Molecular cloning and characterization of 4-hydroxyphenylpyruvate dioxygenase gene from Lactuca sativa

• Published in: Journal of plant physiology 2011-07, Vol.168 (10), p.1076-1083
• Main Authors: Ren, Weiwei, Zhao, Lingxia, Zhang, Lida, Wang, Yuliang, Cui, Lijie, Tang, Yueli, Sun, Xiaofen, Tang, Kexuan
• Format: Article
• Language: English
• Subjects: 4-Hydroxyphenylpyruvate Dioxygenase - genetics; 4-Hydroxyphenylpyruvate Dioxygenase - isolation & purification; 4-Hydroxyphenylpyruvate Dioxygenase - metabolism; 4-Hydroxyphenylpyruvate Dioxygenase - radiation effects; Agroinfiltration; Amino Acid Sequence; Base Sequence; Biological and medical sciences; Chlorophyll - metabolism; Cloning, Molecular; Dehydration - metabolism; DNA, Complementary - chemistry; DNA, Complementary - genetics; DNA, Complementary - isolation & purification; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Plant; HPPD; Lactuca - enzymology; Lactuca - genetics; Lactuca - radiation effects; Lactuca sativa; Light - adverse effects; Medicago truncatula; Molecular Sequence Data; Phylogeny; Plant Leaves - genetics; Plant Leaves - metabolism; Plant Leaves - radiation effects; Plant physiology and development; Plant Roots - genetics; Plant Roots - metabolism; Plant Roots - radiation effects; Plant Stems - genetics; Plant Stems - metabolism; Plant Stems - radiation effects; Real-time fluorescent quantitative PCR (qPCR); Real-Time Polymerase Chain Reaction; RNA, Plant - genetics; Sequence Alignment; Sequence Analysis, DNA; Stress, Physiological; Vitamin E - biosynthesis; Vitamin E - chemistry; Vitamin E biosynthetic pathway
• ISSN: 0176-1617; 1618-1328
• DOI: 10.1016/j.jplph.2010.12.017

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.