Photosynthetic organisms synthesize the amphipathic antioxidants called tocopherols which are essential components of the human diet. To increase the α-tocopherol (vitamin E) content, Arabidopsis genes encoding homogentisate phytyltransferase (HPT) and tocopherol cyclase (TC) were constitutively expressed individually and in combination (HPT:TC) in tobacco plant by Agrobacterium mediated transformation. The transgene was confirmed by polymerase chain reaction (PCR), transgene expression was studied by reverse transcriptase (RT)-PCR, integration of the transgene in the plant genome was confirmed by Southern blot, and α-tocopherol content was quantified using high performance liquid chromatography (HPLC). The α-tocopherol content in transgenic tobacco plants expressing HPT, TC, and HPT:TC was increased by 5.4-, 4.0-, and 7.1-fold, respectively, when compared to the wild type (WT). These results indicate that, the HPT and TC activities are critical for enhancing the vitamin E content in tobacco plants.
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ferric reducing antioxidant power
high performance liquid chromatography
Murashige and Skoog
reverse transcriptase — polymerase chain reaction
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Acknowledgements: The authors are very thankful to Defence Research and Developmental Organization — Centre for Life Sciences, Bharathiar University for financial assistance and support.
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Harish, M.C., Dachinamoorthy, P., Balamurugan, S. et al. Overexpression of homogentisate phytyltransferase (HPT) and tocopherol cyclase (TC) enhances α-tocopherol content in transgenic tobacco. Biol Plant 57, 395–400 (2013). https://doi.org/10.1007/s10535-012-0298-5
Additional key words
- Agrobacterium mediated transformation
- Southern blot