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STANDARDIZATION OF PROTOCOL FOR REGENERATION AND GENETIC TRANSFORMATION OF OKRA (Abelmoschus esculentus [L.] Moench)

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Title STANDARDIZATION OF PROTOCOL FOR REGENERATION AND GENETIC TRANSFORMATION OF OKRA (Abelmoschus esculentus [L.] Moench)
 
Creator Rekha Rani, M
 
Contributor MANORAMA, K
 
Subject STANDARDIZATION, PROTOCOL, REGENERATION, GENETIC, TRANSFORMATION, OKRA
 
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.
 
Date 2016-08-08T10:58:18Z
2016-08-08T10:58:18Z
2007
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/71432
 
Language en
 
Relation D8148;
 
Format application/pdf
 
Publisher ACHARYA N.G. RANGA AGRICULTURAL UNIVERSITY, RAJENDRANAGAR, HYDERABAD