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通过高通量鉴定矮牵牛和喇叭水仙转录组,揭示 an11 突变体中受损的途径。

Revealing impaired pathways in the an11 mutant by high-throughput characterization of Petunia axillaris and Petunia inflata transcriptomes.

机构信息

Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy.

出版信息

Plant J. 2011 Oct;68(1):11-27. doi: 10.1111/j.1365-313X.2011.04661.x. Epub 2011 Jul 14.

Abstract

Petunia is an excellent model system, especially for genetic, physiological and molecular studies. Thus far, however, genome-wide expression analysis has been applied rarely because of the lack of sequence information. We applied next-generation sequencing to generate, through de novo read assembly, a large catalogue of transcripts for Petunia axillaris and Petunia inflata. On the basis of both transcriptomes, comprehensive microarray chips for gene expression analysis were established and used for the analysis of global- and organ-specific gene expression in Petunia axillaris and Petunia inflata and to explore the molecular basis of the seed coat defects in a Petunia hybrida mutant, anthocyanin 11 (an11), lacking a WD40-repeat (WDR) transcription regulator. Among the transcripts differentially expressed in an11 seeds compared with wild type, many expected targets of AN11 were found but also several interesting new candidates that might play a role in morphogenesis of the seed coat. Our results validate the combination of next-generation sequencing with microarray analyses strategies to identify the transcriptome of two petunia species without previous knowledge of their genome, and to develop comprehensive chips as useful tools for the analysis of gene expression in P. axillaris, P. inflata and P. hybrida.

摘要

矮牵牛是一个极好的模式系统,特别适合进行遗传、生理和分子研究。然而,到目前为止,由于缺乏序列信息,全基因组表达分析应用很少。我们应用下一代测序技术,通过从头组装读取,生成了大量的腋花矮牵牛和膨果矮牵牛转录本目录。基于这两个转录组,我们建立了综合的微阵列芯片,用于基因表达分析,并用于分析腋花矮牵牛和膨果矮牵牛的全局和器官特异性基因表达,以及探索在缺乏 WD40 重复(WDR)转录调节因子的矮牵牛杂种突变体花青素 11(an11)中种皮缺陷的分子基础。在 an11 种子与野生型相比差异表达的转录本中,发现了许多预期的 AN11 靶标,但也发现了几个有趣的新候选者,它们可能在种皮形态发生中发挥作用。我们的结果验证了将下一代测序与微阵列分析策略相结合,用于鉴定两个矮牵牛物种的转录组,而无需事先了解它们的基因组,并开发综合芯片作为分析腋花矮牵牛、膨果矮牵牛和矮牵牛杂种基因表达的有用工具。

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