Molecular cloning, characterization and expression analysis of three aldehyde oxidase genes from Pisum sativum L.

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Aldehyde oxidase (AO, EC 1.2.3.1) is a molybdenohydroxylase that is considered to catalyze the last step of abscisic acid (ABA) and indole-3-acetic acid (IAA) synthesis. Three cDNAs encoding aldehyde oxidase proteins in Pisum sativum (cv. Little Marvel) were obtained based on RT-PCR (reverse transcriptase-polymerase chain reaction) strategy. The cloned genes, designated as PsAO1, PsAO2 and PsAO3, are 4630, 4347, 4600 bp in length, respectively, and show high sequence identity to each other and to aldehyde oxidases from other plant species. The deduced PsAO1, PsAO2, and PsAO3 proteins are 1373, 1367, 1367 amino acids in length, respectively, and contain consensus sequences for two iron-sulfur centers, a FAD binding domain, and a molybdenum cofactor (Moco) binding domain. PsAO1 and PsAO2 were mainly expressed in leaves of seedlings and young leaves of adult plants, while the highest PsAO3 transcript level was observed in aging leaves and matured seeds. PsAO2 mRNA was not affected by salinity or ammonium treatment, whereas the transcript level of PsAO3 increased significantly under both stress conditions, with the most pronounced changes in aging leaves, fully expanded leaves and roots. The PsAO1 transcript level was enhanced only in the presence of ammonium in the nutrient medium, but not under salinity. Based on the molecular mass of the deduced proteins and on organ-specific gene expression, studied both under control and stress conditions, the contribution of each PsAO cDNA in the formation of the previously described three dimeric pea AO isoforms and the possible involvement of the PsAO3 in abscisic acid (ABA) synthesis is discussed.