干旱区生态环境知识资源中心(Arid): 小麦指纹图谱数据库的建立及SSR分子标记试剂盒的研发

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	小麦指纹图谱数据库的建立及SSR分子标记试剂盒的研发
其他题名	Development of a Fingerprinting Database and Assembling an SSR Reference Kit for Genetic Diversity Analysis of Wheat
	李根英; Susanne Dreisigacker; Marilyn L Warburton; 夏先春; 何中虎; 孙其信
来源期刊	作物学报
ISSN	0496-3490
出版年	2006
卷号	32 期号:12 页码:1771-1778
中文摘要	本研究以国际玉米小麦改良中心(CIMMYT)、国际干旱地区农业研究中心(ICARDA)、法国Agropolis研究所和中国农业科学院作物科学研究所提供的数据为基础,建立了包括134个SSR引物、2 457个普通小麦基因型的指纹图谱数据库.利用荧光标记法的分析结果,用代表性基因型在某一位点的扩增片段作为银染法的读带依据,开发了SSR分子标记试剂盒,包括46对SSR引物及其PCR反应程序、代表性基因型的DNA样品、592个等位变异的代表性基因型清单及试剂盒使用说明等.该数据库的建立和试剂盒的开发,为利用SSR分子标记技术进行小麦种质遗传多样性研究,实现小麦遗传资源和信息资源全球共享提供了重要工具和技术平台.
英文摘要	Understanding of the current and expanded genetic diversity is very important for raising the yield of wheat. Genetic diversity based on molecular markers has been studied in plants for over two decades. SSR is the currently most popular marker system in wheat. In order to utilize the diversity held in NARS (National Agricultural Research Station) and CGIAR (Consultative Group on International Agricultural Research) germplasm collections, one of the GCP's (Generation Challenge Program) premier capacity building activities is to build databases that contain traditional and molecular data on germplasm so that scientists all over the world can access information with relevance to their region on traits, genes, and sequences. In the present study, a fingerprinting database was established containing 134 SSR primers and 2457 wheat genotypes with the data from CIMMYT and three collaborators: ICARDA, Agropolis, and CAAS. On the base of the database, a SSR reference kit for wheat genetic diversity analysis was developed, which will facilitate the use of this data in new projects and cross-laboratory comparisons. In total, 46 SSR primers with comparatively high polymorphism were selected as the reference markers to constitute the standard allele kit, 334 genotypes fingerprinted within the GCP tier 1 project "Genotyping a composite germplasm set in wheat" were chosen to represent the SSR allele kit consisting of 794 alleles amplified by 46 SSR markers. Genotypes were originally selected and UNA extracted by CIMMYT and 3 additional collaborators: ICARDA, Agropolis, and CAAS. These 334 genotypes, when taken as a group, amplify eveiy allele seen for the wheat genotypes for all the 46 SSRs. The Genotypes were originally amplified with 26 SSRs at CIMMYT, with 8 SSRs at INRA, France, and with 12 SSRs at CAAS. Genotypes were amplified with all SSRs al CIMMYT again to confirm the results. For reamplification at CIMMYT, forward primers were labeled at the 5' end with either one of three phosphoramidite fluorescent dyes 6-carboxyflouresein (6-Fam), tetrachloro-6-carboxyflouresein (Tet) or hexachloro-6-carboxyflouresein (Hex). PCR-reactions were carried out in an MJ-Research thermocycler model FIX225. Amplification products were separated on an ABI-Prism Sequencer?377 using 4.5% polyacrylamide denaturing gels. Fragment sizes were calculated semi-aulomatically with the computer software GeneScan 3.1 by comparing to fragments of an internal size standard (GeneScan 350 or 500) labeled with N,N,N,N,-tetrarnelhyl-6-oarboxyrhodamine (Tamra). GeneScan fragments were assigned to alleles using the categoiy function of the softwai'e Genotyper 2.1. The two genotypes, Opata and Synthetic were run in each gel as controls. Finally, an SSR reference kit was assembled which includes 46 pairs of SSR primers, protocols (PCR condition, detection of polymorphisms, etc), description of polymorphism in the reference samples, reference DNA samples for a complete range of repeatable 592 alleles, methods for comparing new to preexisting data, and classification of various genotypes. The database and the reference kit will provide a powerful tool for genetic diversity studies of wheat germplasm worldwide.
中文关键词	小麦 ; 指纹图谱 ; 数据库 ; SSR引物 ; 标准基因型 ; 试剂盒
英文关键词	Wheat Fingerprinting Database SSR marker Reference genotype Reference kit
类型	Article
语种	中文
收录类别	CSCD
WOS类目	Agriculture
CSCD记录号	CSCD:2622151
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/378527
作者单位	李根英, 中国农业大学农学与生物技术学院, 北京 100094, 中国.; 孙其信, 中国农业大学农学与生物技术学院, 北京 100094, 中国.; 夏先春, 中国农业科学院作物科学研究所/国家农作物基因资源与基因改良重大科学工程, 国家小麦改良中心, 北京 100081, 中国.; 何中虎, 中国农业科学院作物科学研究所/国家农作物基因资源与基因改良重大科学工程, 国家小麦改良中心, 北京 100081, 中国.
推荐引用方式 GB/T 7714	李根英,Susanne Dreisigacker,Marilyn L Warburton,等. 小麦指纹图谱数据库的建立及SSR分子标记试剂盒的研发[J],2006,32(12):1771-1778.
APA	李根英,Susanne Dreisigacker,Marilyn L Warburton,夏先春,何中虎,&孙其信.(2006).小麦指纹图谱数据库的建立及SSR分子标记试剂盒的研发.作物学报,32(12),1771-1778.
MLA	李根英,et al."小麦指纹图谱数据库的建立及SSR分子标记试剂盒的研发".作物学报 32.12(2006):1771-1778.