Uptake and translocation of imidacloprid, clothianidin and flupyradifurone in seed-treated soybeans.

BACKGROUND: Seed treatment insecticides have become a popular management option for early-season insect control. This study investigated the total uptake and translocation of seed-applied [(14) C]imidacloprid, [(14) C]clothianidin and [(14) C]flupyradifurone into different plant parts in three soybean vegetative stages (VC, V1 and V2). The effects of soil moisture stress on insecticide uptake and translocation were also assessed among treatments. We hypothesized that (1) uptake and translocation would be different among the insecticides owing to differences in water solubility, and (2) moisture stress would increase insecticide uptake and translocation. RESULTS: Uptake and translocation did not follow a clear trend in the three vegetative stages. Initially, flupyradifurone uptake was greater than clothianidin uptake in VC soybeans. In V1 soybeans, differences in uptake among the three insecticides were not apparent and unaffected by soil moisture stress. Clothianidin was negatively affected by soil moisture stress in V2 soybeans, while imidacloprid and flupyradifurone were unaffected. Specifically, soil moisture stress had a positive effect on the distribution of flupyradifurone in leaves. This was not observed with the neonicotinoids. CONCLUSIONS: This study enhances our understanding of the uptake and distribution of insecticides used as seed treatments in soybean. The uptake and translocation of these insecticides differed in response to soil moisture stress. © 2015 Society of Chemical Industry.

Transcriptional response of soybean to thiamethoxam seed treatment in the presence and absence of drought stress.

BACKGROUND: Neonicotinoid insecticides are widely known for their broad-spectrum control of arthropod pests. Recently, their effects on plant physiological mechanisms have been characterized as producing a stress shield, which is predicted to enhance tolerance to adverse conditions. Here we investigate the molecular underpinnings of the stress shield concept using the neonicotinoid thiamethoxam in two separate experiments that compare gene expression. We hypothesized that the application of a thiamethoxam seed treatment to soybean would alter the expression of genes involved in plant defensive pathways and general stress response in later vegetative growth. First, we used next-generation sequencing to examine the broad scale transcriptional effects of the thiamethoxam seed treatment at three vegetative stages in soybean. Second, we selected ten target genes associated with plant defense pathways in soybean and examined the interactive effects of thiamethoxam seed treatment and drought stress on expression using qRT-PCR. RESULTS: Direct comparison of thiamethoxam-treated and untreated soybeans revealed minor transcriptional differences. However, when examined across vegetative stages, the thiamethoxam seed treatment induced substantial transcriptional changes that were not observed in untreated plants. Genes associated with photosynthesis, carbohydrate and lipid metabolism, development of the cell wall and membrane organization were uniquely upregulated between vegetative stages in thiamethoxam-treated plants. In addition, several genes associated with phytohormone and oxidative stress responses were downregulated between vegetative stages. When we examined the expression of a subset of ten genes associated with plant defense and stress response, the application of thiamethoxam was found to interact with drought stress by enhancing or repressing expression. In drought stressed plants, thiamethoxam induced (upregulated) expression of a thiamine biosynthetic enzyme (THIZ2) and gibberellin regulated protein (GRP), but repressed (downregulated) the expression of an apetala 2 (GmDREB2A;2), lipoxygenase (LIP), and SAM dependent carboxyl methyltransferase (SAM). CONCLUSIONS: We found evidence that a thiamethoxam seed treatment alters the expression soybean genes related to plant defense and stress response both in the presence and absence of drought stress. Consistent with the thiamethoxam stress shield concept, several genes associated with phytohormones showed enhanced expression in drought stressed plants.

A molecular key for the identification of blow flies in southeastern Nebraska.

Immature blow flies (Calliphoridae) are typically the first colonizers of cadavers. Identification of the early instars using traditional, morphology-based keys is difficult because of their small size, similarity, and simplicity in external morphology. Information derived from molecular genetic data would augment the accurate identification of immature flies. Nine species of blow flies commonly found in southeastern Nebraska were used to examine the utility of molecular-based keys. Polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) were investigated with 10 common, inexpensive, restriction enzymes from an amplicon of approximately 1500 bp spanning the mitochondrial cytochrome oxidase I gene. A simple molecular taxonomic key, comprising RFLP from the restriction enzymes HinfI and DraI, enabled the differentiation of all species used. Further development of PCR-RFLP, including more extensive and intensive examination of blow flies, would benefit forensic laboratories in the accurate identification of evidence consisting of immature blow flies.

Effect of distribution and concentration of topically applied neonicotinoid insecticides in buffalograss, Buchloe dactyloides, leaf tissues on the differential mortality of Blissus occiduus under field conditions.

BACKGROUND: Neonicotinoid insecticides are generally efficacious against many turfgrass pests, including several important phloem-feeding insects. However, inconsistencies in control of western chinch bugs, Blissus occiduus, have been documented in field efficacy studies. This research investigated the efficacy of three neonicotinoid insecticides (clothianidin, imidacloprid and thiamethoxam) against B. occiduus in buffalograss under field conditions and detected statistically significant differences in B. occiduus numbers among treatments. A subsequent study documented the relative quantity and degradation rate of these insecticides in buffalograss systemic leaf tissues, using HPLC. RESULTS: Neonicotinoid insecticides initially provided significant reductions in B. occiduus numbers, but mortality diminished over the course of the field studies. Furthermore, while all three neonicotinoids were present in the assayed buffalograss leaf tissues, imidacloprid concentrations were significantly higher than those of clothianidin and thiamethoxam. Over the course of the 28 day study, thiamethoxam concentrations declined 700-fold, whereas imidacloprid and clothianidin declined only 70-fold and 60-fold respectively. CONCLUSIONS: Field studies continued to verify inconsistencies in B. occiduus control with neonicotinoid insecticides. This is the first study to document the relative concentrations of topically applied neonicotinoid insecticides in buffalograss systemic leaf tissues.

Contact and fumigant toxicity of a botanical-based feeding deterrent of the stable fly, Stomoxys calcitrans (Diptera: Muscidae).

The stable fly, Stomoxys calcitrans (L.), has been considered one of the most serious biting flies of confined and pastured livestock. The economic losses caused by the stable fly to the cattle industry in the United States exceed $2 billion annually. Current practices for managing stable flies using insecticides provide only marginal control. Insecticide resistance has also been recently reported in stable flies. The present study reports the use of plant-based insecticides, for example, essential oils, as alternatives for managing this fly pest. The toxicity of several plant essential oils and selected ingredient compounds was evaluated by contact and fumigant toxicity bioassays. Catnip oil (20 mg dosage) showed the highest toxicity against stable flies, the shortest knock-down time (∼7 min), and the quickest lethal time (∼19 min). Toxicity levels similar to catnip oil were found among three insect repellent compounds (N,N-diethyl-3-methylbenzamide, 2-methylpiperidinyl-3-cyclohexene-1-carboxamide, (1S,2'S)-2-methylpiperidinyl-3-cyclohexene-1-carboxamide). No differences in knock-down and lethal times were found among the catnip oil and its two active ingredient compounds. Similar stable fly mortality was observed using a 20 mg dose of catnip oil in a modified K&D system and a fumigant jar. When catnip oil was topically applied to stable flies, the least lethal dose was 12.5 µg/fly, and a 50 µg/fly dose resulted in 100% mortality. The blood-feeding behavior of stable flies was also negatively affected by the topical application of catnip oil, and the effect was dose-dependent. This study demonstrated that catnip oil has both contact and fumigant toxicity against the stable fly and thus has the potential as an alternative for stable fly control.

Levels of tolerance, antibiosis, and antixenosis among resistant buffalograsses and zoysiagrasses.

The western chinch bug, Blissus occiduus Barber, has been documented as one of the most serious pests of buffalograss, Buchloë dactyloides (Nuttall) Engelmann, and zoysiagrass, Zoysia japonica Steudel, grown for turf in midwestern states. Resistance to the western chinch bug has been identified in both buffalograsses and zoysiagrasses. Choice and no-choice studies were conducted to determine the categories (antibiosis, antixenosis, and tolerance) of three resistant buffalograsses (PX3-5-1', 196', and 184') and three resistant zoysiagrasses (El Toro, Emerald, and Zorro). Antibiosis studies found no significant differences in survival, nymphal development, or fecundity among the resistant and susceptible buffalograsses or zoysiagrasses, indicating that antibiosis is not an important factor in the resistance. Based on chinch bug damage ratings, 184, 196, and PX3-5-1 have comparable levels of tolerance to the known tolerant buffalograss 'Prestige', and Zorro was the most tolerant zoysiagrass. Choice studies indicated the presence of antixenosis in the buffalograss selection 196 and the zoysiagrass Emerald.

Maggot development during morgue storage and its effect on estimating the post-mortem interval.

When insect evidence is obtained during autopsy, forensic entomologists make decisions regarding the effects of low-temperature (-1 degrees C to 4 degrees C) storage of the body and associated insects when estimating the post-mortem interval (PMI). To determine the effects of storage in a morgue cooler on the temperature of maggot masses, temperatures inside and outside of body bags containing a human cadaver and porcine cadavers (seven replicates) were measured during storage. Temperatures remained significantly higher (p<0.05) inside of the body bags relative to the cooler, and remained at levels sufficient for maggot feeding and development. If the assumption that no insect development takes place during preautopsy refrigeration is made, potential error rates in PMI estimation of 8.6-12.8% occur. The potential for blow fly larvae to undergo significant development while being stored in the morgue is a possibility that forensic entomologists should consider during an investigation involving samples collected from autopsy. Case and experimental evidence also demonstrate that substantial tissue loss can occur from maggot feeding during morgue storage.

Chinch bug (Hemiptera: Blissidae) mouthpart morphology, probing frequencies, and locations on resistant and susceptible germplasm.

Chinch bugs are common pests of many agronomic and horticulturally important crops and turfgrasses. Previous research has indicated that some grasses exhibit resistance to multiple chinch bug species, whereas others are resistant to only one species. The objectives of this research were to document differences in the probing frequencies and locations among Blissus species as well as differences in mouthpart morphology as a first step in understanding the differential responses of grasses to chinch bug feeding. Scanning electron microscopy detected differences in the total lengths of proboscises as well as individual mouthpart segments among the four species studied. Blissus occiduus Barber probed significantly more often on buffalograss, Buchloë dactyloides (Nuttall) Engelmann, than any other plant material. Probing locations of B. occiduus and Blissus leucopterus leucopterus (Say) were similar on both B. occiduus-resistant and susceptible buffalograsses and KS94 sorghum, Sorghum bicolor (L.) Moench (B. occiduus-resistant, B. l. leucopterus-resistant). However, on 'Wheatland' sorghum (B. occiduus-resistant, B. l. leucopterus-susceptible), stylet tracts of B. l. leucopterus most often terminated in the bundle sheath cells, whereas those of B. occiduus generally terminated in the vascular tissues.

Evaluation of cool- and warm-season grasses for resistance to multiple chinch bug (Hemiptera: Blissidae) species.

Chinch bugs are common pests of many agronomic and horticulturally important crops and turfgrasses. The extensive overlap of plant hosts and geographic distribution of Blissus leucopterus leucopterus (Say), Blissus leucopterus hirtus Montandon, Blissus insularis Barber, and Blissus occiduus Barber underscores the importance of identifying resistant germplasm. Cool- and warm-season turfgrasses and sorghum, Sorghum bicolor (L.) Moench, were evaluated for resistance to chinch bugs in the Blissus complex, and the presence of multiple resistance was documented. Greenhouse studies established that B. occiduus-resistant ('Prestige', formerly NE91-118) and -susceptible ('378') buffalograsses,, Buchloë dactyloides (Nuttall) Engelmann, were susceptible to all other chinch bug species. KS94 sorghum exhibited resistance to both B. occiduus and B. l. leucopterus, whereas B. insularis-resistant St. Augustinegrass, Stenotaphrum secundatum (Walter) Kuntze ('Floratam'), was also resistant to B. occiduus. B. l. leucopterus-susceptible sorghum ('Wheatland') and B. insularis-susceptible St. Augustinegrasses ('Raleigh' and 'Amerishade') were highly resistant to B. occiduus. Endophyte-free and -enhanced fine fescues (Festuca spp.) were moderately to highly susceptible to B. l. hirtus but moderately to highly resistant to B. occiduus. The results of this research showed the buffalograsses evaluated, including B. occiduus-resistant Prestige, are moderately to highly susceptible to the three other chinch bug species. In contrast, B. occiduus did not cause considerable damage to any of the turfgrasses or sorghum cultivars evaluated, other than buffalograss, irrespective of whether or not they are resistant to another chinch bug species. This information is increasingly important as various grasses become adapted to regions that may possess chinch bug species other than those with which they are typically associated. These levels of Blissus resistance should be included when selecting resistant germplasm for managing Blissus species pests.

Trapping Phyllophaga spp. (Coleoptera: Scarabaeidae: Melolonthinae) in the United States and Canada using sex attractants.

The sex pheromone of the scarab beetle, Phyllophaga anxia, is a blend of the methyl esters of two amino acids, L-valine and L-isoleucine. A field trapping study was conducted, deploying different blends of the two compounds at 59 locations in the United States and Canada. More than 57,000 males of 61 Phyllophaga species (Coleoptera: Scarabaeidae: Melolonthinae) were captured and identified. Three major findings included: (1) widespread use of the two compounds [of the 147 Phyllophaga (sensu stricto) species found in the United States and Canada, males of nearly 40% were captured]; (2) in most species intraspecific male response to the pheromone blends was stable between years and over geography; and (3) an unusual pheromone polymorphism was described from P. anxia. Populations at some locations were captured with L-valine methyl ester alone, whereas populations at other locations were captured with L-isoleucine methyl ester alone. At additional locations, the L-valine methyl ester-responding populations and the L-isoleucine methyl ester-responding populations were both present, producing a bimodal capture curve. In southeastern Massachusetts and in Rhode Island, in the United States, P. anxia males were captured with blends of L-valine methyl ester and L-isoleucine methyl ester.

Host preference of the chinch bug, Blissus occiduus.

The chinch bug, Blissus occiduus Barber (Hemiptera: Blissidae), is an important pest of buffalograss, Buchloë dactyloides (Nutall) Engelmann and potentially other turfgrass, crop, and non-crop hosts. Choice studies documented the number of B. occiduus present on selected turfgrasses, crops and weeds, and provided important insights into the host preferences of this chinch bug. Grasses with the most chinch bugs present included the warm-season turfgrasses B. dactyloides , zoysiagrass, Zoysia japonica Steudel, bermudagrass, Cynodon dactylon (L.) Pers., and St. Augustinegrass, Stenotaphrum secundatum (Walt.) Kuntze. The other grasses tested, green foxtail, Setaria viridis (L.) Beauv, Kentucky bluegrass, Poa pratensis L., perennial ryegrass, Lolium perenne L., rye, Secale cereale L., sorghum, Sorghum bicolor (L.) Moench, tall fescue, Festuca arundinacea Schreb. and wheat Tritium aestivum L. had significantly fewer chinch bugs. Buffalograss and zoysiagrass had the highest numbers of chinch bugs among the warm-season grasses and the buffalograss cultivars '86-120' and 'PX-3-5-1' had more chinch bugs than the zoysiagrass cultivars 'Meyers' and 'El Toro' after the two hour evaluation time.

Turfgrass, crop, and weed hosts of Blissus occiduus (Hemiptera: Lygaeidae).

Blissus occiduus Barber is an important pest of buffalograss, Buchloë dactyloides (Nuttall) Engelmann, turf. No-choice studies documented the susceptibility of selected turfgrasses, crops, and weeds to B. occiduus feeding. Highly to moderately susceptible grasses included buffalograss; yellow Setaria glauca (L.) and green foxtail Setaria viridis (L.); Kentucky bluegrass, Poa pratensis L.; perennial ryegrass, Lolium perenne L.; brome, Bromus spp. Leyss.; zoysiagrass, Zoysia japonica Steudel; Bermuda grass, Cynodon dactylon (L.) Pers.; sorghum, Sorghum bicolor (L.) Moench; tall fescue, Festuca arundinacea Schreb.; and barley Hordeum vulgare (L.). Slightly to nonsusceptible grasses included fine fescue, Festuca ovina hirtula L.; rye, Secale cereale L.; crabgrass Digitaria sanguinalis (L.); bentgrass, Agrostis palustris Huds.; wheat, Tritium aestivun L.; corn, Zea mays L.; fall panicum Panicum dichotomiflorum Michx.; and St. Augustinegrass, Stenotaphrum secundatum (Walt.) Kuntze. The reproductive potential of B. occiduus was also investigated on these same grasses. B. occiduus produced offspring on 15 of the 18 turfgrass, crop, and weed species evaluated. No reproduction occurred on either Bermuda grass or St. Augustinegrass, and buffalograss plants were killed by B. occiduus feeding before offspring could be produced.

Chinch bug-resistant buffalograss: an investigation of tolerance, antixenosis, and antibiosis.

Choice and no-choice studies were conducted to determine the categories (antibiosis, antixenosis, and tolerance) of resistance of four buffalograsses (NE91-118, 'Bonnie Brae', 'Cody', and 'Tatanka') previously identified as resistant to the western chinch bug, Blissus occiduus Barber. Antibiosis studies found no significant differences in western chinch bug fecundity, nymphal development, or survival among the resistant and susceptible buffalograsses. Tolerance studies indicated that NE91-118, Cody, and Tatanka exhibited moderate-to-high levels of tolerance based on western chinch bug damage ratings and plant height, whereas Bonnie Brae exhibited moderate-to-low levels of tolerance. Choice studies indicated the presence of antixenosis in NE91-118, whereas Cody and Tatanka showed little or no antixenosis. Scanning electron microscopy was used to disclose morphological differences between NE91-118 (resistant) and '378' (susceptible). The epicuticular wax structures and trichome densities were similar between 378 and NE91-118, suggesting that morphological structures do not contribute to NE91-118 antixenosis.

Evaluation of buffalograss germplasm for resistance to Blissus occiduus (Hemiptera: Lygaeidae).

Blissus occiduus Barber has emerged as an important insect pest of buffalograss, Buchloë dactyloides (Nuttall) Engelmann, in Nebraska. This research evaluated selected buffalograss germplasm for resistance to B. occiduus. Eleven buffalograss selections were screened for chinch bug resistance in three greenhouse studies and two field evaluations. Based on chinch bug damage, NE91-118, 'Tatanka', 'Bonnie Brae', and 'Cody' were rated highly to moderately resistant. These four buffalograsses exhibited minimal damage, even though all were heavily infested with chinch bugs. NE84-45-3 and '378' were highly susceptible to B. occiduus. Field evaluations confirmed chinch bug resistance ratings under field conditions. NE91-118 displayed high levels of resistance in the field screening evaluations, whereas Cody and Tatanka showed moderate levels of resistance, and 378 was highly susceptible.