Preliminary Studies Of Genetic Transformation System Of Jatropha Curcas L.

Important presupposition for selecting proper medium and antibiotics concentration is the plant genetic engineering using Argobacterium tumefaciens.The cotyledon, leaf, root and the corresponding callus of Jatropha curcas L. were selected to be experimental materials to carry out research on effects of different antibiotics concentration on their growth. Different concentrations of various antibiotics were added into MS culture media with hormone combinations and then their influences on the growth of the explants and callus were compared. After variance analysis and overall observation of tissue culture, it was concluded that the best antibiotic for selecting transformed explants and callus was kanamycin and the best resistance selection concentration was between 10~25mg/L. Ampicillin and Tementin are two ablastin. Generally, the concentration of them were 500mg/L. Because the concentration varied in different bacteria, considering the both effect of inhibiting the bacteria and negative influence of inducing callus by antibiotics. We decreased the concentration of antibiotics. As the results, Ampicillin had little influence on explant, and the effect of inhibiting the bacteria should be considered in advance.High-frequency regenerative and transformation system is build tofound genetic transformation system of Jatropha curcas L. transient expression test is the most fastest and effectivest method to optimize the condition of transformation system.Many factors can influence transient expression.The research mainly tests four factors, that is, the number of days of preparative culture (one day, two days), differentiation culture using hormones with different concentration ratios (IBA 1.0mg/L + BA 0.4 mg/L, IBA 0.2mg/L + BA 2mg/L), with AS or without AS and the type of explants (cotyledon, leaf, dowm hypocotyl and root).The ratio of transient expression is the number of explants that express- instaneously gus gene divides that of all the explants. Concerned with the extent of transient expression of explatns, the results show that the effectiveness of transient expression is rather high(60%) after the cotyledons of Jatropha curcas L. are preparatively cultured in the differentiation culture mediums(IBA 1 mg/L, BA0.4 mg/L) for two days, and AS is added in preparative culture medium and MSo bacterial liquid that has been centrifugated. The extents of colouration are not identical. This may be due to the conformity degrees of T-DNA. Some questions will be resolved, for example, slices are not so ideal that they influence observation and photograph of inner colouration condition, another thing, discoloring time is so long and is not ideal.