The Identification Of Genuineness And Purity Of Pepper Hybrid Cultivars By Molecular Markers

In this study, DNA extraction method was simplified firstly, and thenthe SSR-polymerase chain reaction (PCR) system was optimizated by orthogonaldesign. Identified the purity and authenticity of a hybrid cultivar of Pepper, the inbredparents (JP-6and H-4)and other three varieties（JP-6、JP-7and JP-9） provided byHenan YuYi SEEDS CO., LTD, through SRAP and SSR. Designing for germplasmresources identification and breeding hybrid provide certain theoretical basis, andprovides a new direction for modern biological technology application from traditionalseed production. The results of this study ware as follows:Firstly, extracting chilli DNA using CTAB method and simple alkaline lyses.Results showed that the DNA quality, which was extracted by the method of CTAB,was batter than that extracted by a simple method of alkaline lyses method. But PCRresults are consistent, Two methods all can meet the need of SSR-PCR andSRAP-PCR amplification reaction system. The method of Alkali lyses can be used forPepper PCR analysis because the operation of the method was simple, taking shorttime, at the same time, the result was stable.Secondly, establishing a high efficient and low cost of SSR-PCR optimizationthrough orthogonal design. The results show that: The most suitable for PepperSSR-PCR reaction system is the combination of NO.2.Taq DNA enzyme0.25U、dNTP0.2mmol/L、primer0.3μmol/L、cycle-index30times。Thirdly, SSR primer screening.21pairs SSR primers were used to Pepper thehybrid varieties and its parents for PCR amplification.There were a couple of primers(CA885) display exist polymorphism in both parents and hybrid.The primer CA885can distinguish hybrid and its parents. A primer on, The results of the repeatedamplification testing on those primers were stable.Fourthly, Purity test of hybrid cultivars. The primers CA885were used to test thepurity of hybrid cultivars JP-6and H-4, The results were90.6%and86.0%,respectively. Seeding the seeds JP-6and H-4in the field, through the morphologicalobservation getting the results were90.6%and91.3%respectively. The purity gettingby molecular markers similar or lower than that by the field appraisal.Fifthly, to identify the genuineness for the three Pepper varieties by SRAPMolecular Marker. The experiment using16pair of high polymorphism SRAPprimers to expanse the parents DNA of three pepper varieties of JP-6, JP-7and JP-9.Through the comparison with spectrum we found that JP-6and JP-7were the same species, and JP-9and JP-6, JP-7were different species.The experiment using high polymorphism of16SRAP primers to expanse theparents DNA of three Pepper varieties JP-6, JP-7and JP-9.The result showedthat:JP-6and JP-7was the same variety, and the JP-9was different with JP-6andJP-9.