High-throughput method for detection and quantification of lesions on leaf scale based on trypan blue staining and digital image analysis.

Autor: Mulaosmanovic E; 1Department of Biosystems and Technology, Microbial Horticulture Unit, Swedish University of Agricultural Sciences, PO Box 103, 23053 Alnarp, Sweden., Lindblom TUT; 2Department of Crop Production Ecology, Plant Ecology Unit, Swedish University of Agricultural Sciences, PO Box 7043, 75007 Uppsala, Sweden., Bengtsson M; 3Department of Plant Protection Biology, Chemical Ecology Unit, Swedish University of Agricultural Sciences, PO Box 102, 23053 Alnarp, Sweden., Windstam ST; 4Department of Biological Sciences, State University of New York at Oswego, 7060 NY-104, Oswego, NY 13126 USA., Mogren L; 1Department of Biosystems and Technology, Microbial Horticulture Unit, Swedish University of Agricultural Sciences, PO Box 103, 23053 Alnarp, Sweden., Marttila S; 5Department of Plant Protection Biology, Resistance Biology Unit, Swedish University of Agricultural Sciences, PO Box 102, 23053 Alnarp, Sweden., Stützel H; 6Institute of Horticultural Production Systems, Gottfried Wilhelm Leibniz University Hannover, Herrenhäuser Str. 2, 30419 Hannover, Germany., Alsanius BW; 1Department of Biosystems and Technology, Microbial Horticulture Unit, Swedish University of Agricultural Sciences, PO Box 103, 23053 Alnarp, Sweden.
Jazyk: angličtina
Zdroj: Plant methods [Plant Methods] 2020 May 04; Vol. 16, pp. 62. Date of Electronic Publication: 2020 May 04 (Print Publication: 2020).
DOI: 10.1186/s13007-020-00605-5
Abstrakt: Background: Field-grown leafy vegetables can be damaged by biotic and abiotic factors, or mechanically damaged by farming practices. Available methods to evaluate leaf tissue damage mainly rely on colour differentiation between healthy and damaged tissues. Alternatively, sophisticated equipment such as microscopy and hyperspectral cameras can be employed. Depending on the causal factor, colour change in the wounded area is not always induced and, by the time symptoms become visible, a plant can already be severely affected. To accurately detect and quantify damage on leaf scale, including microlesions, reliable differentiation between healthy and damaged tissue is essential. We stained whole leaves with trypan blue dye, which traverses compromised cell membranes but is not absorbed in viable cells, followed by automated quantification of damage on leaf scale.
Results: We present a robust, fast and sensitive method for leaf-scale visualisation, accurate automated extraction and measurement of damaged area on leaves of leafy vegetables. The image analysis pipeline we developed automatically identifies leaf area and individual stained (lesion) areas down to cell level. As proof of principle, we tested the methodology for damage detection and quantification on two field-grown leafy vegetable species, spinach and Swiss chard.
Conclusions: Our novel lesion quantification method can be used for detection of large (macro) or single-cell (micro) lesions on leaf scale, enabling quantification of lesions at any stage and without requiring symptoms to be in the visible spectrum. Quantifying the wounded area on leaf scale is necessary for generating prediction models for economic losses and produce shelf-life. In addition, risk assessments are based on accurate prediction of the relationship between leaf damage and infection rates by opportunistic pathogens and our method helps determine the severity of leaf damage at fine resolution.
Competing Interests: Competing interestsThe authors declare that they have no competing interests.
(© The Author(s) 2020.)
Databáze: MEDLINE
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