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利用HEGS(高效基因组扫描)/AFLP技术对大麦裸粒颖果基因(nud)进行高效精细定位。

Efficient fine mapping of the naked caryopsis gene ( nud) by HEGS (High Efficiency Genome Scanning)/AFLP in barley.

作者信息

Kikuchi S, Taketa S, Ichii M, Kawasaki S

机构信息

Faculty of Agriculture, Kagawa University, Ikenobe, Miki-cho, Kita-gun, 761-0795, Kagawa, Japan.

出版信息

Theor Appl Genet. 2003 Dec;108(1):73-8. doi: 10.1007/s00122-003-1413-y. Epub 2003 Aug 27.

Abstract

The hulled or naked caryopsis character of barley ( Hordeum vulgare L.) is an important trait for edibility and to follow its domestication process. A single recessive gene, nud, controls the naked caryopsis character, and is located on the long arm of chromosome 7H. To develop a fine map around the nud locus efficiently, the HEGS (High Efficiency Genome Scanning) electrophoresis system was combined with amplified fragment length polymorphism (AFLP). From bulked segregant analysis of 1,894 primer combinations, 12 AFLP fragments were selected as linked markers. For mapping, an F(2 )population of 151 individuals derived from a cross between Kobinkatagi (naked type) and Triumph (hulled type) was used. Seven AFLP markers were localized near the nud region. A fine map was developed with one-order higher resolution than before, along with the seven anchor markers. Among the seven linked AFLP markers (KT1-7), KT1, KT2 and KT6 were co-dominant, and the former two were detected for their single-nucleotide polymorphisms (SNPs) in the same length of fragments after electrophoresis with the non-denaturing gels of HEGS. The nud locus has co-segregated with KT3 and KT7, and was flanked by KT2 and KT4, at the 0.3-cM proximal and the 1.2-cM distal side, respectively. Four of these AFLP markers were converted into sequence-characterized amplified region (SCAR) markers, one of which was a dominant marker co-segregating with the nud gene.

摘要

大麦(Hordeum vulgare L.)的带壳或裸粒颖果特征是关乎可食性及追踪其驯化过程的重要性状。单个隐性基因nud控制裸粒颖果性状,该基因位于7H染色体长臂上。为高效构建nud基因座周围的精细图谱,将高效基因组扫描(HEGS)电泳系统与扩增片段长度多态性(AFLP)相结合。通过对1894对引物组合进行混合分离群体分析,选择了12个AFLP片段作为连锁标记。为进行图谱绘制,使用了由小滨香茅(裸粒类型)和胜利(带壳类型)杂交产生的151个个体的F2群体。7个AFLP标记定位在nud区域附近。利用7个锚定标记构建了分辨率比之前高一阶的精细图谱。在7个连锁AFLP标记(KT1 - 7)中,KT1、KT2和KT6是共显性的,其中前两个在使用HEGS非变性凝胶电泳后,在相同长度片段中检测到单核苷酸多态性(SNP)。nud基因座与KT3和KT7共分离,分别在KT2的0.3厘摩近端和KT4的1.2厘摩远端侧翼。这些AFLP标记中有4个被转化为序列特征化扩增区域(SCAR)标记,其中一个是与nud基因共分离的显性标记。

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