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Transient expression of the homogentisate phytyltransferase gene from Clitoria ternatea causes metabolic enhancement of α-tocopherol biosynthesis and chlorophyll degradation in tomato leaves

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Abstract

Homogentisate prenyltransferase (HPT) is an important enzyme involved in the α-tocopherol (vitamin E) biosynthetic pathway of all plant taxa. Tocopherol biosynthesis and chlorophyll degradation are related, but more information is needed to explain their relationship. In this study, a candidate gene for HPT from Clitoria ternatea (CtHPT) was isolated and identified via a phylogeny-based approach, and its hypothetical protein sequence was analyzed. Transient expression of CtHPT with Agrobacterium-mediated infiltration into tomato leaves was then performed and observed for the metabolic relationship between the α-tocopherol biosynthesis and chlorophyll degradation by gas chromatography–mass spectrometry. In silico analysis showed that CtHPT contained a chloroplast signal peptide and nine-transmembrane α-helixes. The results showed that, the content of α-tocopherol increased in transient expression of CtHPT, with the increased pool sizes of its biosynthetic intermediates: 2-methyl-6-phythylbenzoquinol and 2,3-dimethyl-5-phythylbenzoquinol, and the increased levels of phytol and various fatty acids. Moreover, the CtHPT transient expression was observed to cause chlorophyll deficiency in the tomato leaves with simultaneous increase of phytol and fatty acids, presumably the degradative products of chlorophyll and chloroplast membranes, respectively. It was concluded that the overexpression of CtHPT may enhance the metabolic flow of the α-tocopherol biosynthetic pathway, causing the degradation of chlorophylls, thereby increasing the supply of the precursor phytol for the α-tocopherol biosynthetic pathway.

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Abbreviations

HGA:

Homogentisic acid

PDP:

Phytyl diphosphate

DMPBQ:

2,3-dimethyl-5-phythylbenzoquinol

MPBQ:

2-methyl-6-phytyl-1,4-benzoquinol

HPT:

Homogentisate phytyltransferase

GC–MS:

Gas chromatography–mass spectrometry

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Acknowledgements

We would also like to thanks Professor Tsuyoshi Nakagawa for supporting pGWB6 vector in transient expression experiment. We would also like to thank the 90th Anniversary of Chulalongkorn University Fund for providing a scholarship. This work was supported financially by Chulalongkorn University and the Office of Higher Education Commission.

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Authors and Affiliations

  1. Department of Biochemistry and Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, 10330, Thailand

    Thaniya Wunnakup

  2. Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, 10330, Thailand

    Sornkanok Vimolmangkang & Wanchai De-Eknamkul

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  1. Thaniya Wunnakup
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  2. Sornkanok Vimolmangkang
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  3. Wanchai De-Eknamkul
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Contributions

TW, SV and WD (as PI) designed experiments; TW performed most experiments and collected data; TW, SV and WD analyzed data; SV introduced the method of plant transient expression experiments; TW and WD wrote the manuscript.

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Correspondence to Wanchai De-Eknamkul.

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The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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Wunnakup, T., Vimolmangkang, S. & De-Eknamkul, W. Transient expression of the homogentisate phytyltransferase gene from Clitoria ternatea causes metabolic enhancement of α-tocopherol biosynthesis and chlorophyll degradation in tomato leaves. J. Plant Biochem. Biotechnol. 27, 55–67 (2018). https://doi.org/10.1007/s13562-017-0415-9

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  • DOI: https://doi.org/10.1007/s13562-017-0415-9

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