Data from: Investigating a semiochemical-based mechanism for the beneficial activity of sorghum intercropped with soybeans on management of <i>Popillia japonica</i>
InsectsPopillia japonica were collected daily from wild populations in Riley and Pottawatomie Co., Kansas, during Jul–Aug 2025, with no-kill Japanese/Oriental beetle pheromone traps (Rescue International, Spokane, WA, USA). Traps were hung near home gardens, to avoid capturing beetles from agricultural fields. Beetles were kept in 25 × 25 cm plastic tubs with a bedding of moistened peat and River birch (Betula nigra) leaves and Virginia creeper (Parthenocissus quinquefolia) vines as food. Food was maintained ad libitum prior to use in the experiments below.PlantsSoybean (var. Pioneer P42A84E) and sorghum (var. Pioneer 85P58) were planted in plots at Kansas State University’s North Farm fields (location = 39.206550, -96.593667) on May 13, 2025. Row spacing was 30” (=76.2 cm) for both species.Plant Semiochemical ExtractsExtracts were taken from fresh samples, in the field on 5 August 2025. Intercropped soybean samples were taken from soybean planted directly next to sorghum, while monocropped soybean samples were taken from soybean plants in the middle of the field. Media storage jars (250-mL capacity) were cleaned with methanol, hexane, and dichloromethane. Afterwards, 100 ml of dichloromethane was added to each jar (MilliporeSigma, Burlington, MA, USA), and the jars were brought to the field on ice. For each soybean plant, 3 top leaves and 2 new trifolds were removed and placed into dichloromethane. For each sorghum plant, 1 apicol, approximately 50% of the leaf, and 5 racemes were removed and placed into 100 mL of dichloromethane. Samples were vigorously shaken for 30 seconds, then placed on ice for 1 h before filtration. Extracts were decanted through filter paper 85 mm (Grade 1, Whatman, Global Life Sciences Solutions, Buckinghamshire, UK) into fresh, clean mason jars and then stored at −20 °C until use. Leaves were left overnight to evaporate DCM. Leaves were placed in the oven in the morning to dry, and a dry weight was taken with a balance (Sartorius AG, Göttingen, Germany).Chemical analysis with GC-MSSolid-phase microextraction (SPME) employing a DVB/Carboxen/PDMS fiber (57329-U, Supelco, MilliporeSigma, St. Louis, MO, USA) was conducted to quantify the volatiles present in each solvent extract. A total of 50 μL aliquots of each sample were applied to a 1-cm diameter filter paper disc (Ahlstrom-Munksjö, Mt. Holly Springs, PA, USA) along with 2 μL of tetradecane as an internal standard and allowed to dry. Discs were then transferred into 20 ml headspace vials (093640-036-00, Gerstel Inc., Linthicum Heights, MD, USA) and sealed with a magnetic screw cap with a PTFE-backed septum (093640-040-00, Gerstel Inc.). Headspace vials were incubated at [temperature] for [time] without agitation to facilitate movement of compounds into the vial headspace before the SPME fiber was exposed to the headspace above the sample for 60 min. Samples were analyzed on an Agilent 7890B gas chromatograph (GC) equipped with an Agilent Durabond HP-5 column (30 m length, 0.250 mm diameter, 0.25 μm film thickness) and He as the carrier gas at a constant 1.2 mL min−1 flow rate and 40 cm s−1 velocity, which was coupled with a single-quadrupole Agilent 5977B mass spectrometer (MS). SPME fibers were desorbed in the inlet operated in splitless mode of the GC at 250°C for 3 min. The oven was initially maintained at 35°C for 3 min before increasing to 180°C at a rate of 10°C min−1. The oven temperature then increased from 180–250°C at 20°C min-1, where it was held for 2 min. Mass spectra were scanned from 35 to 550 m/z for the entire length of the run. A mixture containing C8–C20 alkanes was employed to calculate Kovats index for all peaks. Compounds were preliminarily identified by comparison of spectral data with those from the NIST 14 library and by GC retention index and Kovats index. The identities of several compounds were subsequently confirmed by comparing their retention times and mass spectra to authentic commercial standards from MilliporeSigma.Video-tracking Assay with EthovisionVideo-tracking coupled with Ethovision software v.14.0 (Noldus, Inc., Leesburg, VA, USA) was used to analyze P. japonica responses to the plant extracts. Six arenas consisting of Petri dishes (100 × 20 mm; VWR International, Radnor, PA, USA) with an 85-mm filter paper (Grade 1, Whatman; GE Healthcare, Chicago, IL, USA) adhered to the bottom using double-sided sticky tape were arranged below a network video camera (GigE, Basler AG, Ahrensburg, Germany). The entire setup was situated in a walk-in environmental chamber (Percival Scientific, Perry, IA, USA) set to constant 27.5C, and 60% relative humidity during the photophase when beetles are most active. Treatment extracts consisted of (1) 50 μL of monoculture soy extract, (2) 50 μL of intercropped soy (soy grown alongside sorghum) extract, and (3) a mixture of 50 μL of monoculture soy and 50 μL sorghum extract placed on a 1-cm diameter filter paper and allowed to evaporate prior to use. The control consisted of 50 μL of neat dichloromethane on a 1-cm diameter filter paper and allowed to evaporate for the same period. A filter paper with a given treatment was placed on one side of the arena, and a filter paper of neat solvent (DCM) was placed on the opposite side of the arena. Treatment sides were alternated, and the position of treatments was systematically rotated to avoid positional bias.Within each Petri dish, four zones were monitored, including the two halves of the Petri dish arena (i.e. treatment half versus control half) and the two 1 cm diameter stimulus zones nested in the center of each half where stimuli were applied (treatment stimulus zone and control stimulus zone). Popillia japonica were starved separately for 1 h prior to testing. The movement of individual beetles within each arena was simultaneously recorded on an adjacent computer, and after the experiment was finished, loaded into Ethovision with the appropriate detection and arena settings. We used Ethovision to record the distance moved and instantaneous velocity, as well as frequency entering, duration spent in, and latency to finding each half of the arena and the stimulus zones. For each assay, a single beetle was placed at the bottom center of an arena and allowed to interact with the extract and control with movement recorded over 10 min. A total of n = 24 replicate adults were tested per treatment.Release-Recapture Preference AssayPopillia japonica response to fresh plant cuttings was tested in in large plastic containers (25 × 25 × 26 cm) with moistened peat placed in large walk-in environmental chambers (Percival Scientific). Chambers were set to constant 28°C and 65% relative humidity with 16:8 L:D to mimic summer conditions. Soybean leaf cuttings consisted of one trifold and its petiole, while sorghum cuttings consisted of one raceme and two leaf portions weighing approximately 100 g each. Cuttings were placed directly on ice from the field and remained on ice for 1 h before the trial began. Plant petioles were placed into water in vaccine vials (Becton Dickinson, Franklin Lakes, NJ, USA) and closed with parafilm to maintain freshness then inserted into the peat flush at ground level to stand up at opposite sides of the chamber. Large plastic containers were divided into 5 equally-sized (5 cm) zones with a treatment on opposite sides of each container. Specifically, the central release zone was zone 3, while zone 1 and zone 5 contained stimuli. The treatment combinations were as follows; (i) monoculture soy versus intercropped soy (intersoy; soy next to sorghum); (ii) monoculture soy versus a mix of mono soy and sorghum; (iii) monoculture soy versus sorghum; (iv) intercropped soy versus a mix of mono soy and sorghum. Soybean trifolds were paired with similarly-sized trifolds within each container. A total of four unsexed beetles were placed at the center of each container to account for group dynamics. After 1 h and 24 h, beetle locations were recorded by zone. Beetles that did not leave the central zone (zone 3) after the response time were recorded as non-responsive, but still included in the analysis.
Resources
9 resources available
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ethovision_kucherov_etal_agdata_commons.csv
CSV -
dual_choice_treatment4_intersoy_mix_kucherov_etal_agdata_commons.csv
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dual_choice_treatment1_monosoy_intersoy_kucherov_etal_agdata_commons.csv
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dual_choice_treatment2_monosoy_mix_kucherov_etal_agdata_commons.csv
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dual_choice_treatment3_monosoy_sorghum_kucherov_etal_agdata_commons.csv
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ethovision_r_script_kucherov_etal_agdata_commons.R
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dual_assay_choice_r_script_kucherov_etal_agdata_commons.R
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extracts_r_script_kucherov_etal_agdata_commons.R
R -
extracts_soy_kucherov_etal_agdata_commons.csv
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| @type | dcat:Dataset |
|---|---|
| accessLevel | public |
| accrualPeriodicity | irregular |
| bureauCode |
[ "005:18" ] |
| contactPoint |
{ "fn": "Morrison, William", "hasEmail": "mailto:william.morrison@usda.gov" } |
| description | <p dir="ltr"><b><i>Insects</i></b></p><p dir="ltr"><i>Popillia japonica</i> were collected daily from wild populations in Riley and Pottawatomie Co., Kansas, during Jul–Aug 2025, with no-kill Japanese/Oriental beetle pheromone traps (Rescue International, Spokane, WA, USA). Traps were hung near home gardens, to avoid capturing beetles from agricultural fields. Beetles were kept in 25 × 25 cm plastic tubs with a bedding of moistened peat and River birch (<i>Betula nigra</i>) leaves and Virginia creeper (<i>Parthenocissus quinquefolia</i>) vines as food. Food was maintained ad libitum prior to use in the experiments below.</p><p dir="ltr"><b><i>Plants</i></b></p><p dir="ltr">Soybean (var. Pioneer P42A84E) and sorghum (var. Pioneer 85P58) were planted in plots at Kansas State University’s North Farm fields (location = 39.206550, -96.593667) on May 13, 2025. Row spacing was 30” (=76.2 cm) for both species.</p><p dir="ltr"><b><i>Plant</i></b><b><i> </i></b><b><i>Semiochemical</i></b><b><i> </i></b><b><i>Extracts</i></b></p><p dir="ltr">Extracts were taken from fresh samples, in the field on 5 August 2025. Intercropped soybean samples were taken from soybean planted directly next to sorghum, while monocropped soybean samples were taken from soybean plants in the middle of the field. Media storage jars (250-mL capacity) were cleaned with methanol, hexane, and dichloromethane. Afterwards, 100 ml of dichloromethane was added to each jar (MilliporeSigma, Burlington, MA, USA), and the jars were brought to the field on ice. For each soybean plant, 3 top leaves and 2 new trifolds were removed and placed into dichloromethane. For each sorghum plant, 1 apicol, approximately 50% of the leaf, and 5 racemes were removed and placed into 100 mL of dichloromethane. Samples were vigorously shaken for 30 seconds, then placed on ice for 1 h before filtration. Extracts were decanted through filter paper 85 mm (Grade 1, Whatman, Global Life Sciences Solutions, Buckinghamshire, UK) into fresh, clean mason jars and then stored at −20 °C until use. Leaves were left overnight to evaporate DCM. Leaves were placed in the oven in the morning to dry, and a dry weight was taken with a balance (Sartorius AG, Göttingen, Germany).</p><p dir="ltr"><b><i>Chemical analysis with GC</i></b><b><i>-</i></b><b><i>MS</i></b></p><p dir="ltr">Solid-phase microextraction (SPME) employing a DVB/Carboxen/PDMS fiber (57329-U, Supelco, MilliporeSigma, St. Louis, MO, USA) was conducted to quantify the volatiles present in each solvent extract. A total of 50 μL aliquots of each sample were applied to a 1-cm diameter filter paper disc (Ahlstrom-Munksjö, Mt. Holly Springs, PA, USA) along with 2 μL of tetradecane as an internal standard and allowed to dry. Discs were then transferred into 20 ml headspace vials (093640-036-00, Gerstel Inc., Linthicum Heights, MD, USA) and sealed with a magnetic screw cap with a PTFE-backed septum (093640-040-00, Gerstel Inc.). Headspace vials were incubated at [temperature] for [time] without agitation to facilitate movement of compounds into the vial headspace before the SPME fiber was exposed to the headspace above the sample for 60 min. Samples were analyzed on an Agilent 7890B gas chromatograph (GC) equipped with an Agilent Durabond HP-5 column (30 m length, 0.250 mm diameter, 0.25 μm film thickness) and He as the carrier gas at a constant 1.2 mL min<sup>−1</sup> flow rate and 40 cm s<sup>−1</sup> velocity, which was coupled with a single-quadrupole Agilent 5977B mass spectrometer (MS). SPME fibers were desorbed in the inlet operated in splitless mode of the GC at 250°C for 3 min. The oven was initially maintained at 35°C for 3 min before increasing to 180°C at a rate of 10°C min<sup>−1</sup>. The oven temperature then increased from 180–250°C at 20°C min<sup>-1</sup>, where it was held for 2 min. Mass spectra were scanned from 35 to 550 m/z for the entire length of the run. A mixture containing C<sub>8</sub>–C<sub>20</sub> alkanes was employed to calculate Kovats index for all peaks. Compounds were preliminarily identified by comparison of spectral data with those from the NIST 14 library and by GC retention index and Kovats index. The identities of several compounds were subsequently confirmed by comparing their retention times and mass spectra to authentic commercial standards from MilliporeSigma.</p><p dir="ltr"><b><i>Video-tracking Assay with</i></b><b><i> </i></b><b><i>Ethovision</i></b></p><p dir="ltr">Video-tracking coupled with Ethovision software v.14.0 (Noldus, Inc., Leesburg, VA, USA) was used to analyze <i>P. japonica</i><i> </i>responses to the plant extracts. Six arenas consisting of Petri dishes (100 × 20 mm; VWR International, Radnor, PA, USA) with an 85-mm filter paper (Grade 1, Whatman; GE Healthcare, Chicago, IL, USA) adhered to the bottom using double-sided sticky tape were arranged below a network video camera (GigE, Basler AG, Ahrensburg, Germany). The entire setup was situated in a walk-in environmental chamber (Percival Scientific, Perry, IA, USA) set to constant 27.5C, and 60% relative humidity during the photophase when beetles are most active. Treatment extracts consisted of (1) 50 μL of monoculture soy extract, (2) 50 μL of intercropped soy (soy grown alongside sorghum) extract, and (3) a mixture of 50 μL of monoculture soy and 50 μL sorghum extract placed on a 1-cm diameter filter paper and allowed to evaporate prior to use. The control consisted of 50 μL of neat dichloromethane on a 1-cm diameter filter paper and allowed to evaporate for the same period. A filter paper with a given treatment was placed on one side of the arena, and a filter paper of neat solvent (DCM) was placed on the opposite side of the arena. Treatment sides were alternated, and the position of treatments was systematically rotated to avoid positional bias.</p><p dir="ltr">Within each Petri dish, four zones were monitored, including the two halves of the Petri dish arena (i.e. treatment half versus control half) and the two 1 cm diameter stimulus zones nested in the center of each half where stimuli were applied (treatment stimulus zone and control stimulus zone). Popillia japonica were starved separately for 1 h prior to testing. The movement of individual beetles within each arena was simultaneously recorded on an adjacent computer, and after the experiment was finished, loaded into Ethovision with the appropriate detection and arena settings. We used Ethovision to record the distance moved and instantaneous velocity, as well as frequency entering, duration spent in, and latency to finding each half of the arena and the stimulus zones. For each assay, a single beetle was placed at the bottom center of an arena and allowed to interact with the extract and control with movement recorded over 10 min. A total of n = 24 replicate adults were tested per treatment.</p><p dir="ltr"><b><i>Release-Recapture Preference Assay</i></b></p><p dir="ltr"><i>Popillia japonica</i><i> </i>response to fresh plant cuttings was tested in in large plastic containers (25 × 25 × 26 cm) with moistened peat placed in large walk-in environmental chambers (Percival Scientific). Chambers were set to constant 28°C and 65% relative humidity with 16:8 L:D to mimic summer conditions. Soybean leaf cuttings consisted of one trifold and its petiole, while sorghum cuttings consisted of one raceme and two leaf portions weighing approximately 100 g each. Cuttings were placed directly on ice from the field and remained on ice for 1 h before the trial began. Plant petioles were placed into water in vaccine vials (Becton Dickinson, Franklin Lakes, NJ, USA) and closed with parafilm to maintain freshness then inserted into the peat flush at ground level to stand up at opposite sides of the chamber. Large plastic containers were divided into 5 equally-sized (5 cm) zones with a treatment on opposite sides of each container. Specifically, the central release zone was zone 3, while zone 1 and zone 5 contained stimuli. The treatment combinations were as follows; (i) monoculture soy versus intercropped soy (intersoy; soy next to sorghum); (ii) monoculture soy versus a mix of mono soy and sorghum; (iii) monoculture soy versus sorghum; (iv) intercropped soy versus a mix of mono soy and sorghum. Soybean trifolds were paired with similarly-sized trifolds within each container. A total of four unsexed beetles were placed at the center of each container to account for group dynamics. After 1 h and 24 h, beetle locations were recorded by zone. Beetles that did not leave the central zone (zone 3) after the response time were recorded as non-responsive, but still included in the analysis.</p> |
| distribution |
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| identifier | 10.15482/USDA.ADC/31661440.v1 |
| keyword |
[ "1-octanol", "1-octen-3-ol", "Japanese Beetle", "Kansas", "Kansas State University", "Manhattan", "North Agronomy Farm", "Popillia japonica", "SPME GC-MS.", "Semiochemicals", "Solid phase microextraction SPME", "USDA Center for Grain and Animal Health Research", "dual choice", "extracts", "gas chromatography coupled with mass spectrometry", "host cues", "intercropping", "monocropping", "orientation", "sorghum", "source code", "soybean", "taxis" ] |
| license | https://creativecommons.org/publicdomain/zero/1.0/ |
| modified | 2026-07-11 |
| programCode |
[ "005:040" ] |
| publisher |
{ "name": "Agricultural Research Service", "@type": "org:Organization" } |
| temporal | 2025-04-01/2025-09-18 |
| title | Data from: Investigating a semiochemical-based mechanism for the beneficial activity of sorghum intercropped with soybeans on management of <i>Popillia japonica</i> |
