Molecular cloning and expression of a jasmonate biosynthetic gene allene oxide cyclase from Camellia sinensis

As a family of signaling plant hormone, jasmonic acid plays an important role in coordinating plant defense responses to pests and pathogen attack through transcriptional and metabolic changes. In the jasmonate biosynthetic pathway of plants, allene oxide cyclase (AOC) is an essential enzyme. Here w...

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Bibliographic Details
Published in:Canadian journal of plant science 2016-02, Vol.96 (1), p.109-116
Main Authors: Wang, Meng-xin, Ma, Qing-ping, Han, Bao-yu, Li, Xing-hui
Format: Article
Language:English
Subjects:
acide jasmonique
allene
allene oxide cyclase
allène oxyde cyclase
amino acids
biochemical pathways
bioinformatics
Camellia sinensis
clonage moléculaire
complementary DNA
E coli
Empoasca vitis
Escherichia coli
expression
Gene expression
genes
Ipomoea nil
Jasmonic acid
leaves
messenger RNA
Methyl jasmonate
Molecular Biology
molecular cloning
open reading frames
pathogens
Pests
Phylogeny
plant hormones
Plant sciences
quantitative polymerase chain reaction
Salicylic acid
Tea
tea plant
théier, Camellia sinensis
Transcription
transcription (genetics)
Wounding
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Summary:As a family of signaling plant hormone, jasmonic acid plays an important role in coordinating plant defense responses to pests and pathogen attack through transcriptional and metabolic changes. In the jasmonate biosynthetic pathway of plants, allene oxide cyclase (AOC) is an essential enzyme. Here we cloned a cDNA from tea plant (Camellia sinensis), named as CsAOC (GenBank: HQ889679), which was 916 bp, containing an open reading frame (738 bp) encoding 245 amino acids. Comparative and bioinformatic analyses revealed that the deduced protein of CsAOC was highly homologous to AOC from other plant species. Phylogenetic analysis indicated that CsAOC was clustered in a closely related subgroup with AOC of Ipomoea nil. The full-length coding region of CsAOC was ligated with pET-32a and successfully expressed in E. coli BL21 (DE3), and purified. Real-time quantitative PCR analysis revealed that methyl jasmonate (MeJA) treatment on its own potently enhanced its expression over the control (healthy leaves), suggesting feedback activation of CsAOC. The expression under salicylic acid (SA) and wounding treatments was up-regulated. The mRNA expression of CsAOC could be induced by tea geometrids and tea green leafhoppers.
ISSN:0008-4220
1918-1833
1918-1833
DOI:10.1139/cjps-2015-0164