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A  Lycopersicon esculentum  Mill. (tomato) cDNA clone with high similarity to a  Nicotiana plumbaginifolia  Viv. (tobacco) cytochrome P450 gene was isolated using 5' and 3' rapid amplification of cDNA ends (RACE). The isolated cDNA (GenBank Accession No. AF249329) has an open reading frame of 1494 base pairs (bp) and encodes a protein of 498 amino acids with 75% identity to the  N. plumbaginifolia  cytochrome P450 ( CYP72A2 ) and 45% to a  Catharanthus roseus  G. Don (Madagaskar periwinkle)  CYP72A1  protein sequence. By Southern-blot analysis, one or two highly homologous genes were detected in the  L. esculentum  genome. Expression of the cloned P450 gene was regulated by circadian rhythm and enhanced by wounding. Leaf transcripts were detected in the light but not dark. Highest transcript levels were observed 3 hours after mechanical wounding. No increase in expression was seen in response to applications of zeatin as with the  N. plumbaginifolia  gene. Of the tissues analyzed, shoot tips and young leaves and fruit had the highest detectable transcript levels. Attempts to transform more than 1400 cotyledon explants of  L. esculentum  with sense or antisense  CYP72A2  gene constructs produced no transgenic plants.

journal of the american society for horticultural science

Cloning of a Wound Inducible Lycopersicon esculentum Cytochrome P450 Gene and Lack of Regeneration of Transgenic Plants with Sense or Antisense Constructs