Methylated DNA changes associated with the initiation and maintenance of <i>Vitis vinifera in vitro</i> shoot and callus cultures: A possible mechanism for age-related changes

Authors

  • K. Harding
  • Erica E. Benson
  • Kalliope A. Roubelakis-Angelakis

DOI:

https://doi.org/10.5073/vitis.1996.35.79-85

Keywords:

biotechnology, molecular biology, tissue culture, grapevine

Abstract

Tissue culture technologies are an important aspect of the genetic modification of grapevine (Vitis vinifera). The molecular basis of this phenomenon is not well understood, however, the extent of DNA methylation is recognised as a factor in the control of gene expression. This study explores the possibility that DNA methylation may have a role in grapevine culture responses. DNA methylation profiles were constructed for Vitis vinifera, cv. Sultanina, during plantlet micropropagation, callus induction and proliferation.
Methylation of genomic DNA and ribosomal RNA genes (rDNA) was found in glasshouse-grown plants, micropropagated plantlets and callus cultures. An analysis of rDNA showed that glasshouse-grown plants had 74.6% of the recognition sequences for Hpa II methylated at the internal cytosine position CmCGG, whereas 7.7% of the recognition sequences appeared to be methylated at the external cytosine (mCmCGG). The rDNA profiles of micropropagated subcultures S0 (initial subcultures) and S4 (fourth subcultures), representing one year of in vitro growth, showed that the percentage of recognition sequences containing a methylated external cytosine increased from 7.7% in glasshouse-grown plants to 64.5% for S-0 and 72.5% for the S4 subculture. The implications of these findings for the in vitro manipulation of grapevine used in genetic modifications are discussed.

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Published

2015-08-13

Issue

Vol. 35 No. 2 (1996): Vitis

Section

Article

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