An objective high‐throughput screening method for thrips damage quantitation using Ilastik and ImageJ

Visschers, Isabella G. S.; Dam, Nicole M. van; Peters, Janny L.

doi:10.1111/eea.12682

Cited by 20 publications

(23 citation statements)

References 36 publications

(38 reference statements)

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“…This is why we fill the cages with 1% agarose the day before the experiment [23], and sandwich them between two glass plates as shown in Fig 1. Lastly, leaf disks are mechanically excised, thus wounding the tissues, especially along the border of the disk. As reported in [9], several studies found no differences between detached/attached leaves and leaf disc assays. The results obtained with leaf disks ought to be considered as a proxy precisely because of these limitations.…”

Section: Discussionsupporting

confidence: 51%

“…Leaf-disk assays are commonly used to evaluate plant resistance against insects with chewing mouthparts [7,8] by measuring the area of tissues consumed. They can be used also on mites, thrips, aphids, whiteflies, or even fungi by monitoring visual changes related to the damaged areas of the leaves [9]. While disk assays have drawbacks such as the damage inflicted to the tissue, several studies demonstrate that resistance scores obtained using this approach are comparable to those completed on detached or attached leaves [9].…”

Section: Introductionmentioning

confidence: 99%

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A semi-automated design for high-throughput Lepidoptera larvae feeding bioassays

Zoungrana

Legrand

Dillmann

et al. 2020

Preprint

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Lepidopteran pests cause considerable damage to all crops over the world. As larvae are directly responsible for these damages, many research efforts are devoted to find plant cultivars which are resistant against them. However, such studies take time, efforts and are costly, especially when one wants to not only find resistance traits but also evaluate their heritability. We present here a high throughput approach to screen plants for resistance or chemicals for their deterrence, using a leaf-disk consumption assay, which is both suitable for large scale tests and economically affordable. To monitor larvae feeding on leaf disks placed over a layer of agar, we designed 3D models of 50 cages arrays. One webcam can sample simultaneously 3 of such arrays at a rate of 1 image/min, and follow individual feeding activities in each cage as the movements of 150 larvae. The resulting image stacks are first processed with a custom program running under an open-source image analysis package (Icy) to measure the surface of each leaf disk over time. We further developed statistical procedures running under the R package, to analyze the time course of the feeding activities of the larvae and to compare them between treatments. As a test case, we compared how European corn borer larvae respond to quinine, considered as a bitter alkaloid for many organisms, and to Neemazal containing azadirachtin, which is a common antifeedant against pest insects. We found that increasing doses of azadirachtin reduce and delay feeding. However, contrary to our expectation, quinine was found poorly effective at the range of concentrations tested. The 3D printed model of the cage, of the camera holder, the plugins running under Icy, and the R procedures are freely available, and can be modified according to the particular needs of the users.

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Section: Discussionsupporting

confidence: 51%

Section: Introductionmentioning

confidence: 99%

A semi-automated design for high-throughput Lepidoptera larvae feeding bioassays

Zoungrana

Legrand

Dillmann

et al. 2020

Preprint

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“…Per accession 8–10 plants were used for this choice experiment. Leaf disc experiments were performed as described by Visschers et al ( 2018a ). Briefly, using a cork borer, two leaf discs (1.5 cm diameter) were punched from each leaf, thereby avoiding the mid-vein.…”

Section: Methodsmentioning

confidence: 99%

Metabolomics of Thrips Resistance in Pepper (Capsicum spp.) Reveals Monomer and Dimer Acyclic Diterpene Glycosides as Potential Chemical Defenses

et al. 2019

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The development of pesticide resistance in insects and recent bans on pesticides call for the identification of natural sources of resistance in crops. Here, we used natural variation in pepper ( Capsicum spp.) resistance combined with an untargeted metabolomics approach to detect secondary metabolites related to thrips ( Frankliniella occidentalis ) resistance. Using leaf disc choice assays, we tested 11 Capsicum accessions of C. annuum and C. chinense in both vegetative and flowering stages for thrips resistance. Metabolites in the leaves of these 11 accessions were analyzed using LC-MS based untargeted metabolomics. The choice assays showed significant differences among the accessions in thrips feeding damage. The level of resistance depended on plant developmental stage. Metabolomics analyses showed differences in metabolomes among the Capsicum species and plant developmental stages. Moreover, metabolomic profiles of resistant and susceptible accessions differed. Monomer and dimer acyclic diterpene glycosides (capsianosides) were pinpointed as metabolites that were related to thrips resistance. Sucrose and malonylated flavone glycosides were related to susceptibility. To our knowledge, this is the first time that dimer capsianosides of pepper have been linked to insect resistance. Our results show the potential of untargeted metabolomics as a tool for discovering metabolites that are important in plant – insect interactions. Electronic supplementary material The online version of this article (10.1007/s10886-019-01074-4) contains supplementary material, which is available to authorized users.

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“…Our results provide experimental evidence that leaf disc assays are a suitable method for screening resistance to thrips in Capsicum (Maharijaya et al, 2011;Visschers et al, 2018a). In longer-term whole plant screening assays, herbivore-induced defences may play a role when screening for resistance to insects (Dillon, Chludil, Reichelt, Mithöfer, & Zavala, 2018).…”

Section: Whole Plant Versus Leaf Disc Assaysmentioning

confidence: 66%

Resistance to three thrips species in Capsicum spp. depends on site conditions and geographic regions

Visschers

Peters

Timmermans

et al. 2019

J Applied Entomology

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This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made. AbstractCapsicum species are commercially grown for pepper production. This crop suffers severely from thrips damage and the identification of natural sources of thrips resistance is essential for the development of resistant cultivars. It is unclear whether resistance to Frankliniella occidentalis as assessed in a specific environment holds under different conditions. Additionally, other thrips species may respond differently to the plant genotypes. Screening for robust and general resistance to thrips encompasses testing different Capsicum accessions under various conditions and with different thrips species. We screened 11 Capsicum accessions (C. annuum and C. chinense) for resistance to F. occidentalis at three different locations in the Netherlands. Next, the same 11 accessions were screened for resistance to Thrips palmi and Scirtothrips dorsalis at two locations in Asia. This resulted in a unique analysis of thrips resistance in Capsicum at five different locations around the world. Finally, all accessions were also screened for resistance to F. occidentalis in the Netherlands using a leaf disc choice assay, allowing direct comparison of whole plant and leaf disc assays. Resistance to F. occidentalis was only partially consistent among the three sites in the Netherlands.The most susceptible accessions were consistently susceptible, but which accession was the most resistant differed among sites. In Asia, one C. chinense accession was particularly resistant to S. dorsalis and T. palmi, but this was not the most resistant accession to F. occidentalis. Overall, resistance to F. occidentalis correlated with S. dorsalis but not with T. palmi resistance in the C. annuum accessions. Damage inflicted on leaf discs reflected damage on the whole plant level. Our study showed that identifying broad spectrum resistance to thrips in Capsicum may prove to be challenging.Breeding programmes should focus on developing cultivars suitable for growing in defined geographic regions with specific thrips species and abiotic conditions.

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An objective high‐throughput screening method for thrips damage quantitation using Ilastik and ImageJ

Cited by 20 publications

References 36 publications

A semi-automated design for high-throughput Lepidoptera larvae feeding bioassays

A semi-automated design for high-throughput Lepidoptera larvae feeding bioassays

Metabolomics of Thrips Resistance in Pepper (Capsicum spp.) Reveals Monomer and Dimer Acyclic Diterpene Glycosides as Potential Chemical Defenses

Resistance to three thrips species in Capsicum spp. depends on site conditions and geographic regions

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