Publications by authors named "Gerrie L Wiegers"

9 Publications

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An Integrated System for the Automated Recording and Analysis of Insect Behavior in T-maze Arrays.

Front Plant Sci 2019 29;10:20. Epub 2019 Jan 29.

Wageningen Plant Research, Wageningen University and Research, Wageningen, Netherlands.

Host-plant resistance to insects like thrips and aphids is a complex trait that is difficult to phenotype quickly and reliably. Here, we introduce novel hardware and software to facilitate insect choice assays and automate the acquisition and analysis of movement tracks. The hardware consists of an array of individual T-mazes allowing simultaneous release of up to 90 insect individuals from their individual cage below each T-maze with choice of two leaf disks under a video camera. Insect movement tracks are acquired with computer vision software (EthoVision) and analyzed with EthoAnalysis, a novel software package that allows for automated reporting of highly detailed behavior parameters and statistical analysis. To validate the benefits of the system we contrasted two accessions that were previously analyzed for differential resistance to western flower thrips. Results of two trials with 40 T-mazes are reported and we show how we arrived at optimized settings for the different filters and statistics. The statistics are reported in terms of frequency, duration, distance and speed of behavior events, both as sum totals and event averages, and both for the total trial period and in time bins of 1 h. Also included are higher level analyses with subcategories like short-medium-long events and slow-medium-fast events. The time bins showed how some behavior elements are more descriptive of differences between the genotypes during the first hours, whereas others are constant or become more relevant at the end of an 8 h recording. The three overarching behavior categories, i.e., choice, movement, and halting, were automatically corrected for the percentage of time thrips were detected and 24 out of 38 statistics of behavior parameters differed by a factor 2-6 between the accessions. The analysis resulted in much larger contrasts in behavior traits than reported previously. Compared to leaf damage assays on whole plants or detached leaves that take a week or more to complete, results were obtained in 8 h, with more detail, fewer individuals and higher significance. The potential value of the new integrated system, named EntoLab, for discovery of genetic traits in plants and insects by high throughput screening of large populations is discussed.
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http://dx.doi.org/10.3389/fpls.2019.00020DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6361829PMC
January 2019

SIEVE ELEMENT-LINING CHAPERONE1 Restricts Aphid Feeding on Arabidopsis during Heat Stress.

Plant Cell 2017 Oct 28;29(10):2450-2464. Epub 2017 Sep 28.

Bioscience, Wageningen University & Research, 6708 PB Wageningen, The Netherlands.

The role of phloem proteins in plant resistance to aphids is still largely elusive. By genome-wide association mapping of aphid behavior on 350 natural accessions, we identified the small heat shock-like (). Detailed behavioral studies on near-isogenic and knockout lines showed that SLI1 impairs phloem feeding. Depending on the haplotype, aphids displayed a different duration of salivation in the phloem. On mutants, aphids prolonged their feeding sessions and ingested phloem at a higher rate than on wild-type plants. The largest phenotypic effects were observed at 26°C, when expression is upregulated. At this moderately high temperature, mutants suffered from retarded elongation of the inflorescence and impaired silique development. Fluorescent reporter fusions showed that SLI1 is confined to the margins of sieve elements where it lines the parietal layer and colocalizes in spherical bodies around mitochondria. This localization pattern is reminiscent of the clamp-like structures observed in previous ultrastructural studies of the phloem and shows that the parietal phloem layer plays an important role in plant resistance to aphids and heat stress.
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http://dx.doi.org/10.1105/tpc.16.00424DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5774557PMC
October 2017

Genetic architecture of plant stress resistance: multi-trait genome-wide association mapping.

New Phytol 2017 Feb 4;213(3):1346-1362. Epub 2016 Oct 4.

Wageningen University and Research Plant Breeding, Wageningen University and Research, PO Box 386, 6700 AJ, Wageningen, the Netherlands.

Plants are exposed to combinations of various biotic and abiotic stresses, but stress responses are usually investigated for single stresses only. Here, we investigated the genetic architecture underlying plant responses to 11 single stresses and several of their combinations by phenotyping 350 Arabidopsis thaliana accessions. A set of 214 000 single nucleotide polymorphisms (SNPs) was screened for marker-trait associations in genome-wide association (GWA) analyses using tailored multi-trait mixed models. Stress responses that share phytohormonal signaling pathways also share genetic architecture underlying these responses. After removing the effects of general robustness, for the 30 most significant SNPs, average quantitative trait locus (QTL) effect sizes were larger for dual stresses than for single stresses. Plants appear to deploy broad-spectrum defensive mechanisms influencing multiple traits in response to combined stresses. Association analyses identified QTLs with contrasting and with similar responses to biotic vs abiotic stresses, and below-ground vs above-ground stresses. Our approach allowed for an unprecedented comprehensive genetic analysis of how plants deal with a wide spectrum of stress conditions.
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http://dx.doi.org/10.1111/nph.14220DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5248600PMC
February 2017

AtWRKY22 promotes susceptibility to aphids and modulates salicylic acid and jasmonic acid signalling.

J Exp Bot 2016 05 23;67(11):3383-96. Epub 2016 Apr 23.

Plant Research International, Business Unit Bioscience, Wageningen University and Research Centre, PO Box 16, 6700 AA Wageningen, The Netherlands.

Aphids induce many transcriptional perturbations in their host plants, but the signalling cascades responsible and the effects on plant resistance are largely unknown. Through a genome-wide association (GWA) mapping study in Arabidopsis thaliana, we identified WRKY22 as a candidate gene associated with feeding behaviour of the green peach aphid, Myzus persicae The transcription factor WRKY22 is known to be involved in pathogen-triggered immunity, and WRKY22 gene expression has been shown to be induced by aphids. Assessment of aphid population development and feeding behaviour on knockout mutants and overexpression lines showed that WRKY22 increases susceptibility to M. persicae via a mesophyll-located mechanism. mRNA sequencing analysis of aphid-infested wrky22 knockout plants revealed the up-regulation of genes involved in salicylic acid (SA) signalling and down-regulation of genes involved in plant growth and cell-wall loosening. In addition, mechanostimulation of knockout plants by clip cages up-regulated jasmonic acid (JA)-responsive genes, resulting in substantial negative JA-SA crosstalk. Based on this and previous studies, WRKY22 is considered to modulate the interplay between the SA and JA pathways in response to a wide range of biotic and abiotic stimuli. Its induction by aphids and its role in suppressing SA and JA signalling make WRKY22 a potential target for aphids to manipulate host plant defences.
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http://dx.doi.org/10.1093/jxb/erw159DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4892728PMC
May 2016

Automated video tracking of thrips behavior to assess host-plant resistance in multiple parallel two-choice setups.

Plant Methods 2016 18;12. Epub 2016 Jan 18.

PRI-Bioscience, Wageningen University and Research Center, P.O. Box 16, 6700 AA Wageningen, The Netherlands.

Background: Piercing-sucking insects cause severe damage in crops. Breeding for host-plant resistance can significantly reduce the yield losses caused by these insects, but host-plant resistance is a complex trait that is difficult to phenotype quickly and reliably. Current phenotyping methods mainly focus on labor-intensive and time-consuming end-point measurements of plant fitness. Characterizing insect behavior as a proxy for host-plant resistance could be a promising time-saving alternative to end-point measurements.

Results: We present a phenotyping platform that allows screening for host-plant resistance against Western flower thrips (WFT, Frankliniella occidentalis (Pergande)) in a parallel two-choice setup using automated video tracking of thrips behavior. The platform was used to establish host-plant preference of WFT with a large plant population of 345 wild Arabidopsis accessions and the method was optimized with two extreme accessions from this population that differed in resistance towards WFT. To this end, the behavior of 88 WFT individuals was simultaneously tracked in 88 parallel two-choice arenas during 8 h. Host-plant preference of WFT was established both by the time thrips spent on either accession and various behavioral parameters related to movement (searching) and non-movement (feeding) events.

Conclusion: In comparison to 6-day end-point choice assays with whole plants or detached leaves, the automated video-tracking choice assay developed here delivered similar results, but with higher time- and resource efficiency. This method can therefore be a reliable and effective high throughput phenotyping tool to assess host-plant resistance to thrips in large plant populations.
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http://dx.doi.org/10.1186/s13007-016-0102-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4717623PMC
January 2016

High-throughput phenotyping of plant resistance to aphids by automated video tracking.

Plant Methods 2015 30;11. Epub 2015 Jan 30.

Plant Research International, Wageningen University and Research Center, P.O. Box 16, 6700 AA Wageningen, The Netherlands.

Background: Piercing-sucking insects are major vectors of plant viruses causing significant yield losses in crops. Functional genomics of plant resistance to these insects would greatly benefit from the availability of high-throughput, quantitative phenotyping methods.

Results: We have developed an automated video tracking platform that quantifies aphid feeding behaviour on leaf discs to assess the level of plant resistance. Through the analysis of aphid movement, the start and duration of plant penetrations by aphids were estimated. As a case study, video tracking confirmed the near-complete resistance of lettuce cultivar 'Corbana' against Nasonovia ribisnigri (Mosely), biotype Nr:0, and revealed quantitative resistance in Arabidopsis accession Co-2 against Myzus persicae (Sulzer). The video tracking platform was benchmarked against Electrical Penetration Graph (EPG) recordings and aphid population development assays. The use of leaf discs instead of intact plants reduced the intensity of the resistance effect in video tracking, but sufficiently replicated experiments resulted in similar conclusions as EPG recordings and aphid population assays. One video tracking platform could screen 100 samples in parallel.

Conclusions: Automated video tracking can be used to screen large plant populations for resistance to aphids and other piercing-sucking insects.
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http://dx.doi.org/10.1186/s13007-015-0044-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4318543PMC
February 2015

Spatial separation of semiochemical Lurem-TR and entomopathogenic fungi to enhance their compatibility and infectivity in an autoinoculation system for thrips management.

Pest Manag Sci 2016 Jan 17;72(1):131-9. Epub 2015 Feb 17.

International Centre of Insect Physiology and Ecology, Nairobi, Kenya.

Background: The effect of spatial separation of the semiochemical Lurem-TR, which has been found to inhibit conidia of entomopathogenic fungi when put together, on the persistence of conidia of Metarhizium brunneum and M. anisopliae was evaluated in the greenhouse and field in order to develop an autodissemination strategy for the management of Megalurothrips sjostedti on cowpea crop. Influence of spatial separation of the semiochemical on thrips attraction and conidial acquisition by thrips from the autoinoculation device was also investigated in the field.

Results: Persistence of conidia of M. brunneum and M. anisopliae increased with distance of separation of Lurem-TR. Direct exposure of fungus without separation from Lurem-TR recorded the lowest conidial germination as compared with the other treatments. Attraction of thrips to the device also varied significantly according to distance between device and semiochemical, with a higher number of thrips attracted when Lurem-TR was placed in a container below the device and at 10 cm distance. There was no significant difference in conidial acquisition between spatial separation treatments of conidia and Lurem-TR. Attraction of other insect pests to the device did not significantly vary between treatments. Positive correlations were found between conidial acquisition and thrips attraction.

Conclusion: This study suggests that spatial separation of fungal conidia from Lurem-TR in an autoinoculation device could provide a low-cost strategy for effective management of thrips in grain legume cropping systems.
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http://dx.doi.org/10.1002/ps.3979DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5024074PMC
January 2016

Engineered multidomain cysteine protease inhibitors yield resistance against western flower thrips (Frankliniella occidentalis) in greenhouse trials.

Plant Biotechnol J 2004 Sep;2(5):449-58

Plant Research International, PO Box 16, NL-6700 AA Wageningen, The Netherlands.

Western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), cause very large economic damage on a variety of field and greenhouse crops. In this study, plant resistance against thrips was introduced into transgenic potato plants through the expression of novel, custom-made, multidomain protease inhibitors. Representative classes of inhibitors of cysteine and aspartic proteases [kininogen domain 3 (K), stefin A (A), cystatin C (C), potato cystatin (P) and equistatin (EIM)] were fused into reading frames consisting of four (K-A-C-P) to five (EIM-K-A-C-P) proteins, and were shown to fold into functional inhibitors in the yeast Pichia pastoris. The multidomain proteins were expressed in potato and found to be more resistant to degradation by plant proteases than the individual domains. In a time span of 14-16 days, transgenic potato plants expressing EIMKACP and KACP at a similar concentration reduced the number of larvae and adults to less than 20% of the control. Leaf damage on protected plants was minimal. Engineered multidomain cysteine protease inhibitors thus provide a novel way of controlling western flower thrips in greenhouse and field crops, and open up possibilities for novel insect resistance applications in transgenic crops.
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http://dx.doi.org/10.1111/j.1467-7652.2004.00089.xDOI Listing
September 2004

Oral toxicity of Photorhabdus toxins against thrips species.

J Invertebr Pathol 2005 Mar 25;88(3):207-11. Epub 2005 Feb 25.

Plant Research International, P.O. Box 16, 6700AA Wageningen, The Netherlands.

The oral toxicity of excretion products of several Photorhabdus and Xenorhabdus strains was tested on two thrips species: Frankliniella occidentalis and Thrips tabaci. Out of 46 Photorhabdus isolates and six Xenorhabdus isolates only six North American P. temperata isolates were toxic to the thrips species. After 7 days of drinking from P. temperata supernatant a mortality of 90% could be reached. Thrips were also killed after sucking from leaves covered with the toxins. Toxins have a negative effect on thrips fecundity. Possibilities of using P. temperata in the control of thrips will be discussed.
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http://dx.doi.org/10.1016/j.jip.2005.01.009DOI Listing
March 2005