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Clones identification of Sequoia sempervirens (D.Don) Endl.in Chile by using PCR-RAPDs technique

摘要A protocol of polymerase chain reaction-random amplified polymorphic DNAs (PCR-RAPDs) was established to analyse the gene diversity and genotype identification for clones of Sequoia sempervirens (D.Don) Endl.in Chile.Ten (out of 34) clones from introduction trial located in Voipir-Viilarrica,Chile,were studied.The PCR-RAPDs technique and a modified hexadecyltrimethylammonium bromide (CTAB) protocol were used for genomic DNA extraction.The PCR tests were carried out employing 10-mer random primers.The amplification products were detected by electrophoresis in agarose gels.Forty nine polymorphic bands were obtained with the selected primers (BG04,BF07,BF 12,BF13,and BF 14) and were ordered according to their molecular size.The genetic similarity between samples was calculated by the Jaccard index and a dendrogram was constructed using a cluster analysis of unweighted pair group method using arithmetic averages (UPGMA).Of the primers tested,5 (out of 60) RAPD primers were selected for their reproducibility and high polymorphism.A total of 49 polymorphic RAPD bands were detected out of 252 bands.The genetic similarity analysis demonstrates an extensive genetic variability between the tested clones and the dendrogram depicts the genetic relationships among the clones,suggesting a geographic relationship.The results indicate that the RAPD markers permitted the identification of the assayed clones,although they are derived from the same geographic origin.

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DOI 10.1631/jzus.B0820162
发布时间 2009-04-21(万方平台首次上网日期,不代表论文的发表时间)
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浙大学报(英文版)(B辑:生物医学和生物技术)

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