Department of Microbiology and Plant Biology, University of Oklahoma Norman, OK, USA.
Front Plant Sci. 2013 Nov 26;4:484. doi: 10.3389/fpls.2013.00484. eCollection 2013.
Plant root is an organ composed of multiple cell types with different functions. This multicellular complexity limits our understanding of root biology because -omics studies performed at the level of the entire root reflect the average responses of all cells composing the organ. To overcome this difficulty and allow a more comprehensive understanding of root cell biology, an approach is needed that would focus on one single cell type in the plant root. Because of its biological functions (i.e., uptake of water and various nutrients; primary site of infection by nitrogen-fixing bacteria in legumes), the root hair cell is an attractive single cell model to study root cell response to various stresses and treatments. To fully study their biology, we have recently optimized procedures in obtaining root hair cell samples. We culture the plants using an ultrasound aeroponic system maximizing root hair cell density on the entire root systems and allowing the homogeneous treatment of the root system. We then isolate the root hair cells in liquid nitrogen. Isolated root hair yields could be up to 800 to 1000~mg of plant cells from 60 root systems. Using soybean as a model, the purity of the root hair was assessed by comparing the expression level of genes previously identified as soybean root hair specific between preparations of isolated root hair cells and stripped roots, roots devoid in root hairs. Enlarging our tests to include other plant species, our results support the isolation of large quantities of highly purified root hair cells which is compatible with a systems biology approach.
植物根是由具有不同功能的多种细胞类型组成的器官。这种多细胞的复杂性限制了我们对根生物学的理解，因为在整个根水平上进行的“组学”研究反映了构成器官的所有细胞的平均反应。为了克服这一困难，需要一种方法来关注植物根中的一种单一细胞类型。由于其生物学功能（即吸收水和各种养分；豆科植物固氮细菌感染的主要部位），根毛细胞是研究根细胞对各种胁迫和处理的反应的理想单一细胞模型。为了充分研究它们的生物学特性，我们最近优化了获得根毛细胞样本的程序。我们使用超声气雾化系统培养植物，最大限度地提高整个根系上根毛细胞的密度，并允许对根系进行均匀处理。然后我们在液氮中分离根毛细胞。从 60 个根系中可以获得多达 800 到 1000 毫克的植物细胞。以大豆为例，通过比较先前鉴定为大豆根毛特有的基因在分离的根毛细胞和剥离的根、没有根毛的根中的表达水平，评估根毛的纯度。将我们的测试扩大到包括其他植物物种，我们的结果支持大量高度纯化的根毛细胞的分离，这与系统生物学方法兼容。
