Cloning and expression of the 1-aminocyclopropane-1-carboxylic oxidase gene from Agrostis stolonifera.
A gene encoding 1-aminocyclopropane-1-carboxylic oxidase (ACO), which catalyzes the terminal step in ethylene biosynthesis, was isolated from Agrostis stolonifera. The AsACO gene is composed of 975 bp, encoding 324 amino acids. Three exons interspersed by two introns form AsACO gDNA. A BLAST search of the nucleotide sequence revealed a high level of similarity (79-91%) between AsACO and ACO genes of other plants. A phylogenetic tree was constructed via BLAST in the NCBI, and revealed the highest homology with wheat TaACO. The calculated molecular mass and predicted isoelectric point of AsACO were 36.25 and 4.89 kDa, respectively. Analysis of subcellular localization revealed that AsACO is located in the nucleus and cytoplasm. The Fe(II)-binding cofactors and cosubstrate were identified, pertaining to the ACO family. The expression patterns of AsACO were determined by quantitative real time PCR. AsACO expression was highest in the stem, and was strongly up-regulated in response to ethephon, methyl jasmonate, salicylic acid, and cold temperature, but down-regulated in response to drought and NaCl treatment. The protein encoded by AsACO exhibited ACC oxidase activity in vitro. Taken together, these findings suggest that AsACO contains domains common to the ACO family, and is induced in response to exogenous hormones. Conversely, some abiotic stress conditions can inhibit AsACO expression.
Turfgrass Research Institute, College of Forestry, Beijing Forestry University, Beijing, China.|College of Horticulture and Garden, Yangtze University, Jingzhou, China.
G Z
GZ
Xiao
Turfgrass Research Institute, College of Forestry, Beijing Forestry University, Beijing, China.
L J
LJ
Li
Turfgrass Research Institute, College of Forestry, Beijing Forestry University, Beijing, China.
K
K
Teng
Turfgrass Research Institute, College of Forestry, Beijing Forestry University, Beijing, China chaoyuehui@163.com.
Y H
YH
Chao
Turfgrass Research Institute, College of Forestry, Beijing Forestry University, Beijing, China chaoyuehui@163.com.
L B
LB
Han
D031703:Agrostis / Q000201:enzymology* / Q000235:genetics*; D000594:Amino Acid Oxidoreductases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D019295:Computational Biology; D018076:DNA, Complementary / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D005822:Genetic Vectors / Q000378:metabolism; D008970:Molecular Weight; D010802:Phylogeny; D018515:Plant Leaves / Q000201:enzymology; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D013347:Subcellular Fractions / Q000201:enzymology; D014158:Transcription, Genetic
D018076:DNA, Complementary; D010940:Plant Proteins; D000594:Amino Acid Oxidoreductases; C059460:1-aminocyclopropane-1-carboxylic acid oxidase