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in vitro Agrobacterium-mediated Transformation and Regeneration of Transgenic Plants with LMV-CP gene in Chinese Cabbage(Brassica campestris ssp.pekinensis)

Author: ChengXiHua
Tutor: CaoMingQing
School: Capital Normal University
Course: Genetics
Keywords: Target sprouts are Cabbage Explants Tissue culture Regeneration , gNO3 Genotype Transgenic Agrobacterium tumefaciens bar LMV-CP
CLC: S634.903.5
Type: Master's thesis
Year: 2000
Downloads: 112
Quote: 3
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Abstract


In Brassica cultivars , cabbage belongs vitro regeneration ability of poor category. By differentiation medium supplemented with 3mg / L of AgNO 3 , taking the ball significantly increased differentiation rate of adventitious buds cabbage . Study found that cabbage vitro regeneration capacity affected by genotype ; 20 parts in the tested material in cabbage to cotyledon - sub- petiole explants, 17 genotypes have the ability adventitious buds , of which 10 genes type bud differentiation rate of 80 % or more. Different explants ( cotyledon - petiole , cotyledon slices , hypocotyls , leaf slices ) regenerative capacity compared and found cotyledon - petiole and leaf slices with a strong ability to regenerate . Leaf slices hope to become the ideal conversion of certain genotypes receptors. Media and culture conditions from the two aspects of Chinese cabbage plantlets vitrification causes were studied. Within a certain range , lower hormone levels (BA from 2-1mgL -1 , NAA from 1-0.5mgL -1 ), increase the concentration of agar ( 0.8% -1.4 % ) , does not affect the rate of bud differentiation , there is conducive to normal buds form ; while reducing AgNO 3 concentration , sucrose concentration increased , adding activated charcoal or use good ventilation flasks , glass for suppression buds occurred without significant effect. Preliminary view that the hormone concentrations and agar concentration is affected cabbage vitrified main reason. To reproduce high frequency No.80 ,92-2, No.3, No.10 of conversion material . In cotyledon - petiole or leaf sections as explants for Agrobacterium transformation experiments . Agrobacterium strain C 58 , a plasmid vector is pV11, carrying LMV-CP gene and the selectable marker gene bar. To glufosinate (Basta, the active ingredient Phosphinothricin) for the selection of agents to cotyledon - petiole is transformed receptors from No.80 ,92-2 two genotypes resistant plants were screened , frequencies were 2-6 % and 3.7% respectively. Leaf slices to be transformed receptors No.80 obtained from transgenic plants , the frequency of 2.0%. Randomly selected six resistant strains by PCR and dot blot hybridization PCRSouthern prove LMV-CP gene was integrated into the genome of which 3 . Anti- virus detection in progress.

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CLC: > Agricultural Sciences > Gardening > Vegetable gardening > Chinese cabbage > Other
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