Aggregation Behavior and a Putative Aggregation Pheromone in Sugar Beet Root Maggot Flies (Diptera: Ulidiidae).

Male-biased aggregations of sugar beet root maggot, Tetanops myopaeformis (Röder) (Diptera: Ulidiidae), flies were observed on utility poles near sugar beet (Beta vulgaris L. [Chenopodiaceae]) fields in southern Idaho; this contrasts with the approximately equal sex ratio typically observed within fields. Peak observation of mating pairs coincided with peak diurnal abundance of flies. Volatiles released by individual male and female flies were sampled from 08:00 to 24:00 hours in the laboratory using solid-phase microextraction and analyzed using gas chromatography/mass spectrometry (GC/MS). Eleven compounds were uniquely detected from males. Three of these compounds (2-undecanol, 2-decanol, and sec-nonyl acetate) were detected in greater quantities during 12:00-24:00 hours than during 08:00-12:00 hours. The remaining eight compounds uniquely detected from males did not exhibit temporal trends in release. Both sexes produced 2-nonanol, but males produced substantially higher (ca. 80-fold) concentrations of this compound than females, again peaking after 12:00 hours. The temporal synchrony among male aggregation behavior, peak mating rates, and release of certain volatile compounds by males suggest that T. myopaeformis flies exhibit lekking behavior and produce an associated pheromone. Field assays using synthetic blends of the putative aggregation pheromone showed evidence of attraction in both females and males.

Field-Evolved Resistance of Northern and Western Corn Rootworm (Coleoptera: Chrysomelidae) Populations to Corn Hybrids Expressing Single and Pyramided Cry3Bb1 and Cry34/35Ab1 Bt Proteins in North Dakota.

Northern, Diabrotica barberi Smith & Lawrence, and western, D. virgifera virgifera LeConte, corn rootworms (Coleoptera: Chrysomelidae) are major economic pests of corn, Zea mays L., in North America. Corn hybrids expressing Bacillus thuringiensis Berliner (Bt) toxins are commonly used by growers to manage these pests. Several cases of field-evolved resistance to insecticidal proteins expressed by Bt corn hybrids have been documented in many corn-producing areas of North America, but only for D. v. virgifera. In 2016, beetles of both species were collected from five eastern North Dakota corn fields and reared in a growth chamber. In 2017, larvae reared from those populations were subjected to single-plant bioassays to screen for potential resistance to Cry3Bb1, Cry34/35Ab1, and pyramided Cry3Bb1 + Cry34/35Ab1 Bt toxins. Our results provide the first documented report of field-evolved resistance in D. barberi to corn hybrids expressing Cry3Bb1 (Arthur problem population) and Cry34/35Ab1 (Arthur and Page problem populations, and the Ransom and Sargent populations) proteins in North America. Resistance to Cry3Bb1 was also observed in the Ransom population of D. v. virgifera. Increased larval survival on the pyramided Cry3Bb1 + Cry34/35Ab1 hybrid was observed in both species. No cross-resistance was evident between Cry3Bb1 and Cry34/35Ab1 in any of the D. barberi populations tested. Our experiments identified field-evolved resistance to Bt toxins in some North Dakota populations of D. barberi and D. v. virgifera. Thus, more effective control tools and improved resistance management strategies are needed to prolong the durability of this technology for managing these important pests.

Multi-year survival of sugarbeet root maggot (Tetanops myopaeformis) larvae in cold storage.

To test the hypothesis that long-term survival of sugarbeet root maggot in storage is facilitated by larvae undergoing prolonged diapause, respiration and gene expression patterns of field-collected diapausing larvae were compared with those of 1-, 2-, and 5-year laboratory-stored larvae. Additional assessments were made on post-storage survival, emergence, and reproductive fitness of stored larvae. Respirometry, carried out at 5 and 20 degrees C revealed no differences among respiration rates of initially diapausing and long-term stored larvae. A 15 degrees increase in temperature elevated respiration in both diapausing and stored larvae, with levels of CO2 release ranging between 8- and 14-fold higher at 20 degrees C than at 5 degrees C. Similarly, 6-10-fold increases in O2 consumption levels were observed at the higher temperature. A transcript with sequence similarity to the fat body protein 2 (Fbp2) gene was highly expressed in diapausing larvae, and trace levels were expressed in some samples of 1-year stored larvae. However, no expression was detected in 2- and 5-year stored larvae. Survival and emergence studies of stored larvae revealed mixed populations of diapausing (i.e., the 5-17% of larvae that did not pupate) and post-diapausing (62-84% of larvae pupated) insects, with a high incidence of pupation (62%) and emergence (47%), even after 4 years in cold storage. Therefore, extended survival of Tetanops myopaeformis larvae in long-term cold storage is facilitated by two mechanisms, with a majority of larvae in post-diapause quiescence and a smaller fraction in a state of prolonged diapause.

Cold storage of adult Gonatocerus ashmeadi (Hymenoptera: Mymaridae) and effects on maternal and progeny fitness.

Storage of Conatocerus ashmeadi Girault (Hymenoptera: Mymaridae) adults at 2, 5, and 10 degrees C showed that these parasitoids do not survive at 2 degrees C for 5 d, and exposure to 5 and 10 degrees C shortens their life span. The lethal time (LT) 50 (i.e., length of storage time for 50% wasp survival) at 5 degrees C was 14 d for males and approximately 29 d for females, whereas at 10 degrees C was 32 and 39 d, respectively. Effects of adult storage at 10 degrees C on other factors indicating fitness, such as fecundity, developmental time, parasitism, emergence, and sex ratio, were examined on female wasps and their progeny at 10-d intervals for up to 60 d. Glassy-winged sharpshooter, Homalodisca vitripennis (Germar), eggs were used as hosts for propagation of this wasp and for assessing its oviposition and fecundity. Increasing adult storage time decreased the length of the ovipositional period for the maternal generation, and oviposition was decreased by 90% after 60-d storage. A significant reduction in maternal lifetime fecundity occurred after 20-d storage and in the incidence of parasitism after 40 d. We also found a carryover effect caused by storage of the maternal generation that was expressed in the F1 generation. When cold storage of the adult parents was > or = 20 d, we observed delayed development, decreased fecundity, reduced longevity, and increased male production occurring in the F1 generation. Reduced fitness of the F1 generation was also expressed as a decrease in net reproductive rate (R(o)) and an increase in mean generation time (T(c)). However, none of these deleterious effects were evident in the F2 progeny that descended from grandparents that had experienced cold storage. Damage caused by indirect chilling injury and/or induced maternal aging occurring during storage can account for the decreased fitness of maternal and F1 generations. Providing that the limits of cold tolerance of G. ashmeadi as defined in this study are not exceeded, our results show that short-term cold storage of adults could be used in a mass-rearing program.

Oviposition Preference and Larval Host Range of the Sugarbeet Root Maggot (Diptera: Ulidiidae).

The sugarbeet root maggot, Tetanops myopaeformis Röder (Diptera: Ulidiidae), is native to North America. However, its primary crop host, sugarbeet, Beta vulgaris L., was introduced to the continent from Europe in the late 19th century. This field and greenhouse research was conducted to compare the relative attractiveness of eight cultivated and wild plant species for oviposition by T. myopaeformis, and the suitability of these potential host plants for larval development to elucidate the potential native and current host range of this pest. Results indicated that females preferred ovipositing in soil immediately adjacent to or on the following plant species: sugarbeet; spinach, Spinacia olerocea L.; common lambsquarters, Chenopodium album L.; redroot pigweed, Amaranthus retroflexus L.; Palmer amaranth, Amaranthus palmeri S. Wats.; and to a lesser extent, spear saltbush, Atriplex patula L. Larval survival was greatest on spinach, sugarbeet, and spear saltbush, which all belong to the family Chenopodiaceae. Larval survival on these plants suggests that T. myopaeformis could have exploited wild chenopodiaceous plants or others within the order Caryophyllales before sugarbeet was introduced to North America. Low larval survival on common lambsquarters, redroot pigweed, and Palmer amaranth suggests that these species are suboptimal hosts, despite demonstrated attractiveness for oviposition. A general lack of oviposition preference by T. myopaeformis females was observed for sunflower, Helianthus annuus L., and common ragweed, Ambrosia artemisiifolia L. These results provide insights regarding the successful and somewhat rapid host preference shift by this insect to sugarbeet after cultivation of the crop began in the continent.

Transgenic Bt Corn, Soil Insecticide, and Insecticidal Seed Treatment Effects on Corn Rootworm (Coleoptera: Chrysomelidae) Beetle Emergence, Larval Feeding Injury, and Corn Yield in North Dakota.

Northern, Diabrotica barberi Smith & Lawrence (Coleoptera: Chrysomelidae), and western, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), corn rootworms are economic pests of corn, Zea mays L. in North America. We measured the impacts of corn hybrids incorporated with Cry3Bb1, Cry34/35Ab1, and pyramided (Cry3Bb1 + Cry34/35Ab1) Bacillus thuringiensis Berliner (Bt) proteins, tefluthrin soil insecticide, and clothianidin insecticidal seed treatment on beetle emergence, larval feeding injury, and corn yield at five locations from 2013 to 2015 in eastern North Dakota. In most cases, emergence was significantly lower in Bt-protected corn than in non-Bt corn hybrids. Exceptions included Wyndmere, ND (2013), where D. barberi emergence from Cry34/35Ab1 plots was not different from that in the non-Bt hybrid, and Arthur, ND (2013), where D. v. virgifera emergence from Cry3Bb1 plots did not differ from that in the non-Bt hybrid. Bt hybrids generally produced increased grain yield compared with non-Bt corn where rootworm densities were high, and larval root-feeding injury was consistently lower in Bt-protected plots than in non-Bt corn. The lowest overall feeding injury and emergence levels occurred in plots planted with the Cry3Bb1 + Cry34/35Ab1 hybrid. Time to 50% cumulative emergence of both species was 5-7 d later in Bt-protected than in non-Bt hybrids. Tefluthrin and clothianidin were mostly inconsequential in relation to beetle emergence and larval root injury. Our findings could suggest that some North Dakota populations could be in early stages of increased tolerance to some Bt toxins; however, Bt corn hybrids currently provide effective protection against rootworm injury in eastern North Dakota.

Emergence of adult northern and western corn rootworms (Coleoptera: Chrysomelidae) following reduced soil insecticide applications.

A 3-yr investigation was conducted in commercial corn, Zea mays (L.), fields in eastern South Dakota to determine how reduced application rates of planting-time soil insecticides would influence temporal emergence patterns and survival of northern and western corn rootworms, Diabrotica barberi Smith and Lawrence, and D. virgifera virgifera LeConte, respectively. Beetle emergence was monitored at 2-d intervals throughout the entire adult emergence period of three growing seasons from corn plots treated with planting-time applications of labeled (1X) and reduced (0.5 and 0.75X) application rates of terbufos, tefluthrin, and chlorethoxyfos. No consistent insecticide- or rate-related impacts on mean total emergence per trap were recorded for any of the compounds investigated. However, terbufos applications resulted in a 52% reduction in the number of beetles captured per trap, 53% reduction in maximum rate of adult emergence, and a 59% reduction in overall rate of emergence over time for male D. virgifera during 1994. Terbufos also significantly extended the time required for emergence to peak and linear emergence of female D. virgifera to end in 1994. Tefluthrin applications delayed onset, end, and time of maximum emergence of female D. barberi by 9.9, 14.1, and 12 d, respectively, during 1993. Tefluthrin also reduced emergence rates over time for male (38%) and female (46%) D. barberi during 1994. Overall, application rate was inconsequential regarding total emergence, seasonal emergence pattern, or level of plant protection provided for all insecticides we tested in this 3-yr investigation. Our findings demonstrate that, if properly applied, the reduced application rates used in this study provide adequate root protection and will not significantly impact the biology of these pest species.

Oat cover cropping and soil insecticides in an integrated sugarbeet root maggot (Diptera: Otitidae) management program.

Sugarbeet, Beta vulgaris L., producers occasionally establish cereal cover crops to minimize early-season soil erosion, wind abrasion, and mechanical injury of seedlings. We evaluated the use of living oat, Avena sativa L., cover cropping as a cultural tactic to minimize feeding injury from sugarbeet root maggot, Tetanops myopaeformis (Röder), larvae at five field sites during 1996, 1998, and 1999. Sweep-net sampling yielded 4.8-, 11.2-, and 7.2-fold more flies from oat cover-cropped chlorpyrifos, terbufos, and untreated control plots, respectively, than in noncover counterparts. However, larval feeding injury in terbufos-treated plots was reduced when cover-cropped (383 seeds/m2) at St. Thomas in all years. A reduced oat seeding rate (224 seeds/m2) also enhanced root protection in terbufos-treated plots at St. Thomas in 1999. Less root injury was sustained in cover-cropped chlorpyrifos plots than in noncover counterparts at St. Thomas in 2 study yr. Oat cover cropping also frequently resulted in reduced T. myopaeformis feeding injury in the absence of a soil insecticide. Although trends toward increased yields were often evident, significant yield benefits were limited to a 6.8% root yield increase in oat cover plots when compared with noncover treatments overall at St. Thomas in 1996 and an 18.4% sucrose yield increase in terbufos-treated plots at St. Thomas in 1999. These findings suggest that beneficial interactions between planting-time soil insecticides and cereal cover crops are achievable in areas infested by T. myopaeformis. Demonstrated reductions in root feeding injury, combined with additional agronomic benefits, may warrant use of this production practice as part of an integrated management program for this key insect pest of sugarbeet.

Host plant effects on development and reproduction of the glassy-winged sharpshooter, Homalodisca vitripennis (Homoptera: Cicadellidae).

Development, survivorship, longevity, reproduction, and life table parameters of the glassy-winged sharpshooter, Homalodisca vitripennis (Germar), were examined in the laboratory using three host plants, sunflower (Helianthus annuus L.), Chrysanthemum morifolium L., and euonymus (Euonymus japonica Thurb.). Females deposited similar-sized egg masses on all three plants. Hatching was highest with eggs deposited on euonymus and lowest for those deposited on sunflower. Embryonic development time among host plants was similar while nymph development time was shortest on sunflower and longest on euonymus. Nymph survival to adulthood ranged from 32% on euonymus to 82% for those reared on sunflower. Adult females had similar life spans on sunflower and chrysanthemum. H. vitripennis completed a lengthy egg-to-adult development on euonymus, however, mating did not occur. The onset of mating was contingent on maturation of adult females. The majority of mating activity occurred within the first three days after onset. Premating periods ranged from 6 to 7 d on sunflower to 27 d on chrysanthemum, with overall mating rates of 77.4 and 19.8%, respectively. Females typically mated more than once and they had the longest oviposition period and highest egg production on sunflower; ≈ 50 and 67% of total number of eggs were deposited within first 45 d after the start of oviposition on sunflower and chrysanthemum, respectively. Adult size and weight related to which host plant was consumed throughout development. Greater intrinsic and finite rates of increase and net reproduction rate, and shorter population doubling time occurred when the sharpshooters were allowed to develop on sunflower. The overall developmental and reproductive parameters obtained in this study indicate that a mixed host plant system, composed of sunflower and euonymus or chrysanthemum plants, is an efficient means for optimizing egg production and colony maintenance of the glassy-winged sharpshooter.

Internal lipids of sugarbeet root maggot (Tetanops myopaeformis) larvae: effects of multi-year cold storage.

Sugarbeet root maggots, Tetanops myopaeformis (Diptera, Ulidiidae), survive more than five years of laboratory cold (6 degrees C) storage as mature third-instar larvae. To quantify energy costs associated with prolonged storage, internal lipids of larvae stored for 1, 2, 3, and 5 years were compared and characterized with those of field-collected diapausing larvae. Internal lipid concentration was highest (21.8% wet wt. and 29.8% dry wt.) in diapausing larvae. Lipids decreased progressively over storage time with greater than 70% reductions for 5-year stored larvae. Thin-layer chromatographic analysis revealed that triacylglycerols (TAGs) were the most predominant class of internal lipids, with trace amounts of diacylglycerols and hydrocarbons also being present. Gas chromatography-mass spectrometry (GC-MS) analyses of TAG fractions identified ten major fatty acids (FAs). The proportion of unsaturated FAs was higher (73 to 78%) than saturated FAs in diapausing and stored larval groups. Palmitoleic acid (16:1) was the predominant FA, constituting 40-50% of total unsaturated FAs with lesser amounts of myristoleic (14:1), oleic (18:1), lauroleic (12:1), gadoleic (20:1), and the saturated FAs, palmitic (16:0), myristic (14:0), lauric (12:0), stearic (18:0), and arachidic (20:0) being detected at much lower concentrations. Characterization of intact TAGs by high performance liquid chromatography and GC-MS revealed the presence of more than 40 TAG constituents. In conclusion, TAGs are utilized as an important energy source for T. myopaeformis larvae during diapause and long-term cold storage with no observed impact of multi-year storage on the TAG composition and distribution of their fatty acids.

Discovery of Fusarium solani as a naturally occurring pathogen of sugarbeet root maggot (Diptera: Ulidiidae) pupae: prevalence and baseline susceptibility.

The fungus Fusarium solani (Mart.) Sacc. was discovered as a native entomopathogen of the sugarbeet root maggot, Tetanops myopaeformis (Röder), in the Red River Valley of North Dakota during the 2004 sugarbeet production season. This is the first report of a native pathogen affecting the pupal stage of T. myopaeformis. Forty-four percent of larvae collected from a field site near St. Thomas (Pembina Co.) in northeastern North Dakota during May and June of 2004 were infected with the entomopathogen. The mean LC(50) of F. solani, assessed by multiple-dose bioassays with laboratory-reared pupae, was 1.8x10(6)conidia/ml. After isolation and confirmation of pathogenicity, a pure isolate of the fungus was deposited in the ARS Entomopathogenic Fungal Collection (ARSEF, Ithaca, NY) as ARSEF 7382. Symptoms of F. solani infection included rapid pupal tissue atrophy and failure of adults to emerge. Transverse dissections of infected pupae revealed dense hyphal growth inside puparia, thus suggesting fungal penetration and pathogenicity. Mycelia emerged from pupae after host tissues were depleted. Exposure of older pupae to lethal concentrations caused rapid mortality of developing adults inside puparia. A second, more extensive field survey was conducted during the 2005 cropping season, and F. solani infection was observed in root maggots at most locations, although at lower levels (1-10%) of prevalence than in 2004. Aberrant timing or amounts of rainfall received could have caused asynchrony between pathogen and host during the second year of the experiment.