Study On Gladiolus RAPD Analysis

12 common Gladiolus cultivars were systemically studied and discussed from three aspects of biological character, isoenzyme marks and RAPD molecular marks. The results showed:(1) Through studying on biological characters studying of 12 Gladiolus cultivars, the phenomena of homonym, unscientific classification and genetic relation confusion.(2) The analysis of Isoenzyme extracted from Gladiolus corm showed: the effect of superoxide isoenzyme band is best, but polymorphism is worse; hydrogen peroxide isoenzyme is following , its polymorphism is better then superoxide isoenzyme band; the effect of esterase isoenzyme band is bad.(3) The modified SDS method which was used to extract DNA got good results. During RAPD , RNA in the DNA template had no effects on the amplification results. SDS, chloroform, isopropanol disturbed amplification results. The optimalized RAPD systems were: total reaction volume 25ul, including 10 times amplification buffer solution 2.5ul, Mg +2m mol/1, dNTP 150umol/l, primer 0.4u mol/1, genome DNA 20ng,Taq enzyme lU.other components is Super-purified water. Optimizing program : 94 C 10min, one cycle; 94 C 30s, 37'C 60s, 72C 90s, forty cycles ;72C 5min one cycle ; 4C keeping. 12 gladiolus were analysis by RAPD through selected 26 primers, getting 164 bands in all, 151 bands are polymorphism. The ratio of polymorphism is 92.1%. the genetic similar degree 0.660 threshold, the germplasm resources were classified into four groups with cluster analysis, the result was different from that derived from traditional method.