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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

Behavioural parameters ofN. ribisnigrion two lettuce cultivars, Corbana (resistant) and Terlana (susceptible). (a) Percentage of the time spent on long probes (>25 min), and (b) distance moved (cm) were measured by automated video tracking on leaf discs. (c) Percentage of the time spent on phloem feeding (waveform 5), and (d) percentage of time spent on other probes (pathway, phloem salivation and xylem feeding) were measured by EPGs on intact plants (Mann–Whitney U test per time bin, *P < 0.05; **P < 0.01; ***P < 0.001, video tracking: Corbana n = 27, Terlana n = 28, EPG recording: n = 19).
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Fig8: Behavioural parameters ofN. ribisnigrion two lettuce cultivars, Corbana (resistant) and Terlana (susceptible). (a) Percentage of the time spent on long probes (>25 min), and (b) distance moved (cm) were measured by automated video tracking on leaf discs. (c) Percentage of the time spent on phloem feeding (waveform 5), and (d) percentage of time spent on other probes (pathway, phloem salivation and xylem feeding) were measured by EPGs on intact plants (Mann–Whitney U test per time bin, *P < 0.05; **P < 0.01; ***P < 0.001, video tracking: Corbana n = 27, Terlana n = 28, EPG recording: n = 19).

Mentions: Apart from a study system with partial resistance, an example of near-complete resistance was tested with the video tracking platform. The behaviour of black-currant lettuce aphids, N. ribisnigri, biotype Nr:0 was recorded on two near-isogenic lettuce cultivars, the resistant ‘Corbana’ and susceptible ‘Terlana’. Previous studies showed that the Nr gene is responsible for near-complete resistance in Corbana against this biotype of aphids, mainly due to a phloem-located mechanism [34,46]. Our video tracking observations on leaf discs were compared to EPG recording data by ten Broeke et al. [47]. Seven out of nine video tracking variables confirmed that cultivar Corbana was more resistant than cultivar Terlana (Table 1). Aphids on Corbana spent less time on long probes and more time on shorter probes and other activities. In addition, aphids increased their walking activity over time on both cultivars, but generally covered larger distances on Corbana leaf discs (mixed linear model: time effect: P = 0.00, cultivar effect: P = 0.03, time × cultivar interaction: P = 0.77, Figure 8). Yet, the resistance effect was less pronounced in video tracking compared to EPG recording on intact plants: only 11% of the aphids in EPG recordings showed phloem ingestion on Corbana plants, while 78% of the aphids in the video assay performed long probes on Corbana. These long probing events could include other activities, such as water ingestion from xylem vessels, since EPGs showed that on Corbana plants more aphids penetrated xylem sieve elements (12 aphids on Corbana versus 2 aphids on Terlana).Figure 8


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)

Behavioural parameters ofN. ribisnigrion two lettuce cultivars, Corbana (resistant) and Terlana (susceptible). (a) Percentage of the time spent on long probes (>25 min), and (b) distance moved (cm) were measured by automated video tracking on leaf discs. (c) Percentage of the time spent on phloem feeding (waveform 5), and (d) percentage of time spent on other probes (pathway, phloem salivation and xylem feeding) were measured by EPGs on intact plants (Mann–Whitney U test per time bin, *P < 0.05; **P < 0.01; ***P < 0.001, video tracking: Corbana n = 27, Terlana n = 28, EPG recording: n = 19).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig8: Behavioural parameters ofN. ribisnigrion two lettuce cultivars, Corbana (resistant) and Terlana (susceptible). (a) Percentage of the time spent on long probes (>25 min), and (b) distance moved (cm) were measured by automated video tracking on leaf discs. (c) Percentage of the time spent on phloem feeding (waveform 5), and (d) percentage of time spent on other probes (pathway, phloem salivation and xylem feeding) were measured by EPGs on intact plants (Mann–Whitney U test per time bin, *P < 0.05; **P < 0.01; ***P < 0.001, video tracking: Corbana n = 27, Terlana n = 28, EPG recording: n = 19).
Mentions: Apart from a study system with partial resistance, an example of near-complete resistance was tested with the video tracking platform. The behaviour of black-currant lettuce aphids, N. ribisnigri, biotype Nr:0 was recorded on two near-isogenic lettuce cultivars, the resistant ‘Corbana’ and susceptible ‘Terlana’. Previous studies showed that the Nr gene is responsible for near-complete resistance in Corbana against this biotype of aphids, mainly due to a phloem-located mechanism [34,46]. Our video tracking observations on leaf discs were compared to EPG recording data by ten Broeke et al. [47]. Seven out of nine video tracking variables confirmed that cultivar Corbana was more resistant than cultivar Terlana (Table 1). Aphids on Corbana spent less time on long probes and more time on shorter probes and other activities. In addition, aphids increased their walking activity over time on both cultivars, but generally covered larger distances on Corbana leaf discs (mixed linear model: time effect: P = 0.00, cultivar effect: P = 0.03, time × cultivar interaction: P = 0.77, Figure 8). Yet, the resistance effect was less pronounced in video tracking compared to EPG recording on intact plants: only 11% of the aphids in EPG recordings showed phloem ingestion on Corbana plants, while 78% of the aphids in the video assay performed long probes on Corbana. These long probing events could include other activities, such as water ingestion from xylem vessels, since EPGs showed that on Corbana plants more aphids penetrated xylem sieve elements (12 aphids on Corbana versus 2 aphids on Terlana).Figure 8

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