郁金香ISSR-PCR体系建立与优化  

Establishment and Optimization of ISSR-PCR System for Tulip

作　　者：巨秀婷 阿啟兰 

机构地区：[1]青海大学高原花卉研究中心,西宁810016

出　　处：《分子植物育种》2016年第4期973-979,共7页Molecular Plant Breeding

基　　金：青海大学中青年科研基金项目(2013-QNY-4);2015青海省园林植物研究重点实验室(2015-Z-Y17)共同资助

摘　　要：本研究以22个郁金香品种为材料,采用正交设计试验和单因素试验对ISSR-PCR反应中的5个影响因素:Mg^(2+)、d NTPs、Taq酶、引物、模板DNA进行4个水平的优化试验。结果表明最佳反应体系为:25μL体系中Mg^(2+)1.0 mmol/L、d NTPs 0.15 mmol/L、Taq酶0.25U、引物0.2μmol/L、模板DNA 100 ng。退火温度为58℃。该体系的建立有利于ISSR分子标记技术研究郁金香种质资源遗传多样性。In this study the orthogonal test and single-factor test was used to optimize the ISSR-PCR reaction procedures of 22 Tulip cultivars in five factors including the Mg^(2+), d NTPs, Taq DNA polymerase, primers, and concentration of DNA at four levels respectively. The result showed that, the best ISSR-PCR amplification system for Tulip was 1.0 mmol/L Mg^(2+), 0.15 mmol/L d NTPs, 0.25 U Taq polymerase, 0.2 μmol/L primer and 100 ng DNA template in a 25 μL reaction system. Annealing temperature was 58℃. The establishment of the ISSR-PCR reaction conditions could be useful for the further study on the genetic diversity of Tulip by using ISSR molecular marker techniques.

关 键 词：郁金香 ISSR-PCR 正交设计 单因素试验 

分 类 号：S682.263[农业科学—观赏园艺]