[1]练云,冯文强,雷晨芳,等.一种从大豆胞囊线虫中快速提取DNA的方法[J].大豆科学,2016,35(06):1043-1046.[doi:10.11861/j.issn.1000-9841.2016.06.1043]
　LIAN Yun,FENG Wen-qiang,LEI Chen-fang,et al.A Method for Rapid Extraction of DNA from Soybean Cyst Nematode[J].Soybean Science,2016,35(06):1043-1046.[doi:10.11861/j.issn.1000-9841.2016.06.1043]
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一种从大豆胞囊线虫中快速提取DNA的方法

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第35卷 期数: 2016年06期 页码: 1043-1046 栏目: 出版日期: 2016-11-20
Title:
A Method for Rapid Extraction of DNA from Soybean Cyst Nematode
作者:
练云1; 冯文强1; 2; 雷晨芳1; 李金英1; 武永康1; 王庭峰1; 陆莉莉1; 卢为国1
（1.河南省农业科学院 经济作物研究所/国家大豆改良中心郑州分中心/农业部黄淮海油料作物重点实验室，河南 郑州 450002； 2.山东农业大学 生命科学学院/作物生物学国家重点实验室，山东 泰安 271018）
Author(s):
LIAN Yun1;  FENG Wen-qiang1; 2;  LEI Chen-fang1;  LI Jin-ying1;  WU Yong-kang1;  WANG Ting-feng1;  LU Li-li1;  LU Wei-guo1
（1. Institute of Industrial Crops, Zhengzhou National Subcenter for Soybean Improvement/Key Laboratory of Oil Crops in Huang-huaihai Plains, Ministry of Agriculture, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China; 2.State Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural University, Taian 271018, China）
关键词:
大豆; 大豆胞囊线虫; DNA提取; 生理小种
Keywords:
Soybean;  Soybean cyst nematode;  DNA extraction;  Race
DOI:
10.11861/j.issn.1000-9841.2016.06.1043
文献标志码:
A
摘要:
由于大豆胞囊线虫（soybean cyst nematode，SCN）特殊的生长环境，用细菌专用DNA试剂盒提取方法提取DNA的质量不理想，而且耗时长，本文介绍了一种快速提取线虫DNA的方法。通过比较细菌专用DNA试剂盒提取法、质粒试剂盒提取法和CTAB法，发现利用质粒提取试剂盒，改良提取步骤，可以快速的提取质量较好的线虫DNA，rDNA-ITS区域序列扩增可有效扩增出目的基因，该方法可以直接用于线虫基因获取试验。
Abstract:
In this paper, we reported a method for rapid extraction of DNA from soybean cyst nematode(SCN). It is difficult and time consuming to obtain high quality DNA by using bacterial genomic DNA extraction kit for the special growth environment of SCN.Three methods for extraction of DNA were compared in this study, including Bacterial genomic DNA extraction kit, San Prep column plasmid DNA Kit and CTAB method.Results showed that DNA isolated by improved San Prep column plasmid DNA Kit was high quality with time saving. We verified the quality by amplification sequence effectively from rDNA-ITS region. The method could be used for DNA obtaining from cyst and the later molecular manipulations of genes cloning.

参考文献/References:

［1］Peng D L, Peng H, Wu D Q, et al. First report of soybean cyst nematode (Heterodera glycines) on soybean from Gansu and Ningxia China［J］. Plant Disease, 2015, 100(1): 229。

［2］Wrather J A, Koenning S R. Estimates of disease effects on soybean yields in the United States 2003 to 2005［J］. Journal of nematology, 2006, 38(2): 173-180
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［4］Zheng J W, Subbotin S A, Waeyenberge L, et al. Molecular characterisation of Chinese Heteroderaglycines and H. Avenae- populations based on RFLPs and sequences of rDNA-ITS regions［J］. Russian Journal of Nematology, 2000, 8(2): 109-113.
［5］王暄, 梁中伟, 裴世安, 等. 江苏省小麦禾谷孢嚢线虫群体核糖体DNA-ITS区序列分析［J］.南京农业大学学报, 2010, 33(6): 55-62. (Wang X, Liang Z W, Pei S A, et al. Sequences analysis of rDNA-ITS region of cereal cyst nematodes on wheat from Jiangsu Province［J］. Journal of Nanjing Agricultural University, 2010, 33(6): 55-62.)
［6］牛雯雯, 王暄, 李红梅, 等. 基于线粒体DNA-COI序列的禾谷孢嚢线虫和菲利普孢嚢线虫双重PCR检测［J］. 中国农业科学, 2016, 49(8): 1499-1509. (Niu W W, Wang X, Li H M, et al. Duplex-PCR detection for Heteroderaavenae and Heteroderafilipjevi-based on mtDNA-COL sequences［J］. Scientia Agricultura Sinica, 2016, 49(8): 1499-1509.)
［7］郑雅楠, 段玉玺, 孙晶双, 等.大豆胞囊线虫不同虫态DNA提取方法及ITS-rRNA扩增［J］.植物保护学报, 2009, 36(5): 477-478. (Zheng Y N, Duan Y X, Sun J S, et al. DNA extraction methods and ITS-rRNA analysis from different growth stages of Heterodera glycine.［J］.Acta Phytophylacica Sinica, 2009, 36(5): 477-478.）
［8］练云, 魏荷, 王金社, 等. 影响大豆胞囊线虫生理小种鉴定因素探讨［J］. 分子植物育种, 2015, 13(6): 1259-1264 (Lian Y, Wei H, Wang J S, et al. A study on the factors that influence the race-identification of soybean cyst nematode［J］. Molecular Plant Breeding, 2015, 13(6): 1259-1264。）

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备注/Memo

基金项目：国家自然科学基金（31371652）；河南省农业科学院科研发展专项资金（20137905）；国家现代农业产业技术体系建设专项（CARS-004）；河南省科技攻关项目（162102110141）。

第一作者简介：练云（1978-），女，博士，副研究员，主要从事大豆遗传育种研究。E-mail: lianyun262@126.com。
通讯作者：卢为国（1971-），男，博士，研究员，主要从事大豆遗传育种研究。E-mail: 123bean@163.com。

更新日期/Last Update: 2016-12-09