Novel Broccoli 1-Aminocyclopropane-1-carboxylate Oxidase Gene (Bo-ACO3) Associated with the Late Stage of Postharvest Floret Senescence
2003
Yang, Jerry | Chu, Fang Hua | Wang, Yuh Tai | Chen, Yu Ting | Yang, Shang-Fa | Shaw, Jei-Fu
A novel 1-aminocyclopropane-1-carboxylate (ACC) oxidase gene (Bo-ACO3) was first isolated from senescing broccoli florets by subtractive hybridization. The cDNA clone comprised a 963 bp open reading frame encoding a protein of 321 amino acids. The predicted molecular mass and pI were 36 kDa and 5.42, respectively. Bo-ACO3 shares 68% identity in the coding region with Bo-ACO1 (ACC Ox1) and Bo-ACO2 (ACC Ox2) and is quite divergent from the 3‘ untranslated regions. Bo-ACO3 transcript was accumulated to high levels only at the late stage of senescence after harvest. Southern blot hybridization using full-length cDNA as a probe suggested that the Bo-ACO3 gene is a single-copy gene in the broccoli genome. The deduced 321 amino acid sequence of Bo-ACO3 shares 70% identity with either Bo-ACO1 or Bo-ACO2. The BO-ACO3 gene was expressed in Escherichia coli as a 38 kDa active ACO enzyme. It was concluded that Bo-ACO3 is a senescence-associated gene involved in the late-phase senescence of postharvest broccoli.
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