[1]李 强,苏二虎,高聚林,等.大豆CDDP分子标记技术体系的优化、验证及引物筛选[J].大豆科学,2013,32(03):310-315.[doi:10.11861/j.issn.1000-9841.2013.03.0310]
　LI Qiang,SU Er-hu,GAO Ju-lin,et al.Optimization,Validation and Primer Screening of CDDP Molecular Marker System in Soybean[J].Soybean Science,2013,32(03):310-315.[doi:10.11861/j.issn.1000-9841.2013.03.0310]
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大豆CDDP分子标记技术体系的优化、验证及引物筛选

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第32卷 期数: 2013年03期 页码: 310-315 栏目: 出版日期: 2013-06-25
Title:
Optimization,Validation and Primer Screening of CDDP Molecular Marker System in Soybean
作者:
?李 强1; 2; 苏二虎2; 高聚林1; 李丽君3; 谢 岷1; 于晓芳1
?1.内蒙古农业大学 农学院，内蒙古 呼和浩特 010019；2.内蒙古自治区农牧业科学院 玉米研究所，内蒙古 呼和浩特 010031；3.内蒙古自治区种子管理站，内蒙古 呼和浩特 010010
Author(s):
?LI Qiang1; 2;  SU Er-hu2;  GAO Ju-lin1;  LI Li-jun3;  XIE Min1;  YU Xiao-fang1
?1.College of Agronomy,Inner Mongolia Agricultural University,Huhhot 010019,China;2.Maize Research Institute,Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences,Huhhot 010031,China;3.Seeds Management Station of Inner Mongolia,Hohhot 010010,China
关键词:
大豆; CDDP; 正交设计; 体系验证; 引物筛选
Keywords:
Soybean; CDDP; Orthogonal design; System validation; Primers screening
分类号:
S565.1
DOI:
10.11861/j.issn.1000-9841.2013.03.0310
文献标志码:
A
摘要:
?CDDP分子标记是一种新型目的基因分子标记技术。采用L16（45）正交试验设计对影响大豆CDDP-PCR反应的Mg2+浓度、Taq DNA聚合酶用量、引物浓度、dNTPs浓度和模板DNA用量等因素进行优化。优化后的大豆CDDP-PCR体系为：Mg2+浓度2.0 mmol·L-1、Taq聚合酶用量1.5 U、引物浓度0.375 μmol·L-1、dNTPs浓度0.3 mmol·L-1、DNA模板用量40 ng。该反应体系在16个大豆品种的验证中表现稳定可靠。利用大豆品种吉育75、本地黑豆及其9株F2代对该反应体系进行了初步的遗传验证，结果显示，杂交后代植株中出现了双亲的位点及亲本位点的缺失。并从21条引物中筛选出条带清晰、多态性较好的13条引物。该反应体系的建立为大豆种质遗传多样性分析、遗传连锁图谱构建及分子标记辅助育种奠定了基础。
Abstract:
?Conserved DNADerived Polymorphism(CDDP)is a novel gene targeted marker technique.L16（45）orthogonal experiment design were applied to optimize CDDP-PCR amplification system of soybean in five factors of Mg2+, Taq DNA polymerase,primer,dNTPs and template DNA.The suitable reaction system was obtained,that was 20 μL containing 2.0 mmol·L-1 Mg2+,1.5 U Taq polymerase,0.375 μmol·L-1 primer,0.3 mmol·L-1 dNTPs,40 ng DNA template.The optimized CDDPPCR system was tested on sixteen soybeans,and the result was stable and reliable.Amplifications and genetic verification were carried out on Jiyu 75,local black soybean and 9 F2 progenies from Jiyu 75×local black soybean.The results showed that the site in all progenies were derived from their parents and had their parents’ deletion.From the 21 primer combinations tested,thirteen were selected with clear band patterns and abundant polymorphism.The system provides the basis for evaluation of genetic diversity,construction of genetic linkage map and molecular marker assisted breeding for the soybean.

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更新日期/Last Update: 2014-03-04