Hammond John P, Bowen Helen C, White Philip J, Mills Victoria, Pyke Kevin A, Baker Alan J M, Whiting Steven N, May Sean T, Broadley Martin R
Warwick HRI, University of Warwick, Wellesbourne, Warwick CV35 9EF, UK.
New Phytol. 2006;170(2):239-60. doi: 10.1111/j.1469-8137.2006.01662.x.
Whole-genome transcriptome profiling is revealing how biological systems are regulated at the transcriptional level. This study reports the development of a robust method to profile and compare the transcriptomes of two nonmodel plant species, Thlaspi caerulescens, a zinc (Zn) hyperaccumulator, and Thlaspi arvense, a nonhyperaccumulator, using Affymetrix Arabidopsis thaliana ATH1-121501 GeneChip arrays (Affymetrix, Santa Clara, CA, USA). Transcript abundance was quantified in the shoots of agar- and compost-grown plants of both species. Analyses were optimized using a genomic DNA (gDNA)-based probe-selection strategy based on the hybridization efficiency of Thlaspi gDNA with corresponding A. thaliana probes. In silico alignments of GeneChip probes with Thlaspi gene sequences, and quantitative real-time PCR, confirmed the validity of this approach. Approximately 5000 genes were differentially expressed in the shoots of T. caerulescens compared with T. arvense, including genes involved in Zn transport and compartmentalization. Future functional analyses of genes identified as differentially expressed in the shoots of these closely related species will improve our understanding of the molecular mechanisms of Zn hyperaccumulation.
全基因组转录组分析正在揭示生物系统在转录水平上是如何被调控的。本研究报告了一种可靠方法的开发,该方法使用Affymetrix拟南芥ATH1-121501基因芯片阵列(美国加利福尼亚州圣克拉拉市的Affymetrix公司)对两种非模式植物物种——锌(Zn)超积累植物天蓝遏蓝菜和非超积累植物田野遏蓝菜的转录组进行分析和比较。在琼脂培养和堆肥培养的两种植物的地上部分对转录本丰度进行了定量分析。基于天蓝遏蓝菜基因组DNA(gDNA)与相应拟南芥探针的杂交效率,使用基于gDNA的探针选择策略对分析进行了优化。基因芯片探针与天蓝遏蓝菜基因序列的电子比对以及定量实时PCR证实了该方法的有效性。与田野遏蓝菜相比,天蓝遏蓝菜地上部分约有5000个基因差异表达,包括参与锌转运和区室化的基因。对在这些近缘物种地上部分差异表达的基因进行未来的功能分析,将增进我们对锌超积累分子机制的理解。