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小麦中国春背景中长穗偃麦草染色体特异分子标记研究

发表时间:2014-03-15  浏览量:1473  下载量:463
全部作者: 陈士强,朱雪,张超,曹文广,GEORGEFedak,陈秀兰,陈建民
作者单位: 扬州大学生物科学与技术学院;江苏里下河地区农业科学研究所;加拿大东部谷物与油料研究中心
摘 要: 根据大麦反转录转座子BARE-1的长末端重复序列(long terminal repeated,LTR)保守区域及水稻反转录转座子RIRE-1的反转录酶中心序列自行设计11条引物,组成52个引物组合,对中国春(Chinese spring,CS)-长穗偃麦草二体附加系及代换系、普通小麦CS和二倍体长穗偃麦草基因组DNA扩增筛选,得到145条覆盖长穗偃麦草所有7条染色体的特异简单重复序列间区(inter-simple sequence repeat, ISSR)、反转录转座子扩增多态性间区(inter-retrotransposon amplified polymorphism, IRAP)及反转录转座子微卫星间多态性(retrotransposon microsatellite amplified polymorphism, REMAP)标记。任意选择60个特异片段,经扩增、克隆、测序,并与已知小麦序列进行比对,发现其中34个片段的序列为长穗偃麦草特异性序列。根据其DNA序列设计新引物,将其中12个成功转化为长穗偃麦草染色体特异序列特异性扩增区(sequence characterized amplified region, SCAR)标记。结果表明:根据ISSR,IRAP及REMAP技术发展的12个SCAR标记具有很好的稳定性与重复性,可用于小麦与长穗偃麦草远缘杂交后代中长穗偃麦草染色质的快速检测和特定染色体的快速跟踪检测。
关 键 词: 遗传学;长穗偃麦草;染色体特异分子标记;序列特异性扩增区
Title: Research of chromosome specific molecular markers for Thinopyrum elongatum in a wheat background
Author: CHEN Shiqiang, ZHU Xue, ZHANG Chao, CAO Wenguang, GEORGE Fedak, CHEN Xiulan, CHEN Jianmin
Organization: College of Bioscience and Biotechnology, Yangzhou University; Lixiahe Region Agricultural Scientific Research Institute of Jiangsu; Eastern Cereal and Oilseed Research Centre
Abstract: 11 primers were synthesized according to the reverse transcriptase and long terminal repeated (LTR) conserved regions of retrotransposons BARE-1 from barley and RIRE-1 from rice. 52 pairs of primer combinations based on these 11 primers were used for DNA amplification of Chinese spring (CS)-Thinopyrum elongatum addition lines, substitution lines plus the two parents. The results showed that 145 specific framents of inter-simple sequence repeat (ISSR), inter-retrotransposon amplified polymorphism (IRAP) and retrotransposon microsatellite amplified polymorphism (REMAP) were obtained which distributed over all the 7E genome chromosomes of Th. elongatum. 60 specific fragments of ISSR, IRAP and REMAP were randomly selected for cloning and sequencing. 34 sequences were found not to be homologous with wheat sequences, which were considered to be the specific sequences of Th. elongatum. Thirty-four primers acording to the 34 specific sequences were synthesized, and 12 chromosome-specific markers of Th. elongatum were obtained which were furtherly converted into sequence-characterized amplified regions (SCAR) markers. The results indicated that ISSR, IRAP and REMAP techniques can be used to develop chromosome-specific SCAR markers with good stability and repeatability. These specific SCAR markers can now be used to detect and identify Th.elongatum chromosomes in a wheat background and possibly even some introgressed segments.
Key words: genetics; Thinopyrum elongatum; chromosome-specific molecular markers; sequence characterized amplified region
发表期数: 2014年3月第5期
引用格式: 陈士强,朱雪,张超,等. 小麦中国春背景中长穗偃麦草染色体特异分子标记研究[J]. 中国科技论文在线精品论文,2014,7(5):476-486.
 
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