Plant Genetics, TUM School of Life Sciences Weihenstephan, Technical University of Munich (TUM), Freising, Germany.
Methods Mol Biol. 2021;2309:157-177. doi: 10.1007/978-1-0716-1429-7_13.
Arbuscular mycorrhiza is an ancient symbiosis between most land plants and fungi of the Glomeromycotina, in which the fungi provide mineral nutrients to the plant in exchange for photosynthetically fixed organic carbon. Strigolactones are important signals promoting this symbiosis, as they are exuded by plant roots into the rhizosphere to stimulate activity of the fungi. In addition, the plant karrikin signaling pathway is required for root colonization. Understanding the molecular mechanisms underpinning root colonization by AM fungi, requires the use of plant mutants as well as treatments with different environmental conditions or signaling compounds in standardized cocultivation systems to allow for reproducible root colonization phenotypes. Here we describe how we set up and quantify arbuscular mycorrhiza in the model plants Lotus japonicus and Brachypodium distachyon under controlled conditions. We illustrate a setup for open pot culture as well as for closed plant tissue culture (PTC) containers, for plant-fungal cocultivation in sterile conditions. Furthermore, we explain how to harvest, store, stain, and image AM roots for phenotyping and quantification of different AM structures.
丛枝菌根是大多数陆地植物与 Glomeromycotina 真菌之间的古老共生关系，其中真菌为植物提供矿物质营养，以换取光合作用固定的有机碳。独脚金内酯是促进这种共生关系的重要信号分子，因为它们由植物根系分泌到根际中，以刺激真菌的活性。此外，植物卡里酮信号通路是根定殖所必需的。为了理解丛枝菌根真菌定殖的分子机制，需要使用植物突变体以及在标准化共培养系统中用不同的环境条件或信号化合物进行处理，以允许可重复的根定殖表型。在这里，我们描述了如何在模式植物百脉根和拟南芥中，在受控条件下建立和量化丛枝菌根。我们展示了一种在开放盆式培养和封闭植物组织培养（PTC）容器中进行的设置，用于在无菌条件下进行植物-真菌共培养。此外，我们还解释了如何收获、储存、染色和成像 AM 根，以对不同的 AM 结构进行表型和量化分析。
