Articles
ISOLATION AND CHARACTERIZATION OF FLAVONOID 3′ HYDROXYLASE (F3’H) GENE AND GENETIC TRANSFORMATION IN BUTTERFLY PEA (CLITORIA TERNATEA LINN.) VIA AGROBACTERIUM TUMEFACIENS
Article number
836_35
Pages
247 – 253
Language
English
Abstract
The flavonoid 3 hydroxylase (F3H) gene denoted as an AsF3H was isolated and characterized from petals of Ascocenda sp.
The partial cDNA encoded a 264 amino acid polypeptide which showed a high homology to known F3H genes, especially, F3H from Gerbera hybrida. F3H is cytochrome P450 and it is a key enzyme in the flavonoid pathway leading to the production of the red colored anthocyanins.
The AsF3H transcript was the most abundant in petals from flowers at an early stage of development and levels declined as the flower matures.
No transcript levels were detected in leaves and stem.
In butterfly pea (Clitoria ternatea Linn.) a genetic transformation system via Agrobacterium tumefaciens was developed, therefore the CuDFR gene was constructed in pBI121 and transformed into butterfly peas leaves, petunia (Petunia axillaris) and tobacco (N. tabacum) by A. tumefaciens strain AGLO. After 4 months in tissue culture, the transgenic shoots were detected on selective media and the efficiency of transformation was 20, 25 and 23% of GUS assay respectively.
Moreover PCR technique reveals the positive bands of GUS gene and 35S promoter.
The partial cDNA encoded a 264 amino acid polypeptide which showed a high homology to known F3H genes, especially, F3H from Gerbera hybrida. F3H is cytochrome P450 and it is a key enzyme in the flavonoid pathway leading to the production of the red colored anthocyanins.
The AsF3H transcript was the most abundant in petals from flowers at an early stage of development and levels declined as the flower matures.
No transcript levels were detected in leaves and stem.
In butterfly pea (Clitoria ternatea Linn.) a genetic transformation system via Agrobacterium tumefaciens was developed, therefore the CuDFR gene was constructed in pBI121 and transformed into butterfly peas leaves, petunia (Petunia axillaris) and tobacco (N. tabacum) by A. tumefaciens strain AGLO. After 4 months in tissue culture, the transgenic shoots were detected on selective media and the efficiency of transformation was 20, 25 and 23% of GUS assay respectively.
Moreover PCR technique reveals the positive bands of GUS gene and 35S promoter.
Publication
Authors
P. Buddharak, R. Chundet
Keywords
flavonoid, anthocyanins, Ascosenda sp., Clitoria ternatea Linn., DFR, F3¿H
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