSTS2 and VaSTS3 was increased, while expression of VaSTS5, VaSTS8, VaSTS9, and VaSTS10 was significantly decreased. These results indicate that CA increased resveratrol accumulation in V. amurensis calli via selective enhancement of expression of individual STS genes.
  Applied biochemistry and biotechnology 09/2011; 165(5-6):1427-36. · 1.94 Impact Factor
• Article: Mutation of Panax ginseng genes during long-term cultivation of ginseng cell cultures.

  Konstantin V Kiselev, Olga A Shumakova, Galina K Tchernoded
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  ABSTRACT: It has previously been shown that the nucleotide sequences of the Agrobacterium rhizogenes rolC locus and the selective marker nptII developed mutations during the long-term cultivation of transgenic cell cultures of Panax ginseng. In the present report, we analyzed the nucleotide sequences of selected plant gene families in the 20-year-old P. ginseng 1c cell culture and in leaves of cultivated P. ginseng plants. We sequenced the Actin genes, which are a family of house-keeping genes; the phenylalanine ammonia-lyase (PAL) and dammarenediol synthase genes (DDS), which actively participate in the biosynthesis of ginsenosides; and the somatic embryogenesis receptor kinase (SERK) genes, which control plant development. We demonstrate that the plant genes also developed mutations during long-term cultivation. The highest level of nucleotide substitution was detected in the sequences of the SERK genes (2.00±0.11 nt per 1000 nt), and the level was significantly higher when compared with the cultivated P. ginseng plant. Interestingly, while the diversity of Actin genes was similar in the P. ginseng cell culture and the cultivated plants, the diversity of the DDS and SERK genes was less in the 20-year-old cell culture than in the cultivated plants. In this work, we detail the level of nucleotide substitutions in different plant genes during the long-term culture of plant cells.
  Journal of plant physiology 07/2011; 168(11):1280-5. · 2.50 Impact Factor