Expression of ACC Oxidase Gene from Sugarcane Induced by Hormones and Environmental Force

In the present study, a full-length cDNA encoding 1-aminocyclopropane-1-carboxylic acid oxidase gene has been cloned from sugarcane （named GZ-ACO）. Two primers were designed for coding the ORF in the full-length cDNA of GZ-ACO gene from sugarcane. PCR amplification was performed with sugarcane DNA template, and a fragment of 1 104 bp （GZ34） was obtained. GZ34 was labeled with [α-32P] dCTP as the probe and used for hybridization after cloning and sequencing. Southern blotting analysis indicated that there were at least three other sequences, which weakly hybridized with the GZ34. Northern analysis showed that GZ34 was strongly induced by treatment with IAA, BA, ethephon, LiC1 and cold stress, respectively. As a contrast, the mRNA for ACO gene was at lower levels for both the light-grown and dark-grown plants without additional treatment. There were two transcripts in the dark-grown plants and three transcripts in the treatments with IAA, BA and cold stress, but there was only one transcript in ethephon treatment. It showed that GZ-ACO might be a gene connected with ethylene formation and take part in response to the induction of plant hormone and environmental stress.

Isolation and Characterization of Cucumber ACC Oxidase Gene and Its Upstream

Ethylene has been implicated as a sex-determining hormone in cucumber. Its exogenous application increases femaleness,and gynoecious genotypes were reported to produce more ethylene. 1-aminocyclopropane-1-carboxylate (ACC) oxidase(ACO) is the key enzyme in ethylene biosynthesis. In this study, a 1200 base pair (bp) candidate fragment was amplifiedfrom the cucumber genome with degenerated primers derived from the ACO amino acid consensus sequence amongdifferent plant species. The coding region and its upstream (1 155 bp) were obtained by vector-mediated inverse PCR. Thenovel gene was analyzed by bioinformatics tools. Four exons and three introns were identified in the coding sequence.The spliced length of mRNA was 933 nucleotides (nts) and it encoded 311 amino acids. Phylogenic analysis result of thenew gene (CsACO4, GenBank accession number AY450356) was in accordance with the evolution relationship of geneticsamong various plant species. Northern blotting showed that the gene expressed among female flowers of gynoecious andmonoecious genotypes, it could not express in other organs. This implied that the gene might be correlated with the femalebehavior positively. Further work is on the way to demonstrate the complexity of the relationship between the endogenousethylene and the sex determination.

An ACC Oxidase Gene Essential for Cucumber Carpel Development

Sex determination in plants gives rise to unisexual flowers that facilitate outcrossing and enhance genetic diversity. In cucumber and melon, ethylene promotes carpel development and arrests sta- men development. Five sex-determination genes have been identified, including four encoding 1-aminocyclopropane-l-carboxylate （ACC） synthase that catalyzes the rate-limiting step in ethylene biosynthesis, and a transcription factor gene CmWIP1 that corresponds to the Mendelian locus gynoecious in melon and is a negative regulator of femaleness. ACC oxidase （ACO） converts ACC into ethylene; how- ever, it remains elusive which ACO gene in the cucumber genome is critical for sex determination and how CmWIP1 represses development of female flowers. In this study, we discovered that mutation in an ACO gene, CsAC02, confers androecy in cucumber that bears only male flowers. The mutation disrupts the enzymatic activity of CsAC02, resulting in 50% less ethylene emission from shoot tips. CsAC02 was ex- pressed in the carpel primordia and its expression overlapped with that of CsACS11 in female flowers at key stages for sex determination, presumably providing sufficient ethylene required for proper CsACS2 expression. CmAC03, the ortholog of CsACO2, showed a similar expression pattern in the carpel region, suggesting a conserved function of CsACO2/CmACO3. We demonstrated that CsWlP1, the ortholog of CmWlP1, could directly bind the promoter of CsAC02 and repress its expression. Taken together, we propose a presumably conserved regulatory module consisting of WlP1 transcription factor and ACO controls unisexual flower development in cucumber and melon.

Propylene Effects on Ethylene Biosynthesis in Relation to Gene Expression of ACS,ACO in Persimmon Fruit

Influences of propylene treatment on fruit softening and ethylene biosynthesis of persimmon genotype, 'Hiratanenashi' were investigated. The treatment with propylene could accelerate the softening and enhance ethylene biosynthesis, with higher sensitivity of immature fruit to the propylene was consistent with mRNA increase derived from transcription of ACS and ACO genes. Furthermore, ethylene synthesis increase in immature fruits was controlled and regulated mainly by DK-ACS1, DK-ACS2, DK-ACO1 and DK-ACO2, but regulated only by DK-ACS1 and DK-ACO1 in mature fruits.