Molecular cloning, characterization and expression of a jasmonate biosynthetic pathway gene encoding allene oxide cyclase from Camptotheca acuminata.
AOC (allene oxide cyclase; EC 5.3.99.6), an essential enzyme in jasmonic acid and its methyl ester biosynthesis, was cloned from Camptotheca acuminata (named as CaAOC), a native medicinal plant species in China. CaAOC had significant similarity at the amino-acid level with AOCs from other plant species. Comparison between the sequences of the full-length cDNA and genomic DNA of CaAOC revealed that the genomic DNA of CaAOC contained an 89-bp intron and a 240-bp intron. Southern-blot analysis indicated that CaAOC was a multiple-copy gene, and real-time quantitative PCR analysis showed that CaAOC was expressed constitutively in all organs tested, with the highest expression level in leaves. The results from treatment experiments using different signalling components, including methyl jasmonate, abscisic acid, salicylic acid and H(2)O(2), revealed that expression of CaAOC had a prominent diversity. Heavy metal (copper) significantly enhanced CaAOC expression, whereas wounding (induced by UV-B) was not so effective.
State Key Laboratory of Genetic Engineering, School of Life Sciences, Morgan-Tan International Center for Life Sciences, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, Fudan University, Shanghai 200433, People's Republic of China.
Yan
Y
Pi
Zhihua
Z
Liao
Keji
K
Jiang
Beibei
B
Huang
Zhongxiang
Z
Deng
Dongli
D
Zhao
Hainian
H
Zeng
Xiaofen
X
Sun
Kexuan
K
Tang
D001483:Base Sequence; D029769:Camptotheca / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods; D003300:Copper / Q000494:pharmacology; D003517:Cyclopentanes / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000502:physiology*; D018506:Gene Expression Regulation, Plant / Q000187:drug effects / Q000502:physiology*; D019746:Intramolecular Oxidoreductases / Q000096:biosynthesis* / Q000235:genetics; D008969:Molecular Sequence Data; D054883:Oxylipins / Q000378:metabolism*; D010940:Plant Proteins / Q000096:biosynthesis* / Q000235:genetics
D003517:Cyclopentanes; D054883:Oxylipins; D010940:Plant Proteins; C011006:jasmonic acid; D003300:Copper; D019746:Intramolecular Oxidoreductases; C021377:hydroperoxide isomerase