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Studies on Markers-Free Transformation Tomato with Isoflavone Synthase Gene

Author: ZhangLuXia
Tutor: NiuJianXin；MaBingGang
School: Shihezi University
Course: Pomology
Keywords: Processing Tomato Unmarked Isoflavone synthase In vitro regeneration Genetic transformation Cre / lox gene deletion system
CLC: S641.2
Type: Master's thesis
Year: 2008
Downloads: 120
Quote: 1
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Abstract


Need to use a selectable marker gene, because the conversion efficiency, the majority of plant genetic transformation operation, such as antibiotic or herbicide resistance genes to selection of transformants, although still no direct research results show that the selectable marker gene affect human health or environmental safety However, in recent years, people are growing concerns about genetically modified food security, marker-free transgenic plants came into being in order to eliminate the concerns of the safety of genetically modified foods. Initially Cre / lox recombination system can effectively remove the marker gene with building good the Cre / loxp composition type carrier by Agrobacterium-mediated transformation of tobacco, to obtain a total transformation of tobacco plants; plant isoflavones are a class of enhanced plant disease resistance and induce nodule formation and prevention of hormone-related cancer, relieve menopausal syndrome bioactive secondary metabolites, the synthesis of a key enzyme isoflavone synthase (IFS); characteristics of this study in order to improved the quality of processing tomato enable people to better health while eating processed tomato products, optimized processing tomato genetic transformation regeneration system, to build a plant expression vector containing the IFS gene pBI121-ifs, and Agrobacterium tumefaciens-mediated transformation The processing tomato cotyledons and transgenic plants; fruit trees because of their own characteristics, genetic transformation with great difficulty and particularity. So this article to strawberries and apples as test materials, the establishment of efficient genetic transformation system for transgenic strawberries and apples. The main conclusions of the study are as follows: the test the 22 co-transformed tobacco leaves to extract DNA for PCR analysis, the results show that there are 10 deleted successfully gus gene to delete rate was 45.5%; 2. Agrobacterium-mediated transformation processing tomato filter out the most appropriate medium: pre-culture medium MS B5 IAA1.0mg / L 6-BA2.0mg / L, the screening transformation Miao medium for MS B5 IAA1.0mg / L 6-BA2.0 mg / L Cef500mg / L Kan50mg / L, transformed seedlings rooting medium of l/2MS B5 IBA2.0mg / L Cef300mg / L Kan50mg / L 11 conversion processing tomato extract DNA and RNA PCR and RT-PCR detection The results prove that 4 ifs gene has been successfully transferred to a conversion efficiency of 36.4%; 3. strawberry vitro regeneration system for establishing best medium screened: the shoot tip micropropagation medium 1/2MS B5 NAA 0.5 mg / L TDZ0.5mg / L, adventitious buds induced the medium MS B5 NAA0.3mg / L TDZ3.0 mg / L; Apple vitro regeneration system to establish the best medium screened: shoot tips micropropagation medium l/2MS B5 NAA0.2mg / L 6-BA 1.0 mg / L, adventitious bud induction medium for MS B5 NAA0.1mg / L ZT1.0 mg / L. The above study may lay a technical foundation for further security transgenic fruit.

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CLC: > Agricultural Sciences > Gardening > Vegetable gardening > Solanaceous > Tomatoes ( tomatoes )
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