Colorado potato beetle resistance management strategies for transgenic potatoes

NewLeaf potatoes could provide substantial ecological and economic benefits to potato growers. A concern with NewLeaf potatoes, however, is that Colorado potato beetle may develop resistance to the Bt endotoxin because of the intense selection pressure imposed by its constant presence in the transgenic crop. Before these cultivars were released, however, entomologists throughout North America conducted biological research needed for a strategy to prevent or delay resistance in Colorado potato beetle. This paper describes the considerations and research behind the proposed strategy. Because NewLeaf potatoes express a relatively constant and high concentration of Bt endotoxin throughout the foliage of each plant, reducing selection for resistance must come from planting standard potatoes as a refuge for susceptible beetles. Avoiding resistance through random mixtures of NewLeaf and standard potatoes was evaluated in laboratory and field studies. The conclusion from this research was that the seed mixture would not provide an effective refuge. The most effective alternative refuge was judged to be a block of standard potatoes planted in the same field as the NewLeaf potatoes. By allowing susceptible beetles to survive in their fields, potato growers might sustain the benefits of very effective controls.     

Population dynamics of the Colorado potato beetle (coleoptera:chrysomelidae) in western Massachusetts

Population trends for first-and second-generation Colorado potato beetle (CPB),Leptimtarsa decemlineata (Say), on potato in western Massachusetts were examined using density estimates obtained from direct counts of eggs, early and late instars, and adults. There was a range of phenologies and relative sizes of first-and second-generation populations. The size of the first generation was largely determined by the density of colonizing adults; however, even very low numbers of colonizing adults were sufficient to initiate damaging populations. The number of adults produced one year was not a good predictor of the number of colonizing adults the following year. This study indicated that dispersal was an important factor driving CPB population dynamics. Because dispersal and overwintering mortality greatly reduce the number of adults surviving from one year to the next within the same field, we suggest that reduction of end-of-season adult populations is not efficacious, and that control efforts should focus on reducing the number of colonizing adults by rotating fields, delaying planting dates or using a trap crop.     

Theoretical and empirical assessment of a seed mix refuge in corn for southwestern corn borer

Larval movement in seed mixes of Bt and non-Bt corn (Zea mays L.) can potentially increase the rate at which resistance evolves compared to a structured refuge. Pyramided Bt crops with multiple efficacious Bt toxins make it less likely that interplant larval movement will be successful within a seed mix field but high levels of larval movement may decrease refuge value as larvae moving from non-Bt plants die on neighboring Bt plants. Estimates of refuge productivity obtained from multi-site field studies evaluating larval movement and survival of the southwestern corn borer (SWCB), Diatraea grandiosella Dyar, in MON 89034 seed mix refuge fields were compared to predictions from model simulations of larval movement within a seed mix refuge. Results from field studies and modeling simulations of interplant movement show that seed mix refuge was a productive source of susceptible SWCB and that successful interplant movement by larvae within a seed mix refuge was unlikely to occur at frequency that would increase the fitness of resistant heterozygotes or low level resistance mechanisms. Combined, these results indicate that MON 89034 with a 5% seed mix refuge is a durable IRM tactic for SWCB across a range of larval movement and refuge assumptions and show that a 5% seed mix refuge can delay resistance longer than a similar-sized structured refuge given the nature of grower compliance with refuge requirements.     

Susceptibility of imidacloprid-resistant Colorado potato beetles to non-neonicotinoid insecticides in the laboratory and field trials

Repeated use of neonicotinoid insecticides has resulted in the first reported cases of Colorado potato beetle (Leptinotarsa decemlineata (Say)) resistance to imidacloprid. In the laboratory we determined susceptibility of the imidacloprid-resistant Colorado potato beetles from a population in Southern Maine to other insecticides currently registered for use on potato. This population was about 30-fold resistant to imidacloprid and could not be effectively controlled by its applications. Control mortality was significantly higher for the imidacloprid-resistant larvae than for the susceptible larvae, suggesting that fitness disadvantages may be associated with the resistance trait. Resistant larvae exhibited significantly less mortality than susceptible larvae when exposed to cyfluthrin, carbaryl, azinphosmethyl, and methamidophos. Their susceptibility to oxamyl was also somewhat reduced, although it did provide nearly 100% mortality at the highest concentration tested. Disulfoton was highly toxic to the resistant larvae. Oxamyl killed about 40% of the adults in greenhouse assays with potted potato plants, altered their feeding behavior (fewer adults up on plants), and reduced defoliation by more than 90%. Disulfoton was not lethal to adults, but significantly suppressed their feeding. In field trials with the resistant population, oxamyl and imidacloprid + spinosad provided the best beetle control. Novaluron had no detectable effect on beetle densities. There was little difference between the plots treated with imidacloprid or thiamethoxam and the untreated control. Our results suggest that insecticide rotation may be a valuable option for managing imidacloprid-resistant Colorado potato beetle populations. We also had a good consistency between the results of the Petri dish, greenhouse, and field experiments, indicating that screening under laboratory confinement may be useful when developing initial recommendations to potato growers in areas affected by resistance to neonicotinoids.     

ScreeningSolanum germplasm for resistance to Colorado potato beetle

TuberousSolanum accessions were screened for resistance to Colorado potato beetle (CPB),Leptinotarsa decemlineata (Say), in terms of CPB-caused defoliation in field trials with natural infestations of CPB. Cluster and multivariate discriminant analyses were used to distinguish among species and accessions. Several highly CPB-resistant and relatively homogeneous accessions were identified.     

An on-farm insecticide resistance test kit for colorado potato beetle (Coleoptera: chrysomelidae)

Test kits were developed to enable growers to assess insecticide resistance in Colorado potato beetle populations. Kits included tests for resistance to: carbofuran (Furadan® 4F), phosmet (Imidan® 50WP), esfenvalerate (Asana® 1.9EC), and a combination of esfenvalerate & piperonyl butoxide (Butacide® 8EC). Tests were petri dishes containing filter paper treated with a discriminating concentration of insecticide (i.e., kills susceptible beetles but not resistant beetles). Discriminating concentrations were determined by testing susceptible and resistant strains of beetle across a range of insecticide concentration. About 200 test kits were distributed during the summer of 1988. Results received indicate: 1) Michigan beetle populations were either susceptible or highly resistant to carbofuran, 2) phosmet resistance was more widespread than previously thought, 3) many populations showed moderate to high levels of resistance to esfenvalerate, and 4) adding piperonyl butoxide reduced resistance to esfenvalerate, in many, but not all populations. Resistance kits help growers determine if insecticide resistance is present in beetle populations (either prior to or after an insecticide application), and also generate data useful to research and extension personnel in documenting the extent and distribution of resistance.     

Use of reciprocal grafts to elucidate mode of resistance to Colorado potato beetle (Leptinotarsa decemlineata (Say)) and potato aphid (Macrosiphum euphorbiae (Thomas)) in six wildSolanum species

Reciprocal grafts were done between potato (Solanum tuberosum) and six wildSolanum species known to be resistant to the Colorado potato beetle (Leptinotarsa decemlineata (Say)). Potato or wildSolanum foliage from each reciprocal graft was fed to first instar beetles or adult potato aphids (Macrosiphum euphorbiae (Thomas)). The proportion of second instar larvae after 48 h and survival and fecundity (number of nymphs per female) of the aphids were used to i estimate the resistance level of the foliage. Survival and fecundity of potato aphid were reduced on wildSolanum foliage (except withS. circaeifolium ssp.capsicibaccatum andS. okadae) used as scion or rootstock, but were not different from the control (potato-potato graft) on potato foliage. Development of Colorado potato beetle onSolanum foliage was reduced. In some cases, potato foliage grafted to wildSolanum became resistant to the beetle. Results suggest that a chemical factor or factors providing resistance to Colorado potato beetle are translocated from foliage of some wildSolanum species to potato foliage. They also indicate that mode of resistance to the two insects differs quantitatively or qualitatively.     

Influence of Colorado potato beetle sample counts and plant defoliation on potato tuber production

Field experiments were designed to subject Superior potato plants to various levels of defoliation by the Colorado potato beetle (CPB)Leptinotarsa decemlineata (Say), Coleoptera: Chrysomelidae. Defoliation occurred during each of five consecutive plant growth periods and CPB population, plant leaf area, and tuber weight data were recorded for each period. A visual defoliation rating scheme provided an accurate estimate of actual potato plant leaf area of defoliated plants. Data generated from regression analysis demonstrated a significant dependence of leaf area on CPB numbers per plant during some plant growth periods, but numbers of CPB accounted for very little of the total variation in tuber weight. Plant leaf area was the most important independent variable in the tuber weight regression model.     

Bacillus thuringiensis a potential control agent for the Colorado potato beetle

In laboratory tests conducted to determine the effectiveness of the exotoxin ofBacillus thuringiensis against the Colorado potato beetle,Leptinotarsa decemlineata (Say), applications sprayed on tomato foliage at dilutions ranging from 10^?1 through 10^?4 killed 90% of the 1st and 2nd stage beetle larvae. As the dosage decreased, the time to achieve this level of mortality increased from 4 to 9 days. Adult beetles were not killed by these treatments; however they did cease feeding. In field tests, the highest dosages ofB. t. sprayed on either tomato or potato plants provided adequate or nearly complete protection when applied weekly. Young larvae that ingested sub-lethal doses developed into adults with atrophied mouthparts and antennae, transformations of the paired parts of the labial palps into an unpaired part, the appearance of a tarsal-like claw on a palp, and malformation of the cranial sclerities.     

Horizontal resistance insolanum tuberosum to colorado potato beetle (leptinotarsa decemlineata say)

Horizontal resistance (HR) was assessed as a strategy to reduce potato defoliation by the Colorado potato beetle (CPB). Hybrids amongSolanum tuberosum varieties grown in Iowa were first evaluated with a detached leaflet feeding bioassay. Beginning in 1998, resistance was increased annually over three years by intermating and selecting the most resistant l%–2% of each generation of 300–400 plants using the bioassay. When C (= mg leaf tissue consumed_* neonate larva^-1 over 48 h) was measured for the 1% most-resistant genotypes of all generations grown side by side in 2000, it ranged from 6.1 mg_* larva^-1 in the parents to 2.3 for progeny of the F₃ generation compared to 2.7 in ND2858-1, a resistant check. Field trials in Wisconsin showed less defoliation in the F₁ and F₂ selected genotypes than in the parents. A second breeding program using moreS. tuberosum parents with greater genetic diversity, and much larger breeding populations, resulted in significantly faster accumulation of resistance, further suggesting that recurrent mass selection is effective. Foliar glycoalkaloid analyses revealed low levels of solanine and chaconine in the F₃ selections; no leptines or leptinines were detected. Because the parents wereS. tuberosum varieties in which vertical resistance to CPB is unknown, and because the resistance to feeding from parents to the F₃ resistant plants was linear and gradual while ranging from initially very low to considerably higher in the third year, horizontal resistance is strongly implicated. The development of a significant degree of resistance after three rounds of breeding using relatively small breeding populations suggests that HR can be developed from varieties ofSolanum tuberosum lacking initial high levels of resistance to CPB.     

Identification of late blight, Colorado potato beetle, and blackleg resistance in three Mexican and two South American wild 2x (1EBN)Solanum species

Wild potatoes are important sources of genes for resistance to disease and insect pests. A collection of wild Mexican and South AmericanSolarium species from the US potato Genebank was evaluated under laboratory and/or field conditions for their reaction to late blight (Phytophthora infestans), Colorado potato beetle (CPB,Leptinotarsa decemlineata Say), and blackleg (Erwinia carotovora subsp.atroseptica (van Hall) Dye) in order to identify individual genotypes with multiple resistance genes. Late blight inoculations using aggressive isolates (US-8/A2 and US-11/A1 mating types) of P.infestans revealed a wide range of variation for resistance between and within the accessions of the wild species tested. For late blight, susceptible as well as moderately to highly resistant genotypes were observed in all the species tested. However, at least one accession from the three Mexican and one South American wild diploid species tested showed a relatively uniform high level of resistance toP. infestans. These includedS. bulbocastanum, S. pinnatisectum, S. cardiophyllum, andS. circaeifolium. Two accessions from South American speciesS. commersonii were highly susceptible to late blight. For the Colorado potato beetle test, only one species,S. pinnatisectum appeared uniformly resistant to CPB under field conditions. Results of screening for blackleg resistance showed that there were major differences between genotypes in the wild species. Accessions ofS. circaeifolium PI 498119 andS. bulbocastanum PI 243504 were identified as having significantly higher blackleg resistance than cultivated potato and the other wild species tested. However, genotypes from these two accessions were more susceptible to late blight and CPB. Characterization of theP. infestans isolate P1801C.16 used for late blight evaluation and multi-locus isolate tests using US-8/A2 and US-11/A1 races revealed that the resistance inS. pinnatisectum genotypes tested corresponded to a race-non-specific genetic system, which was different from any existing R genes.Solanum pin-natisectum genotypes with both high levels of late blight and CPB resistance as well as blackleg resistance genotypes identified in the present study represent a diverse gene pool that may be useful for development of new potato cultivars with multiple disease and insect resistance. The potential utilization of these valuable sources for improvement of cultivated potato is discussed.     

Epidemiology of potato leaf roll virus in the Fraser River delta of British Columbia

The spread of potato leaf roll virus (PLRV) and the resulting phloem necrosis in the tubers has been a limiting factor in the production of the Netted Gem (Russet Burbank) variety in southwestern British Columbia, Canada. Steckling sugar beets are an important overwintering host for nymphs and adults of the green peach aphid,Myzus persicae (Sulzer). By June, aphids are widely disseminated to the florets and new leaves of sugar beet and other hosts, including potato. Colonies with alates are produced on summer hosts which are not treated with insecticides. The peak of the flight of alates from the colonies on the summer hosts occurs during the first half of August. Those alates that were produced on PLRV-infected potato and which travel to other potatoes transmit the virus as soon as they feed. By the end of August, the sugar beet seed crops and the early and mid-season potato crops have been defoliated or harvested. Thus many colonies ofM. persicae are destroyed and the threat of PLRV spread diminishes. Crop and weed plants which serve as overwintering hosts become infested during late summer with alate aphids. Their offspring survive the winter in numbers which are determined by the weather and survival of host plants.     

Evaluation of European corn borer Ostrinia nubilalis (Lepidoptera: Crambidae) larval movement and survival in structured and seed blend refuge plantings

Blending Bacillus thuringiensis (Bt) corn and non-Bt corn seeds in the same bag is a convenient and easier compliance solution to satisfy refuge regulations. However, there has been considerable debate and concern about larval movement of the target insects in a seed blend because of its potential effect on the development of resistance. Reported here are studies to determine the larval movement of European corn borers, Ostrinia nubilalis (Hübner), in a seed blend compared to a structured refuge and their effects on survival, feeding injury and larval fitness. Also evaluated is the relative performance of a purple-seeded corn hybrid as a surrogate host plant for tracking gene flow. Seed blend and structured refuge arrangements of Agrisure GT/CB/LL Bt corn expressing Cry1Ab and isoline plants were evaluated at two locations in 2013 and 2014 by manually infesting refuge plants with first and second generation corn borer larvae. The majority of leaf and stalk injury was recorded on the refuge plant, which amounted to 66–83% of the total tunnel length. In both refuge arrangements, plant injury and the number of larvae recovered significantly decreased on neighboring plants with increasing distance away from the refuge plant in the same row. The relative decline in injury was much more pronounced in the seed blend, with neighbor Bt plants experiencing very minor tunneling or no injury at all due to the high dose trait. Eighty-nine percent fewer live larvae were also found on neighbor Bt plants in the seed blend compared to those found in the structured refuge plots. Larvae that moved to Bt plants as well as older instars that were manually placed on Bt plants failed to complete development. Comparison of the number of tunnels and larvae recovered per refuge plant indicated that the seed blend may not produce as many susceptible individuals as those produced in a structured refuge arrangement. Although the purple-seeded hybrid showed lower levels of ear and stalk injury, it could serve as an appropriate surrogate refuge plant to track gene flow.     

Progress report: Resistance to the Colorado potato beetle and potato leafhopper inSolanum tuberosum subsp.andigena

An extensive collection of 1527 clones ofS. tuberosum subsp.andigena was screened in 1973 in unreplicated plots to test for resistance to the Colorado potato beetle and potato leafhopper. All clones were susceptible to the beetle. Of 17 clones selected in the 1973 tests for resistance to the potato leafhopper, 7 selections were significantly more resistant than the control (Katahdin) in a replicated experiment in 1974.     

Direct application of reduced rates of aldicarb to potato seedpieces

Aldicarb was applied directly to potato seedpieces at 0.3, 0.6, 1.1, 2.2 and 3.4 kg AI/ha at planting and at 2.24 kg Al/ha over the plants at crop emergence. All seedpiece treatments resulted in superior early season protection from Colorado potato beetle,Leptinotarsa decemlineata (Say). Lower seedpiece rates lost effectiveness by early July, while higher rates protected foliage into early August. In a second study, plots treated with 1.1 kg AI/ha to the seedpiece produced crop yields as high as those treated with additional aldicarb at crop emergence. It is suggested that low rates of aldicarb applied directly to the seedpiece at planting could offer effective early season insect control while reducing the risk of groundwater contamination.     

Field selection for esfenvalerate resistance by the Colorado potato beetle

Five insecticide application regimes were evaluated for two years to determine their influence on the development of esfenvalerate resistance by the Colorado potato beetle (CPB),Leptinotarsa decemlineata (Say). These included sequential applications of esfenvalerate; two alternated uses of esfenvalerate, azinphosmethyl, endosulfan, oxamyl, andBacillus thuringiensis or cryolite; an IPM program; and one early season esfenvalerate application. Both CPB adults and first instars were tested for esfenvalerate resistance before and after insecticide applications. The LC₅₀’s in the early 1992 season were correlated with values in the late 1991 season. However, over the winter, esfenvalerate resistance of CPB adults decreased, but was relatively stable for first instars. Esfenvalerate LC₅₀’s late in the season were generally higher than values early in the season. Increases in esfenvalerate LC₅₀’s were correlated with the number of esfenvalerate applications made in these fields during the growing season, with the greatest increase found in the sequential esfenvalerate treatment.     

The effect of light intensity on Colorado potato beetle resistance and foliar glycoalkaloid concentration of fourSolanum chacoense clones

Controlled environment experiments were used to determine the effect of light intensity on levels of glycoalkaloids and resistance to the Colorado potato beetle (CPB),Leptinotarsa decemlineata Say, inSolanum chacoense Bitter clones that synthesized acetylated glycoalkaloids, the leptines. Plants were grown under two light intensities, 50 or 250 ?mol m^-2S^-1, and bioassayed for CPB resistance by placing neonate larvae on excised foliage. Larval development stage and mortality were recorded after four days. The leptine-synthesizing clones responded to the increase in light intensity with a 2–4 fold increase in level of foliar leptines. Larval development rate on foliage from plants grown under high light intensity was reduced and larval mortality increased. Results showed a significant (p>0.05) effect of light on glycoalkaloid synthesis and CPB resistance.     

The germplasm release of NYL 235–4, a clone with resistance to the Colorado potato beetle

Clones selected from the wild diploid species,Solanum berthaultii, have been shown to possess valuable levels of resistance to the Colorado potato beetle (Leptinotarsa decemlineata), as well as to insects such as aphids, flea beetles, leafhoppers and the potato tuber moth. Resistance is associated with the presence of glandular trichomes on the foliage ofS. berthaultii (2). Six generations after producing a hybrid between the diploid species and a tetraploid variety, a clone with good beetle and leafhopper resistance, as well as reasonable agronomic characteristics has been produced.     

Impacts of Dryland Cropping Systems on Ground Beetle Communities (Coleoptera: Carabidae) in the Northern Great Plains

Ground beetles are natural predators of insect pests and small seeds in agroecosystems. In semiarid cropping systems of the Northern Great Plains, there is a lack of knowledge to how ground beetles are affected by diversified cover crop rotations. In a 2-yr study (2018 and 2019), our experiment was a restricted-randomization strip-plot design, comprising summer fallow, an early-season cover crop mixture (five species), and a mid-season cover crop mixture (seven species), with three cover crop termination methods (i.e., herbicide, grazing, and haying). Using pitfall traps, we sampled ground beetles in five 48-h intervals throughout the growing season (n = 135 per year) using growing degree day (GDD) accumulations to better understand changes to ground beetle communities. Data analysis included the use of linear mixed-effects models, perMANOVA, and non-metric multidimensional scaling ordinations. We did not observe differences among cover crop termination methods; however, activity density in the early-season cover crop mixture decreased and in summer fallow increased throughout the growing season, whereas the mid-season cover crop mixture peaked in the middle of the summer. Ground beetle richness and evenness showed a nonlinear tendency, peaking in the middle of the growing season, with marginal differences between cover crops or fallow after the termination events. Also, differences in ground beetle composition were greatest in the early- and mid-season cover crop mixtures earlier in the growing season. Our study supports the use of cover crop mixtures to enhance ground beetle communities, with potential implications for pest management in dryland cropping systems.     

Fine screeningSolanum (potato) germplasm accessions for resistance to Colorado potato beetle

This study was conducted to investigate the utility of systematic screening at the genotype level withinSolanum accessions highly resistant to the Colorado potato beetle. Evaluations of clonally replicated genotypes showed that most accessions reported to be uniform when screened as populations contained small but significant variation among genotypes for resistance to oviposition, larvae, and defoliation, differences for numbers of larvae being most common. Adult counts and percentage defoliation were not as useful in evaluating among-genotype variability in beetle resistance. Genotypes ofS. pinnatisectum WRF 343 andS. tarijense PI 473227 were the most uniformly and highly resistant to Colorado potato beetle. Genotypes of S.berthaultii PI 473331,S. chacoense PI 473405, andS. tarijense PI 473336 were moderately to highly resistant, and genotypes ofS. bukasovii PI 473494 andS. canasense PI 230511 were uniformly susceptible to Colorado potato beetle. Nonparametric correlation analyses indicated that number of egg masses, small larvae, large larvae, and defoliation scores were positively correlated, negatively correlated, or not correlated, depending on the species. One generation of selection attempting to segregate resistance and susceptibility in nearly uniform and highly resistantS. pinnatisectum WRF 343 resulted in “divergent” populations that could not be distinguished from each other or the base population. Thus, through genotype (“fine”) screening and selection, we showed that some existing populations are virtually pure for extreme resistance. Use of individuals from such families would make screening breeding populations more efficient, and reduce the risk of losing resistance genes that have non-dominant effects. Fine screening, recurrent selection, and maintenance of such elite populations is recommended as an extension of population-based evaluation usually done by genebanks.