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开发匿名cDNA微阵列以研究千里光属植物在杂交物种形成过程中花转录组的变化。

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

作者信息

Hegarty Matthew J, Jones Joanna M, Wilson Ian D, Barker Gary L, Coghill Jane A, Sanchez-Baracaldo Patricia, Liu Guoqing, Buggs Richard J A, Abbott Richard J, Edwards Keith J, Hiscock Simon J

机构信息

School of Biological Sciences, University of Bristol, Woodland Road, Bristol BS8 1UG, UK.

出版信息

Mol Ecol. 2005 Jul;14(8):2493-510. doi: 10.1111/j.1365-294x.2005.02608.x.

Abstract

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.

摘要

种间杂交是一个重要过程,通过该过程可能会发生突然的物种形成。近年来,已通过多种技术鉴定出与杂交物种形成相关的基因变化,包括AFLP/SSR图谱分析、基因组原位杂交/荧光原位杂交以及cDNA-AFLP差异显示。然而,由于许多杂交物种缺乏广泛的序列数据,且难以将来自模式物种的商业可用微阵列的结果外推到非模式杂交类群上,利用微阵列技术分析与杂交物种形成相关的全基因组/转录组变化的进展一直有限。因此,越来越多研究非模式系统的研究人员开始转向开发“匿名”cDNA微阵列,通过打印未测序的cDNA克隆来降低生产微阵列的时间和成本,并且只对那些显示出有趣表达模式的克隆进行测序。在此,我们描述了匿名cDNA微阵列的创建、测试及初步应用,以研究千里光属中与异源多倍体物种形成相关的花转录组变化。我们报告了异源六倍体杂种千里光、其亲本类群滇千里光(二倍体)和欧洲千里光(四倍体)以及中间三倍体(不育)杂种巴氏千里光之间基因表达的比较。匿名微阵列分析揭示了这四个类群之间花基因表达的显著差异,并证明了该技术在研究非模式开花植物杂交的遗传影响方面的强大作用。

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