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簇毛麦基因组特异DNA序列标记的建立和应用

Development and Application of A Dasypyrum Genome-specific DNA Marker

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【作者】 迟世华杨足君冯娟周建平刘成任正隆

【Author】 CHI Shi-hua,YANG Zu-jun,FENG Juan,ZHOU Jian-ping,LIU Cheng,REN Zheng-long(College of Life Science and Technology,University of Electronic Science and Technology,Chengdu,Sichuan 610054,China)

【机构】 电子科技大学生命科学与技术学院电子科技大学生命科学与技术学院 四川成都610054四川成都610054

【摘要】 为建立簇毛麦基因组特异DNA序列的分子标记,以普通小麦中国春、绵阳11、川农17和R111以及多年生簇毛麦、硬粒小麦-多年生簇毛麦双二倍体TDB-3为材料,用120条随机引物进行RAPD分析,筛选出一个多年生簇毛麦的特异性RAPD片段,克隆测序表明该片段全长为937 bp(记为OPM2937)。以OPM2937的DNA序列为基础设计了一对特异引物,并用该特异引物对多年生簇毛麦、二倍体簇毛麦以及中国春-二倍体簇毛麦附加系CSDA1V~CSDA7V与小麦-多年生簇毛麦衍生后代进行了扩增,结果表明,凡具有簇毛麦染色体的材料均可扩增出目标DNA片段,而不具簇毛麦染色体的材料均未能得到扩增。将OPM2937在NCBI中进行序列比对,发现它是一个新的序列,说明OPM2937为簇毛麦基因组的一个特异序列,该DNA序列标记可用于快速跟踪检测小麦背景中的簇毛麦染色体。

【Abstract】 Random amplified polymorphic DNA(RAPD) analysis was performed on common wheat lines Chinese Spring,Mianyang11,Chuannong 17,and R111,as well as Dasypyrum breviaristatum,Triticum durum-D.breviaristatum amphiploid with 120 random primers,and a D.breviaristatum specific RAPD fragment was obtained.The fragment of 937 bp was revealed by cloning and sequencing,which was thus designated as OPM2937.Based on the sequence of OPM2937,A pair of primer was developed,and used to amplify the D.breviaristatum,D.villosum,T.aestivum Chinese Spring-D.villosum addition lines CSDA1V to CSDA7V,and wheat-D.breviaristatum derivatives.The results indicated that the target product of 900bp was produced only from the lines with the Dasypyrum chromosomes,while it was not observed in the lines belong to the common wheat.The OPM2937 was a new sequence when Blast it in NCBI gene bank,which further confirmed that it was a Dasypyrum genome-specific sequence. It could be used a novel molecular marker for detecting and tracing the Dasypyrum chromatin introduced in wheat background.

【基金】 国家自然科学基金项目(30170502);国家自然科学基金项目(30170579)
  • 【文献出处】 麦类作物学报 ,Journal of Triticeae Crops , 编辑部邮箱 ,2006年04期
  • 【分类号】S512.9
  • 【被引频次】11
  • 【下载频次】85
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