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Citation:

Optimization of SRAP-PCR System for Camellia oleifera

  • Camellia oleifera is one of the important oil tree species in south China, and C. oleifera industry is quickly developed with the support of the national policies in recent years. The disorder of C. oleifera varieties is one of the key issues restricting the development of C. oleifera industry. Because of high polymorphism, good repeatability, less use of DNA and so on, SRAP as a new marker was used in identification of cultivars, analysis of genetic resources and genetic diversity in recent years. In this paper, the orthogonal design was used to optimize SRAP-PCR system for C. oleifera by 5 factors (Mg2+, dNTPs, primer, Taq polymerase, DNA template) and 4 levels, respectively. The data were analyzed by software SPSS V13.0. A suitable SRAP-PCR system (20 μL) was established as: 75 ng DNA template, 1.5 mmol·L-1 Mg2+, 0.15 mmol·L-1 dNTPs, 1U Taq DNA polymerase, 0.4 μmol·L-1 primer, 1×PCR buffer. The result of optimal SRAP-PCR system will provide a foundation for the identification of C. oleifera cultivars.
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  • [1] 胡冬南, 游美红, 袁生贵, 等. 不同配方施肥对幼龄油茶的影响[J]. 西北林学院学报, 2005, 20(1): 94-97

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    [4] 张智俊. 油茶优良无性系组织培养、RAPD分子鉴别和cDNA文库构建的研究 . 长沙:中南林学院, 2003

    [5] 阙生全, 张 芳. 油茶DNA提取及RAPD分析最佳反应体系的建立[J]. 安徽农业科学, 2008, 36(3): 902-903, 905

    [6] 张国武, 钟文斌, 乌云塔娜, 等. 油茶优良无性系ISSR分子鉴别[J]. 林业科学研究, 2007, 20(2): 278-282

    [7] 温 强, 雷小林, 叶金山, 等. 油茶高产无性系的ISSR分子鉴别[J]. 中南林业科技大学学报, 2008,28(1): 39-43

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    [9] Li G, Quiros C F. Sequence-related amplified polymorpgism (SRAP) a new marker system based on a simple PCR reaction: its application to mapping and gene tagging in Brassica [J]. Theor Appl Genet, 2001, 103: 455-461
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    [13] [13] 张四普, 汪良驹, 曹尚银, 等. 23个石榴基因型遗传多样性的SRAP分析[J]. 果树学报, 2008, 25(5): 655-660

    [14] [14] 刘建丰, 王志德, 刘艳华, 等. 应用SRAP标记研究烟草种质资源的遗传多样性[J]. 中国烟草科学, 2007,28(5): 49-53

    [15] [15] 王大一, 吴 洁, 谭文芳, 等. 四川省高淀粉甘薯品种资源亲缘关系SRAP分析[J]. 西南农业学报,2007,20(3): 506-509

    [16] [16] 陈万胜, 王元英, 罗成刚, 等. 利用正交设计优化烟草SRAP反应体系[J]. 分子植物育种, 2008, 6(1): 177-182

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    [19] [19] 李 莉, 彭建营, 郑宝强. 枣SRAP-PCR体系的正交优化[J]. 农业生物技术学报, 2008, 16(2): 361-362

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Optimization of SRAP-PCR System for Camellia oleifera

Abstract: Camellia oleifera is one of the important oil tree species in south China, and C. oleifera industry is quickly developed with the support of the national policies in recent years. The disorder of C. oleifera varieties is one of the key issues restricting the development of C. oleifera industry. Because of high polymorphism, good repeatability, less use of DNA and so on, SRAP as a new marker was used in identification of cultivars, analysis of genetic resources and genetic diversity in recent years. In this paper, the orthogonal design was used to optimize SRAP-PCR system for C. oleifera by 5 factors (Mg2+, dNTPs, primer, Taq polymerase, DNA template) and 4 levels, respectively. The data were analyzed by software SPSS V13.0. A suitable SRAP-PCR system (20 μL) was established as: 75 ng DNA template, 1.5 mmol·L-1 Mg2+, 0.15 mmol·L-1 dNTPs, 1U Taq DNA polymerase, 0.4 μmol·L-1 primer, 1×PCR buffer. The result of optimal SRAP-PCR system will provide a foundation for the identification of C. oleifera cultivars.

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