The Activity Determination And Anti-fungal Activity Analysis Of The GAFP Expressed In Transgenic Gene Pepper

Objective:In this research, the propose was to determin the expression of Gastrodia Antifungal Protein (GAFP) expressed in transgenic Gene "ZunJiao No.1" Pepper, analyses the anti-fungal activity of the expression protein of the transgenic Gene Pepper and obtain the new fuchsin pepper which has the anti-fungal ability. Methods:(1) Abstraction and purification the GAFP of transgenic pepper using the cation exchange chromatography;(2) Identify the obtained GAFP by using the SDS-PAGE gel electrophoresis;(3)Separated the fungi from the infected pepper of the indoor pepper and the wild pepper by using the microorganisms diluted Separation method;(4) Analyses the anti-fungal ability of GAFP expressed in transgenic pepper to the G.capitatus, the Fusarium and the Trichoderma viride fungi by the growth rate method, and the results will showed with inhibition rate (%),said that as the experimental group (A group);(5)The control groups are the Wild Gastrodia GAFP (B1 group) and non-transgenic pepper extract (B2 group); Results:(1)The result of the SDS-PAGE gel electrophoresis showed that the transgenic pepper has the Specific bands at the GAFP point;(2) We separated the fungi the G.capitatus and the Fusarium from the infected pepper of the indoor pepper and the wild pepper;(3) The transgenic pepper expression protein GAFP has the varying degrees of inhibition to the G.capitatus,the Fusarium and the Trichoderma viride fungi on the different concentrations at 0.0125mg/ml,0.025mg/ml,0.05mg/ml,0.1mg/ml,0.2mg/ml,and at the 0.2mg/ml concentration, the inhibition rate of the transgenic pepper expression protein GAFP to the G.capitatus,the Fusarium had exceed 50%, and reached to 61.38%,54.52% respectively, the inhibition rate to the Trichoderma viride had reached to 45.65%. Compared with the control group, they have the significant differences (P< 0.05);(4) The Wild Gastrodia GAFP has the varying degrees of inhibition to the G.capitatus,the Fusarium and the Trichoderma viride fungi on the different concentrations at 0.0125mg/ml,0.025mg/ml,0.05mg/ml, 0.1mg/ml,0.2mg/ml, and at the 0.2mg/ml concentration, all of the inhibition rate of the Wild Gastrodia GAFP to the Gcapitatus,the Fusarium and the Trichoderma viride had exceed 50%, and reached to 61.39%,50.19% and 54.73% respectively. Compared with the control group, they have the significant differences (P< 0.05); (5) Compared the inhibition of the non-transgenic pepper extract to the G.capitatus, the Fusarium and the Trichoderma viride fungi with the control group's, they have not the significant differences;(6) We analysised and compared the date of the two anti-fungal groups the transgenic pepper extract group and the Wild Gastrodia extract group by Statistics software, the results showed that they have not the significant differences. Conclusion:The GAFP has the similar anti-fungal ability to the wild Gastrodia GAFP,which has expressed in transgenic Gene Pepper system successfully.