Analysis The Genetic Relationship Of22Inbred Lines Of Hot Pepper

The genetic relationship of22inbred lines of hot pepper materials were analyzed through three methods of the morphological analysis, ester enzyme (EST) isozyme analysis and ISSR molecular marker analysis. The results are as follows:1. Through analyzing nine morphological traits of the22hot peppers by the SPSS19.0,22materials could be divided into eight categories breeding excellent inbred lines.2. Through the breeding excellent inbred lines22young leaves esterase electrophoresis found, the esterase isozymes of22hot pepper were separated out six isozyme bands. When the sterase isozyme electrophoresis information chang into a0,1matrix, with NTSYS (2.10) clustering analysis, the22hot pepper materials could be divided into eight categories in a0.82degrees.3. This experiment used CTAB method to extract DNA, it could remove polysaccharide and phenolic substances better and got a better quality of DNA, accord with ISSR molecular marker requirements4. Through the orthogonal design optimization method, set up the optimal reaction system ISSR:reaction should be20μl total system, of which10by PCR buffer solution2.0μl;25μmol/L MgCl2solution1.6μl,2.5mmol/L of dNTP solution1.6μl,10μmol/L random primers0.8μl;40ng formwork DNA1μl;2.5U/μl Taq DNA polymerase0.5μl, add deionized water or steam up to20double waterμl, finally add15～20μl mineral oil covered. Because ISSR primer is different, the annealing temperature is different, elected by screening for annealing temperature56℃. PCR amplification program for94℃the degeneration3min,94℃degeneration45S,56℃90S resilience,72℃extensions2min,35circulation; Finally in72℃extensions10min.5. The30ISSR primers from Shanghai ShengGong production were studied and selected the5high polymorphism, stability good ISSR primer, for the DNA expansion of22chili breeding excellent inbred lines. There were total51 amplification bands by the5primer. There were38polymorphism bands. The ratio of polymorphism was60%to90%, and the average polymorphism ratio is74.5%.6. Using the NTSYS, the ISSR amplification map of22breeding excellent inbred line hot peppers were analyzed. The22breeding excellent inbred lines could be divided into eight categories in a0.78degrees.7. The clustering results of between the esterase isozymes and the ISSR molecular markers has better similar than the morphological analysis and there were significant difference between the former two methods and the morphological analysis. It may result from the environment affecting the morphological characters.