Studies On Transformation Of Resveratrol Synthase Gene Into Strawberry (Fragaria×Ananassa Duch)

Strawberry is an important commercial fruit crop cultivated throughout the world. However, the production and quality are seriously affected by plant microbial pathogens. Currently, genetic transformaton is one of the most attractive means for plant breeding since this technology allows the introduction of desired traits into plants. Resveratrol plays a important role in phytochemical of human health benefits. Much attention has been focused on resveratrol and RS gene because of their implication as an important phytoalexin. Most of the crops are lack of RS, so it is interesting to obtain pathogens resistance and high quality strawberry with RS by genetic engineering. In the study of effect of many factors on the regeneration of leaf discs from Allstar and Tudla,it was found that during the 45 days of leaf discs culture, differentiation medium (MS+B5) was replaced by fresh medium every other 15 days,and was supplemented with corresponding hormore combinations : TDZ 2.0 mg/L + IAA 0.1mg/L ,TDZ 1.5 mg/L + IBA 0.1mg/L,TDZ 1.0 mg/L in three phases respectively ,which gave a high and steady shoot regeneration frequency of more 75%. Plant expression vector of pC2PRS was constructed by inserting the the fragment of RScDNA from C1PRS to pCAMBIA2301-pTΩ4A.. The pC2PRS was then mobilized into EHA105 by using freeze-thaw method , which can be used for transformation to strawberry. The RS gene was introduced into strawberry by the simplest and most successful Agrobacterium-mediated transfer using the established transformation system. In the couse of experiment, the procedures of transformation was studied, including infecting time,different types of explant and selection methods.The optimized condition was that infection time was 4-5min and post-selection was two weeks. Furthermore, co-culture time was determined, according to the signal of bacterium appearing from co-culture leaf discs kept directly on medium. Ultimately ,two individual transgenic plants have been obtained , characterization of rooted transformed plants in the presence of screening medium and PCR analysis with the target fragment of 1200bp revealed that the integration of the transferred foreign RS gene into the the strawberry genome.