Wu Chengcang, Sun Shuku, Nimmakayala Padmavathi, Santos Felipe A, Meksem Khalid, Springman Rachael, Ding Kejiao, Lightfoot David A, Zhang Hong-Bin
Department of Soil and Crop Sciences and Institute for Plant Genomics and Biotechnology, Texas A&MUniversity, College Station, Texas 77843-2123, USA.
Genome Res. 2004 Feb;14(2):319-26. doi: 10.1101/gr.1405004. Epub 2004 Jan 12.
Genome-wide physical maps are crucial to many aspects of advanced genome research. We report a genome-wide, bacterial artificial chromosome (BAC) and plant-transformation-competent binary large-insert plasmid clone (hereafter BIBAC)-based physical map of the soybean genome. The map was constructed from 78001 clones from five soybean BAC and BIBAC libraries representing 9.6 haploid genomes and three cultivars, and consisted of 2905 BAC/BIBAC contigs, estimated to span 1408 Mb in physical length. We evaluated the reliability of the map contigs using different contig assembly strategies, independent contig building methods, DNA marker hybridization, and different fingerprinting methods, and the results showed that the contigs were assembled properly. Furthermore, we tested the feasibility of integrating the physical map with the existing soybean composite genetic map using 388 DNA markers. The results further confirmed the nature of the ancient tetraploid origin of soybean and indicated that it is feasible to integrate the physical map with the linkage map even though greater efforts are needed. This map represents the first genome-wide, BAC/BIBAC-based physical map of the soybean genome and would provide a platform for advanced genome research of soybean and other legume species. The inclusion of BIBACs in the map would streamline the utility of the map for positional cloning of genes and QTLs, and functional analysis of soybean genomic sequences.
全基因组物理图谱对于高级基因组研究的许多方面都至关重要。我们报告了基于细菌人工染色体(BAC)和具备植物转化能力的二元大插入质粒克隆(以下简称BIBAC)构建的大豆基因组全基因组物理图谱。该图谱由来自五个大豆BAC和BIBAC文库的78001个克隆构建而成,这些文库代表9.6个单倍体基因组以及三个品种,图谱包含2905个BAC/BIBAC重叠群,估计物理长度跨越1408 Mb。我们使用不同的重叠群组装策略、独立的重叠群构建方法、DNA标记杂交以及不同的指纹识别方法评估了图谱重叠群的可靠性,结果表明重叠群组装正确。此外,我们使用388个DNA标记测试了将物理图谱与现有的大豆复合遗传图谱整合的可行性。结果进一步证实了大豆古老四倍体起源的性质,并表明尽管需要付出更多努力,但将物理图谱与连锁图谱整合是可行的。该图谱是首个基于BAC/BIBAC构建的大豆基因组全基因组物理图谱,将为大豆及其他豆科物种的高级基因组研究提供一个平台。图谱中包含BIBAC将简化图谱在基因和QTL定位克隆以及大豆基因组序列功能分析中的应用。