[1]胡兴华,李景剑,王燕,等.茶花品种ISSR指纹图谱反应体系建立与优化[J].北方园艺,2014,38(22):84-88.
　HU Xing-hua,LI Jing-jian,WANG Yan,et al.Reaction System Optimization and Primer Screening of ISSR Fingerprint Map for Camellia Cultivars[J].Northern Horticulture,2014,38(22):84-88.
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茶花品种ISSR指纹图谱反应体系建立与优化

《北方园艺》[ISSN:1001-0009/CN:23-1247/S] 卷: 第38卷 期数: 2014年22期 页码: 84-88 栏目: 出版日期: 2014-11-30
Title:
Reaction System Optimization and Primer Screening of ISSR Fingerprint Map for Camellia Cultivars
文章编号:
1001-0009（2014）22-0084-05
作者:
胡兴华; 李景剑; 王燕; 黄仕训
广西壮族自治区中国科学院 广西植物研究所，广西 桂林 541006
Author(s):
HU Xing-hua; LI Jing-jian; WANG Yan; HUANG Shi-xun
Guangxi Institute of Botany,Guangxi Zhuang Autonomous Region and the Chinese Academy of Sciences,Guilin,Guangxi 541006
关键词:
茶花品种; 分子指纹图谱; ISSR体系优化; 引物筛选
Keywords:
Camellia cultivar; molecular fingerprint map; ISSR reaction system optimizing; primer screening
分类号:
S 796
文献标志码:
A
摘要:
以30个茶花品种为试材，采用5因素4水平正交实验以及单因素优化试验方法，研究Mg2+浓度、dNTP浓度、引物浓度、Taq DNA聚合酶浓度和模板DNA浓度对ISSR-PCR指纹图谱条带清晰度的影响，以进行茶花品种ISSR-PCR反应体系优化及引物筛选。结果表明：茶花品种ISSR-PCR最适扩增条件为25 μL反应体系中，Mg2+3.0 mmol/L、dNTPs 0.2 mmol/L、引物0.3 mmol/L、Taq DNA聚合酶0.5 U、模板DNA 80 ng以及52.1℃退火温度；试验从100个ISSR引物中筛选出12个适用引物；并对12个ISSR引物的多态性和稳定性进行了检验。
Abstract:
Taking 30 Camellia cultivars as test materials,the effects of concentrations of Mg2+,dNTP,Primers,Taq DNA polymerase,template DNA,and the annealing temperature for primers on the ISSR-PCR molecular fingerprint map were examined by orthogonal design test and single factor modified.The results showed that the optimal ISSR-PCR conditions in the experiments were as following:in 25 μL reaction system containing Mg2+ 3.0 mmol/L、dNTPs 0.2 mmol/L、primers 0.3 mmol/L、Taq DNA polymerase 0.5 U、template DNA 80 ng and annealing temperature 52.1℃.Base on this optimal reaction system,100 ISSR primers were used to screen for the suitable primers with 1 cultivar samples,of which 12 ISSR primers with high resolution and multiple polymorphic bands were screened.Further amplification of 30 Camellia japonica cultivars using these 12 primers demonstrated that this optimized reaction system had specialties of good repeatability and stability.

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备注/Memo

第一作者简介：胡兴华(1974-)，男，博士，副研究员，研究方向为植物资源保护利用及动植物关系。E-mail:huxh773@163.com.

责任作者：黄仕训(1966-)，男，本科，研究员，研究方向为珍稀濒危植物资源保护利用。E-mail:hsx@gxib.cn.基金项目：广西科技攻关资助项目（桂科攻10100012-4）。
收稿日期：2014-07-21

更新日期/Last Update: 2014-12-12