Bulk Sorting Plant Protoplasts for Secondary Cell Wall Biosynthesis

This application note describes the use of protoplasts in investigating the regulatory mechanisms of secondary cell wall deposition in plants. Protoplasts are spherical plant cells lacking cell walls, and they can be derived from leaf tissue or cell culture. To identify regenerating protoplasts, fluorescent stains against cell wall components such as cellulose can be applied. However, the expression and staining can be variable, and thus, sorting can be a useful method to select target populations to accelerate downstream applications. The application of bulk sorting with fluorescent stains against cellulose can serve as a critical step to expedite isolation of healthy cell wall regenerating protoplasts, making downstream applications such as cryo-EM or continued culturing more robust.

Fast and gentle cell sorters from Bio-Techne are ideal for this purpose as they can rapidly isolate and dispense positive protoplasts in bulk by processing 50 protoplasts/second and yielding 80-90% output purity in a single step. The findings demonstrate the versatility of potential sorting methodologies utilizing the Pala platform’s bulk sorting feature, empowering plant scientists to streamline efficient isolation of cell wall regenerating protoplasts.