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利用BAC-FISH技术对异源四倍体棉花同源染色体组中的同源和部分同源区段进行检测与定位。

Detection and mapping of homologous and homoeologous segments in homoeologous groups of allotetraploid cotton by BAC-FISH.

作者信息

Wang Kai, Guo Wangzhen, Zhang Tianzhen

机构信息

National Key Laboratory of Crop Genetics and Germplasm Enhancement, Cotton Research Institute, Nanjing Agricultural University, Nanjing, China.

出版信息

BMC Genomics. 2007 Jun 19;8:178. doi: 10.1186/1471-2164-8-178.

Abstract

BACKGROUND

Cotton, as an allopolyploid species, contains homoeologous A and D subgenomes. The study of the homoeologous (duplicated) segments or chromosomes can facilitate insight into the evolutionary process of polyploidy and the development of genomic resources. Fluorescence in situ hybridization (FISH) using bacterial artificial chromosome (BAC) clones as probes has commonly been used to provide a reliable cytological technique for chromosome identification. In polyploids, it also presents a useful approach for identification and localization of duplicated segments. Here, two types of BACs that contained the duplicated segments were isolated and analyzed in tetraploid cotton by FISH.

RESULTS

Homologous and homoeologous BACs were isolated by way of SSR marker-based selection and then used to develop BAC-FISH probes. Duplicated segments in homoeologous chromosomes were detected by FISH. The FISH and related linkage map results followed known reinforced the relationships of homoeologous chromosomes in allotetraploid cotton, and presented a useful approach for isolation of homoeologous loci or segments and for mapping of monomorphic loci. It is very important to find that the large duplicated segments (homologous BACs) do exist between homoeologous chromosomes, so the shot-gun approach for genome sequencing was unavailable for tetraploid cotton. However, without doubt, it will contain more information and promote the research for duplicated segments as well as the genome evolution in cotton.

CONCLUSION

These findings and the analysis method by BAC-FISH demonstrated the powerful nature and wide use for the genome and genome evolutionary researches in cotton and other polyploidy species.

摘要

背景

棉花作为一种异源多倍体物种,包含同源的A和D亚基因组。对同源(重复)片段或染色体的研究有助于深入了解多倍体的进化过程以及基因组资源的开发。使用细菌人工染色体(BAC)克隆作为探针的荧光原位杂交(FISH)通常被用于提供一种可靠的染色体识别细胞学技术。在多倍体中,它也是鉴定和定位重复片段的一种有用方法。在此,通过FISH在四倍体棉花中分离并分析了两种包含重复片段的BAC。

结果

通过基于SSR标记的选择方法分离出同源和部分同源BAC,然后用于开发BAC-FISH探针。通过FISH检测到部分同源染色体中的重复片段。FISH及相关连锁图谱结果与已知情况相符,加强了异源四倍体棉花中部分同源染色体的关系,并为分离部分同源基因座或片段以及单态性基因座的定位提供了一种有用的方法。发现部分同源染色体之间确实存在大的重复片段(同源BAC)非常重要,因此对于四倍体棉花而言,鸟枪法基因组测序方法并不适用。然而,毫无疑问,它将包含更多信息,并促进对棉花中重复片段以及基因组进化的研究。

结论

这些发现以及BAC-FISH分析方法证明了其在棉花和其他多倍体物种的基因组及基因组进化研究中的强大作用和广泛用途。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc04/1906772/74b942578cd1/1471-2164-8-178-1.jpg

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