title: Molecular Cloning and Characterization of Hydroperoxide Lyase Gene in the Leaves of Tea Plant (Camellia sinensis).
abstract: Hydroperoxide lyase (HPL, E.C. 4.1.2.) is the major enzyme in the biosynthesis of natural volatile aldehydes and alcohols in plants, however, little was known about HPL in tea plants (Camellia sinensis). A unique cDNA fragment was isolated by suppressive subtractive hybridization (SSH) from a tea plant subjected to herbivory by tea geometrid Ectropis obliqua. This full length cDNA acquired by RACE was 1476 bp and encoded 491 amino acids. DNA and protein BLAST searches showed high homology to HPL sequences from other plants. The His-tag expression vector pET-32a(+)/CsHPL was constructed and transferred into Escherichia coli Rosetta (DE3). The expression product of recombinant CsHPL in E. coli was about 60 kDa. The enzyme activity of CsHPL was 0.20 μmol·min(-1)·mg(-1). Quantitative RT-PCR analysis indicated CsHPL was strongly up-regulated in tea plants after Ectropis obliqua attack, suggesting that it may be an important candidate for defense against insects in tea plants.
authors.affiliation: State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University , 130 Changjiang West Road, Hefei, Anhui 230036, China.
authors.forename: Wei-Wei
authors.initials: WW
authors.lastname: Deng
authors.affiliation: State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University , 130 Changjiang West Road, Hefei, Anhui 230036, China.
authors.forename: Yi-Lin
authors.initials: YL
authors.lastname: Wu
authors.affiliation: State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University , 130 Changjiang West Road, Hefei, Anhui 230036, China.
authors.forename: Ye-Yun
authors.initials: YY
authors.lastname: Li
authors.affiliation: State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University , 130 Changjiang West Road, Hefei, Anhui 230036, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Tan
authors.affiliation: State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University , 130 Changjiang West Road, Hefei, Anhui 230036, China.
authors.forename: Chao-Ling
authors.initials: CL
authors.lastname: Wei
mesh_terms: D000446:Aldehyde-Lyases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D028241:Camellia sinensis / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000469:parasitology; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D008969:Molecular Sequence Data; D009036:Moths / Q000502:physiology; D018515:Plant Leaves / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D016415:Sequence Alignment
chemical_list: D010940:Plant Proteins; D003577:Cytochrome P-450 Enzyme System; D000446:Aldehyde-Lyases; C054540:hydroperoxide lyase
keywords: Camellia sinensis; Ectropis obliqua; hydroperoxide lyase; induced defense response; suppressive subtractive hybridization (SSH)