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| 山核桃AFLP实验技术体系的建立 | |
| 引用本文： | 郭金剑,池伟,赵鑫珠,王丽,曾燕如,张秋月.山核桃AFLP实验技术体系的建立[J].浙江林学院学报,2008,25(4):532-537. |
| 作者姓名： | 郭金剑 池伟 赵鑫珠 王丽 曾燕如 张秋月 |
| 作者单位： | 1. 浙江林学院浙江省现代森林培育技术重点实验宦,浙江临安,311300 2. 浙江世界贸易中心长乐实业有限公司,浙江临安,311300 3. 浙江林学院天则山核桃科技开发有限公司,浙江临安,311300 |
| 基金项目： | 浙江省自然科学基金资助项目 , 浙江省现代森林培育技术重点实验室开放基金资助项目 , 国家自然科学基金资助项目 |
| 摘 要： | 以充分展开的山核桃Carya cathayensis嫩叶为材料、利用改良CTAB（十六烷基三甲基溴化铵）法与CTAB裂解一硅珠吸附法提取山核桃总DNA，对适合于山核桃叶片的AFLP（扩增片段长度多态性）分析体系进行了摸索。结果表明：①改良CTAB法适用于从山核桃叶中提取高质量的DNA，提取的DNA可用于AFLP的分析；②采用EcoR I/Mse I 酶切组合，酶切连接一步法与两步法的效果没有明显的差异，但一步法操作更为简便；③适合于山核桃叶AFLP分析的引物组合为E-ACT＋M-CAT，E-ACT＋M-CTG，E-ACG＋M-CAT，E-ACG＋M-CTG。扩增产物经60g·L^-1变性聚丙烯酰胺凝胶电泳后进行银染，条带信号强度一致性好，条带分布均匀、清晰，分辨率较高。 |
| 关 键 词： | 植物学 山核桃 DNA提取 扩增片段长度多态性(AFLP) |
An AFLP analysis system for Carya cathayensis |
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| Institution: | GUO Jin-jian, CHI Wei, ZHAO Xin-zhu, WANG Li, ZENG Yan-ru, ZHANG Qiu-yue (1. The Key Laboratory for Modern Silvicultural Technology of Zhejiang Province, Zhejiang Forestry College Lin＇an 311300, Zhejiang, China; 2. Changle Co. Ltd, Zhejiang World Trade Center, Lin＇an 311300 Zhejiang, China; 3. Tianze Science and Technology Development Co. Ltd. for Hickory, Zhejiang Forestry College, Lin＇an 311300, Zhejiang, China) |
| Abstract: | Total DNA was extracted from young, fully opened leaves of Carya cathayensis （hickory） by an improved cetyl trimethylammonium bromide （CTAB） method and a CTAB-silicon method, based on which an amplified fragment length polymorphism （AFLP）analysis system for hickory was developed. An electrophoresis was used on the amplified products in a 6% denatured polyacrylamide gel, followed by silver staining, with clearly identified bands shown. Specifically： （1）the improved CTAB method was appropriate for DNA extraction from hickory leaves with extracted DNA qualified for AFLP analysis; （2）with a combination of EcoR I and Mse I for enzyme cutting, the results were the same for the one-step method combining enzyme cutting and adaptor ligation and the two-step method which separated them with the one-step method being much easier; and （3）there were four primer pairs suitable for this AFLP analysis, namely E-ACT ＋ M-CAT, E-ACG ＋ M-CAT, E-ACG ＋ M-CAT, and E-ACG ＋ M-ATG. |
| Keywords: | botany Carya cathayensis （hickory） extraction of DNA amplified fragment length poly morphism （AFLP） |
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