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Title: Cytosine methylation occurs in a CDC48 homologue and a MADS-box gene during adventitious shoot induction in Petunia leaf explants
Authors: Prakash, A.P.
Kush, A.
Lakshmanan, P.
Kumar, P.P. 
Keywords: CDC48 homologue
Demethylating agents
DNA methylation
MADS-box gene
Petunia hybrida
Shoot regeneration
Issue Date: 1-May-2003
Citation: Prakash, A.P., Kush, A., Lakshmanan, P., Kumar, P.P. (2003-05-01). Cytosine methylation occurs in a CDC48 homologue and a MADS-box gene during adventitious shoot induction in Petunia leaf explants. Journal of Experimental Botany 54 (386) : 1361-1371. ScholarBank@NUS Repository.
Abstract: The DNA methylase inhibitors, 5-azacytidine and 5-aza-2′-deoxycytidine inhibited adventitious shoot induction in Petunia leaf cultures. Cytosine methylation status at CCGG sites in shoot- and callus-inducing culture treatments was analysed by coupled restriction enzyme digestion (Hpall or Mspl) and random amplification. Two differentially methylated genomic DNA bands from the PCR products were cloned (OPU9-1 and OPU9-2) and sequenced. The open reading frames contained in OPU9-1 and OPU9-2 showed similarity to CDC48 and MADS-box genes, respectively. Cytosine methylation was restored at CCGG sites when the leaf explants were transferred from medium containing the drugs to medium without the drugs, simultaneously recovering the ability to develop adventitious shoot buds. Furthermore, combined bisulphite treatment and restriction analysis revealed differential methylation of CGCG sites in the drug-treated and control cultures. These results demonstrate that cytosine methylation at CCGG and CGCG sites within a MADS-box gene and a CDC48 homologue, among others, shows strong positive correlation with adventitious shoot bud induction in Petunia leaf explants.
Source Title: Journal of Experimental Botany
ISSN: 00220957
DOI: 10.1093/jxb/erg155
Appears in Collections:Staff Publications

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