Study On Regeneration In Vitro And HAL1 Gene Transformation Of Strawberry

Shoot in vitro regeneration of strawberry cultivars."Selva", "Pink Panda", "Gongsimei yihao" and "Adi" were investigated. The results indicated that several factors affected regeneration rate of strawberry including cultivars, hormone concentrations and combinations, the types of the culture medium, explants types, leaf age and dark treatment. The regeneration system of strawberry was optimized. A rapid and efficient regeneration system of strawberry cultivar "Selva" was established. By using leaf discs co-cultivation with Agrobacterium tumefaciens, glyphosate-resistance plants were obtained, the genetic transformation system of strawberry cultivar "Selva" was established. The main results are as shown:1 Preliminary selection of gene types and regeneration culture mediumStrawberry leaves from field were used as experiment material, gene types and regeneration culture medium were preliminarily selected. At the same time, 4 cultivar sterile test-tube plantlets were acquired. The results showed that leaf regeneration efficiency of strawberry cultivar "Selva" was apparently higher then the other three cultivar. Accordingly, "Selva" was selected as experiment material to do the next regeneration and transgene transformation experiment.2 Effect of different factors on leaf regeneration of strawberry test tube plant(l)The efficiency of regeneration depended on the combination of hormones and the type of the nutrient medium.The combination of hormones and the type of the nutrient medium affected the regeneration directly. MS produced the greatest shoot regeneration rate compared with MS minerals plus B₅ vitamins and 1/2MS medium. The effect of cytokinin KT and 6-BA separately are not apparent, but by using both cytokinins leaf regeneration rate and regeneration bud number per leaf increased obviously, and regeneration adventitious buds grew very well. Explants regenerated on MS medium supplemented with KT1.0mg/L, BA 1.5mg/L and NAA0.1mg /L . Regenerated shoots propagation medium was MS+KT1.0mg/L+BA0.5mg/L+NAA0.02mg/L, and the suitable for root-promoting medium was 1/2 MS.