PDF(Pubmed)
Abstract：
The integration of data from multiple sources and analytical techniques to obtain novel insights and answer challenging questions is a hallmark of modern science. In arthropods, exocrine secretions may act as pheromones, defensive substances, antibiotics, as well as surface protectants, and as such they play a crucial role in ecology and evolution. Exocrine chemical compounds are frequently characterized by gas chromatography-mass spectrometry. Technological advances of recent years now allow us to routinely characterize the total gene complement transcribed in a particular biological tissue, often in the context of experimental treatment, via RNAseq. We here introduce a novel methodological approach to successfully characterize exocrine secretions and full transcriptomes of one and the same individual of oribatid mites. We found that chemical extraction prior to RNA extraction had only minor effects on the total RNA integrity. De novo transcriptomes obtained from such combined extractions were of comparable quality to those assembled for samples that were subject to RNA extraction only, indicating that combined chemical/RNA extraction is perfectly suitable for phylotranscriptomic studies. However, in-depth analysis of RNA expression analysis indicates that chemical extraction prior to RNAseq may affect transcript degradation rates, similar to the effects reported in previous studies comparing RNA extraction protocols. With this pilot study, we demonstrate that profiling chemical secretions and RNA expression levels from the same individual is methodologically feasible, paving the way for future research to understand the genes and pathways underlying the syntheses of biogenic chemical compounds. Our approach should be applicable broadly to most arachnids, insects, and other arthropods.
摘要：
整合来自多个来源的数据和分析技术以获得新颖的见解并回答具有挑战性的问题是现代科学的标志。在节肢动物中，外分泌分泌物可以作为信息素，防御性物质,抗生素,以及表面保护剂，因此，它们在生态和进化中起着至关重要的作用。外分泌化学化合物通常通过气相色谱-质谱法表征。近年来的技术进步现在使我们能够常规地表征在特定生物组织中转录的总基因补体，通常在实验治疗的背景下，通过RNAseq.我们在这里介绍了一种新的方法学方法，可以成功地表征一种和同一个体的外分泌分泌物和完整的转录组。我们发现在RNA提取之前的化学提取对总RNA完整性仅有较小的影响。从这种组合提取获得的从头转录组具有与仅进行RNA提取的样品组装的质量相当的质量。这表明联合化学/RNA提取完全适合于页转录组学研究。然而,RNA表达分析的深入分析表明，RNAseq之前的化学提取可能会影响转录物降解率，与之前比较RNA提取方案的研究报告的效果相似.通过这项试点研究，我们证明了来自同一个个体的化学分泌物和RNA表达水平的分析在方法上是可行的，为未来的研究铺平了道路，以了解生物化合物合成的基础基因和途径。我们的方法应该广泛适用于大多数蜘蛛，昆虫,和其他节肢动物。
