细叶桉ISSR-PCR体系的建立与优化  

Establishment and Optimization of ISSR-PCR System of Eucalyptus Teretticornis

作　　者：田华[1] 张党权[1] 谭晓风[1] 黄青云[1] 邓顺阳[1] 樊绍刚[1] 谷振军[1] 

机构地区：[1]中南林业科技大学经济林育种与栽培国家林业局重点实验室,湖南长沙410004

出　　处：《经济林研究》2009年第1期13-16,79,共5页Non-wood Forest Research

基　　金：湖南省长沙市科技计划重点项目"转基因抗寒桉树新品种的培育与应用"(K069083-22)

摘　　要：细叶桉是我国近年来成功引种的耐寒桉树种类之一,但其种源关系难以区分,不利于后续新品种的培育。为给后续的分子水平上鉴定细叶桉种源关系打基础,以细叶桉叶片基因组DNA为材料,分析了能够影响细叶桉ISSR-PCR的主要参数如模板DNA、引物浓度、dNTPs、Mg2+浓度、TaqDNA聚合酶用量、退火温度等6个因素对扩增结果的影响情况,建立了适用于细叶桉ISSR-PCR的反应体系和扩增条件。Eucalyptus teretticornis is one cold-resistant species of Eucalyptus which had been successfully introduced into China in recent years, but its provenance relations were difficult to distinguish, which was disadvantageous to the breeding of follow-up new variety. In order to lay the foundation for identification of Eucalyptus teretticornis provenance relations on molecular level, effect of six ISSR-PCR parameters of Eucalyptus teretticornis on amplification results were analyzed using genomic DNA of Eucalyptus teretticornis leaves, such as dosage of template DNA and TaqDNA polymerase, concentration of primer, Mg^2＋ and dNTPs, and annealing temperature. Suitable reaction system and amplification conditions of Eucalyptus teretticornis ISSR-PCR were finally established.

关 键 词：细叶桉 ISSR—PCR 反应体系 扩增条件 

分 类 号：S727.1[农业科学—林木遗传育种]