Fiers Martijn, Hoogenboom Jorin, Brunazzi Alice, Wennekes Tom, Angenent Gerco C, Immink Richard G H
Bioscience, Wageningen University and Research, 6700 AP Wageningen, The Netherlands.
Laboratory of Molecular Biology, Wageningen University and Research, 6708 PB Wageningen, The Netherlands.
Plant Methods. 2017 Oct 3;13:78. doi: 10.1186/s13007-017-0230-2. eCollection 2017.
Floral timing is a carefully regulated process, in which the plant determines the optimal moment to switch from the vegetative to reproductive phase. While there are numerous genes known that control flowering time, little information is available on chemical compounds that are able to influence this process. We aimed to discover novel compounds that are able to induce flowering in the model plant Arabidopsis. For this purpose we developed a plant-based screening platform that can be used in a chemical genomics study.
Here we describe the set-up of the screening platform and various issues and pitfalls that need to be addressed in order to perform a chemical genomics screening on Arabidopsis plantlets. We describe the choice for a molecular marker, in combination with a sensitive reporter that's active in plants and is sufficiently sensitive for detection. In this particular screen, the firefly Luciferase marker was used, fused to the regulatory sequences of the floral meristem identity gene , which is an early marker for flowering. Using this screening platform almost 9000 compounds were screened, in triplicate, in 96-well plates at a concentration of 25 µM. One of the identified potential flowering inducing compounds was studied in more detail and named Flowering1 (F1). F1 turned out to be an analogue of the plant hormone Salicylic acid (SA) and appeared to be more potent than SA in the induction of flowering. The effect could be confirmed by watering Arabidopsis plants with SA or F1, in which F1 gave a significant reduction in time to flowering in comparison to SA treatment or the control.
In this study a chemical genomics screening platform was developed to discover compounds that can induce flowering in Arabidopsis. This platform was used successfully, to identify a compound that can speed-up flowering in Arabidopsis.
花期调控是一个受到严格调控的过程，在此过程中植物决定从营养生长阶段转换到生殖生长阶段的最佳时机。虽然已知有许多基因控制开花时间，但关于能够影响这一过程的化合物的信息却很少。我们旨在发现能够诱导模式植物拟南芥开花的新型化合物。为此，我们开发了一个基于植物的筛选平台，可用于化学基因组学研究。
在这里，我们描述了筛选平台的设置以及在对拟南芥幼苗进行化学基因组学筛选时需要解决的各种问题和陷阱。我们描述了分子标记的选择，以及与一种在植物中具有活性且对检测足够敏感的灵敏报告基因相结合的情况。在这个特定的筛选中，使用了与花分生组织特性基因的调控序列融合的萤火虫荧光素酶标记，该基因是开花的早期标记。使用这个筛选平台，以25 μM的浓度在96孔板中对近9000种化合物进行了一式三份的筛选。对其中一种鉴定出的潜在开花诱导化合物进行了更详细的研究，并将其命名为开花素1（F1）。结果表明，F1是植物激素水杨酸（SA）的类似物，并且在诱导开花方面似乎比SA更有效。通过用SA或F1浇灌拟南芥植物可以证实这种效果，与SA处理或对照相比，F1显著缩短了开花时间。
在本研究中，开发了一个化学基因组学筛选平台来发现能够诱导拟南芥开花的化合物。该平台已成功用于鉴定一种能够加速拟南芥开花的化合物。
