Structure and expression of three genes encoding ACC oxidase homologs from melon (Cucumis melo L.)

Abstract

The enzyme ACC oxidase catalyses the last step of ethylene biosynthesis in plants, converting 1-aminocyclopropane-1-carboxylic acid (ACC) to ethylene. We have previously described the isolation and characterization of a cDNA clone (pMEL1) encoding an ACC oxidase homolog from melon (Cucumis melo L.). Here we report the isolation and characterization of three genomic clones, corresponding to three putative members of the ACC oxidase gene family in melon. All are transcriptionally active. The sequences of these genes have been determined. One genomic clone (CM-ACO1), corresponding to the cDNA previously isolated, presents a coding region interrupted by three introns. Its transcription initiation site has been defined with RNA from ripe fruit and ethylene-treated leaves. The other two genes (CM-ACO2, CM-ACO3) have only two introns, at positions identical to their counterparts inCM-ACO1. The degree of DNA homology in the coding regions ofCM-ACO2 andCM-ACO3 relative toCM-ACO1 is 59% and 75%, respectively.CM-ACO2 andCM-ACO3 are 59% homologous in their coding regions. These three genes have close homology toPH-ACO3, a member of the ACC oxidase multigene family of petunia. The predicted amino acid sequences of CM-ACO1 and CM-ACO3 are 77% to 81% identical to those encoded by the tomato and petunia genes, while the deduced amino acid sequence ofCM-ACO2 shows only 42% to 45% homology. RT-PCR analysis using gene-specific primers shows that the three genes are differentially expressed during development, ethylene treatment and wounding.CM-ACO1 is induced in ripe fruit and in response to wounding and to ethylene treatment in leaves.CM-ACO2 is detectable at low level in etiolated hypocotyls.CM-ACO3 is expressed in flowers and is not induced by any of the stimuli tested.