Detecting the orientation of newly-deposited crystalline cellulose with fluorescent CBM3.

ABSTRACT: Cellulose microfibril patterning influences many of the mechanical attributes of plant cell walls. We developed a simple, fluorescence microscopy-based method to detect the orientation of newly-synthesized cellulose microfibrils in epidermal peels of onion and Arabidopsis. It is based on Alexa Fluor 488-tagged carbohydrate binding module 3a (CBM3a) from Clostridium thermocellum which displayed a nearly 4-fold greater binding to cell walls at pH 5.5 compared with pH 8. Binding to isolated cellulose did not display this pH dependence. At pH 7.5 fibrillar patterns at the surface of the epidermal peels were visible, corresponding to the directionality of surface cellulose microfibrils, as verified by atomic force microscopy. The fibrillar pattern was not visible as the labeling intensit

SUBMITTER: Pfaff SA 

PROVIDER: S-EPMC9678952 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature