M. D. Stamm

Waveform Library for Chinch Bugs (Hemiptera: Heteroptera: Blissidae): Characterization of Electrical Penetration Graph Waveforms at Multiple Input Impedances

ABSTRACT Electrical penetration graph (EPG) monitoring has been used extensively to elucidate mechanisms of resistance in plants to insect herbivores with piercing-sucking mouthparts. Characterization of waveforms produced by insects during stylet probing is essential to the application of this technology. In the studies described herein, a four-channel Backus and Bennett AC-DC monitor was used to characterize EPG waveforms produced by adults of two economically important chinch bug species: southern chinch bug, Blissus insuhris Barber, feeding on St. Augustinegrass, and western chinch bug, Blissus occiduus Barber, feeding on buffalograss. This is only the third time a heteropteran species has been recorded by using EPG; it is also the first recording of adult heteropterans, and the first of Blissidae. Probing of chinch bugs was recorded with either AC or DC applied voltage, no applied voltage, or voltage switched between AC and DC mid-recording, at input impedances ranging from 106 to 1010 &OHgr;, plus 1013 &OHgr;, to develop a waveform library. Waveforms exhibited by western and southern chinch bugs were similar, and both showed long periods of putative pathway and ingestion phases (typical of salivary sheath feeders) interspersed with shorter phases, termed transitional J wave and interruption. The J wave is suspected to be an &KHgr; wave, that is, in EPG parlance, a stereotypical transition waveform that marks contact with a preferred ingestion tissue. The flexibility of using multiple input impedances with the AC-DC monitor was valuable for determining the electrical origin (resistance vs. electromotive force components) of the chinch bug waveforms. It was concluded that an input impedance of 107 &OHgr;, with either DC or AC applied voltage, is optimal to detect all resistanceand electromotive force-component waveforms produced during chinch bug probing. Knowledge of electrical origins suggested hypothesized biological meanings of the waveforms, before time-intensive future correlation experiments by using histology, microscopy, and other techniques.

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

BackgroundNeonicotinoid 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.ResultsDirect 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).ConclusionsWe 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.

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.

Dose-Response Relationships of Clothianidin, Imidacloprid, and Thiamethoxam to Blissus occiduus (Hemiptera: Blissidae)

ABSTRACT The western chinch bug, Blissus occiduus Barber (Hemiptera: Blissidae), has emerged as a serious pest of buffalograss, Buchloë dactyloides (Nuttall) Engelmann. In general, neonicotinoid insecticides effectively control a variety of turfgrass insects, particularly phloem-feeding pests. However, because of well documented inconsistencies in control, these compounds are generally not recommended for chinch bugs. This study was designed to document the contact and systemic toxicity of three neonicotinoid insecticides (clothianidin, imidacloprid, and thiamethoxam) to B. occiduus. In contact bioassays, thiamethoxam was ≈20-fold less toxic than clothianidin or imidacloprid to B. occiduus nymphs and three-fold more toxic to adults. In adult systemic bioassays, thiamethoxam was up to five-fold more toxic than clothianidin or imidacloprid. Interestingly, thiamethoxam was significantly more toxic to adults than to nymphs in both contact and systemic bioassays. This was not observed with clothianidin or imidacloprid. Bifenthrin, used for comparative purposes, exhibited 1,844-fold and 122-fold increase in toxicity to nymphs and adults, respectively. These results provide the first documentation of the relative toxicity of these neonicotinoid insecticides to B. occiduus.

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.

Feeding Behavior of Aphis glycines (Hemiptera: Aphididae) on Soybeans Exhibiting Antibiosis, Antixenosis, and Tolerance Resistance

Abstract Aphis glycines Matsumura (Hemiptera: Aphididae) is a serious pest of soybean in North America. Plant resistance is a valuable tool for the management of this pest, and a better understanding of the interactions between aphid and soybeans expressing varying levels and different categories of resistance can assist in the development of aphid resistant or tolerant genotypes. The electrical penetration graph (EPG) technique was used to evaluate the feeding behavior of A. glycines (biotype 1) on 4 soybean genotypes: (1) ‘Dowling’ (contains Rag1 gene and exhibits antibiosis); (2) PI 200538 (contains Rag2 gene and exhibits antixenosis); (3) KS4202 (exhibits tolerance); and (4) ‘SD76RR’ (susceptible to aphid). Aphids spent shorter periods in the sieve element phase on ‘Dowling’ and exhibited a greater number of pathway phases, non-probing events, and a longer time in nonprobing events in PI 200538 and ‘Dowling.’ For ‘SD76RR’ and KS4202, aphids demonstrated more sustained phloem ingestion, spent shorter time in non-probing events, and exhibited fewer pathway phases and potential drops. These results indicate that resistance factors are present in the phloem of ‘Dowling.’ For PI 200538, it is suggested that antixenotic factors are involved in resistance to A. glycines. Because KS4202 is tolerant to biotype 1 of A. glycines, the suitability of this genotype was expected already. This study provides important data that contribute to the understanding of how soybean aphids (biotype 1) feed on soybean genotypes with various aphid resistant genes and categories. In addition to assisting in the distinction between resistance categories, these results are useful in soybean breeding programs focusing on developing genotypes with greater resistance to insects.