Sugarbeet Root Maggot Research Articles

Optimizing a pheromone lure for the sugar beet root maggot fly, Tetanops myopaeformis

AbstractWe previously described a putative aggregation pheromone in adults of the sugar beet root maggot, Tetanops myopaeformis (von Röder) (Diptera: Ulidiidae), comprising nine compounds identified from males. Here, we conducted a series of experiments aimed at simplifying the blend of compounds necessary to achieve attraction as well as determining the dose that maximizes captures when formulated into an attractant lure. In all experiments, females showed stronger and more consistent evidence of attraction than males. White sticky traps baited with different blends of pheromone compounds that included the major component, (R)‐(−)‐2‐nonanol, showed significantly higher female captures relative to those baited with blends that excluded the major component. (R)‐(−)‐2‐nonanol alone was at least as effective as any blend that included this compound with other minor pheromone components. Lures using racemic 2‐nonanol were as effective as the (R) enantiomer for both females and males, with some evidence of weak attraction to the (S) enantiomer (which is not produced by males) observed as well. Maximum capture rates using racemic 2‐nonanol were estimated to occur with doses of ca. 795.5 and 621.6 mg for females and males, respectively. Addition of 2‐nonanol lures to standard orange sticky stake traps currently used to monitor flies increased captures of both sexes. The pheromone lure developed here could improve trapping efficiency of current monitoring programs for T. myopaeformis and may also be used to develop other management tools for this important pest of sugar beet.

Genetic differentiation and diversity of sugarbeet germplasm resistant to the sugarbeet root maggot

AbstractGermplasm lines with resistance to the sugarbeet root maggot (SBRM) have been developed and released to the public, providing a means to generate hybrids with resistance against the most devastating insect pest of sugarbeet in North America. Effective use of this germplasm, however, requires knowledge of relative strengths of SBRM resistance between lines and knowledge of the diversity and genetic relationships between germplasm. Therefore, field studies comparing SBRM resistance of four released SBRM-resistant germplasm lines (F1015, F1016, F1024 and F1043), a SBRM-resistant parent (PI 179180) and an unreleased SBRM-resistant population (F1055) were performed, and genetic analysis of the diversity and relationships between SBRM-resistant germplasm and their available parents was conducted using simple sequence repeat (SSR) markers. Under natural SBRM infestations, resistant germplasm exhibited significantly less SBRM damage than a susceptible control, with similar, high levels of resistance in F1016, F1024, F1043, F1055 and PI 179180 and lower resistance in F1015. SSR analysis revealed genetic similarities between F1016, F1024 and F1055, while F1015 and F1043 were genetically distinct from these lines. Among resistant genotypes, F1015 and F1043 exhibited greatest and least within-line genetic diversity, indicating greater and lesser inbreeding for F1043 and F1015, respectively. Similarities in damage ratings and genetics for F1016, F1024 and F1055 indicate that these lines are likely to be equally effective at introducing SBRM resistance into elite populations and in combining ability. In contrast, F1043, with its unique parentage and genetic dissimilarity from other resistant lines, provides a genetically distinct, but similarly effective, source of SBRM resistance.

Screening for Resistance Against the Sugarbeet Root Maggot, Tetanops myopaeformis (Diptera: Ulidiidae), Using a Greenhouse Bioassay.

The sugarbeet root maggot, Tetanops myopaeformis (von Röder) (Diptera: Ulidiidae), is a major pest of sugar beet Beta vulgaris L. (Carophyllales: Amaranthaceae)in the United States and Canada. Larval feeding on roots can reduce both stand and yield. Current management practices are heavily reliant on chemical control. However, the carbamate and organophosphate insecticides that are commonly used against T. myopaeformis are being phased out of use. Host plant resistance against this pest shows promise, but difficulties with maintaining T. myopaeformis in culture have largely limited such studies to the field. A primary objective of this study was to develop protocols for rearing a laboratory colony of T. myopaeformis that would expedite assays aimed at screening for host plant resistance. Third (final) instar larvae were collected from the field and reared to the adult stage. These laboratory-reared adults laid eggs and ultimately produced a second generation of third-instar larvae in the lab. Adult flies reared from field-collected larvae were used to examine the modality of resistance of a known resistant variety by performing no-choice and paired-choice experiments alongside a susceptible variety in the greenhouse. Paired-choice tests showed no difference in oviposition rates between the two varieties, whereas no-choice tests showed significantly greater feeding damage and abundance of larvae on the susceptible variety. For the resistant variety examined here, we observed evidence of antibiosis, not antixenosis, as the putative modality of resistance. Our laboratory and greenhouse protocols can be used to expedite the process of developing varieties with resistance to this key pest of sugar beet.

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.

Relative Performance of Resistant and Susceptible Sugabeet Hybrids in Environments with and without Sugarbeet Root Maggot

Relative Performance of Resistant and Susceptible Sugabeet Hybrids in Environments with and without Sugarbeet Root Maggot

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.

Transcriptome analysis of sugar beet root maggot (Tetanops myopaeformis) genes modulated by the Beta vulgaris host.

Sugar beet root maggot (SBRM, Tetanops myopaeformis von Röder) is a major but poorly understood insect pest of sugar beet (Beta vulgaris L.). The molecular mechanisms underlying plant defense responses are well documented, however, little information is available about complementary mechanisms for insect adaptive responses to overcome host resistance. To date, no studies have been published on SBRM gene expression profiling. Suppressive subtractive hybridization (SSH) generated more than 300 SBRM ESTs differentially expressed in the interaction of the pest with a moderately resistant (F1016) and a susceptible (F1010) sugar beet line. Blast2GO v. 3.2 search indicated that over 40% of the differentially expressed genes had known functions, primarily driven by fruit fly D. melanogaster genes. Expression patterns of 18 selected EST clones were confirmed by RT-PCR analysis. Gene Ontology (GO) analysis predicted a dominance of metabolic and catalytic genes involved in the interaction of SBRM with its host. SBRM genes functioning during development, regulation, cellular process, signaling and under stress conditions were annotated. SBRM genes that were common or unique in response to resistant or susceptible interactions with the host were identified and their possible roles in insect responses to the host are discussed.

Cover crop and conidia delivery system impacts on soil persistence of Metarhizium anisopliae (Hypocreales: Clavicipitaceae) in sugarbeet

The sugarbeet root maggot, Tetanops myopaeformis (Röder), is a major North American pest of sugarbeet, Beta vulgaris L. Previous research suggests that moderate T. myopaeformis control is possible with the entomopathogen Metarhizium anisopliae (Metch.) Sorok. We conducted a three-year (2002–2004) experiment to assess impacts of oat, Avena sativa L. and rye, Secale cereale L., cover crops on persistence of corn grit-based granular or spray formulations of M. anisopliae isolate ATCC 62176 (i.e. MA 1200) applied at 8×1012 viable conidia/ha in sugarbeet. More colony forming units (CFUs) were detected immediately after application [0 days after treatment (DAT)] in spray plots than granule-treated plots. However, 76–92% declines in CFUs per gram of soil occurred in spray plots within 30 DAT. Substantially (i.e. 83–560%) more rainfall occurred in June 2002 than during June of any other year. Subsequently, 71–670% increases in CFU concentrations occurred by 60 DAT in M. anisopliae granule-treated plots with oat or rye cover crops that year. CFU density increases were higher in cover crops in 2002, but no significant cover crop effects were detected. Conidia persisted for up to 30 DAT in M. anisopliae spray plots and 60 DAT in granule-treated plots in 2002; however, no increases occurred in the years with less June rainfall. Trends suggest that M. anisopliae aqueous sprays result in greater conidia concentrations than granules at sugarbeet plant bases in June during T. myopaeformis oviposition and larval establishment on host plants. Increases are possible when delivering conidia via granules, but high post-application rainfall could be necessary for conidia production.

Water balance in the sugarbeet root maggot Tetanops myopaeformis, during long‐term low‐temperature storage and after freezing

The sugarbeet root maggot Tetanops myopaeformis Röder (Diptera: Ulidiidae) can be stored in moist sand at 4–6 °C for up to 5 years and is freeze‐tolerant. The majority of stored larvae survive in a state of post‐diapause quiescence and the remainder are in a multi‐year diapause. The present study aims to determine larval water content and water loss rates in diapausing and low‐temperature stored larvae. Body water content ranges from 57% to 70.1%. Two distinct groupings of larvae are revealed based on dry weights. The first group consists of the diapausing larvae and larvae stored for 1 year. This group has significantly higher dry weights than the second grouping, which consists of the larvae stored for 2 and 3 years. There are no significant differences within each group. Larval water losses follow a first‐order kinetic relationship with time. Larvae stored for 2 years lose water at a significantly higher rate than diapausing larvae. Larvae exhibit no active water uptake at storage temperatures. A freezing event does not induce a significant decrease in wet weights, nor does it increase larval water loss rates. These results indicate that metabolic water and the microclimate during storage are key factors enabling the long‐term survival of T. myopaeformis larvae during low‐temperature storage, and may provide insights for maintaining other insect species under similar conditions.

Beta vulgaris L. serine proteinase inhibitor gene expression in insect resistant sugar beet

Expression pattern of a sugar beet serine proteinase inhibitor gene, BvSTI, was characterized in response to mechanical and fall armyworm (FAW; Spodoptera frugiperda J.E. Smith) induced wounding. BvSTI expression was analyzed in three breeding lines moderately resistant to sugar beet root maggot (Tetanops myopaeformis Roder) and in a susceptible line, F1010. Increased mechanical wound induced levels of BvSTI expression were observed in all resistant lines as compared to F1010 during the first 24 h. The most intensive response to wounding was observed in one of the resistant lines, F1016, with a maximum 5- and 2.5-fold increase of BvSTI transcript levels over non-wounded roots and leaves, respectively. In contrast, slight increase of BvSTI transcript levels in leaves and even an initial decrease in roots were observed in F1010. BvSTI transcript accumulation in F1016 and F1010 tissues wounded by FAW showed a similar gene expression pattern, but it was delayed and less intense than the response incited by abiotic wounding. On the protein level, BvSTI specific antibody confirmed increased accumulation of the 30 kDa BvSTI protein in wounded leaves but not in roots of F1016 and F1010. Using trypsin inhibition assays, the activity of BvSTI was confirmed in F1016 roots and leaves and F1010 leaves. In F1010 roots BvSTI activity was completely lacking. We conclude that BvSTI gene expression was wound induced in the insect resistant germplasm suggesting that BvSTI can be used in biotechnological approaches or in breeding programs for improving insect resistance.

Registration of F1024 Sugarbeet Germplasm with Resistance to Sugarbeet Root Maggot

F1024 (Reg. No. GP‐272, PI 658654) sugarbeet (Beta vulgaris L.) germplasm with resistance to sugarbeet root maggot (Tetanops myopaeformis von Röder) was released by the USDA‐ARS and the North Dakota Agricultural Experiment Station, North Dakota State University, Fargo, ND on 15 Dec. 2009. F1024 was selected from a population formed by crossing F1016, a germplasm line resistant to root maggot, with a breeding line resistant to Cercospora leaf spot (CLS; caused by Cercospora beticola Sacc.). The population was subjected to three cycles of mass selection for resistance to root maggot followed by three cycles of selection among half‐sib families. Under natural root maggot infestations, F1024 had a damage rating of 2.1 (on a scale of 1 to 9, where 0 = no maggot feeding, and 9 = >75% of root surface with feeding scars), compared with an average of 6.1 for two susceptible commercial hybrids. In a 2009 CLS evaluation, F1024 had a significantly lower disease rating than the susceptible check, and the difference between F1024 and the resistant checks was not significant. The performance of testcross hybrids between the component half‐sib families of F1024 and a susceptible cytoplasmic‐male‐sterile line provided additional validation of the potential of root maggot–resistant hybrids in areas where root maggot is a perpetual threat.

Effects of Steam-Distilled Shoot Extract ofTagetes minuta(Asterales: Asteraceae) and Entomopathogenic Fungi on LarvalTetanops myopaeformis

Interactions of a biopesticidal formulation of steam distilled shoot extract of Mexican marigold, Tagetes minuta, and entomopathogenic fungi were evaluated for management of the sugarbeet root maggot, Tetanops myopaeformis (Röder). Shoot extract plus surfactant (E-Z Mulse) (=T. minuta oil) was used in a 65:35 ratio to test the hypothesis that this fungicidal and nematocidal biopesticide causes dose-dependent mortality and developmental arrest of T. myopaeformis but does not interfere with the action of entomopathogenic fungi when applied together. A soil-petri dish bioassay system was developed to test the hypothesis. For diapausing, nonfeeding but active 12-mo-old third-instar larvae, 0.5% T. minuta oil treatment (=0.325% active ingredient [AI]) was sufficient to prevent pupation without mortality, but 0.75% T. minuta oil treatment (=0.458% AI) was lethal for 93% of the test insects. The effect of T. minuta oil on fungal efficacy under simultaneous use was studied using a model system of two entomopathogenic fungi, Beauveria bassiana (Bals.) Vuillemin. TM28 and Metarhizium anisopliae variety anisopliae (Metsch.) Sorokin MA 1200, in a soil-based bioassay with larval sugarbeet root maggots. No adverse effects of T. minuta oil on action of entomopathogenic fungi and no synergy were found; an additive effect of the T. minuta oil and each fungal isolate separately was found.

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.

Economic Benefits of Additive Insecticide Applications for Root Maggot Control in Replanted Sugarbeet

Multiple factors can necessitate replanting of sugarbeet, Beta vulgaris (L.). Later-established plants in replanted fields will typically be smaller and more vulnerable to herbivory by insect pests than those established at earlier, more conventional planting dates. A field study was conducted during 2004, 2007, and 2008 in Pembina County, North Dakota to determine if additive insecticide protection is warranted to manage sugarbeet root maggot (SBRM), Tetanops myopaeformis (Roder), in replanted sugarbeet fields initially protected by conventional at-plant insecticide applications. Replanting with supplemental at-plant applications of terbufos 15G and chlorpyrifos 15G reduced SBRM feeding injury and increased root yields by 8.4 to 11.9 Mg/ha. Replant applications of terbufos 15G, applied to plots initially established with at-plant chlorpyrifos 15G, resulted in 32.2% and 34.8% more root and sucrose yield, respectively, and generated $405 more gross economic return per hectare than the stand-alone at-plant application of chlorpyrifos 15G. Postemergence liquid insecticides provided slightly less consistent control and yield benefits than granular insecticides applied at replanting, although the disparities rarely involved significant differences in gross economic return. The results of this investigation show that additive insecticide applications in replanted sugarbeet provide significant reductions in SBRM feeding injury that translate to major increases in yield and economic return. This study also demonstrates the economic

Supercooling point plasticity during cold storage in the freeze-tolerant sugarbeet root maggotTetanops myopaeformis

The sugarbeet root maggot Tetanops myopaeformis (Roder) overwinters as a freeze-tolerant third-instar larva. Although most larvae are considered to over- winter for only 1 year, some may exhibit prolonged diapause in the field. In the lab- oratory, they can live for over 5 years using a combination of diapause and post-diapause quiescence. In the present study, the cold survival strategies of these larvae during storage is investigated by measuring their supercooling points in com- bination with survival data. Supercooling points (SCPs) change significantly during storage, highlighted by a marked increase in the range of SCPs recorded, although the ability to tolerate freezing is not affected. Additionally, a freezing event 're- focuses' the SCPs of aged larvae to levels similar to those seen at diapause initia- tion. This change in SCPs is dependant not only on the initial freezing event, but also on the parameters of the incubation period between freezing events. Finally, the temperatures of larval overwintering microhabitats are monitored during the 2007 - 2008 boreal winter. The results indicate that, although overwintering larva are physiologically freeze-tolerant, they may essentially be freeze avoidant during overwintering via microhabitat selection.

Production of microsclerotia of the fungal entomopathogen Metarhizium anisopliae and their potential for use as a biocontrol agent for soil-inhabiting insects

Microsclerotia (MS), overwintering structures produced by many plant pathogenic fungi, have not been described for Metarhizium anisopliae. Three strains of M. anisopliae – F52, TM109, and MA1200 – formed MS in shake flask cultures using media with varying carbon concentrations and carbon-to-nitrogen (C:N) ratios. Under the conditions of this study, all strains produced MS, compact hyphal aggregates that become pigmented with culture age, in addition to more typical blastospores and mycelia. While all strains formed desiccation tolerant MS, highest concentrations (2.7–2.9 × 10 8 L −1 liquid medium) were produced in rich media with C:N ratios of 30:1 and 50:1 by strain F52. All three strains of M. anisopliae produced similar biomass concentrations when media and growth time were compared. Strain MA1200 produced higher concentrations of blastospores than the other two strains of M. anisopliae with highest blastospore concentrations (1.6 and 4.2 × 10 8 blastospores ml −1 on days 4 and 8, respectively) in media with the highest carbon and nitrogen concentrations. Microsclerotial preparations of M. anisopliae containing diatomaceous earth survived air-drying (to <5 % moisture) with no significant loss in viability. Rehydration and incubation of air-dried MS granules on water agar plates resulted in hyphal germination and sporogenic germination to produce high concentrations of conidia. Bioassays using soil-incorporated, air-dried MS preparations resulted in significant infection and mortality in larvae of the sugar beet root maggot, Tetanops myopaeformis. This is the first report of the production of sclerotial bodies by M. anisopliae and provides a novel approach for the control of soil-dwelling insects with this entomopathogenic fungus.

Efficacy of Metarhizium anisopliae microsclerotial granules1

The entomopathogenic fungus Metarhizium anisopliae has very recently been shown to produce microsclerotia (MS) – compact, heavily melanised, hyphal aggregates – in liquid media. Soil incorporation bioassays of dried MS preparations of three isolates of M. anisopliae were conducted using third instar Tetanops myopaeformis (sugarbeet root maggot) in clay and/or clay loam field soils as a model system to demonstrate efficacy. At rates as low as 23 mg MS granules/100 g dry soil, the biocontrol efficacy of MS granules of M. anisopliae Strain F52 produced in liquid media with a high carbon concentration (36 g/L) and high C:N ratios (30:1, 50:1) were superior to MS preparations produced in low carbon (8 g carbon/L) media and a high carbon medium with a 10:1 C:N ratio. Bioassays using MS formulations of M. anisopliae strains MA1200 and TM109 produced in high carbon and high C:N ratio media were superior in efficacy to the other MS production media tested. MS preparations of M. anisopliae F52 showed superior efficacy against the sugarbeet root maggot in comparison with more conventional, conidia-covered nutritive (corn grit) granules in a clay and clay soil. The MS granules were also highly efficacious against the sugarbeet root maggot at soil moisture levels as low as 0.983 A w (−2.33 MPa). Granular preparations incorporating Metarhizium MS can serve as a viable formulation for the use of this fungus against soil insects.

Recent advances in functional genomics for sugar beet (Beta vulgaris L.) improvement: progress in determining the role of BvSTI in pest resistance in roots

To gain knowledge of root resistance mechanisms in sugar beet, Beta vulgaris L., our laboratory has been studying the interaction of sugar beet with its most devastating insect pest, the sugar beet root maggot (SBRM; Tetanops myopaeformis Roder). Damage from SBRM infestations is a serious problem and current control measures rely on environmentally damaging insecticides. We recently reported root-specific gene expression incited by SBRM feeding in a moderately resistant F1016 and a susceptible parental F1010 line. AcDNA expressed sequence tag (EST) coding for a serine (trypsin-type) protease inhibitor (BvSTI) was identified and investigated further here. BvSTI shares sequence similarity with a root-specific tomato gene whose expression is induced by insect feeding. Since serine proteases comprise the major digestive enzymes in root maggot midguts, we hypothesize BvSTI may be involved in resistance. To elucidate the functional role of BvSTI, its coding region was fused to the CaMV 35S promoter and constitutively expressed in sugar beet hairy roots and N. benthamiana plants. In BvSTI-transformed F1010 hairy roots, trypsin inhibitory activity increased 2 to 4-fold. Using a polyacrylamide gel assay, new trypsin-like PI activity was detected in BvSTI-N. benthamiana plants. Since SBRM cannot be reared in vitro, two other insects that utilize serine digestive proteases, fall armyworm (Spodoptera frugiperda) and tobacco hornworm (Manduca sexta), were screened for resistance. To date, we demonstrated that 1) fall armyworm will feed on sugar beet hairy roots and 2) tobacco hornworm fed BvSTI-N. benthamiana leaves had reduced weights and pupal sizes. These results suggest that BvSTI may contribute to the moderate resistance of F1016 roots to SBRM. Functional analysis of additional ESTs will further support efforts to characterize the components of sugar beet root resistance mechanisms.

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 °C revealed no differences among respiration rates of initially diapausing and long-term stored larvae. A 15° increase in temperature elevated respiration in both diapausing and stored larvae, with levels of CO 2 release ranging between 8- and 14-fold higher at 20 °C than at 5 °C. Similarly, 6–10-fold increases in O 2 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.

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 1.8 × 10 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.