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Study on Transformation of Malus Spectabilis with Insect-resistant Gene Cry1Ab

Author: TanLiXia
Tutor: WangDun
School: Northwest University of Science and Technology
Course: Agricultural Entomology and Pest Control
Keywords: Cry1Ab Tissue culture Begonia flowers Blade Shoot regeneration Agrobacterium
CLC: S682.19
Type: Master's thesis
Year: 2011
Downloads: 23
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Abstract


In this experiment, Begonia plantlets as material for different concentrations of hormones on the growth and proliferation of Begonia flower bud leaf disc regeneration were studied, while recombinant plant expression vector pBI121-cry1Ab, and Agrobacterium Agrobacterium-mediated transformation of Begonia flowers Cry1Ab gene were studied to establish Begonia flower efficient regeneration system in vitro and transgenic system for subsequent in-depth study of transgenic Begonia flowers foundation. The following is the main results obtained: (1) to Begonia flowers and leaves of tissue culture, the application of orthogonal experiment method, based on MS medium were added to different concentrations of TDZ combinations and proportions of 6-BA/IBA / IBA combinations as proliferation medium and leaf disc adventitious shoot regeneration medium, the effects of different plant hormones and their value ratio on tissue culture, leaf shoot regeneration and rooting of plants and explores the different placement of leaf discs its regeneration. The results showed that: Add 6-BA 1.0 mg. L -1 IBA 0.1 mg. L -1 MS medium most appropriate Begonia plantlets proliferation and growth, multiplication up to 8.11, the average height of Begonia flowers plantlets 2.14 cm; adding TDZ 1.0 mg. L -1 IBA 0.5 mg. L -1 on MS medium leaf regeneration efficiency highest regeneration efficiency was 100%, the average number of regenerated buds per leaf was 6.73; adding 0.3 mg. L -1 IBA's 1/2 MS rooting efficiency of 65% or more; polarity test results show that Begonia flower to leaf disc regeneration medium is far better contact with the shaft end. (2) was cloned by PCR from the strain of Bt C3 has been Cry1Ab gene coding sequences in prokaryotic Bioassay Cry1Ab insecticidal activity to determine their expression products, the constructed plant expression vector pBI121-cry1Ab, for transgenic research. (3) By kanamycin (kanamycin, Kan) resistance testing and cefotaxime (Cefotaxime Na, Cef) resistance test study to determine the optimal induction of adventitious buds Begonia mosaic set of critical screening concentration Kan and Cef , respectively, 25 mg. L -1 and 200 mg. L -1 . (4) the establishment of regeneration system and build Begonia mosaic expression vectors based on Agrobacterium-mediated method for the genetic transformation of Begonia flowers. GUS gene containing the empty vector infection after flowering crabapple, right out of the callus induced GUS histochemical staining results showed: GUS positive rate was 17.58%; the Cry1Ab gene transformation Begonia flowers, after initially identified by PCR testing two were positive transgenic plants resistant plants.

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CLC: > Agricultural Sciences > Gardening > Ornamental Horticulture ( flowers and ornamental trees) > Perennial Flowers > Perennial Flowers Class > Other
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