Analysis On The Genetic Diversity Of Liquidambar Formosana Of Zheng Jiang Province

Liquidambar formosana Hance. is Hamame lidaceae deciduous trees. Distributed in the Qinling Mountains area to the south of Huaihe, Cross the North tropical and south, in the tropical north, 4 climate zones, is a fast-growing tree species in our country important indigenous broadleaf, ornamental, medicinal and timber and other values in a body. In this experiment, The leaves of Liquidambar for materials, through the comparison of different DNA extraction methods, Extraction of genomic DNA of optimal Liquidambar formosana, At the same time, Liquidambar genomic SRAP system were optimized, and the system is applied to the 7 groups of Liquidambar materials were amplified by SRAP analysis. The results are as follows:1.In this experiment, by using the modified CTAB method and kit method(developed by Bioteke) the two methods were compared for t Liquidambar formosana leaves DNA extraction results, Found that the two kinds of extraction methods of Liquidambar leaves DNA content and purity are not significant, which can meet the requirements of the subsequent PCR amplification. For old leaves, The yield and purity of the modified CTAB method to extract genomic DNA of old leaves were relatively high kit method. The latter not only content is low, and contain other impurities, degradation of DNA is more serious, will adversely affect the response to the downstream of PCR and other DNA molecules. Using the modified CTAB extraction method effectively solves the problem of Liquidambar genomic DNA extraction of browning and polyphenol, polysaccharide and protein secondary metabolites interference and other issues. From the detection of the purity of the DNA template can be used for SRAP analysis; in addition, the method also has the advantages of low cost, simple operation, time-saving features.2.By the orthogonal experiment design Ll6(45), five factors including Mg2+ concentration, d NTPs concentration, primer concentration, Taq DNA polymerase and template DNA amount in SRAP-PCR reaction system were optimized, he optimized SRAP-PCR reaction system was as follow: total volume 20 μL, containing 1×PCR Buffer, 1.5 mmol?L-1 Mg2+, 0.16 mmol?L-1 d NTPs, 0.9 U Taq DNA polymerase, 0.5 μmol?L-1 primer, 40 ng template DNA. SRAP-PCR amplification procedure: pre-denaturation at 95℃for 5 min, 94 ℃ degeneration for 30 sec, annealed at 35 ℃ for 1 min, 72 ℃extension of 2 min, 5 cycles; 94 ℃ degeneration for 30 sec, annealed at 50 ℃ for 30 sec, 72 ℃extension of 1 min,30 cycles; 72 ℃extension of 10 min; kept at 4℃. The results showed that the primers were selected on random selection of genomic DNA could be amplified polymorphic and clear bands of DNA fragments. The description of the SRAP-PCR reaction system is stable and reliable, suitable for Liquidambar genomic DNA for SRAP-PCR amplification reaction.3. Genetic diversity of different populations of Liquidambar formosanaThe application of a molecular markers by 16 primer combinations in seven collected from zhe jiang province maple tree group(hang zhou, lis hui, wen zhou, qu zhou, tai zhou, zhou shan) SRAP- PCR analysis of samples of 120 individuals, total of 902 amplified bands, of which 897 bands were polymorphic.Percentage of polymorphic bands(PPB)and Shannon ’ s phenotypic diversity index(Ho)were respectively 82.18%and 0.4935 at the species level, at the population level, Percentage of polymorphic bands(PPB) excursion is 54.29~74.29. Shannon’s phenotypic diversity index(Ho) excursion is 0.3199~0.3928.In natural populations of Liquidambar formosana within and between groups had significant genetic variation(P<0.001). The genetic variation among populations accounted for 15.04% of the total variance, Genetic variation within the groups account for 84.96% of the total variance.4. The genetic distance of the natural populations of Liquidambar formosanaThe genetic similarity and genetic distance calculated Liquidambar group by Nei ’index, The results showed that the group genetic similarity between the changes in the 0.8999~0.9685, The mean genetic similarity of 0.9396. The genetic distance of 0.1055~0.0320, Mean genetic distance was 0.0624. Chun An and Tong Lu the smallest genetic distance is 0.0320, While Cun An and Wen Chen into the largest genetic distance was 0.1055.5. The dendrogram of the natural population of Liquidambar formosanaAccording to the genetic distance of Liquidambar formosana species groups, Cluster analysis was performed on 7 groups of Liquidambar formosana Hance by UPGMA clustering method, The dendrogram of genetic relationship of reflection between the various groups. The clustering results of Liquidambar formosana populations found their geographic distribution pattern of roughly coincide.