[1]陈名红,李玉,刘多,等.适于ISSR-PCR扩增的百合基因组DNA的提取[J].北方园艺,2012,36(24):108-110.
　CHEN Ming-hong,LI Yu,LIU Duo,et al.Extraction of Genomic DNA for ISSR-PCR Reaction in Lilium[J].Northern Horticulture,2012,36(24):108-110.

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适于ISSR-PCR扩增的百合基因组DNA的提取

《北方园艺》[ISSN:1001-0009/CN:23-1247/S] 卷: 第36卷期数: 2012年24期页码: 108-110 栏目: 出版日期: 2012-11-30

Title:: Extraction of Genomic DNA for ISSR-PCR Reaction in Lilium

文章编号:: 1001-0009（2012）24-0108-03

作者:: 陈名红¹; 2; 李玉¹; 刘多¹; 熊华斌¹; 李成云²; 1.云南民族大学民族药资源化学国家民委-教育部重点实验室，云南昆明 650031；
2.云南农业大学农业生物多样性与病虫害控制教育部重点实验室，云南昆明 650201

Author(s):: CHEN Ming-hong¹; 2; LI Yu1; LIU Duo¹; XIONG Hua-bin¹; LI Cheng-yun²; 1.Key Laboratory of Ethnic Medicine Resource Chemistry，State Ethnic Affairs Commission and Ministry of Education，Yunnan University of Nationalities，Kunming，Yunnan 650031;
2.Key Laboratory of the Ministry of Educational for Agro-biodiversity and Pests Control，Yunnan Agricultural University，Kunming，Yunnan 650201

关键词:: 百合; 基因组DNA; DNA提取; ISSR

Keywords:: Lilium; genomic DNA; DNA extraction; inter-simple sequence repeat

分类号:: S 682.2+9

文献标志码:: A

摘要:: 为了方便稳定地获得适合于ISSR-PCR扩增的高质量百合基因组DNA，以卷丹百合为试材，在传统的CTAB法基础上进行了改良和优化，筛选出较理想的DNA提取方法。结果表明：用改良CTAB法提取的百合基因组DNA电泳条带清晰，无降解现象，OD_260/OD₂₈₀值在1.7～2.0之间，用于ISSR-PCR扩增时其稳定性和可重复性都很好，说明DNA纯度和产率完全满足百合ISSR-PCR扩增分析的要求，为百合属植物遗传多样性分析和品种分子鉴别等研究奠定了基础。

Abstract:: In order to obtain high quality genomic DNA suitable for ISSR-PCR reaction conveniently and stably in Lilium，taking Lilium tigrinm ?as matrials,the course of DNA extraction was improved and optimized based on the traditional CTAB method and an ideal method was obtained here.The results showed that the OD₂₆₀/OD₂₈₀of genomic DNA extracted by the modified CTAB method ranged from 1.7 to 2.0 with clear electrophoresis strip and its stability and repeatability were very good when used for ISSR-PCR reaction，accounting for its purity and content were all up to the standards for ISSR-PCR analysis.The research laid the foundation for the study on genetic polymorphism analysis，cultivar identification，etc.of Lilium at the molecular level.

参考文献/References:

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[5]Tsumura Y，Ohba K，Straus S H.Diversity and inheritance of inter-simple sequence repeat polymorphisms in Douglas-fir (Pseudotsuga menziesii) and sugi (Cryptom eria japonica)[J].Theor Appl Genet,1996,92:40-45.

[6]Devarumath R M，Nandy S，Rani V.RAPD，ISSR and RFLP fingerprintings as useful markers to evaluate genetic integrity of micropropagated plants of three diploid and triploid elite tea clones representing Camellia sinensis(China type) and C.assamicassp.Assamica (Assam-India type)[J].Plant Cell Rep,2002,21:166-173.

[7]Fernandez M E，Figueiras A M，Benito C.The use of ISSR and RAPD markers for detecting DNA polymorphism，genotype identification and genetic diversity among barley cultivars with known origin[J].Theor App l Genet,2002,104:845-851.

[8]王关林,方宏筠.植物基因工程[M].北京:科学出版社,2002:742-744.

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备注/Memo

第一作者简介：陈名红（1974-），男，土家族，在读博士，副教授，研究方向为植物分子生物学。E-mail:cmhkm@yahoo.com.cn.

责任作者：李成云（1964-），男，博士，教授，研究方向为植物分子生物学。E-mail:li.chengyun@gmail.com.

基金项目：国家自然科学基金资助项目（30860161）；云南省教育厅科学研究基金资助项目（2011Y212）。

更新日期/Last Update: 2014-08-31