Kudoh Hiroshi, Honjo Mie N, Nishio Haruki, Sugisaka Jiro
Center for Ecological Research, Kyoto University, Otsu, Shiga, Japan.
Methods Mol Biol. 2018;1830:41-57. doi: 10.1007/978-1-4939-8657-6_3.
The majority of organismal phenomena show functional significance in the context of natural environments. However, we know little about how dynamic gene expression is controlled under natural complex conditions. One of the most attractive challenges in current biology is to understand organismal functions in natural environments. We established and have developed long-term "in natura" study sites of Arabidopsis halleri to evaluate precise control of gene expression in natural environments. At the sites, we monitored meteorological factors, recorded plant growth and phenology, and collected RNA and chromatin samples to investigate dynamics of transcription and epigenetic modifications. Here, we introduce the in natura study sites, especially with the emphasis on methodologies for setting up study sites in natural plant populations and collecting samples used in transcriptomics and epigenetics in natural environments. Although the methods introduced here need to be modified depending on situations of one's study systems, our case can be a model for planning new in natura studies.
大多数生物体现象在自然环境中都具有功能意义。然而,我们对自然复杂条件下动态基因表达是如何被调控的却知之甚少。当前生物学中最具吸引力的挑战之一就是了解生物体在自然环境中的功能。我们建立并发展了长期的拟南芥“自然状态下”的研究地点,以评估自然环境中基因表达的精确调控。在这些地点,我们监测气象因素,记录植物生长和物候,并收集RNA和染色质样本,以研究转录和表观遗传修饰的动态变化。在此,我们介绍自然状态下的研究地点,尤其着重于在自然植物种群中建立研究地点以及在自然环境中收集转录组学和表观遗传学所用样本的方法。尽管此处介绍的方法需要根据自身研究系统的情况进行修改,但我们的案例可为规划新的自然状态下的研究提供一个范例。