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High-throughput method for detection and quantification of lesions on leaf scale based on trypan blue staining and
Emina Mulaosmanovic1, Tobias U T Lindblom2, Marie Bengtsson3
11Department of Biosystems and Technology, Microbial Horticulture Unit, Swedish University of Agricultural Sciences, PO Box 103, 23053 Alnarp, Sweden.
Plant Methods
|May 12, 2020
Summary
A new trypan blue staining method accurately detects and quantifies leaf damage in vegetables, including microlesions, enabling better yield loss prediction and risk assessment.
Area of Science:
- Plant pathology
- Agricultural science
- Horticulture
Background:
- Leafy vegetables face damage from various biotic, abiotic, and mechanical factors.
- Current damage assessment methods often rely on visible color changes, which may not always occur or appear late.
- Accurate detection of both macro and microlesions is crucial for effective plant health management.
Purpose of the Study:
- To develop and validate a novel, sensitive method for visualizing and quantifying leaf damage at the leaf scale.
- To enable automated detection and measurement of damaged areas, including single-cell lesions.
- To provide a tool for accurate plant damage assessment in leafy vegetables.
Main Methods:
- Whole leaves were stained with trypan blue dye, which selectively stains compromised cell membranes.
- An automated image analysis pipeline was developed to identify and measure stained (lesion) areas.
- The method was tested on field-grown spinach and Swiss chard to demonstrate its efficacy.
Main Results:
- A robust, fast, and sensitive method for leaf-scale damage visualization and quantification was established.
- The image analysis pipeline accurately identified leaf areas and individual lesions down to the cellular level.
- The methodology proved effective for damage detection and quantification in spinach and Swiss chard.
Conclusions:
- The novel method allows for the detection and quantification of both macro and microlesions at any stage, irrespective of visible symptoms.
- Accurate quantification of wounded leaf areas is essential for developing economic loss prediction models and shelf-life assessments.
- This fine-resolution damage assessment supports risk assessments related to pathogen infection rates and disease severity.
