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Development of anonymous cDNA microarrays to study changes to the Senecio floral transcriptome during hybrid speciation

机译:开发匿名cDNA微阵列以研究杂种形成过程中千里光花转录组的变化

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Interspecific hybridization is an important process through which abrupt speciation can occur. In recent years, genetic changes associated with hybrid speciation have been identified through a variety of techniques, including AFLP/SSR mapping, GISH/FISH and cDNA-AFLP differential display. However, progress in using microarray technology to analyse whole genome/transcriptome changes associated with hybrid speciation has been limited due to the lack of extensive sequence data for many hybrid species and the difficulties in extrapolating results from commercially available microarrays for model species onto nonmodel hybrid taxa. Increasingly therefore researchers studying nonmodel systems are turning to the development of 'anonymous' cDNA microarrays, where the time and cost of producing microarrays is reduced by printing unsequenced cDNA clones, and sequencing only those clones that display interesting expression patterns. Here we describe the creation, testing and preliminary use of anonymous cDNA microarrays to study changes in floral transcriptome associated with allopolyploid speciation in the genus Senecio. We report a comparison of gene expression between the allohexaploid hybrid, Senecio cambrensis, its parental taxa Senecio squalidus (diploid) and Senecio vulgaris (tetraploid), and the intermediate triploid (sterile) hybrid Senecioxbaxteri. Anonymous microarray analysis revealed dramatic differences in floral gene expression between these four taxa and demonstrates the power of this technique for studies of the genetic impact of hybridization in nonmodel flowering plants.
机译:种间杂交是一个重要的过程,通过它可以发生突然的物种形成。近年来,已通过多种技术鉴定了与杂种形成有关的遗传变化,包括AFLP / SSR定位,GISH / FISH和cDNA-AFLP差异展示。但是,由于缺乏许多杂种的广泛序列数据,并且难以将可商购获得的用于模型物种的微阵列结果推算到非模型杂种单元中,使用微阵列技术分析与杂种形成相关的全基因组/转录组变化的进展受到限制。 。因此,越来越多的研究非模型系统的研究人员转向开发“匿名” cDNA微阵列,通过打印未测序的cDNA克隆并仅对显示有趣表达模式的那些克隆进行测序,可以减少生产微阵列的时间和成本。在这里,我们描述了匿名cDNA微阵列的创建,测试和初步使用,以研究与千里光属中的异源多倍体物种形成有关的花卉转录组的变化。我们报告了同种六倍体杂种,千里光cambrensis,其亲本分类群千里光角鲨(二倍体)和千里光寻常型(四倍体),和中间三倍体(无菌)的千里光混合杂种的基因表达之间的比较。匿名微阵列分析揭示了这四个类群之间花卉基因表达的显着差异,并证明了该技术对研究非模型开花植物中杂交的遗传影响的能力。

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