A fluorescence nanoparticles-based method for visualizing and quantifying root cap mucilage

抄録

The root cap mucilage, a gel-like substance secreted by border cells or border-like cells at the root cap, plays a crucial role in plant root adaptation to soil environments. Despite its importance, methods for analyzing root cap mucilage, especially visualization techniques, remain limited. This study introduces a simple method using fluorescent silica nanoparticles (Quartz Dot) to visualize and quantify the area of secreted mucilage with confocal laser scanning microscopy. The nanoparticles used in this study are embedded with fluorescent probes and large enough to be excluded from the apoplastic mucilage matrix, thereby outlining the boundary of the mucilage region surrounding the root cap. Combined with fluorescent probes for cell wall staining, this approach allows the quantification of the minute mucilage areas surrounding the Arabidopsis root cap. The method was validated by comparing the mucilage region in wild-type Arabidopsis and the smb brn1 brn2 mutant, revealing a significant reduction in mucilage areas in the mutant. Additionally, we investigated the effects of several abiotic stresses, including toxic ions and nutrient starvation, on the area of Arabidopsis root cap mucilage, and further identified soil stresses that promote expansion of the root cap mucilage region, underscoring the sensitivity and precision of our method. Collectively, this study established a simple and robust technique for visualizing and quantifying the root cap mucilage region. The demonstrated utility of this method indicates its potential for broader applications in understanding the physiological and molecular mechanisms underlying root cap functionalization and its role in adaptation to soil environments.