Analysis Study Of Hot Pepper(Capsicum Spp.) With RAPD And ISSR

30 pepper germplasm materials was used in the experiment,and 7 botanical and fruit characters were investigated such as the average height, the first flowering node, single fruit weight, fruit length and diameter. The genomic DNA of pepper germplasm materials was extracted by using improved CTAB, followed by the detection of its quality and purity, and then optimized the reaction system of the RAPD and ISSR by orthogonal design.12 pairs of RAPD primers and 11 pairs of ISSR primers were screened out for amplifying genomic DNA.The genetic diversity of cluster was analyzed by the NTSYS-pc (2.10) software.The main results obtained were as follows:As the 7 morphological characters of pepper germplasm materials were measured, the 30 pepper germplasm were divided into 8 categories with SPSS(11.5) software.By using improved CTAB,the purity and quality of DNA obtained of pepper germplasm was satisfying in the experiment.By the optimized orthogonal design,the analysis of hot pepper germplasm of RAPD and ISSR reaction system has established.The total volume of RAPD optimal system is 20μL,of which 10×PCR buffer 2.0μL,25μmol/L MgCL2 solution 1.5μL,2mmol/L of dNTPs solution 2.0μL,10μmol/L random primers 1.0μL,50ng/μL DNA template 1.0μL,2.5U/μL TaqDNA polymerase 0.4μL, and add 20μL paraffin oil cover.PCR amplification program was 94℃for 3min;94℃denaturation 60S,37℃recovery of 90S,72℃extension 120S,35 cycles;finally 72℃for 10min. ISSR reaction system, the total volume of 20μL, of which 10×PCR buffer solution 2.0μL,25μmol/L MgCl2 solution 1.5μL,2mmol/L of the dNTPs solution 2.0μL,10μmol/L random primers 1.0μL,50ng/μL template DNA 1.0μL,2.5U/μL TaqDNA polymerase 0.5μL,and add 20μL paraffin oil cover.Amplification program was 94℃for 3min;94℃denaturation 45S,48℃-56℃refolding 90S,72℃extension 120S,35 cycles;finally extension at 72℃for 10min.With NTSYS-pc (2.10) software, cluster analysis of RAPD and ISSR amplification pattern of 30 pepper germplasm showed that the genetic polymorphism of amplified bands of ISSR primers of pepper germplasm was significantly higher than RAPD,in other words ISSR molecular marker was more suitable for hot pepper germplasm genetic diversity;and similar conclusion was obtained by synthesize comparison analysis.