Vernet Helena, Fullana Aïda Magdalena, Sorribas Francisco Javier, Gualda Emilio J
Department of Agri-Food Engineering and Biotechnology, Universitat Politècnica de Catalunya, Esteve Terradas 8, 08860 Castelldefels, Spain.
Plants (Basel). 2022 Apr 26;11(9):1165. doi: 10.3390/plants11091165.
Plant-parasitic nematodes are a significant cause of yield losses and food security issues. Specifically, nematodes of the genus can cause significant production losses in horticultural crops around the world. Understanding the mechanisms of the ever-changing physiology of plant roots by imaging the galls induced by nematodes could provide a great insight into their control. However, infected roots are unsuitable for light microscopy investigation due to the opacity of plant tissues. Thus, samples must be cleared to visualize the interior of whole plants in order to make them transparent using clearing agents. This work aims to identify which clearing protocol and microscopy system is the most appropriate to obtain 3D images of tomato cv. Durinta and eggplant cv. Cristal samples infected with to visualize and study the root-nematode interaction. To that extent, two clearing solutions (BABB and ECi), combined with three different dehydration solvents (ethanol, methanol and 1-propanol), are tested. In addition, the advantages and disadvantages of alternative imaging techniques to confocal microscopy are analyzed by employing an experimental custom-made setup that combines two microscopic techniques, light sheet fluorescence microscopy and optical projection tomography, on a single instrument.
植物寄生线虫是造成产量损失和粮食安全问题的重要原因。具体而言，该属线虫可在全球范围内导致园艺作物的重大产量损失。通过对线虫诱导形成的虫瘿进行成像来了解植物根系不断变化的生理机制，可为线虫防治提供深刻见解。然而，由于植物组织不透明，受感染的根系不适合进行光学显微镜观察。因此，必须使用透明剂对样本进行透明处理，以便观察整株植物的内部结构。本研究旨在确定哪种透明处理方案和显微镜系统最适合获取感染了[线虫名称未给出]的番茄品种Durinta和茄子品种Cristal样本的三维图像，以可视化和研究根与线虫的相互作用。为此，测试了两种透明溶液（BABB和ECi）与三种不同脱水溶剂（乙醇、甲醇和1-丙醇）的组合。此外，通过在一台仪器上结合光片荧光显微镜和光学投影断层扫描这两种显微技术的实验定制装置，分析了共聚焦显微镜替代成像技术的优缺点。
