Molecular characterization of CYP73A9 and CYP82A1 P450 genes involved in plant defense in pea

Abstract

Cytochrome P450 monooxygenases (P450s) mediate a wide range of oxidative reactions involved in the biosynthesis of phenylpropanoids, terpenes, and alkaloids. Two pea (Pisum sativum) P450 cDNAs (CYP73A9v1, encoding transcinnamic acid hydroxylase [t-CAH] in the core phenylpropanoid pathway, and CYP82A1v1, possibly encoding an activity in a late branch of the phenylpropanoid pathway) have previously been described. Of three CYP73A9 genes now isolated, the CYP73A9v1 gene is full-length with two introns at positions conserved in other t-CAH genes, and the CYP73A9v2 and CYP73A9v3 gene fragments are 5'-truncated and lack introns. The full-length CYP82A1v2 gene contains a single intron at an alternate position. Nucleotide searches of the CYP73A9v1 and CYP82A1v2 promoters have indicated that the regulatory sequences for these early and late phenylpropanoid transcripts are substantially different. The P-, L-, and H-boxes identified in white light-, ultraviolet light-, and elicitor-induced footprints in early phenylpropanoid promoters (phenylalanine ammonia lyase [PAL], 4-coumarate coenzyme A:ligase [4-CL], and chalcone synthase [CHS]) are conserved in the t-CAH promoter and are absent from the CYP82A1v2 promoter. Both promoters contain TCA motifs identified in stress-responsive promoters, box IV elements identified in elicitor-responsive PAL and CHS promoters, and spatially conserved wound-response elements potentially coordinating regulation of these wound-responsive promoters.