Amos B K, Pook Victoria G, Debolt Seth
Department of Horticulture, University of Kentucky.
Department of Horticulture, University of Kentucky;
J Vis Exp. 2018 Apr 30(134):57393. doi: 10.3791/57393.
Chemical genetics is increasingly being employed to decode traits in plants that may be recalcitrant to traditional genetics due to gene redundancy or lethality. However, the probability of a synthetic small molecule being bioactive is low; therefore, thousands of molecules must be tested in order to find those of interest. Liquid handling robotics systems are designed to handle large numbers of samples, increasing the speed with which a chemical library can be screened in addition to minimizing/standardizing error. To achieve a high-throughput forward chemical genetics screen of a library of 50,000 small molecules on Arabidopsis thaliana (Arabidopsis), protocols using a bench-top multichannel liquid handling robot were developed that require minimal technician involvement. With these protocols, 3,271 small molecules were discovered that caused visible phenotypic alterations. 1,563 compounds induced short roots, 1,148 compounds altered coloration, 383 compounds caused root hair and other, non-categorized, alterations, and 177 compounds inhibited germination.
化学遗传学越来越多地被用于解析植物中的性状，这些性状可能由于基因冗余或致死性而难以用传统遗传学方法研究。然而，合成小分子具有生物活性的概率较低；因此，必须测试数千种分子才能找到感兴趣的分子。液体处理机器人系统旨在处理大量样品，除了将误差最小化/标准化之外，还能提高化学文库的筛选速度。为了对拟南芥上的50000个小分子文库进行高通量正向化学遗传学筛选，开发了使用台式多通道液体处理机器人的方案，这些方案所需的技术人员参与最少。通过这些方案，发现了3271个导致可见表型改变的小分子。1563种化合物诱导根变短，1148种化合物改变颜色，383种化合物导致根毛及其他未分类的改变，177种化合物抑制发芽。
