Production of protocormlike bodies and Agrobacteriummediated Genetic transformation of the orchid spice Vanilla planifolia (Andrews)
DSpice at Indian Institute of Spices Research
View Archive InfoField | Value | |
Creator |
RETHEESH, S T
BHAT, A I |
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Date |
2010-07-29T08:22:18Z
2010-07-29T08:22:18Z 2010-07-29T08:22:18Z |
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Identifier |
http://hdl.handle.net/123456789/528
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Description |
Induction of Protocorm-like bodies (PLB’s) from shoot tips of Vanilla planifolia is found efficient in MS medium containing 0.1 mgl-1 thidiazuron compared to the MS medium containing BAP and NAA, either singly or in combination. Genetic fidelity test of the plantlets regenerated from PLB’s by RAPD using 23 random primers revealed no genetic variation within the progeny and to the parent plant from which they were originated. The PLB’s thus produced were genetically transformed by Agrobacteriummediated genetic transformation with glucoronidase β reporter gene. Agrobacterium strain EHA 105 containing the plant transformation vector pBI121 was used in the study. The use of acetosyringone combined with the three step selection of transgenic plantlets with increasing concentration of antibiotic yielded high frequency (47.3%) of antibiotic resistant plantlets. A total of 134 transgenic lines were obtained and all were positive for the transgene when tested by PCR. GUS histochemical assay also revealed cent per cent of the 15 putative plants tested were positive for the βglucoronidase enzyme activity. Southern hybridization of randomly selected 20 plants revealed that 70% were positive with one to two copies of the transgene. This is a simple and highly efficient protocol for the routine transformation of vanilla with other useful genes. |
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Format |
698481 bytes
application/pdf |
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Language |
en
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Subject |
Vanilla
transgenic plants GUS assay genetic engineering |
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Title |
Production of protocormlike bodies and Agrobacteriummediated Genetic transformation of the orchid spice Vanilla planifolia (Andrews)
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Type |
Preprint
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