Department of Agronomy, University of Wisconsin, Madison, Wisconsin, United States of America.
PLoS One. 2013 Apr 23;8(4):e61005. doi: 10.1371/journal.pone.0061005. Print 2013.
Transcriptome analysis is a valuable tool for identification and characterization of genes and pathways underlying plant growth and development. We previously published a microarray-based maize gene atlas from the analysis of 60 unique spatially and temporally separated tissues from 11 maize organs [1]. To enhance the coverage and resolution of the maize gene atlas, we have analyzed 18 selected tissues representing five organs using RNA sequencing (RNA-Seq). For a direct comparison of the two methodologies, the same RNA samples originally used for our microarray-based atlas were evaluated using RNA-Seq. Both technologies produced similar transcriptome profiles as evident from high Pearson's correlation statistics ranging from 0.70 to 0.83, and from nearly identical clustering of the tissues. RNA-Seq provided enhanced coverage of the transcriptome, with 82.1% of the filtered maize genes detected as expressed in at least one tissue by RNA-Seq compared to only 56.5% detected by microarrays. Further, from the set of 465 maize genes that have been historically well characterized by mutant analysis, 427 show significant expression in at least one tissue by RNA-Seq compared to 390 by microarray analysis. RNA-Seq provided higher resolution for identifying tissue-specific expression as well as for distinguishing the expression profiles of closely related paralogs as compared to microarray-derived profiles. Co-expression analysis derived from the microarray and RNA-Seq data revealed that broadly similar networks result from both platforms, and that co-expression estimates are stable even when constructed from mixed data including both RNA-Seq and microarray expression data. The RNA-Seq information provides a useful complement to the microarray-based maize gene atlas and helps to further understand the dynamics of transcription during maize development.
转录组分析是一种用于鉴定和描述植物生长和发育相关基因和途径的有价值的工具。我们之前发表了一个基于微阵列的玉米基因图谱,该图谱是通过对 11 个玉米器官的 60 个独特的时空分离组织进行分析得到的[1]。为了增强玉米基因图谱的覆盖范围和分辨率,我们使用 RNA 测序(RNA-Seq)分析了代表五个器官的 18 个选定组织。为了直接比较这两种方法,我们使用 RNA-Seq 评估了最初用于我们基于微阵列的图谱的相同 RNA 样本。这两种技术产生了相似的转录组谱,从高 Pearson 相关统计数据(范围从 0.70 到 0.83)和组织的几乎相同聚类中可以明显看出。RNA-Seq 提供了转录组的增强覆盖,在至少一种组织中检测到的过滤后的玉米基因中,有 82.1%通过 RNA-Seq 检测到,而通过微阵列检测到的只有 56.5%。此外,在通过突变分析得到了很好的历史特征的 465 个玉米基因中,有 427 个在至少一种组织中通过 RNA-Seq 显示出显著表达,而通过微阵列分析得到的只有 390 个。与微阵列衍生的图谱相比,RNA-Seq 提供了更高的分辨率,可用于识别组织特异性表达,以及区分密切相关的同源基因的表达谱。从微阵列和 RNA-Seq 数据中得出的共表达分析表明,两种平台都产生了广泛相似的网络,即使从包括 RNA-Seq 和微阵列表达数据的混合数据中构建共表达估计,其稳定性也很好。RNA-Seq 信息为基于微阵列的玉米基因图谱提供了有用的补充,并有助于进一步了解玉米发育过程中转录的动态。