[1]陈丽丽,王明玖,何丽君,等.野生大豆ISSR体系的优化及其在远缘杂交后代鉴定中的利用[J].大豆科学,2013,32(04):459-466.[doi:10.11861/j.issn.1000-9841.2013.04.0459]
　CHEN Li-li,WANG Ming-jiu,HE Li-jun,et al.Optimization for ISSR Reaction System of Wild Soybean and Its Utilization in Distant Hybrid Identification[J].Soybean Science,2013,32(04):459-466.[doi:10.11861/j.issn.1000-9841.2013.04.0459]
点击复制

野生大豆ISSR体系的优化及其在远缘杂交后代鉴定中的利用

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第32卷 期数: 2013年04期 页码: 459-466 栏目: 出版日期: 2013-08-25
Title:
Optimization for ISSR Reaction System of Wild Soybean and Its Utilization in Distant Hybrid Identification
作者:
?陈丽丽1; 王明玖1; 何丽君2; 王连生3
?1.内蒙古农业大学 生态环境学院，内蒙古 呼和浩特 010019；2.内蒙古农业大学 农学院，内蒙古 呼和浩特 010019；3.绿源植物生物质开发利用研究所，内蒙古 通辽 028000
Author(s):
?CHEN Li-li1; WANG Ming-jiu1; HE Li-jun2; WANG Lian-sheng3
?1.College of Ecology and Environmental Science,Inner Mongolia Agricultural University,Huhhot 010019,China;2.College of Agronomy,Inner Mongolia Agricultural University,Huhhot 010019,China;3.LyuYuan Plant Biomass Development and Utilization Institute,Tongliao 028000,China
关键词:
野生大豆; 正交设计; ISSR; 杂种鉴定
分类号:
S565.1
DOI:
10.11861/j.issn.1000-9841.2013.04.0459
文献标志码:
A
摘要:
??? ?以内蒙古野生大豆为材料，运用5因素5水平正交试验设计对ISSRPCR反应体系中DNA模板、Taq酶、引物、dNTPs和Mg2+用量进行优化试验，之后利用优化后的ISSR-PCR体系对内蒙古野生大豆及其与栽培大豆的杂交后代进行遗传关系研究。正交试验结果表明，各因素对野生大豆ISSR-PCR反应影响大小依次为Taq酶＞Mg2+＞引物＞模板DNA=dNTPs，最佳反应体系为模板DNA 50 ng、Taq酶0.5 U、引物（10 μmol·L-1）1.0 μL、Mg2+（25 mmol·L-1）2.0 μL及dNTPs（25 mmol·L-1 each）3.0 μL，2 μL 10×PCR buffer、ddH2O补足至20 μL。以10个引物对供试材料进行扩增，共得到条带109个，多态性百分率为60.23%，其中引物S23可以将所有供试材料进行区分。基于ISSR标记的聚类分析将13个供试材料划分为3类，S001、S002、2、5、8-1和8-4为第一类，29、SF、NY、ZS、0004和PZ为第二类，Y为第三类。
Abstract:
?Genomic DNA of Glycine soja was tested as the template,optimization experiments on some important parameters affecting ISSR-PCR amplification were conducted to establish an optimum system suitable for ISSRPCR of hybrids of Glycine soja×Glycine max.Orthogonal design was used to optimize ISSR amplification system of the hybrids with five factors(including DNA template,Taq polymerase,primer,dNTP and Mg2+)at five levels,respectively.Then using the best system to study the genetic relationship among the lines.According to the results of orthogonal test,the order of factors affecting the ISSRPCR reaction of Glycine soja was:Taq polymerase＞Mg2+)＞primer＞DNA template=dNTPs.A suitable ISSRPCR reaction system was established,its total volume was 20 μL,containing 50 ng DNA template,0.5 U Taq polymerase,1.0 μL primer(10 μmol·L-1),3.0 μL dNTPs(25 mmol·L-1 each),2.0 μL Mg2+(25 mmol·L-1) and 2 μL 10×PCR buffer.A total of 109 loci were amplified with 10 primers,the polymorphic loci percentage was 60.23%,the primer S23 could distinguish all the tested materials.All the tested materials were divided into three groups using cluster analysis.S001,S002,2,5,8-1 and 8-4 were divided into the first group,29,SF,NY,ZS,0004 and PZ were divided into the second group,Y was the third group.

相似文献/References:

[1]高越,刘辉,陶波.抗草甘膦野生大豆筛选及其抗性生理机制研究[J].大豆科学,2013,32(01):76.[doi:10.3969/j.issn.1000-9841.2013.01.018]
　GAO Yue,LIU Hui,TAO Bo.Screening and Physiological Mechanisms of Resistance to Glyphosate in Wild Soybeans(Glycine soja)[J].Soybean Science,2013,32(04):76.[doi:10.3969/j.issn.1000-9841.2013.01.018]
[2]王军卫,侯立江,李? 登,等.野生大豆紫色酸性磷酸酶PAP1基因的克隆及分析[J].大豆科学,2013,32(05):596.[doi:10.11861/j.issn.1000-9841.2013.05.0596]
　WANG Jun-wei,HOU Li-jiang,LI Deng,et al.Cloning and Sequence Analysis of Purple Acid Phosphotase PAP1 Gene in Wild Soybean[J].Soybean Science,2013,32(04):596.[doi:10.11861/j.issn.1000-9841.2013.05.0596]
[3]王军卫,侯立江,李 登,等. 野生大豆紫色酸性磷酸酶PAP1基因的克隆及分析[J].大豆科学,2013,32(05):596.
　WANG Jun-wei,HOU Li-jiang,LI Deng,et al. Cloning and Sequence Analysis of Purple Acid Phosphotase PAP1 Gene in Wild Soybean[J].Soybean Science,2013,32(04):596.
[4]王丽燕.硅对野生大豆幼苗耐盐性的影响及其机制研究[J].大豆科学,2013,32(05):659.[doi:10.11861/j.issn.1000-9841.2013.05.0659]
　WANG Li-yan.Effects of Silicon on Salt Tolerance of Glycine soja Seedlings and Its Mechanism[J].Soybean Science,2013,32(04):659.[doi:10.11861/j.issn.1000-9841.2013.05.0659]
[5]郑世英,萧蓓蕾,金桂芳.NaCl胁迫对野生大豆和栽培大豆叶绿素及光合特性的影响[J].大豆科学,2013,32(04):486.[doi:10.11861/j.issn.1000-9841.2013.04.0486]
　ZHENG Shi-ying,XIAO Bei-lei,JIN Gui-fang.Effect of NaCl Stress on Chlorophyll Content and Photosynthetic Characteristics of Glycine soja and Glycine max[J].Soybean Science,2013,32(04):486.[doi:10.11861/j.issn.1000-9841.2013.04.0486]
[6]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.[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(04):310.[doi:10.11861/j.issn.1000-9841.2013.03.0310]
[7]徐艳平,胡翠美,张文会,等.干旱胁迫对野生大豆幼苗光合作用相关指标的影响[J].大豆科学,2013,32(03):341.[doi:10.11861/j.issn.1000-9841.2013.03.0341]
　XU Yan-ping,HU Cui-mei,ZHANG Wen-hui,et al.Effect of Simulated Drought Stress on Photosynthesis Related Indexes at Seedling Stage of Wild Soybeans[J].Soybean Science,2013,32(04):341.[doi:10.11861/j.issn.1000-9841.2013.03.0341]
[8]胡卫静,何丽君,何劲莉,等.NaCl胁迫对野生与栽培大豆杂交后代株系生理指标的影响[J].大豆科学,2013,32(03):349.[doi:10.11861/j.issn.1000-9841.2013.03.0349]
　HU Wei-jing,HE Li-jun,HE Jin-li,et al.Effects of NaCl Stress on Physiological Characters of Soybean Hybrids from Glycine max × Glycine soja[J].Soybean Science,2013,32(04):349.[doi:10.11861/j.issn.1000-9841.2013.03.0349]
[9]王 旻,梁 玉,王欣欣,等.即墨野生大豆主要成分及其营养价值分析[J].大豆科学,2013,32(03):355.[doi:10.11861/j.issn.1000-9841.2013.03.0355]
　WANG Min,LIANG Yu,WANG Xin-xin,et al.Assessment on Nutritional Compositions and Value of Jimo Wild Soybean[J].Soybean Science,2013,32(04):355.[doi:10.11861/j.issn.1000-9841.2013.03.0355]
[10]程鹏,徐鹏飞,范素杰,等.野生大豆接种大豆疫霉根腐病菌后过氧化物酶（POD）活性变化[J].大豆科学,2013,32(02):197.[doi:10.3969/j.issn.1000-9841.2013.02.013]
　CHENG Peng,XU Peng-fei,FAN Su-jie,et al.Response of POD Activity in Glycine soja ?Inoculated by Phytophthora sojae[J].Soybean Science,2013,32(04):197.[doi:10.3969/j.issn.1000-9841.2013.02.013]

备注/Memo

?基金项目内蒙古应用技术研究与开发项目（20091402）；内蒙古自然科学基金（2010MS0511）。

更新日期/Last Update: 2014-02-14