Pourkheirandish M, Wicker T, Stein N, Fujimura T, Komatsuda T
National Institute of Agrobiological Sciences, Tsukuba, 305-8602, Japan.
Theor Appl Genet. 2007 May;114(8):1357-65. doi: 10.1007/s00122-007-0522-4. Epub 2007 Mar 21.
In cultivated barley (Hordeum vulgare ssp. vulgare), six-rowed spikes produce three times as many seeds per spike as do two-rowed spikes. The determinant of this trait is the Mendelian gene vrs1, located on chromosome 2H, which is syntenous with rice (Oryza sativa) chromosomes 4 and 7. We exploited barley-rice micro-synteny to increase marker density in the vrs1 region as a prelude to its map-based cloning. The rice genomic sequence, covering a 980 kb contig, identified barley ESTs linked to vrs1. A high level of conservation of gene sequence was obtained between barley chromosome 2H and rice chromosome 4. A total of 22 EST-based STS markers were placed within the target region, and the linear order of these markers in barley and rice was identical. The genetic window containing vrs1 was narrowed from 0.5 to 0.06 cM, which facilitated covering the vrs1 region by a 518 kb barley BAC contig. An analysis of the contig sequence revealed that a rice Vrs1 orthologue is present on chromosome 7, suggesting a transposition of the chromosomal segment containing Vrs1 within barley chromosome 2H. The breakdown of micro-collinearity illustrates the limitations of synteny cloning, and stresses the importance of implementing genomic studies directly in the target species.
在栽培大麦(Hordeum vulgare ssp. vulgare)中,六棱穗每穗产生的种子数量是二棱穗的三倍。该性状的决定因素是位于2H染色体上的孟德尔基因vrs1,它与水稻(Oryza sativa)的4号和7号染色体存在共线性。我们利用大麦 - 水稻的微共线性来增加vrs1区域的标记密度,作为基于图谱克隆该基因的前奏。覆盖980 kb重叠群的水稻基因组序列鉴定出了与vrs1连锁的大麦EST。在大麦2H染色体和水稻4号染色体之间获得了高度保守的基因序列。总共22个基于EST的STS标记被定位在目标区域内,这些标记在大麦和水稻中的线性顺序相同。包含vrs1的遗传窗口从0.5 cM缩小到0.06 cM,这有助于通过一个518 kb的大麦BAC重叠群覆盖vrs1区域。对重叠群序列的分析表明,水稻Vrs1的一个直系同源基因存在于7号染色体上,这表明包含Vrs1的染色体片段在大麦2H染色体内发生了转位。微共线性的破坏说明了共线性克隆的局限性,并强调了直接在目标物种中开展基因组研究的重要性。