Construction Of Plant Expression Vector Of Raphanus Satives-Antifungal Protein 2 Gene And Transformation Pepper(Capsicum Annuam L)

The serious plant disease problem of pepper results in yield losses, because of its susceptibility to many pathogens, including fungi, bacteria, viruses, and nematodes, et al. Recently, Breeding new varieties for disease resistance by genetic engineering techniques has been a new strategy. The promoter of prp1-1from potato could be induced by many pathogens, such as late blight disease. The promoter of prp1-1 was cloned from the potato leaf DNA. pBPR-2, the inducible plant expression vector comprising prp1-1 promoter and Rs-AFP2 gene, and pBCR-2, the constructive plant expression vector comprising CaMV 35S promoter and Rs-AFP2 gene, were constructed. Rs-AFP2 genes with pBPR-2 and pBCR-2 were introduced into Bulgariajanjiao pepper, respectively. The main results in this study are indicated as follows:1. The promoter of prp1-1 was cloned from the potato leaf DNA by the method of PCR. pBPR-2, the inducible plant expression vector comprising prp1-1 promoter and Rs-AFP2 gene were constructed, as result Rs-AFP2 gene was induced to express by plant pathogenic fungi under the control of the prp1-1 promoter fragment; At the same time , pBCR-2, the constructive plant expression vector comprising CaMV 35S promoter and Rs-AFP2 gene , were constructed..2. A regeneration system of Bulgariajanjiao pepper plant was established by optimizing the media. The optimal medium for the stage of cotyledons differentiation was comprised of MS, 6-BA(5.0mg/L), IAA(0.5mg/L), AgNO3(5mg/L) and PT(0.3mg/L); the optimal medium for the stage of adventitious buds elongation was comprised of MS, GA₃(1 mg/L), 6-BA(1.5 mg/L) and PT(0.3mg/L); the optimal medium for the stage of plantlets rooting was comprised of MS, IAA(1.0 mg/L) and NAA(0.5 mg/L).