Research On Transformation Of Dendrathema Grandiflorum 'Xiao Huang' Ju Resistant To Beetles

Adventitious shoots are induced from both stem and leaf explants of Dendrathema grandiflorum 'Xiao Huang' Ju in this research. The optimized medium for stem and leaf explants contains Murashige & Skoog basic medium supplemented with 2 mg/L of 6-BA and 0.1 mg/L of NAA , and 1 mg/L of 6-BA and 0.1 mg/L of NAA, respectively. More shoots are generated from leaf than from stem explants. The medium for rooting of shoots is optimized as 1/2 Murashige & Skoog basic medium supplemented with 0.1 mg/L of NAA, with a rooting efficiency of 100%. Kanamycin at 40 mg. L-1 and 10 mg/L in the medium gives effective inhibition to the regeneration of shoots from the stem and leaf explants, respectively. In addition, Kanamycin at 20 mg/L in the medium shows obvious inhibition to the regeneration of roots from the shoots. This dissertation provides the basis for the transformation research on D. grandiflorum.Cry3Aa was cloned from Bacillus Thuringiensis 886 strain, which was toxic to the worm of long horn beetle. Cry3Aa gene was modified according to the charactors of poplus genes and the other features of eukaryotic gene, and was artificially synthesized and named modified cry3. Cry3Aa and modified cry3 were cloned into binary vector pBI121 and pBin438, respectively named pBI121-cry3Aa and pBin438-modified cry3, cry3Aa and modified cry3 under the control of the cauliflower mosaic virus 35s promoter. Dendrathema grandiflorum 'Xiao Huang' Ju was transformed using Agrobacterium tumefaciens-mediated transformation and gene gun shot transformation. Four cry3Aa and Five modified cry3 independently transformed lines were produced, these lines detected by PCR and DNA blotting.