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STUDIES ON THE DEVELOPMENT OF TRANSGENIC TOMATO (Lycopersicon esculentum Mill.) via AGROBACTERIUM AND BIOLISTIC METHODS

KrishiKosh

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Title STUDIES ON THE DEVELOPMENT OF TRANSGENIC TOMATO (Lycopersicon esculentum Mill.) via AGROBACTERIUM AND BIOLISTIC METHODS
 
Creator USHA KIRAN, B
 
Contributor Sokka Reddy, S
 
Subject STUDIES, DEVELOPMENT, TRANSGENIC, TOMATO, AGROBACTERIUM, BIOLISTIC, METHODS
 
Description Tomato is the world’s largest vegetable crop after potato and sweet potato and
sweet potato. It is one of the most important protective foods. It is cultivated in 4.5 m ha
in the world with a production of 125 million metric tones where as in India it is
cultivated in 0.521 m ha with a production of 9.06 million metric tones. In Andhra
Pradesh it is grow in 0.084 m ha with a production of 1.6 million metric tones. There is
an ample scope for increasing the area and production and productivity as the demand for
the tomato is increasing day by day. Although the production and protection technologies
are well developed there are several constraints as crop suffers from biotic and abiotic
stresses. In the past few decades efforts are directed to impart resistance to these stresses,
shelf life improvement, quality and yield improvement through recombinant DNA
technology.
The present investigation is a small piece of work representing standardization of
reproducible highly efficient protocol for regeneration and transformations. Several
hormonal combinations have been tried on MS medium which revealed that Zeatin (0.5
mg/l) and IAA (0.1 mg/l) can produce the best shooting efficiency. MS medium
supplemented with IAA could give profuse rooting in all the shoots. Regenerated plants
have been successfully acclimatized in green house conditions and subsequently
transferred to field. The clonal fidelity of the regenerated plants was studied with RAPD
markers, which indicated their genetic purity without any observable polymorphism.
Genetic transformation through Agrobacterium and Biolistic methods were
initially adopted using GUS reporter gene for standardizing the conditions required for
transformation with Cry 1Ac gene which confers resistance to tomato shoot and fruit
borer. The GUS expression was better in Agrobacterium method as compared to that of
biolistic method. The Cry 1 Ac could be successfully transferred to tomato as evidenced
by PCR studies with Cry 1Ac specific primers. A total of 330 explants were transformed
through Agrobacterium and 156 explants were transformed through biolistic method. 35
transformants were obtained from Agrobacterium mediated transformation with
transformation efficiency of 11% and 13 transformants were obtained from biolistic
method with transformation efficiency of 8%.
Our studies are quite encouraging to take up the following line of research work in
tomato:
1. Growing of transformants to maturity stage and harvesting the transformed seeds.
2. Confirmation for Cry 1Ac by southern blot analysis.
3. Estimating the levels of cry protein in different tissues at different stages of plant
growth.
4. Conducting bioassay studies for resistance confirmation.
5. Conducting inheritance studies.
6. Conducting toxicological and other tests prescribed by biosafety committees.
7. Submitting the proposals to regulatory bodies for commercial release if any plant
is found superior.
8. Gene pyramiding to make the resistance process more effective.
9. Studying the suitability of several cry clones available with us.
10. Conducting suitability of different promoters for over expression of cry protein
and use of fruit specific promoters.
 
Date 2016-08-08T11:00:33Z
2016-08-08T11:00:33Z
2007
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/71433
 
Language en
 
Relation D8149;
 
Format application/pdf
 
Publisher ACHARYA N.G. RANGA AGRICULTURAL UNIVERSITY, RAJENDRANAGAR, HYDERABAD