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

Kloth KJ, Ten Broeke CJ, Thoen MP, Hanhart-van den Brink M, Wiegers GL, Krips OE, Noldus LP, Dicke M, Jongsma MA - Plant Methods (2015)

Bottom Line: Functional genomics of plant resistance to these insects would greatly benefit from the availability of high-throughput, quantitative phenotyping methods.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.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Entomology, Wageningen University, P.O. Box 16, 6700 AA Wageningen, The Netherlands ; Laboratory of Plant Physiology, Wageningen University, P.O. Box 16, 6700 AA Wageningen, The Netherlands ; Plant Research International, Wageningen University and Research Center, P.O. Box 16, 6700 AA Wageningen, The Netherlands.

ABSTRACT

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.

No MeSH data available.


Related in: MedlinePlus

Contribution of salivation to phloem ingestion. Percentage of time spent salivating in the phloem compared to the total phloem phase (salivation + ingestion) of M. persicae aphids on Arabidopsis accessions Co-2 (resistant) and Sanna-2 (susceptible) (Mann–Whitney U test, *P < 0.05; **P < 0.01; ***P < 0.001, left bars: EPG recording intact plants: n = 19, right bars: EPG recording leaf discs: Co-2 n = 9, Sanna-2 n = 8, error bars represent standard error).
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Fig7: Contribution of salivation to phloem ingestion. Percentage of time spent salivating in the phloem compared to the total phloem phase (salivation + ingestion) of M. persicae aphids on Arabidopsis accessions Co-2 (resistant) and Sanna-2 (susceptible) (Mann–Whitney U test, *P < 0.05; **P < 0.01; ***P < 0.001, left bars: EPG recording intact plants: n = 19, right bars: EPG recording leaf discs: Co-2 n = 9, Sanna-2 n = 8, error bars represent standard error).

Mentions: To validate whether automated video tracking delivered a reliable proxy for plant resistance, feeding behaviour of M. persicae was measured during 8 hours continuous recording on two natural accessions of Arabidopsis, Co-2 and Sanna-2 (Additional file 3). These accessions were selected from a population of hundreds of accessions based on preliminary video tracking data. Automated video tracking showed that M. persicae aphids walked larger distances on Co-2 and reduced the mean duration of long probes (Table 1). EPG recordings on intact plants confirmed shorter durations of (sustained) phloem ingestion, and additionally revealed more short probes, non-probing behaviour and a delayed phloem uptake on Co-2 (Table 1). This behaviour is an indication of both epidermis/mesophyll-located and phloem-located resistance in Co-2 against M. persicae. All aphids ingested phloem, but quantitative differences in feeding behaviour between aphids on Co-2 and on Sanna-2 were already apparent in the first hour (Figure 5). A reproduction assay on intact plants confirmed that Co-2 was indeed more resistant than Sanna-2, although the resistance was not absolute. Depending on plant age, aphids either started reproduction later or produced fewer offspring (Figure 6). Although we had been able to correctly identify a quantitative difference in resistance with automated video tracking, the effects were smaller than in EPG recordings on intact plants. To verify whether the plant line effects in the video tracking assay were attenuated due to the use of excised plant tissue, the EPG experiment was repeated with leaf discs. Particularly for the resistant accession, aphid feeding behaviour was different and involved more phloem uptake and fewer short probes on leaf discs compared to intact plants (Additional file 1: Table S2). The only significant difference between the accessions that remained was a reduced duration of phloem uptake events on Co-2 (Table 1). In addition, contribution of salivation to the phloem phase, required to suppress (callose-mediated) sieve-plate occlusion [45], was equal on leaf discs but higher on intact plants of Co-2 (Figure 7). This indicates that the resistance mechanisms in intact plants were partially lost in leaf discs.Table 1


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

Kloth KJ, Ten Broeke CJ, Thoen MP, Hanhart-van den Brink M, Wiegers GL, Krips OE, Noldus LP, Dicke M, Jongsma MA - Plant Methods (2015)

Contribution of salivation to phloem ingestion. Percentage of time spent salivating in the phloem compared to the total phloem phase (salivation + ingestion) of M. persicae aphids on Arabidopsis accessions Co-2 (resistant) and Sanna-2 (susceptible) (Mann–Whitney U test, *P < 0.05; **P < 0.01; ***P < 0.001, left bars: EPG recording intact plants: n = 19, right bars: EPG recording leaf discs: Co-2 n = 9, Sanna-2 n = 8, error bars represent standard error).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
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getmorefigures.php?uid=PMC4318543&req=5

Fig7: Contribution of salivation to phloem ingestion. Percentage of time spent salivating in the phloem compared to the total phloem phase (salivation + ingestion) of M. persicae aphids on Arabidopsis accessions Co-2 (resistant) and Sanna-2 (susceptible) (Mann–Whitney U test, *P < 0.05; **P < 0.01; ***P < 0.001, left bars: EPG recording intact plants: n = 19, right bars: EPG recording leaf discs: Co-2 n = 9, Sanna-2 n = 8, error bars represent standard error).
Mentions: To validate whether automated video tracking delivered a reliable proxy for plant resistance, feeding behaviour of M. persicae was measured during 8 hours continuous recording on two natural accessions of Arabidopsis, Co-2 and Sanna-2 (Additional file 3). These accessions were selected from a population of hundreds of accessions based on preliminary video tracking data. Automated video tracking showed that M. persicae aphids walked larger distances on Co-2 and reduced the mean duration of long probes (Table 1). EPG recordings on intact plants confirmed shorter durations of (sustained) phloem ingestion, and additionally revealed more short probes, non-probing behaviour and a delayed phloem uptake on Co-2 (Table 1). This behaviour is an indication of both epidermis/mesophyll-located and phloem-located resistance in Co-2 against M. persicae. All aphids ingested phloem, but quantitative differences in feeding behaviour between aphids on Co-2 and on Sanna-2 were already apparent in the first hour (Figure 5). A reproduction assay on intact plants confirmed that Co-2 was indeed more resistant than Sanna-2, although the resistance was not absolute. Depending on plant age, aphids either started reproduction later or produced fewer offspring (Figure 6). Although we had been able to correctly identify a quantitative difference in resistance with automated video tracking, the effects were smaller than in EPG recordings on intact plants. To verify whether the plant line effects in the video tracking assay were attenuated due to the use of excised plant tissue, the EPG experiment was repeated with leaf discs. Particularly for the resistant accession, aphid feeding behaviour was different and involved more phloem uptake and fewer short probes on leaf discs compared to intact plants (Additional file 1: Table S2). The only significant difference between the accessions that remained was a reduced duration of phloem uptake events on Co-2 (Table 1). In addition, contribution of salivation to the phloem phase, required to suppress (callose-mediated) sieve-plate occlusion [45], was equal on leaf discs but higher on intact plants of Co-2 (Figure 7). This indicates that the resistance mechanisms in intact plants were partially lost in leaf discs.Table 1

Bottom Line: Functional genomics of plant resistance to these insects would greatly benefit from the availability of high-throughput, quantitative phenotyping methods.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.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Entomology, Wageningen University, P.O. Box 16, 6700 AA Wageningen, The Netherlands ; Laboratory of Plant Physiology, Wageningen University, P.O. Box 16, 6700 AA Wageningen, The Netherlands ; Plant Research International, Wageningen University and Research Center, P.O. Box 16, 6700 AA Wageningen, The Netherlands.

ABSTRACT

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.

No MeSH data available.


Related in: MedlinePlus