title: Molecular cloning and characterization of 4-hydroxyphenylpyruvate dioxygenase gene from Lactuca sativa.
abstract: Vitamin E has been found to be associated with an important antioxidant property in mammals and plants. In photosynthetic organisms, the enzyme 4-hydroxyphenylpyruvate dioxygenase (HPPD; E.C. 1.13.11.27) plays an important role in the vitamin E biosynthetic pathway. The full-length cDNA encoding HPPD was isolated from Lactuca sativa L. by rapid amplification of cDNA ends (RACE). The cDNA, designated as LsHPPD, was 1743 base pairs (bp) long containing an open reading frame (ORF) of 1338 bp encoding a protein of 446 amino acids. Sequence analysis indicated that LsHPPD shared high identity with HPPD from Medicago truncatula L. Real-time fluorescent quantitative PCR (qPCR) analysis revealed that LsHPPD was preferentially expressed in mature leaves compared with other tissues and that the LsHPPD expression was sensitive to high light and drought stress treatments. Transient expression of LsHPPD via agroinfiltration resulted in 12-fold increase in LsHPPD mRNA expression level and 4-fold enhancement in α-tocopherol content compared with the negative control. A decrease in chlorophyll content and inhibition of photosystem II were observed during stress treatments and agroinfiltration.
authors.affiliation: Plant Biotechnology Research Center, School of Agriculture and Biology, Key Laboratory of Urban Agriculture South, Ministry of Agriculture, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, Shanghai Jiao Tong University, Shanghai 200240, PR China.
authors.forename: Weiwei
authors.initials: W
authors.lastname: Ren
authors.forename: Lingxia
authors.initials: L
authors.lastname: Zhao
authors.forename: Lida
authors.initials: L
authors.lastname: Zhang
authors.forename: Yuliang
authors.initials: Y
authors.lastname: Wang
authors.forename: Lijie
authors.initials: L
authors.lastname: Cui
authors.forename: Yueli
authors.initials: Y
authors.lastname: Tang
authors.forename: Xiaofen
authors.initials: X
authors.lastname: Sun
authors.forename: Kexuan
authors.initials: K
authors.lastname: Tang
mesh_terms: D015111:4-Hydroxyphenylpyruvate Dioxygenase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism* / Q000528:radiation effects; D000595:Amino Acid Sequence; D001483:Base Sequence; D002734:Chlorophyll / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D003681:Dehydration / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D018545:Lactuca / Q000201:enzymology* / Q000235:genetics* / Q000528:radiation effects; D008027:Light / Q000009:adverse effects; D008969:Molecular Sequence Data; D010802:Phylogeny; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D018517:Plant Roots / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D018547:Plant Stems / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D018749:RNA, Plant / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013312:Stress, Physiological; D014810:Vitamin E / Q000096:biosynthesis / Q000737:chemistry
chemical_list: D018076:DNA, Complementary; D018749:RNA, Plant; D014810:Vitamin E; D002734:Chlorophyll; D015111:4-Hydroxyphenylpyruvate Dioxygenase