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Molecular analysis of a homogentisate phytyltransferase gene from Lactuca sativa L.

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Abstract

Tocochromanols, usually known as vitamin E, play a crucial role in human and animal nutrition. The enzyme homogentisate phytyltransferase (HPT) performs the first committed step of the vitamin E biosynthetic pathway. The full-length cDNA encoding HPT was isolated from Lactuca sativa L. by rapid amplification of cDNA ends (RACE). The cDNA, designated as LsHPT, was 1,670 bp long containing an open reading frame (ORF) of 1,185 bp which encoded a protein of 395 amino acids. Sequence analysis indicated that the deduced protein, named as LsHPT, shared high identity with other dicotyledonous HPTs. Real-time fluorescent quantitative PCR (qPCR) analysis revealed that LsHPT was preferentially expressed in mature leaves compared with other tissues. When lettuce plants were subjected to drought and high-light stress treatments, LsHPT expression was markedly increased. Expression of LsHPT in Arabidopsis showed that LsHPT could enhance the α-tocopherol biosynthesis in Arabidopsis. Transient expression of LsHPT via agroinfiltration resulted in 9-fold increase in LsHPT mRNA level and nearly 18-fold enhancement in α-tocopherol content compared with the negative controls.

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Abbreviations

HPT:

Homogentisate phytyltransferase

RACE:

Rapid amplification of cDNA ends

ORF:

Open reading frame

FW:

Fresh weight

PFD:

Photon flux density

HPLC:

High performance liquid chromatography

X-gluc:

5-Bromo-4-chloro-3-indolyl-β-d-glucuronic acid

GUS:

β-Glucuronidase

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Acknowledgments

This work was funded by China “973” Program (grant number 2007CB108805) and Shanghai Leading Academic Discipline Project (project number B209).

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Authors and Affiliations

  1. Plant Biotechnology Research Center, School of Agriculture and Biology, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, Shanghai Jiao Tong University, Shanghai, 200240, People’s Republic of China

    Weiwei Ren, Lingxia Zhao, Lida Zhang, Yuliang Wang, Lijie Cui, Yueli Tang & Kexuan Tang

  2. State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, Morgan-Tan International Center for Life Sciences, Fudan University, Handan Road 220, Shanghai, 200433, People’s Republic of China

    Xiaofen Sun

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  1. Weiwei Ren
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  2. Lingxia Zhao
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  3. Lida Zhang
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  4. Yuliang Wang
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  5. Lijie Cui
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  6. Yueli Tang
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  8. Kexuan Tang
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Correspondence to Kexuan Tang.

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Ren, W., Zhao, L., Zhang, L. et al. Molecular analysis of a homogentisate phytyltransferase gene from Lactuca sativa L.. Mol Biol Rep 38, 1813–1819 (2011). https://doi.org/10.1007/s11033-010-0297-6

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  • DOI: https://doi.org/10.1007/s11033-010-0297-6

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