| The objective of this study was to research in regeneration of'Ruby Seedless''Blush Seedless'and'Long Thompson Seedless'from shoot organogenesis and somatic embryogenesis, grape transformation by Agrobacterium infection haboring the novel gene related to ALDH and detection transformed grape. The results of the researches are as follow:1. The optimum media of leaves'bud regeneration of'Ruby Seedless'and'Long Thompson Seedless'are respectively MS+TDZ 2.0mg/L+NAA 0.1mg/L and MS+BAP 5.0mg/L+NAA 0.5mg/L. The frequency of adventitious bud incidence respectively was 33.3% and 25.0%. The rate of adventitious bud regeneration was 31.01%, which from petioles of'Long Thompson Seedless'in MS supplemented with BAP 5.0mg/L and NAA 0.5mg/L. Rate of bud regeneration of'Ruby Seedless'was greater from leaves than petioles. The rate of stems'bud regeneration is lowest. Culturing adventitious buds in 1/2MS medium with BAP 1.0mg/L and IBA 0.5mg/L enhanced shoots growth. Shoots converted to plantlets in 6 weeks. The plantlets were rooted in 1/2MS+IBA 0.2mg/L.2. Somatic embryogenesis from anthers and ovaries of'Blush seedless'(Vitis vinifera L.) were studied. The best combination of hormones for embryogenic callus inducement of'Blush seedless'from anthers and ovarys are MS+BAP 0.5mg/L+2, 4-D 0.5mg/L and MS+BAP 1.0mg/L+2, 4-D 1.0mg/L, with the inducement rate are 60% and 25%, respectively. Somatic embryo from anthers and ovarys are induced in MS supplemented with BAP 0.5 mg/L, with the rate are 84.67% and 41.67%, respectively. The embryos were germinated and proliferated in MS free-hormone medium. 64.0% embryos were converted to plantlets in WPM medium composed of 0.1mg/L IBA.3. The novel gene related to ALDH was introduced into European grapevine by Agrobacterium infection. Transgenic grape were obtained from shoot organogenesis and somatic embryogenesis. 17 lines of'Ruby Seedless'from infected leaves were grown to be normal plantlets. But only 1 in 17 lines (5.88%) of plants were detected to be positive by PCR analysis ALDH gene. 18 lines of'Long Thompson Seedless'from infected petioles were grown to be normal plants. 3 in 18 lines (16.67%) of plants were detected to be positive by PCR analysis. 66 lines from infected petioles'callus of'Long Thompson Seedless'were grown... |
