Analysis Of Genetic Variability Of None Or Little Cones Individuals In Platanus By ISSR And SSR Markers

Platanus are important landscape species in large and medium-sized cites, butevery year their cone shedding and flying pollen bring huge inconvenience to citizen’s dailylife and make them healthy in danger. In order to lower or eliminate their cone shedding, westudy on Platanus mostly from phenotypic variation and follow-up observations, yet rarelyworked at the molecular level. Compare with the traditional identification methods such asmorphology, sporopollen and peroxidase isozyme marker, molecular markers has manybenefits such as fast, accurate, high polymorphism, not influent by external environment,organization category, development period and the rest. The ISSR and SSR markers wereemployed in study to analysis Genetic Variability of none or little cones Individuals inplatanus, provide basis for breeding none-or-little cones platanus acerifolia and spreadingnone-or-little cones platanus acerifolia.Main results of the present study as follows．(1). The method on the basis of the traditional CTAB method was improved, and then usedit to extract DNA of Platanus acerifolia, and then test the quality of DNA, the results showsthat the improved CTAB method is suitable for extracting high quality genomic DNA ofPlatanus acerifolia.(2). ISSR technique is employed to research14platanus acerifolia whose phenotype isnone-or-little cones this study, we can analysis their genetic relatedness at the molecularlevel in order to breeding none-or-little cones platanus acerifolia. The result shows that,18primers were selected for their clear banding patterns and steady consistent amplifications;11primers were employed in this study. The results of PCR amplification in statistical,105amplified DNA fragments for clearly, steady and repeatable amplifications were obtainedfrom11primers and there were99polymorphic bands accounting for93.84％. The range ofvariation of primers was6-12, while9.6DNA fragments were obtained from each primerand the average polymorphic bands were8.9. Polymorphism range of variation is81.82%-100.00%. The clades formed indicated that in a similar level of0.67, the14platanus acerifolia were clustered into five groups. According to the phenotypecharacteristic of platanus and others methods, we tentative recognized6was none-or-littlecones platanus acerifolia. The number1,2,3,13are little conges platanus acerifolia. (3). ISSR technique is employed to research platanus acerifolia and their Breedingseedlings, The results are as follows,13of18pairs primers were selected in this study;ISSR molecular markers reveal much higher polymorphism between platanusacerifolia,which is up to82.35%; clustering three groups of materials, the first set ofmaterials shows that there is no variation between cutting and mother plant, while there issmall variation between grafting and mother plant; the second and the third group ofmaterials shows the variation genetic variation appears normal in Grafting breedingseedlings and cuttings breeding seedlings. Based on the above results of three groups ofmaterials, so we can speculate that roughly, there is variation between platanus acerifoliaand grafting, so does with cutting, while the variation between platanus acerifolia andgrafting is bigger than another.(4). SSR technique is employed to research platanus acerifolia and their Breedingseedlings. The results are as follows,10pairs of primers, which is suited to sycamoreamplication, were screened out by repeated screening amplifying the10pairs of primers, weobtain36alleles,whose numbers ranged from2-5, with an average per primer alleles of3.6.The total polymorphism rate is97.4%, while the distribution of polymorphic ratio of eachprimer is66.7.0%-100.0%. Clustering three groups of materials, The results of these threegroups of materials are consistent,so we can get the conclusion that, there is variationbetween platanus acerifolia and grafting, there may be smaller variation between cuttingand mother plant.（5）.During the prosess of none-or-little cones platanus acerifolia popularize, using themethod of cutting would be better.