In Vitro Cellular & Developmental Biology - Plant

, Volume 49, Issue 2, pp 175–182

DNA mutagenesis in 2- and 20-yr-old Panax ginseng cell cultures

Authors

    • Laboratory of Biotechnology, Institute of Biology and Soil ScienceFar East Branch of Russian Academy of Sciences
  • Alexandra S. Dubrovina
    • Laboratory of Biotechnology, Institute of Biology and Soil ScienceFar East Branch of Russian Academy of Sciences
  • Olga A. Shumakova
    • Laboratory of Biotechnology, Institute of Biology and Soil ScienceFar East Branch of Russian Academy of Sciences
    • Department of Biochemistry and BiotechnologyFar Eastern Federal University
Plant Tissue Culture

DOI: 10.1007/s11627-012-9475-7

Cite this article as:
Kiselev, K.V., Dubrovina, A.S. & Shumakova, O.A. In Vitro Cell.Dev.Biol.-Plant (2013) 49: 175. doi:10.1007/s11627-012-9475-7

Abstract

Previously, Panax ginseng var. Mimaki C.A. Meyer had been shown to accumulate genetic mutations during long-term propagation of a callus culture over a period of 20 yr. In this study, we analyzed the mutation types and frequency in a 2-yr-old P. ginseng callus culture and compared it with the 20-yr-old callus culture, and leaves of cultivated plants. We analyzed the sequence variability between the Actin genes, which are a family of housekeeping genes; phenylalanine ammonia-lyase (PAL) and dammarenediol synthase (DDS), which actively participate in the biosynthesis of ginsenosides; and the somatic embryogenesis receptor kinases (SERK), which control plant development. The frequency of point mutations in the Actin, PAL, DDS, and SERK genes in the 2-yr-old P. ginseng callus culture was markedly higher than in cultivated plants, but lower than in the 20-yr-old callus culture. Most of the mutations in the 2-yr-old P. ginseng calli were A↔G and T↔C transitions, as in the 20-yr-old calli and intact P. ginseng plants. The number of nonsynonymous mutations was higher in the 2- and 20-yr-old callus cultures than the number of nonsynonymous mutations in cultivated P. ginseng. Interestingly, the total number of N→G or N→C substitutions in the analyzed genes was 1.6 times higher than the total number of N→A or N→T substitutions. Using a methylation-sensitive DNA fragmentation assay, we showed the level of methylcytosine to be higher in the DNA of the 20-yr-old P. ginseng calli that than in the DNA of the 2-yr-old cultures.

Keywords

Panax ginseng Cell cultures Mutagenesis Nucleotide substitutions

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© The Society for In Vitro Biology 2012