文献类型：期刊文献

中文题名：桤木属植物AFLP反应体系的建立与优化

英文题名：Establishment of the Optimized AFLP Analysis System for Alnus

作者：饶龙兵[1] 杨汉波[1,2] 郭洪英[3] 段红平[2] 陈益泰[1]

第一作者：饶龙兵

机构：[1]中国林科院亚热带林业研究所;[2]云南农业大学农学与生物技术研究所;[3]四川林业科学研究院

年份：2014

卷号：12

期号：3

起止页码：547-553

中文期刊名：分子植物育种

外文期刊名：Molecular Plant Breeding

收录：CSTPCD;;北大核心:【北大核心2011】；CSCD:【CSCD_E2013_2014】；

基金：十二五国家科技支撑项目桤木育种专题(2012BAD01B0604);中央级公益性科研院所基本科研业务费专项资金(RISF2013010)共同资助

语种：中文

中文关键词：AFLP;欧洲桤木;多态性

外文关键词：AFLP, A. glutinosa, Polymorphism

分类号：Q75

摘要：为了建立桤木属植物AFLP体系,筛选扩增条带丰富的引物,本研究以欧洲桤木(A.glutinsoa)为材料,对AFLP反应过程中的关键因素(DNA质量和浓度,酶切,扩增体系等)进行了研究。结果表明:预扩增稀释产物3.0μL,酶切时间以5 h,连接16 h,连接产物稀释10倍用于预扩增,预扩增产物稀释20倍为宜。确定桤木AFLP-PCR 20μL的反应体系:模板DNA 300 ng,10×Taq DNA聚合酶Buffer(含Mg2+)2.0μL,dNTPs0.3μL,引物1.0μL,Taq DNA聚合酶0.6 U,加ddH2O至20μL,并利用建立的体系筛选出7对扩增条带多、条带清晰、多态性好的引物。本研究建立了适用于桤木属植物的AFLP反应体系,并筛选出适合桤木属植物的引物,为今后利用AFLP标记技术进行桤木属遗传多样性分析提供了一定的实验依据。
The optimized AFLP analysis system was established for A lnus. A. glutinosa was used to comparatively study and analysis their essential factors influencing the results of AFLP, including the quality and concentration of the extracted DNA, digested time and amplification system that influenced the results of the AFLP. The results indicated that the dosage of template DNA was 300 ng; The reaction time of enzyme digestion was 5 h; The production of enzyme digestion copulation 16 h; 20 times of dilution for the products of pre-amplification was suggested for selective amplification; The system of AFLP-PCR that diluted production of selective amplification 3.0 μL, 10×Taq DNA polymerase Buffer （include Mg2＋） 2.0 μL, dNTPs 0.3 μL, primers 1.0 μL, Tat/DNA polymerase 0.6 U, mix in ddH20 to 20 μLo Selected 7 pairs by many amplified bands, screening repeatedly and high polymorphism. By using the AFLP reaction system, primers suitable to Alnus were screened out. Those results provide fundamentals for further molecular studies on A lnus using AFLP marker.