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IJBT Vol.08(4) [October 2009] >


Title: Transformation of tomato using biolistic gun for transient expression of the -glucuronidase gene
Authors: Ruma, D
Dhaliwal, M S
Kaur, Ajinder
Gosal, S S
Keywords: Lycopersicon esculentum
biolistic transformation
GUS assay
cotyledons
shoot tips
hypocotyls
-glucuronidase gene
Issue Date: Oct-2009
Publisher: CSIR
Abstract:  We report for the first time, conditions for the biolistic transformation of tomato for the introduction of -glucuronidase gene (gusA) into different explants viz., shoot tips, hypocotyls and cotyledons of genotype IPA-3. The respective per cent plant regeneration in different explants was 95.16, 79.30 and 90.14% on MS media supplemented with BAP (2.0 mg L-1) and Kn (1.0 mg L-1). The influence of physical parameters of particle gun on rates of transient GUS expression have been investigated viz., quantity of DNA, distance between the microcarrier launch assembly and target tissues; and the biological factors associated with the target tissues i.e., effect of osmoticum (mannitol), pre-bombardment culture period and post-bombardment culture period. Maximum GUS expression of 25.00, 33.00 and 22.20% was respectively recorded when 10-d-old shoot tips, hypocotyls and cotyledons were bombarded with 18 L of DNA suspension. A firing distance of 7.5 cm was found to be most suitable recording 34.12, 36.56 and 22.69% transient GUS expression in shoot tips, hypocotyls and cotyledons, respectively. Addition of osmoticum into the culture medium reduced per cent GUS expression significantly in all the explants even at 1 molar concentration. Pre-culture of explants prior to bombardment also deduced the transformation efficiency, however post bombardment culture period of one day resulted in maximum GUS expression in all the explants.
Page(s): 363-369
ISSN: 0975-0967 (Online); 0972-5849 (Print)
Source:IJBT Vol.08(4) [October 2009]

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