木麻黄AFLP分析体系的建立及引物筛选  

Establishment of the Analytical System for Casuarinaceae AFLP and Its Primer Selection

作　　者：黄桂华[1] 高丽丽[2] 仲崇禄[1] 苏晓华[3] 张勇[1] 陈羽[1] 陈珍[1] 谢冬勇 

机构地区：[1]中国林业科学研究院热带林业研究所,广东广州510520 [2]华南农业大学园艺学院,广东广州510642 [3]中国林业科学研究院林业研究所,北京100091 [4]广州市绿化公司,广东广州510170

出　　处：《中南林业科技大学学报》2007年第1期20-24,34,共6页Journal of Central South University of Forestry & Technology

基　　金：科技部863项目(2002AA241091);广东省自然科学基金(06024658;2006-2008);中澳合作项目(1994-2004);广东省科技计划重点引导项目(2004B20801008)

摘　　要：采用CTAB法改良技术提取木麻黄基因组DNA,在提取液中加入2%β-巯基乙醇防氧化和2%的PVP去除酚类等物质,获得高质量的基因组DNA,以满足AFLP对模板DNA高质量的要求;采用分步酶切和连接后,进行预扩和选扩,摸索建立和优化了木麻黄AFLP分析体系,探讨了木麻黄AFLP分析的影响因素;从64对AFLP核心引物组中筛选出了8对适合木麻黄分析的引物组合.同时探讨了AFLP技术在木麻黄研究领域的应用前景.An in,proved method of CTAB was used in this study to extract the total DNA of Casuarinaceae. Then, high quality genomic DNA was isolated for AFLP analysis with 2% of mercapto-ethanol in the extracting solution to prevent formation of oxides and 2% of PVP to eliminate phenols from genomic DNA. By means of double enzyme restriction, adapter-ligation, pre-amplification and selective amplification, AFLP analytical system of Casuarinaceae was established and optimized. 8 primer combinations were selected from 64 pairs of core combinations of AFLP after a series of tests. The application prospect of AFLP in Casuarinaceae research field was also discussed.

关 键 词：生物技术 AFLP 分析体系 引物 木麻黄 

分 类 号：S792.93[农业科学—林木遗传育种] Q781[农业科学—林学]