A Visual Leaf Zymography Technique for the In Situ Examination of Plant Enzyme Activity under the Stress of Environmental Pollution

Abstract

This study established a high-efficiency fluorescence quenching approach for the in situ visualization and modeling of the spatial distribution of xylanase, β-glucosidase, and phosphatase activities in plant leaves under pollution stress (namely, the leaf zymography technique, LZT). In the LZT, a membrane saturated with an enzyme-specific fluorescent substrate on the leaf surface was incubated and the fluorescence image generated on the membrane under ultraviolet light was recorded. An image-based modeling method for restoring the morphological traits of the true image by reducing noise was developed to ensure the accurate estimation of enzyme activities. The LZT could simultaneously measure 48 samples within 2 h, with good reproducibility. The results obtained by the LZT were comparable to those obtained by a conventional biochemical analysis method and presented lowcost and convenient advantages. This paper explains the theoretical basis required to investigate the realistic application of the LZT for assessing ecotoxicity in large-scale monitoring.

A 1D-SP-Net to Determine Early Drought Stress Status of Tomato (Solanum lycopersicum) with Imbalanced Vis/NIR Spectroscopy Data

“…However, both solutions require deep understanding of the progression of stress induction, for which methodologies capable of detecting stress responses in the early stages are essential [5,6]. Those methodologies include the determination of specific gene expression by polymerase chain reaction technology, quantification of enzyme activity and of active compounds by zymography, spectrometry, and chromatography analysis, and the direct or indirect measurement of other physiological parameters by various technologies and methods [7][8][9][10][11]. Nevertheless, these methods are relatively invasive, time-consuming, and poorly replicable.…”