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Biologia Plantarum

, Volume 57, Issue 2, pp 395–400 | Cite as

Overexpression of homogentisate phytyltransferase (HPT) and tocopherol cyclase (TC) enhances α-tocopherol content in transgenic tobacco

  • M. C. Harish
  • P. Dachinamoorthy
  • S. Balamurugan
  • S. Bala Murugan
  • R. SathishkumarEmail author
Brief Communication

Abstract

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.

Additional key words

Agrobacterium mediated transformation RT-PCR Southern blot 

Abbreviations

BAP

benzylaminopurine

CTAB

cetyltrimethylammonium bromide

DPPH

1,1-diphenyl-2-picrylhydrazyl

FRAP

ferric reducing antioxidant power

HPLC

high performance liquid chromatography

HPT

homogentisate phytyltransferase

IAA

indoleacetic acid

MS

Murashige and Skoog

RT-PCR

reverse transcriptase — polymerase chain reaction

TC

tocopherol cyclase

WT

wild type

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Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • M. C. Harish
    • 1
    • 2
  • P. Dachinamoorthy
    • 1
  • S. Balamurugan
    • 1
  • S. Bala Murugan
    • 1
  • R. Sathishkumar
    • 1
    • 2
    Email author
  1. 1.Plant Genetic Engineering Laboratory, Department of BiotechnologyBharathiar UniversityCoimbatoreIndia
  2. 2.DRDO-BU Centre for Life SciencesBharathiar UniversityCoimbatoreIndia

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