Abstract

BackgroundCotton, 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.ResultsHomologous 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.ConclusionThese 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.

Highlights

  • Cotton, as an allopolyploid species, contains homoeologous A and D subgenomes

  • Homoeologous bacterial artificial chromosome (BAC) derived from homoeologous chromosomes SSR primer pair NAU837 yielded two PCR amplicons from G. barbadense cv

  • DcFoiigsttuorrinbeub1tyioFnISaHnalysis of homoeologous segments in tetraploid Distribution analysis of homoeologous segments in tetraploid cotton by Fluorescence in situ hybridization (FISH). (A) Figure showed that the identification of the BAC clone 75F07 containing the polymorphic locus of NAU837-205, and 68D15 containing the monomorphic locus NAU837-195 between TM-1 and Hai7124 by SSR marker NAU837

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Summary

Introduction

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 presents a useful approach for identification and localization of duplicated segments. When genes are duplicated as a consequence of polyploidization, they may continue to evolve at the same rate as they did in their diploid ancestors, or they may be subject to pressures that lead to differential rates of sequence evolution [2] These duplicated sequences and their functions are maintained intact or undergo long-term evolutionary change via sequence elimination [3,4], sequence rearrangement [5], gene silencing [6], or acquisition of new function [7]. The various aspects of genome evolution involving duplicated sequences in polyploids have been reviewed elsewhere [2]

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