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Studies on Introducing Bt-toxin Gene into Rice Via Agrobacterium Tumefaciens

Author: PiCanHui
Tutor: YiZiLi
School: Hunan Agricultural University
Course: Botany
Keywords: Indica Callus Agrobacterium Transformation Insect-resistant
CLC: Q943
Type: Master's thesis
Year: 2003
Downloads: 120
Quote: 2
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Abstract

Two advantages of to cultivate super hybrid rice is widely used. The pro this Peiai 64S and 9311 as the main object of study, to affect its mature embryo-derived callus induction, differentiation and plant regeneration conditions were optimized; affecting Agrobacterium transforming the conditions of the above indica exploration and optimization; establish a high-frequency Regeneration System for efficient genetic transformation method. Molecular detection and analysis of insect resistance to transgenic plants. 1. Pei'ai 64S and 9311 from the regeneration system optimization study impact of two indica rice callus induction, the second-generation and differentiation into seedlings factors, results show that: the physiological state of the seed itself is the main factors to affect the frequency of callus induction one; N6 medium with a large number of elements with MS trace element combination, while improving VB1 and inositol content can significantly improve callus induction and training effect; appropriate concentrations of 2,4-D, add other things source hormone can not significantly improve the rate of callus induction embryogenic callus growth state, but can be improved; alternate use of different medium composition and the hormone concentration ratio allows long-term subculture embryogenic state to extend Following cultured time; predifferentiated and drying treatment of the plant regeneration frequency and quality greatly improved. We optimized culture conditions and treatment, Peiai 64S and 9311 callus induction frequency, respectively, 62.45% and 85.30%; Following the culture time up to three months or so to maintain a better state of embryonic growth ; Following on behalf of two of embryonic growth state callus frequency can reach 85.5% and 87.7%, respectively. 2. Agrobacterium-mediated transformation of Bt toxic protein gene into rice same as the main material of the two indica compare the pre-cultured immature embryos and young panicles, immature embryo, mature embryo-derived callus as transformed by body of callus transformation frequency, the results show that the pre-culture the most appropriate receptors as Agrobacterium-mediated transformation of immature embryos of 4 to 6 days; followed embryogenic callus derived from immature embryo growth and mature embryos in good condition . YEB medium as a co-culture medium for Agrobacterium EHA 05 ... Ub. m), the bacterial concentration for the OD value of 0.8, the highest conversion rate of callus. Agrobacterium infection process of callus, negative pressure processing can significantly improve the the rice callus conversion frequency. 60ml syringe, pull the distance to determine the most effective 6cm around, pull 40 times the number of rice callus transformation frequency. CaZ + and surfactant treatment to increase the frequency of transformation is the major innovation of this experiment. In After callus disseminated, respectively, to a concentration of 20mmol / L, and 30mmol / L of the solution treated CaC12 Pui dwarf 645 and 93if embryogenic callus in the treatment liquid at the same time to add 0. l% Tween 20 in order to reduce the surface tension of the cells, the transformation and regeneration of the transformed plants to promote callus best. Treated as described above allows the conversion rate of 93 callus and resistant plant regeneration rate reached 58.33% and 33.33% respectively. Select the part of the plants were analyzed by PCR analysis, DNA dot blot, PCR-Southern hybridization. More molecules test results showed that most of the genome of the transformed plants obtained by the Resistance Screening exogenous gene has been integrated.

Studies on Introducing Bt-toxin Gene into Rice Via Agrobacterium Tumefaciens

Abstract

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