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Indian Journal of Biotechnology (IJBT) >
IJBT Vol.08 [2009] >
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.
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| Page(s): | 363-369 |
| ISSN: | 0975-0967 (Online); 0972-5849 (Print) |
| Source: | IJBT Vol.08(4) [October 2009]
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