PARTICLE GUN MEDIATED DNA DELIVERY AND TRANSIENT EXPRESSION OF -GLUCURONIDASE (GUS) GENE IN NIGER (Guizotia Abysinica Cass.)
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Date
2006
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ACHARYA N.G. RANGA AGRICULTURAL UNIVERSITY, RAJENDRANAGAR, HYDERABAD
Abstract
In the present investigation, in vitro studies were carried out to determine the
optimum hormonal combination, to identify the best explant and best genotype for
callus induction and shoot regeneration. Transformation studies were carried out to
determine the frequency of transient GUS expression.
Callus was induced from all the explants and on all the media tested. Roots
recorded highest frequency of callus induction but, failed to regenerate shoots.
Hypocotyls, cotyledons and primary leaves were not significantly different for callus
induction. The medium with 2.0 mg/l TDZ and 0.2 mg/l NAA was found to be
superior over all other combinations tested.
Shoots were induced from all the explants, but cotyledonary explants
recorded highest frequency of shoot induction followed by hypocotyl and primary
leaf. The medium with 0.5 mg/l BA was best for shoot induction followed by
medium with 0.5 mg/l TDZ.
Genotypic differences were observed for both callus induction and shoot
induction. CTP-local responded optimally for callus induction and JNC-6 for shoot
induction. The medium with 0.5 mg/l TDZ and 0.2 mg/l NAA was best for callus
induction while, the medium with 1.0 mg/l BA and 0.2 mg/l NAA was best for shoot
induction.
Based on the results of transformation experiments, hypocotyl were found to
respond well in terms of transient GUS expression followed by primary leaf and
cotyledons. Among the varieties tested, Deomali has shown highest transformability.
Thus, the present study suggested direct regeneration of transgenic plants
from hypocotyl, cotyledon and leaf explants and applicability of particle
bombardment as an effective method of gene transfer.
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Keywords
PARTICLE, GUN, MEDIATED, DNA, DELIVERY, TRANSIENT, EXPRESSION, GLUCURONIDASE, GENE, NIGER