| 厚朴AFLP分子标记体系的建立 |
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| 引用本文： | 蒋燕锋,斯金平,黄华宏,程龙军.厚朴AFLP分子标记体系的建立[J].浙江林学院学报,2010,27(2):304-309. |
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| 作者姓名： | 蒋燕锋 斯金平 黄华宏 程龙军 |
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| 作者单位： | 浙江林学院林业与生物技术学院,浙江,临安,311300 |
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| 基金项目： | 浙江省重大科技计划项目 |
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| 摘 要： | 以厚朴 Magnolia officinalis为材料，通过对扩增片段长度多态性（AFLP）反应体系中的酶切模板用量、扩增条件等几个方面进行优化，建立了较为理想的厚朴AFLP反应体系。结果表明：①采用十六烷基三甲基溴化铵（CTAB）硅珠法提取的DNA质量较好，适用于AFLP分析。②酶切的基因组DNA以400 ng为宜；预扩增产物的稀释倍数以30倍为宜；20.00 μL选扩反应体系中Mg2+最佳浓度为2.00 mmol·L－1，三磷酸脱氧核苷酸（dNTPs）最佳浓度为0.225 mmol·L－1，Taq DNA 聚合酶最佳用量为16.67 nkat时，扩增带清晰且稳定，扩增效果较好。旨在为研究厚朴天然种群的遗传多样性及遗传结构分析、新品种选育及鉴定和遗传图谱构建等奠定理论基础。图7表1参12
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| 关 键 词： | 中草药 厚朴 扩增片段长度多态性 中药材 |
An AFLP molecular labeling technique for Magnolia officinalis |
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| JIANG Yan-feng,SI Jin-ping,HUANG Hua-hong,CHENG Long-jun.An AFLP molecular labeling technique for Magnolia officinalis[J].Journal of Zhejiang Forestry College,2010,27(2):304-309. |
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| Authors: | JIANG Yan-feng SI Jin-ping HUANG Hua-hong CHENG Long-jun |
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| Institution: | (School of Forestry and Biotechnology, Zhejiang Forestry College, Lin＇an 311300, Zhejiang, China) |
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| Abstract: | To establish an efficient and stable amplified fragment length polymorphism(AFLP)analysis system for Magnolia afficinalis,selection and optimization of DNA template treatments and selective amplification conditions were undertaken using the cetyl trimethylammonium bromide(CTAB)-SiO_2 method for mapping.Results indicated that:High quality DNA samples were obtained with the CTAB-SiO_2 method and the DNA template used was 400ng.The best treatments for the analysis were Mg~(2+)of 2.00mmol·L~(-1),de oxyribonucleotide triphosphates(dNTPs)of 0.90mmol·L~(-1),and Taq DNA polymerase of 16.67nkat.This research showed that the CTAB-SiO_2 method was fit for AFLP analysis on Magnolia,that a 30 times dilution for the pre-amplification products could be used for selective amplification,and that the silver-staining AFLP system could provide a method for further study with molecular markers in the field of genetic diversity,for breeding new cultivars,and for genetic relationships with Magnolia afficinalis. |
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| Keywords: | Chinese herbal medicine Magnolia officinalis amplified fragment length polymorphism(AFLP) Chinese medicine |
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