ENTOMOLOGY RESEARCH REPORT - 2013 Field Evaluations Of

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Zsofia Szendrei, Department of Entomology MSU517-974-8610, szendrei@msu.eduENTOMOLOGY RESEARCH REPORT - 2013Field evaluations of registered and experimental insecticides for managingColorado potato beetle on potatoesThe Colorado potato beetle (Leptinotarsa decemlineata, Say, Coleoptera:Chrysomelidae) is the most widespread and destructive insect pest of potato crops in the easternUnited States and Canada. Its ability to develop resistance to insecticides makes it veryimportant to continue testing the efficacy of both new insecticide chemistries and existingcompounds. Such tests provide data on comparative effectiveness of products and data to helpsupport future registrations and use recommendations.METHODSFifteen insecticide treatments and an untreated check (Table 1) were tested at the MSUMontcalm Research Farm, Entrican, MI for control of Colorado potato beetle. ‘Atlantic’ potatoseed pieces were planted 12 in. apart, with 34 in. row spacing on 3 June 2012. Treatments werereplicated four times in a randomized complete block design. Plots were 50 ft. long and threerows wide with untreated guard rows bordering each plot.Brigadier 2SC, Verimark 20SC, Admire Pro 4.6SC, and Platinum 75SG treatments wereapplied as in-furrow sprays at planting on 3 June 2013. Post-plant directed sprays were appliedwhen potatoes first started emerging from the soil, on 10 June 2013. Foliar treatments were firstapplied at greater than 50% Colorado potato beetle egg hatch on 28 June 2013. Based on theeconomic threshold of more than one large larva per plant, additional first generation sprays wereneeded for Gladiator (11 and 18 July), Athena (11 July), Blackhawk (11 July), the low rate ofTorac 15 EC (11 July), and Admire Pro (11 and 18 July); no subsequent applications werenecessary for any of the Dupont treatments. All applications were made using a single-nozzlehand-held boom (30 gallons/acre and 30 psi).stPost-spray counts of first generation Colorado potato beetle adults, small larvae (1 andndrdth2 instars), and large larvae (3 and 4 instars) from five randomly selected plants from themiddle row of each plot were made weekly, starting on 2 July. Plots were visually rated fordefoliation weekly by estimating total defoliation per plot.The numbers of small larvae, large larvae, and adults, as well as the defoliation ratings,were transformed log (x 1) prior to analysis. Analysis of variance was used for data analysisand ad-hoc Tukey means separation was used to compare treatment means (P 0.05).RESULTSExcept for Admire Pro and Athena, all treatments resulted in significantly fewer smalllarvae than the untreated control, while all treatments significantly reduced the number of largelarvae per plant, compared to the untreated (Table 2). There were also significant differences innumbers of large larvae among the insecticide treatments. All three systemic products (AdmirePro, A16901, and Platinum 75 SG) performed well, with A16901 having significantly fewer largelarvae than six of the foliar products. Among the foliar products, Admire Pro required weeklysprays, while F9318 and the low rate of Torac 15 EC were applied three of the four weeks.Athena, Blackhawk, and the high rate of Torac 15 EC required one subsequent application, alltwo weeks after the initial application. Of these, however, only Blackhawk provided reduction inaverage large larvae below the threshold of one per plant. Despite one fewer application for thehigh rate of Torac 15 EC, no significant differences in beetle life stages or defoliation were notedbetween the high and low rates for this product. All three Benevia 10 OD treatments requiredonly the initial foliar application to provide first generation beetle control.The untreated plots had significantly greater defoliation compared to all other treatments.The seasonal defoliation average was 36.6% in the untreated plots, compared to less than 6% forall other treatments. Differences in defoliation among insecticide treated plots ranged from 1.1 to5.9%. Neonicotinoid insecticides are still providing sufficient Colorado potato beetle control forMichigan farmers, but new chemistries like Benevia 10 OD are also proving to be effective.1

Zsofia Szendrei, Department of Entomology MSU517-974-8610, szendrei@msu.eduTable 1. Insecticide treatments for Colorado potato beetle management in a field trial conducted by MSU vegetable entomology in 2013.Trt ctive IngredientChemical ClassRateBrigadier 2SCFMC2 SCbifenthrinpyrethroid25.6 oz/AGladiatorFMC0.25 EWabamectin, z-cypernethrinavermectin, pyrethroid14 oz/A3Capture LFRFMC1.5 SCbifenthrinpyrethroid25.6 oz/AAdmire PROFMC4.6 SCimidaclopridneonicotinoid5.22 oz/AAthenaFMC0.87 EWabamectin, bifenthrinavermectin, pyrethroid16 oz/A4Brigadier 2 SCFMC2 ECbifenthrin, imidaclopridpyrethroid, neonicotinoid25.6 oz/AGladiatorFMC0.25 EWabamectin, z-cypernethrinavermectin, pyrethroid14 oz/A5VerimarkDupont20SCcyazypyrdiamide13.5 oz /AAsana XLDupont0.66 ECesfenvaleratepyrethroid9.6 oz/A6ExirelDupont10 SEcyazypyrdiamide5 oz/A7PlatinumSyngenta2 SLthiamethoxamneonicotinoids8 oz/ABeneviaDupont10 ODcyazypyrdiamide5 oz/A8PlatinumSyngenta2 SLthiamethoxamneonicotinoids8 oz/A9Benevia Dupont10 ODcyazypyrdiamide5 oz/AMSOmethylated seed oilsurfactant0.5% v/v10Benevia Dupont10 ODcyazypyrdiamide5 oz/AMSOmethylated seed oilsurfactant0.5% v/v11ExirelDupont10 SEcyazypyr (cyatraniliprole)diamide6.75 oz/A12Admire PROBayer4.6 SCimidaclopridneonicotinoid1.3 oz/A13BlackhawkDow36spinosadspinosyns3.2fl oz/A14ToracNichino15 ECtolfenpyradpyrazoles14 oz/A15ToracNichino15 ECtolfenpyradpyrazoles21 oz/A16Admire PROBayer4.6 SCimidaclopridneonicotinoid7 oz/A* First foliar application was made at 50% egg-hatch, all subsequent foliar applications are based on 1 large larva per plant threshold.TypeApplicationDates in 2013*at plantingfoliarat plantingpost-plant, directedfoliarpost-plant, directedfoliarat plantingfoliarfoliarat plantingfoliarat plantingfoliar6/36/28, 7/186/36/196/28, 7/116/196/28, rfoliarfoliarfoliarat planting6/286/28, 7/11, 7/186/28, 7/116/28, 7/116/286/32

Zsofia Szendrei, Department of Entomology MSU517-974-8610, szendrei@msu.eduTable 2. Mean Colorado potato beetle (CPB) per plant by date in the 2013 MSU vegetable entomology field trial. The bold numbers in the top rowcorrespond to the treatment numbers in Table 1. Bold letters in the green colored ‘small and large larvae’ rows at the bottom of the table indicatesignificant differences among the treatments, Tukey HSD (α 0.05).dateMean CPB12345678910111213141516 TOTALintact egg masses0.70.70.250.50.50.30 0.15 0.50.70.25 0.90.80.6 0.45 0.60.52hatched egg masses 003.20.5502.45 000.20.33.55 2.400.800.70.922-Julsmall larvae7.100.050.500.7001.15 0.25 0.75 0.45 0.650.9100.79large 0.10.25 0.15 0.55 0.15 0.05 0.15 0.40.20.05 0.05 0.25 0.19intact egg masses0.85 0.550.40.950.60.3000.20.25 0.6510.30.70.21.10.49hatched egg masses 1.05 2.23.90.11.32.75 002.45 2.05 1.95 3.0500.25 0.45 0.05 1.525-Julsmall larvae2.5 0.25 1.25 5.35 0.35 0.15 000.25 0.0503.65 0.41.25 0.30.10.94large larvae2.9500000000000.3 0.05 0.1500.15 0.21adults0.55 0.35 0.35 0.35 0.750.3 0.25 0.1 0.150.20.10.2 0.250.1 0.05 0.10.26intact egg masses0.65 0.35 0.150.50.15 0.15 0.05 00.150.50.15 0.1 0.25 0.25 0.40.40.25hatched egg masses 2.0501.050.20.550.1000.8001.901.25 0.600.6110-Julsmall larvae7.85 0.71.93.15 0.350.3000.81.350.54.71.91.25 1.55 3.45 2.09large larvae10.7 0.31.13.1500.05 00000.05 6.95 1.4520.10.91.66adults00.25 0.35 0.25 0.250.3 0.3 0.1 0.15 0.25 0.25 1.25 0.30.10.10.20.26intact egg masses00000.05 0.05 000.1500.0500.0500.2 0.15 0.04hatched egg masses 00000.250000.700.4000.25 0.5500.1317-Julsmall larvae2.05 0.65 0.15 0.65 0.05 0.45 0 0.10.50.60.6 0.45 0.050.1 0.45 1.35 0.54large larvae4.05 1.650.31.25 0.250.6000.250.80.55 1.3500.75 0.31.10.93adults0000.05 0.050.10 0.150000.05 0.050000.03intact egg masses00000000000000000.00hatched egg masses 000000.2 0.15 000000.25 0.65000.0724-Julsmall larvae0.35 0.40.10.30.45 0.25 0 0.10.50.550.500.0500.200.22large larvae0.8 0.45 0.65 0.65 0.050.1000.550.60.4500.150.20.600.31adults1.35 0.150.10.250.10.15 0.1 0.1500.10.2 0.25 0.20.25 0.20.10.23intact egg masses0.44 0.32 0.16 0.39 0.26 0.16 0.01 0.03 0.20.29 0.22 0.4 0.28 0.31 0.25 0.45 0.26hatched egg masses 0.62 0.44 1.63 0.17 0.421.1 0.03 00.83 0.47 1.18 1.47 0.05 0.64 0.32 0.15 0.65TOTAL small larvae3.97a 0.4bc 0.69bc 1.99b 0.24bc 0.37bc 0c 0.04c 0.64bc 0.56bc 0.47bc 1.85b 0.61bc 0.7bc 0.7bc 0.98bc 0.92large larvae3.97a 0.48bc 0.41bc 1.01bc 0.06c 0.15c 0c 0c 0.16c 0.28bc 0.21c 1.72b 0.33bc 0.62bc 0.22c 0.43bc 0.64adults0.44 0.17 0.18 0.21 0.25 0.22 0.16 0.21 0.09 0.12 0.14 0.43 0.20.1 0.08 0.13 0.193

Zsofia Szendrei, Department of Entomology MSU517-974-8610, A"seasonal"average"of"large"larvae"per"plant"( 2cracking"2"foliar"2"2"ABCD" ABCD"7"Brigadier"Admire" Capture"25.6"oz/A" Pro" 25.6"oz/A"7"oz/A"2"Admire" Gladiator" Torac" Gladiator"Pro" 14"oz/A" 14"oz/A" 14"oz/A"(2)"1.3"oz/A" (2)"(2)"(3)"2"2"Admire"Pro"5"oz/A"ABCD" ABCD" ABC"10" 11"ABC"ABC"8"9"2"2"2"2"2"2"2"2"2"2"2"2"AB"A"A"12" 13" 14" 15" 16"Verimark"Pla5num"Pla5num"13.5"oz/A" 8"oz/A" 8"oz/A"2"Athena" Black2 Benevia" Torac" Exirel" Benevia" Exirel"16"oz/A" hawk" 5"oz/A" 21"oz/A"6.7"oz/A" 5"oz/A" 5"oz/A"2"(2)" 3.2"oz/A" (1)" *" (1)"(1)"(1)" *" "–"not"currently"registered""Summary of 2013 CPB insecticide trial results, Moncalm Potato Research Farm.4

Zsofia Szendrei, Department of Entomology MSU517-974-8610, szendrei@msu.eduSusceptibility of Colorado potato beetle populations to imidacloprid andthiamethoxamImidacloprid (i.e.: Admire Pro) and thiamethoxam (i.e.: Platinum, Actara) continue to be the mostcommon means of Colorado potato beetle management. Today, greater than 75% of thecommercial potato acres in the northeastern and midwestern United States are protected bythese compounds (NASS 2006). Such consistent and heavy dependency on any compound setsthe stage for resistance development. Further complicating the issue is the availability of genericimidacloprid formulations; these formulations drive down product cost, which will likely lead toeven greater field exposure to these compounds. All of these reasons strongly support the needto continue monitoring resistance development and to encourage growers to adopt resistancemanagement strategies.Our objective was to continue gathering data on susceptibility to imidacloprid andthiamethoxam in Colorado potato beetle populations collected from commercial potato fields inMichigan and other regions of the United States. To accomplish this objective, Colorado potatobeetle populations were bioassayed with imidacloprid and/or thiamethoxam.METHODSDuring 2013, 11 Colorado potato beetle populations were collected from Michigan. Cooperatorsalso provided populations from New York (1), Maine (1), and Virginia (3). One susceptiblelaboratory strain was also tested (Table 3). To assure only healthy beetles were tested, newlyreceived beetles were maintained at room temperature and 16:8 L:D photoperiod and fedpesticide-free, greenhouse-grown potato foliage for 3-7 days before they were used in thebioassay.Adult Colorado potato beetles were treated with 1 µl of acetone/insecticide solution of known concentration applied to the ventral surface of the abdomen using a 50 µl Hamiltonmicrosyringe. Two populations with known resistance issues (Jamesport, NY and Tuscola, MI)required two applications of 1 µl of acetone/insecticide solution per beetle to achieve the desireddose (ie., 1 µl of 20.0 µg/µl plus 1 µl of 10.0 µg/µl to get a dose of 30.0 µg/µl). A range of four to11 concentrations, plus an acetone-only control, was selected for each population, depending onthe number of available beetles and known resistance history for each population. In eachbioassay, 27-40 adults were treated with each concentration (nine to 10 beetles per dish andthree to four dishes per concentration). Following treatment, beetles were placed in 100 mm diam. Petri dishes lined with Whatman No. 1 filter paper and provided with fresh potato foliage.They were kept at 25 1 C and the foliage and filter paper were checked daily and changed asneeded.Beetle response was assessed 7 days post treatment. A beetle was classified as dead if itsabdomen was shrunken, it did not move when its legs or tarsi were pinched, and its elytra weredarkened. A beetle was classified as walking and healthy if it was able to grasp a pencil and walkforward normally. A beetle was classified as poisoned if its legs were extended and shaking, itwas unable to right itself or grasp a pencil, and it was unable to walk forward normally at leastone body length. Beetles that had died due to Beauvaria spp. infection were excluded fromanalysis; these beetles were easily recognized by their pale, petrified appearance and/orpresence of white filamentous fungi. Dead and poisoned beetle numbers were pooled foranalysis. Data were analyzed using standard log-probit analysis (SAS Institute, 2009).RESULTSThe LD50 value (dose lethal to 50% of the beetles) for the susceptible laboratory strain was 0.042µg/beetle for imidaclorid and 0.054 µg/beetle for thiamthoxam (Table 4).The LD50 values from the field for imidacloprid ranged from 0.215 µg/beetle (SackettPotatoes Field 2) to 4.435 µg/beetle (Sackett Ranch LJ7) for Michigan populations. Theimidacloprid LD50 values for the out-of-state populations ranged from 0.088 µg/beetle (Aroostook,Maine) to 0.496 (Jamesport, NY). LD50 values for imidacloprid for all populations were5

Zsofia Szendrei, Department of Entomology MSU517-974-8610, szendrei@msu.edusignificantly higher than the susceptible laboratory strain. In 2013, 60% of the Michigan sampleswere greater than 10-fold resistant to imidacloprid, compared to 75% in 2012, 57% in 2011, 60%in 2010, and 85% in 2009.The LD50 values for thiamethoxam in Michigan ranged from 0.044 µg/beetle (Main FarmsH6) to 0.478 µg/beetle (Kalkaska), and from 0.002 µg/beetle (Montgomery, VA) to 0.496µg/beetle (Jamesport, NY) for out-of-state populations. None of the populations were greaterthan 10-fold resistant to thiamethoxam.Table 3. Colorado potato beetle populations tested for susceptibility to imidacloprid andthiamethoxam in 2013.Michigan populationsAnderson Brothers Field 23 Summer adults were collected on 12 Aug 2013 by Mark Otto, AgriBusiness Consultants, Inc., from commercial potato fields in Montcalm County.Kalkaska Summer adults were collected on 31 July 2013 from a commercial potato field nearKalkaska MI.Main Farms Summer adults were collected by Mark Otto, Argi-Business Consultants, Inc. fromcommercial potato fields in Mecosta and Montcalm Counties.Field C10 Adults were collected on Aug 7 2013.Field H6 Adults were collected on June 28 2013.Field R6 Adults were collected on July 30 2013.Sackett PotatoesSackett Potatoes Field 2 Adults were collected on June 14 2013.Sackett Potatoes Field 19 Adults were collected on June 14 2013.Sackett Potatoes Field 26 Adults were collected on Aug 19 2013.Sackett Potatoes Field 150-1 Adults were collected on June 24 2013.Sackett RanchSackett Ranch LJ7 Adults were collected on June 25 2013.Sackett Ranch LJ7 Adults were collected on July 23 2013.Sackett Potatoes Field 26 Adults were collected on Aug 19 2013.Sackett Potatoes Field 150-1 Adults were collected on June 24 2013.6

Zsofia Szendrei, Department of Entomology MSU517-974-8610, szendrei@msu.eduTable 4. LD50 values (µg/beetle) and 95% fiducial limits for Colorado potato beetle populationstreated with imidacloprid and thiamethoxam, 7 days post treatment.IMIDACLOPRIDMichigan PopulationsAnderson Brothers Field 23*Kalkaska*Main Farms C10*Main Farms H6 Main Farms R6*Sackett Potatoes Field 2 Sackett Potatoes Field 19 Sackett Potatoes Field 26*Sackett Potatoes Field 150-1 Sackett Ranch LJ7 Sackett Ranch LJ7 *Out of State PopulationsAroostook, Maine*Jamesport, New York Modestown, VA Painter, VA Montgomery, VA(Whitehorne)*Laboratory strainNew JerseyTHIAMETHOXAMMichigan PopulationsAnderson Brothers Field 23*Kalkaska*Main Farms C10*Main Farms H6 Main Farms R6*Sackett Potatoes Field 2 Sackett Potatoes Field 19 Sackett Potatoes Field 26*Sackett Potatoes Field 150-1 Sackett Ranch LJ7 Sackett Ranch LJ7 *Out of State PopulationsAroostook, Maine*Jamesport, New York Modestown, VA Painter, VA Montgomery, VA(Whitehorne)*Laboratory strainNew Jersey Overwintered generation* Summer generationLD50 10.4930.6654.43595 % confidence intervals0.184 - 0.5931.879 - 16.5050.076 – 4.3440.386 – 0.6971.039 - 2.7910.104 – 0.3530.140 – 0.9260.118 – 0.5700.274 – 0.6930.309 – 1.1023.283 – 6.1030.0885.5770.6550.0810.0890.051 – 0.1293.829 – 6.9710.507 – 0.8570.057 – 0.110n/a0.0420.037 – 0.055LD50 10.0710.1770.37795 % confidence intervals0.085 – 0.1680.291 – 0.8160.094 – 0.4550.020 – 0.0800.051 – 0.3270.132 – 0.236n/a0.054 – 0.1990.041 – 0.1010.133 – 0.225n/a0.0210.4960.2940.0500.0020.014 – 0.030.376 – 0.6200.244 – 0.3340.289 – 0.070n/a0.0540.0357 – 0.09477

Zsofia Szendrei, Department of Entomology MSU517-974-8610, szendrei@msu.eduMolecular genetic mechanisms of CPB insecticide resistanceIn 2013 we identified numerous genes that play a role in CPB insecticide resistance using ourlaboratory CPB populations and RNA sequencing. We developed primers for some of the genesof interest and investigated the level of gene expression using real-time PCR method (MSUResearch and Technology Support Facility). We determined that a selection event with aninsecticide causes over-expression of these genes, which confirmed that these genes are trulyinvolved in the insecticide resistance mechanism, and we also compared these genes in differentpopulations. These results showed that in geographically different populations (MI vs. NY, bothimidacloprid resistant) different genes have evolved to play a role in insecticide resistance (seefigure below).3025relative daclopridresistant strainPre-selectionPre-selection*F1PostPost selection:selection: non-selectedF1non-selectedF1Post-selection:Post selection: Z-F1CYP9Z3CYP6-11CYP9ZCYP9Z2 F2CYP9ZPCYP9Z2F2Imidacloprid resistant strain (NY)180160140Pre-selection120F1 Post selection: non-selected100F1 Post selection: 11* indicates significance at P 0.018

Except for Admire Pro and Athena, all treatments resulted in significantly fewer small larvae than the untreated control, while all treatments significantly reduced the number of large larvae per plant, compared to the untreated (Table 2). There were also significant differences in numbers of large larvae among the insecticide treatments.

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