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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Main Author:	A. Rahman, Siti Suhailah
Format:	Thesis
Language:	English
Published:	2005
Subjects:	Agrobacterium Genetic transformation. Transgenic plants.
Online Access:	http://psasir.upm.edu.my/id/eprint/5951/1/FBSB_2005_27%20IR.pdf
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spelling	my-upm-ir.59512022-02-10T03:33:42Z Development of Agrobacterium-Mediated for Chi2;1 Promoter Analysis of llycopersicon Esculentum on Mt11 2005-05 A. Rahman, Siti Suhailah 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 Agrobacterium Genetic transformation. Transgenic plants. 2005-05 Thesis http://psasir.upm.edu.my/id/eprint/5951/ http://psasir.upm.edu.my/id/eprint/5951/1/FBSB_2005_27%20IR.pdf text en public masters Universiti Putra Malaysia Agrobacterium Genetic transformation. Transgenic plants. Faculty of Biotechnology and Biomokular Sciences Mohd Saleh, Norihan
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English
advisor	Mohd Saleh, Norihan
topic	Agrobacterium Genetic transformation. Transgenic plants.
spellingShingle	Agrobacterium Genetic transformation. Transgenic plants. A. Rahman, Siti Suhailah Development of Agrobacterium-Mediated for Chi2;1 Promoter Analysis of llycopersicon Esculentum on Mt11
description	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
format	Thesis
qualification_level	Master's degree
author	A. Rahman, Siti Suhailah
author_facet	A. Rahman, Siti Suhailah
author_sort	A. Rahman, Siti Suhailah
title	Development of Agrobacterium-Mediated for Chi2;1 Promoter Analysis of llycopersicon Esculentum on Mt11
title_short	Development of Agrobacterium-Mediated for Chi2;1 Promoter Analysis of llycopersicon Esculentum on Mt11
title_full	Development of Agrobacterium-Mediated for Chi2;1 Promoter Analysis of llycopersicon Esculentum on Mt11
title_fullStr	Development of Agrobacterium-Mediated for Chi2;1 Promoter Analysis of llycopersicon Esculentum on Mt11
title_full_unstemmed	Development of Agrobacterium-Mediated for Chi2;1 Promoter Analysis of llycopersicon Esculentum on Mt11
title_sort	development of agrobacterium-mediated for chi2;1 promoter analysis of llycopersicon esculentum on mt11
granting_institution	Universiti Putra Malaysia
granting_department	Faculty of Biotechnology and Biomokular Sciences
publishDate	2005
url	http://psasir.upm.edu.my/id/eprint/5951/1/FBSB_2005_27%20IR.pdf
_version_	1747810515079397376

Development of Agrobacterium-Mediated for Chi2;1 Promoter Analysis of llycopersicon Esculentum on Mt11

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