Gualtieri Gustavo, Kulikova Olga, Limpens Erik, Kim Dong-Jin, Cook Douglas R, Bisselin Ton, Geurts René
Molecular Biology, Wageningen University, Netherlands.
Plant Mol Biol. 2002 Sep;50(2):225-35. doi: 10.1023/a:1016085523752.
The crop legume pea (Pisum sativum) is genetically well characterized. However, due to its large genome it is not amenable to efficient positional cloning strategies. The purpose of this study was to determine if the model legume Medicago truncatula, which is a close relative of pea, could be used as a reference genome to facilitate the cloning of genes identified based on phenotypic and genetic criteria in pea. To this end, we studied the level of microsynteny between the SYM2 region of pea and the orthologous region in M. truncatula. Initially, a marker tightly linked to SYM2 was isolated by performing differential RNA display on near-isogenic pea lines. This marker served as the starting point for construction of a BAC physical map in M. truncatula. A fine-structure genetic map, based on eight markers from the M. truncatula physical map, indicates that the two genomes in this region share a conserved gene content. Importantly, this fine structure genetic map clearly delimits the SYM2-containing region in pea and the SYM2-orthologous region in M. truncatula, and should provide the basis for cloning SYM2. The utility of the physical and genetic tools in M. truncatula to dissect the SYM2 region of pea should have important implications for other gene cloning experiments in pea, in particular where the two genomes are highly syntenic within the region of interest.
豆科作物豌豆(Pisum sativum)在遗传学上已得到充分表征。然而,由于其基因组庞大,不适用于高效的定位克隆策略。本研究的目的是确定豌豆的近缘种、模式豆科植物蒺藜苜蓿(Medicago truncatula)是否可用作参考基因组,以促进基于豌豆表型和遗传标准鉴定的基因的克隆。为此,我们研究了豌豆SYM2区域与蒺藜苜蓿直系同源区域之间的微共线性水平。最初,通过对近等基因豌豆品系进行差异RNA展示,分离出一个与SYM2紧密连锁的标记。该标记作为构建蒺藜苜蓿BAC物理图谱的起点。基于蒺藜苜蓿物理图谱中的八个标记构建的精细结构遗传图谱表明,该区域的两个基因组具有保守的基因含量。重要的是,这个精细结构遗传图谱清楚地界定了豌豆中含SYM2的区域和蒺藜苜蓿中SYM2直系同源区域,应为克隆SYM2提供基础。蒺藜苜蓿中用于剖析豌豆SYM2区域的物理和遗传工具的实用性,对豌豆中的其他基因克隆实验应具有重要意义,特别是在两个基因组在感兴趣区域内高度共线性的情况下。