三角梅ISSR反应体系的建立和优化  

Establishment and Optimization of ISSR System in Bougainvillea

作　　者：李房英[1] 黄彦晶[2] 吴少华[2] 赖钟雄[2] 

机构地区：[1]福建农林大学园林学院,福州350002 [2]福建农林大学园艺学院,福州350002

出　　处：《生物技术通报》2010年第7期142-145,152,共5页Biotechnology Bulletin

基　　金：国家支撑计划(子课题)菊花等闽台特色花卉新品种引进创新和离体保存研究(2007BAD07B03)

摘　　要：对影响三角梅ISSR-PCR扩增反应的各个参数进行优化,建立适合三角梅的ISSR反应体系:PCR反应体积为20μL,其中10×buffer(含Mg2+)2.0μL,dNTP250μmol/L,Taq酶1.0U,引物0.3μmol/L,模板DNA20ng。扩增程序:94℃预变性5min;94℃变性1min,51.6℃退火1min,72℃延伸2min,34个循环;最后72℃延伸7min。该反应体系标记点位清晰、稳定、重复性好,适宜三角梅ISSR分析,为应用ISSR技术鉴定三角梅种质资源、分子标记辅助选择育种及其遗传多样性研究奠定了基础。Factors affecting ISSR-PCR application of Bougainvillea were optimized and ISSR system for Bougainvillea was established. Each 20 μL amplification reaction consisted of 2. 0 μL 10 × buffer（ include Mg^2＋ ） ,250 μ mol/L dNTP, 1.0 U Taq polymerase, 0. 3 p.mol/L ISSR primer,20ng DNA template. Amplication was under the following conditions：5 min at 94℃ for lcycle,followed by 1 min at 94℃ ,1 min at 51.6℃ ,and 2 min at 72℃ for 34 cycles,and 7 min at 72℃ for a final extension. The clear,stable and reproducible optimal ISSR-PCR reaction system was established for ISSR analysis of Bougainvillea,which could laid foundation for ISSR analysis on studies of germplasm resources identification,molecular marker assisted breeding and genetic diversity.

关 键 词：三角梅 ISSR 体系建立 优化 

分 类 号：S685.99[农业科学—观赏园艺]