Studies On The Transformation Of Long Ya Lillium With Chitinase And β-1,3-glucanase Genes

In terms with the principle of Fusarium Oxysporiun caused plant disease:bundles were blocked and Fusarid acid killing cells was formed by hyphae so that caused water metabolism abnormal and plant wilting .In order to find out effective method of anti-Fiisarium Oxysporuin,Long Ya Lillium was taken as material with plant tissue culture and genetic transformation techniques in this paper.Two main contents were concluded in this paper :the plant regeneration and genetic transformation.Some effective factors were studied in tissue culture in order to establish a quick ,high frequency regeneration system of genetic transformation for Long Ya Lillium. The results indicated that the scale leaf was the best appropriate explant .The combination of MS +2.4-D 2mg/L+6-BA 0.2mg/Lwas the optimum medium for the induction of calli.The combination of MS +6-BA lmg/L+NAA0.05mg/L was the best medium for differentiation and elongation.The medium of MS +NAA2mg/L was the best for adventitious root.The engineering bacterium which carried bcih I-Chi and I-Glu cDNA was pCG-II.Two methods of Agrobacterium -mediated and gene gun were used to transformate Long Ya Lillium.The results of PCR analysis and southern dot blotting hybridization demonstrated that the Chi a nd GlucDNA have been intergrated into host genome.At the same time; compared Agrabactenum-mediated method with gene gun method ,the transformation frequency of the former was 16.7%,while the latter was 50%,so gene gun transformation method was suitable for Long Ya Liiliwn.