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

  • Konstantin V. Kiselev
  • Alexandra S. Dubrovina
  • Olga A. Shumakova
Plant Tissue Culture

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 

Notes

Acknowledgments

This work was supported by grants from the Russian Foundation for Basic Research (10-04-00189-а) and from the Far East Division of the Russian Academy of Sciences (12-III-А-06-072).

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Copyright information

© The Society for In Vitro Biology 2012

Authors and Affiliations

  • Konstantin V. Kiselev
    • 1
  • Alexandra S. Dubrovina
    • 1
  • Olga A. Shumakova
    • 1
    • 2
  1. 1.Laboratory of Biotechnology, Institute of Biology and Soil ScienceFar East Branch of Russian Academy of SciencesVladivostokRussia
  2. 2.Department of Biochemistry and BiotechnologyFar Eastern Federal UniversityVladivostokRussia

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