A novel cytochrome P450 gene from Catharanthus roseus cell line C20hi: cloning and characterization of expression

An expressed sequence tag (EST) obtained from a subtractive-suppression hybridization cDNA library constructed using Catharanthus roseus cell line C20hi and its parental cell line C20D was used to clone a full-length cytochrome P450 cDNA of cyp71d1. The encoded polypeptide contained 507 amino acids with 39–56% identity to other CYP71D subfamily members at the amino acid level. Expression characteristics of cyp71d1 were determined using semi-quantitative RT-PCR. The cyp71d1 transcript was expressed in all three cell lines with the highest level in the cell line C20hi. In the mature C. roseus plant, the cyp71d1 cDNA was highly expressed in petals, roots and stems, but very weakly expressed in young leaves. Its transcription level increased with the development of flowers. 2,4-D could down-regulate the transcription of cyp71d1, as did KT, but only to a minor degree. Neither light nor yeast elicitor could induce the transcription of cyp71d1.

The full length cDNA of a cytochrome P450 gene cyp71d1 was cloned based on the EST sequence. Semi-quantitative RT-PCR revealed that cyp71d1 expressed highly in the full habituated cell line C20hi and in the petals, roots, and stems of the mature plant. It could be down-regulated by 2,4-D and KT. Light could not induce the expression of cyp71d1.Figure optionsDownload as PowerPoint slide