The Research Of ISSR On Relationship Of5Species Of Wild Camellia Of Genus Camellia And Fingerprint Construction

As a specie of woody oil plants on unique importance in the Southern part of China,Camellia oleifera is mostly composed of unsaturated fatty acids (UFA),such as oleic acid, linoleic acid and so on, Camellia oil is a high quality edible oil. Camellia oleifera has a high ornamental value,and it can be used as a gardentree species. Camellia oleifera is one of the important species for soil and water conservation in southern China river basin, which has high ecological value and economic value.In this study,5species of wild Camellia oleifera were used to detect genetic relationships and establish their ISSR fingerprints by using inter-simple sequence repeats (ISSR) markers,and this study can provide a molecular basis for selecting the excellent characters of Camellia oleifera, molecular marker assisted selection breeding and hybrid breeding.We selected100samples from5species of wild Camellia as the experimental material. Firstly we optimize5single element by single factor experiment and determined the optimal concentration rang in every single element. Secondly we preliminarily screened out the preferable ISSR-PCR reaction system by orthogonal experiment and used range analysis to optimize the system furtherly to get the optimal ISSR-PCR reaction system for wild Camellia:20μL reaction system,DNA template30ng, primer0.7μmol/L, dNTPs0.25mmol/L, Mg2+2.5mmol/L, Taq DNA polymerase1.75U.Using the ISSR-PCR reaction system, we selected10primers from100ISSR primers, at the same time, determined the optimal annealing temperature.100samples from5species of wild Camellia by ISSR-PCR amplification, a total of166bands were samplified by10ISSR primers,on average, each ISSR primer amplified16.6bands,In166bands with159polymorphic bands, the percentage of polymorphic bands was95.78%,and the length of the bands is between200and2700bp.the proportion of polymorphic loci from high to low of the5species of wild Camellia oleifera Abel from high to low.Camellia yuhsienensis>Camellia meiocarpa>Camellia brevistytd>Camellia magniflpra>Camellia phellocapsa.According to the genetic identity and Nei’s genetic distance,5species of wild Camellia were analyzed by UPGMA cluster analysis, the result indicates the relationship between Camellia meiocarpa and Camellia brevistyta is the closest and the relationship between Camellia meiocarpa and Camellia magniflpra is the farthest.According to ISSR-PCR amplification5species of wild Camellia by10ISSR primers,the5species of wild Camellia could be identifed Individually by UBC844、 UBC873and UBC899,and the other7ISSR primers could not do that.By using UBC899primer amplified21specific bands, the5kinds of wild camellia100samples of specific DNA fingerprinting were constructed using0/1digital matrix, genetic breeding and cross breeding of Camellia provide guidance, enriched the gene bank of China.