Novel broccoli 1-aminocyclopropane-1-carboxylate oxidase gene (Bo-ACO3) associated with the late stage of postharvest floret senescence.
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.
Yang
Chih-Yuan
CY
Life Science Center, Hsing Wu College, No. 11-2 Fen-Liao Road, Lin-Kou, Taipei, Taiwan 11244.
Chu
Fang Hua
FH
Wang
Yuh Tai
YT
Chen
Yu-Ting
YT
Yang
Shang Fa
SF
Shaw
Jei-Fu
JF
D000594:Amino Acid Oxidoreductases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D000595:Amino Acid Sequence; D001483:Base Sequence; D001937:Brassica / Q000201:enzymology* / Q000235:genetics; D018076:DNA, Complementary / Q000737:chemistry; D018744:DNA, Plant / Q000737:chemistry; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D015151:Immunoblotting; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D018744:DNA, Plant; D000594:Amino Acid Oxidoreductases; C059460:1-aminocyclopropane-1-carboxylic acid oxidase