STANDARDIZATION OF PROTOCOL FOR REGENERATION AND GENETIC TRANSFORMATION OF OKRA (Abelmoschus esculentus [L.] Moench)
KrishiKosh
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Title |
STANDARDIZATION OF PROTOCOL FOR REGENERATION AND GENETIC TRANSFORMATION OF OKRA (Abelmoschus esculentus [L.] Moench)
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Creator |
Rekha Rani, M
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Contributor |
MANORAMA, K
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Subject |
STANDARDIZATION, PROTOCOL, REGENERATION, GENETIC, TRANSFORMATION, OKRA
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Description |
Okra (Abelmoschus esculentus [L.] Moench) is an important vegetable crop. India ranks first in okra production with 3.5 million metric tones from an area of 0.35 million hectares (FAO Statistical Database, 2005). Ability to regenerate okra plants in vitro would permit the rapid propagation of this important vegetable crop, and also aid in standardization of genetic transformation protocols. Genetic improvement of crop plants can be achieved by transferring new loci across diverse sources through transformation. For successful development of transgenic plants, identification of suitable target tissue and efficient gene transfer protocols are essential. Hence, it is essential to standardize the protocol for in vitro regeneration of okra. Explants (cotyledons, hypocotyls, cotyledonary nodal segments and shoot tips) were obtained from aseptically grown okra seedlings. Explants were cultured on MS media supplemented with different levels of auxins, cytokinins and auxin-cytokinin combinations. Callusing was observed from cotyledons, hypocotyls and cotyledonary nodal meristems. Greater proliferation of roots was observed with NAA. No shooting was observed at any of the concentrations of auxins. Both Kinetin and Zeatin were also proved ineffective in inducing shooting. Cotyledonary nodal meristems and shoot tips supplemented with BAP and NAA showed shooting and rooting. These shoots were sub-cultured on another medium with concentration of 0.5 mg/l BAP and 1.0 mg/l NAA, in which elongation in shoot and root length was observed. Plantlets transferred to soil grew normally. Genetic transformation with Agrobacterium carrying the plasmid pBI121 with a selectable marker gene npt-II that confers resistance against the antibiotic kanamycin was carried out. Transformed cells were cultured on kanamycin (50 mg/l), cekatoxime (300 mg/l). Proliferation of callus was achieved in this selection medium and Agrobacterium was suppressed completely. About 50-60% calli showed GUS expression, confirming transformation. The present study thus indicates that genetic transformation in okra can be successfully achieved by optimizing various parameters for regeneration and Agrobacterium infection. |
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Date |
2016-08-08T10:58:18Z
2016-08-08T10:58:18Z 2007 |
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Type |
Thesis
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Identifier |
http://krishikosh.egranth.ac.in/handle/1/71432
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Language |
en
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Relation |
D8148;
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Format |
application/pdf
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Publisher |
ACHARYA N.G. RANGA AGRICULTURAL UNIVERSITY, RAJENDRANAGAR, HYDERABAD
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