| More researches on the regeneration system in vitro and the transformation systemof transforming the human hepatitis B virus Large surface antigen gene into apple andtomato had been carried out. We optimized the high efficient regeneration system in vitroand transformation system, obtained the transgenic plant with S1S2S gene identified byGUS staining and PCR assay. The main results were as follows:1 We optimized the high efficient regeneration system in vitro and transformation system;Three kinds of S1S2S gene regulated by different promoters were introduced into 'Ryokano Kisetsu' apple by Agrobacterium tumefaciens-mediated transformation. The transgenicplants were obtained through kanamycin selection and GUS assay. The results indicatedthat the leaf's base was optimal for transformation; the kanamycin selecting pressure ofleaf and young tissue culture plant were 15mg/L and 25mg/L respectively; Carbenicillinwas better than Cefotaxine on the effect of restraining bacteria; 7 positive plants wereObtained by selecting of GUS staining and PCR amplification, 2 plants with AG208, 3plants with AG206 and 2 plants with AT110 among them; these results showed that S1S2Sgene had been conformed into apple genome.2. We used Agrobacterium-mediated regeneration assistant by ultrasonic to carry out S1S2Sgene transformation on 'Beni Aika'. This thesis studied the effects of ultrasonic treat time,treat period, treat power, explants types and acetosyringone concentration in Agrobacterimsuspension solution on S1S2S gene transformation rate by the method of instantaneousexpression of GUS gene. The results showed that we could get the optimal GUS geneinstantaneous expression rate after Agrobacterium dipped 'Beni Aika' for 4 minutes treated30 seconds using ultrasonic with 90w power, then inoculated regeneration medium for 3days co-cultivation. Under the condition of optimal treatment we could get 138 calluses and11 young plants with fastness from 600 transformed 'Beni Aika' leaves, the transformationrate was 1.83ï¼….3.We used tomato cultivars 'Jiangshu No1' as experimental materials and MS as basicmedium, this thesis studied the effects of different time treatment combinations of 70ï¼…alcohol and 20ï¼…NaClO solutions on the sterilization effect of seed surface and bourgeon. In addition to that, this thesis also studied the effects of different gene types, the variety andconcentration combinations of plant growth regulated substance on the regeneration oftomato cotyledon. We introduced three vectors of S1S2S gene into the plant byAgrobacterium-mediated method and got corresponding kanamycin young plants. Thetransgenic plants were tested by GUS staining and PCR assay. These results showed thatusing 70ï¼…alcohol to treat 50 seconds, then used 20ï¼…NaClO solutions to dip 30 minutes,'Jiangsu No.1's germination rate was 98ï¼…and no contamination; Different tomato cultivarshad significant difference in callus inducement rate and infinitive bud differentiation rate atthe same medium. The suited medium for cotyledon regeneration was MS+ZT 0.5mg/L+IAA 0.3mg/L. The suited medium for root generation was MS+NAA 0.5mg/L. Thetransgenic plants of 'Jiangsu No.1' through Agrobacterium transformation (71 plants withAG208, 55 plants with AG206, 39 plants with AT110) contained only one positive plant ofAG208 and AG206 respectively by PCR assay.
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