Development of Agrobacterium-Mediated for Chi2;1 Promoter Analysis of llycopersicon Esculentum on Mt11
Improvement of Agrobacterium-mediated transformation system (AMT) in MTll cultivar is required to achieve higher efficiency of gene transfer to produce transgenic plants of MT11. Various concentrations of kinetin and zeatin were tested for cultivar MTll and VF36 explants, up to 100% of the expla...
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| Format: | Thesis |
| Language: | English |
| Published: |
2005
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| Subjects: | |
| Online Access: | http://psasir.upm.edu.my/id/eprint/5951/1/FBSB_2005_27%20IR.pdf http://psasir.upm.edu.my/id/eprint/5951/ |
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| Summary: | Improvement of Agrobacterium-mediated transformation system
(AMT) in MTll cultivar is required to achieve higher efficiency of
gene transfer to produce transgenic plants of MT11. Various
concentrations of kinetin and zeatin were tested for cultivar MTll
and VF36 explants, up to 100% of the explants are capable of
producing calli from cotyledon and hypocotyls. Kinetin at 5 mg/L
was highly significant (p0.01) in terms of the largest number of
shoot regeneration from cotyledons explants of MTll cultivar. The
use of 2 mg/L zeatin was highly significant (p0.01) for shoot
elongation of American variety, VF36.
The addition of ascorbic acid @re- and post- inoculation with
Agrobacterium) and acetosyringone were significant (p0.01) for cell
recovery and in enhancing the development of putatively transformed
plantlets of MTll and VF36 tomato cultivar. The combined
interactions between pBY4.1 and acetosyringone was found to be
highly significant (pc0.01) in terms of number of putatively
transformed plants obtained. Kanamycin at 100 mg/L is the most
suitable minimal inhibition concentration to be use in the
transformation. Agrobacterium tumefaciens strain LBA4404 is more
suitable to be use in the transformation system compared to GV2260
strain for MTl 1 cultivar.
The PCR amplification technique determined that the full length of
Chi2;l promoter (1336 bp) and the deletions: pBY2.3 (226 bp),
pBY4.1 (435 bp), pBY6.1 (616 bp) and pBY8.1 (865 bp) showed the
1.8 kb GUS genes after gel electrophoresis. The pistils from the
transgenic plants showed the expression of GUS genes blue stained
in the transmitting tract, except for pBYO.5 construct (58 bp) and the
control. Southern blots analysis showed pBY4.1 deletion had the
strongest signal. Seedlings from pBY4.1 flower were tested and
confirmed that the foreign integrated genes is inherited |
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