CSIRO Land and Water, Locked Bag 2007, Kirrawee, NSW 2232, Australia.
Aquat Toxicol. 2013 Aug 15;138-139:12-25. doi: 10.1016/j.aquatox.2013.04.002. Epub 2013 Apr 17.
The transcriptomic profile of the marine diatom, Phaeodactylum tricornutum, exposed to several ecologically relevant stressors, was used to develop toxicity identification evaluation (TIE)-like gene expression assays. Algal growth inhibition was measured by flow cytometry to determine exposure concentrations that elicited a sublethal toxic response. P. tricornutum was exposed to concentrations of copper (2 μg L⁻¹), cadmium (5 μg L⁻¹), silver (20 μg L⁻¹), simazine (75 μg L⁻¹), the water accommodated fraction (WAF) of weathered crude oil (5 mg L⁻¹), 50 μg L⁻¹ ammonia, a decreased salinity treatment (15‰), and a mixture exposure of ammonia, decreased salinity and cadmium (10 μg L⁻¹). Analysis of the gene expression via microarray indicated that unique transcriptomic signals were generated for each of the individual treatments. Transcriptomic profiles of ammonia and the mixture treatment overlapped substantially. Photosynthesis related transcripts were altered in the simazine (herbicide) treatment. A transcript involved in degrading hydrocarbons, dioxygenase, had increased abundance after crude oil exposure. Overall, transcriptomic responses in the different treatments were associated with stress responses, membrane transport, transcription and translation and could be linked to contaminant mode of action. The transcriptomic profiles were used to design real-time (quantitative) polymerase chain reaction (qPCR) assays that would link changes in transcript abundance to a particular stressor in a TIE-based approach. At least one transcript for each contaminant tested (copper, cadmium, silver, salinity and ammonia) responded exclusively to that contaminant. With further development of additional transcriptomic markers for each contaminant, this new approach has potential to enhance traditional toxicology bioassays by providing additional lines of evidence to identify biologically relevant stressors within a contaminated ecosystem based on changes in the transcriptomic profile.
对暴露于几种生态相关胁迫因子的海洋硅藻三角褐指藻的转录组谱进行了研究,以开发毒性鉴定评估(TIE)类似的基因表达分析。通过流式细胞术测量藻类生长抑制来确定引起亚致死毒性反应的暴露浓度。将三角褐指藻暴露于铜(2μg/L)、镉(5μg/L)、银(20μg/L)、西玛津(75μg/L)、风化原油的水容纳部分(5mg/L)、50μg/L 氨、低盐度处理(15‰)和氨、低盐度和镉的混合物暴露(10μg/L)。通过微阵列分析基因表达表明,每种单独处理都产生了独特的转录组信号。氨和混合物处理的转录组谱重叠很大。在西玛津(除草剂)处理中,光合作用相关的转录本发生了改变。一种参与降解烃的双氧酶,在接触原油后丰度增加。总的来说,不同处理中的转录组反应与应激反应、膜转运、转录和翻译有关,并且可以与污染物作用模式相关联。转录组谱用于设计实时(定量)聚合酶链反应(qPCR)分析,该分析将转录本丰度的变化与基于 TIE 的特定胁迫因子联系起来。在所测试的每种污染物(铜、镉、银、盐度和氨)中,至少有一种转录本对该污染物有特异性反应。通过为每种污染物进一步开发更多的转录组标记物,这种新方法有可能通过基于转录组谱的变化为识别受污染生态系统中的生物相关胁迫因子提供额外的证据,从而增强传统毒理学生物测定。