Life Sciences Commons^™

Up-Regulation Of Apoptotic- And Cell Survival-Related Gene Pathways Following Exposures Of Western Corn Rootworm To B. Thuringiensis Crystalline Pesticidal Proteins In Transgenic Maize Roots, Brad S. Coates, Emeline Deleury, Aaron J. Gassmann, Bruce E. Hibbard, Lance Meinke, Nicholas J. Miller, Jennifer L. Petzold-Maxwell, B. Wade French, Thomas W. Sappington, Blair D. Siegfried, Thomas Guillemaud

Department of Entomology: Faculty Publications

Background: Resistance of pest insect species to insecticides, including B. thuringiensis (Bt) pesticidal proteins expressed by transgenic plants, is a threat to global food security. Despite the western corn rootworm, Diabrotica virgifera virgifera, being a major pest of maize and having populations showing increasing levels of resistance to hybrids expressing Bt pesticidal proteins, the cell mechanisms leading to mortality are not fully understood.

Results: Twenty unique RNA-seq libraries from the Bt susceptible D. v. virgifera inbred line Ped12, representing all growth stages and a range of different adult and larval exposures, were assembled into a reference transcriptome. Ten-day exposures …

A Deeper Meaning For Shallow-Level Phylogenomic Studies: Nested Anchored Hybrid Enrichment Offers Great Promise For Resolving The Tiger Moth Tree Of Life (Lepidoptera: Erebidae: Arctiinae), Nicolas J. Dowdy, Shannon Keating, Alan R. Lemmon, Emily M. Lemmon, William E. Conner, Clare H. Scott Chialvo, Susan Weller, Rebecca B. Simmons, Melissa S. Sisson, Jennifer M. Zaspel

Department of Entomology: Faculty Publications

Anchored hybrid enrichment (AHE) has emerged as a powerful tool for uncovering the evolutionary relationships within many taxonomic groups. AHE probe sets have been developed for a variety of insect groups, though none have yet been shown to be capable of simultaneously resolving deep and very shallow (e.g., intraspecific) divergences. In this study, we present NOC1, a new AHE probe set (730 loci) for Lepidoptera specialized for tiger moths and assess its ability to deliver phylogenetic utility at all taxonomic levels. We test the NOC1 probe set with 142 individuals from 116 species sampled from all the major lineages of …

Transcriptional Responses Of Soybean Aphids To Sublethal Insecticide Exposure, Laramy S. Enders, Leslie C. Rault, Tiffany Heng-Moss, Blair Siegfried, Nicholas J. Miller

Department of Entomology: Faculty Publications

Insecticides are a key tool in the management of many insect pests of agriculture, including soybean aphids. The selection imposed by insecticide use has often lead to the evolution of resistance by the target pest through enhanced detoxification mechanisms. We hypothesised that exposure of insecticide-susceptible aphids to sublethal doses of insecticides would result in the up-regulation of genes involved in detoxification of insecticides, revealing the genes upon which selection might act in the field. We used the soybean aphid biotype 1 reference genome, version 6.0 as a reference to analyze RNA-Seq data. We identified multiple genes with potential detoxification roles …

Global Responses Of Resistant And Susceptible Sorghum (Sorghum Bicolor) To Sugarcane Aphid (Melanaphis Sacchari), Hannah M. Tetreault, Sajjan Grover, Erin Scully, Tammy Gries, Nathan A. Palmer, Gautam Sarath, Joe Louis, Scott E. Sattler

Department of Entomology: Faculty Publications

The sugarcane aphid (Melanaphis sacchari) has emerged as a significant pest for sorghum. The use of sugarcane aphid-resistant sorghum germplasm with integrated pest management strategies appears to be an excellent solution to this problem. In this study, a resistant line (RTx2783) and a susceptible line (A/BCK60) were used to characterize the differences in plant responses to the sugarcane aphid through a series of experiments, which examined global sorghum gene expression, aphid feeding behavior and inheritance of aphid resistance. The global transcriptomic responses to sugarcane aphids in resistant and susceptible plants were identified using RNA-seq and compared to the …

Association Of Age, Sex, And Pyrethroid Resistance Status On Survival And Cytochrome P450 Gene Expression In Aedes Aegypti (L.), Leslie C. Rault, Scott T. O'Neal, Ellis J. Johnson, Troy D. Anderson

Department of Entomology: Faculty Publications

Aedes aegypti is a vector of viruses that negatively impact human health. Insecticide resistance complicates mosquito control efforts, but understanding the mechanisms of resistance can help to improve management practices. This study examined different factors that could influence the interpretation of toxicity bioassays and gene expression studies in A. aegypti, including sex and age, in the context of resistance to pyrethroids. Bioassays using a pyrethroid-resistant strain, Puerto Rico (PR), and a pyrethroid-susceptible strain, Rockefeller (Rock), of A. aegypti were conducted with females and males of three age groups to determine differences in mortality induced by deltamethrin. Overall, strain was …

Investigation Of Cry3bb1 Resistance And Intoxication In Western Corn Rootworm By Rna Sequencing, Leslie Catherine Rault, Blair D. Siegfried, Aaron J. Gassmann, Haichuan Wang, Gary J. Brewer, Nicholas J. Miller

Department of Entomology: Faculty Publications

The western corn rootworm (WCR) Diabrotica virgifera virgifera LeConte is a major pest of corn that has evolved resistance to transgenic maize that produces insecticidal Cry toxins. The specific mode of action of Cry3Bb1 and mechanism of resistance in WCR are unknown. This study compared gene expression between Cry3Bb1-susceptible and Cry3Bb1-resistant WCR neonates, in the presence and absence of Cry3Bb1. RNA-Seq data were analyzed to identify differentially expressed transcripts between strains of WCR, providing candidate transcripts for resistance to Cry3Bb1. Constitutive and Cry3Bb1-induced differences between strains caused the differential expression of 608 transcripts after 8 hr. Differentially expressed transcripts between …

Evaluation Of Smartstax And Smartstax Pro Maize Against Western Corn Rootworm And Northern Corn Rootworm: Efficacy And Resistance Management, Graham P. Head, Matthew W. Carroll, Sean P. Evans, Dwain M. Rule, Alan R. Willse, Thomas L. Clark, Nicholas P. Storer, Ronald D. Flannagan, Luke W. Samuel, Lance Meinke

Department of Entomology: Faculty Publications

Background: Cases of western corn rootworm (WCR) field-evolved resistance to Cry3Bb1 and other corn rootworm (CRW) control traits have been reported. Pyramid products expressing multiple CRW traits can delay resistance compared to single trait products. We used field studies to assess the pyramid CRW corn products, SmartStax (expressing Cry3Bb1 and Cry34Ab1/Cry35Ab1) and SmartStax PRO (expressing Cry3Bb1, Cry34Ab1/Cry35Ab1 and DvSnf7), at locations with high WCR densities and possible Cry3Bb1 resistance, and to assess the reduction in adult emergence attributable to DvSnf7 and other traits. Insect resistance models were used to assess durability of SmartStax and SmartStax PRO to WCR resistance.

Results: …

Rnai Induced Knockdown Of A Cadherin-Like Protein (Ef531715) Does Not Affect Toxicity Of Cry34/35ab1 Or Cry3aa To Diabrotica Virgifera Virgifera Larvae (Coleoptera: Chrysomelidae), Sek Yee Tan, Murugesan Rangasamy, Haichuan Wang, Ana Maria Vélez, James Hasler, David Mccaskill, Tao Xu, Hong Chen, Jessica Jurzenski, Matthew Kelker, Xiaoping Xu, Kenneth E. Narva, Blair D. Siegfried

Department of Entomology: Faculty Publications

The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, is an important maize pest throughout most of the U.S. Corn Belt. Bacillus thuringiensis (Bt) insecticidal proteins including modified Cry3Aa and Cry34/35Ab1 have been expressed in transgenic maize to protect against WCR feeding damage. To date, there is limited information regarding the WCR midgut target sites for these proteins. In this study, we examined whether a cadherin-like gene from Diabrotica virgifera virgifera (DvvCad; Gen-Bank accession # EF531715) associated with WCR larval midgut tissue is necessary for Cry3Aa or Cry34/ 35Ab1 toxicity. Experiments were designed to examine the sensitivity of WCR to …

Cry1f Resistance Among Lepidopteran Pests: A Model For Improved Resistance Management?, Ana María Vélez, Neetha Nanoth Vellichirammal, Juan Luis Jurat-Fuentes, Blair Siegfried

Department of Entomology: Faculty Publications

The Cry1Fa protein from the bacterium Bacillus thuringiensis (Bt) is known for its potential to control lepidopteran pests, especially through transgenic expression in maize and cotton. The maize event TC1507 expressing the cry1Fa toxin gene became commercially available in the United States in 2003 for the management of key lepidopteran pests including the European corn borer, Ostrinia nubilalis, and the fall armyworm, Spodoptera frugiperda. A high-dose/refuge strategy has been widely adopted to delay evolution of resistance to event TC1507 and other transgenic Bt crops. Efficacy of this strategy depends on the crops expressing a high dose of the …

Effect Of Cry1f Maize On The Behavior Of Susceptible And Resistant Spodoptera Frugiperda And Ostrinia Nubilalis, Ana María Vélez, Analiza P. Alves, Erin E. Blankenship, Blair Siegfried

Department of Entomology: Faculty Publications

Understanding the behavior of pests targeted with Bacillus thuringiensis Berliner (Bt) crops is important to define resistance management strategies. Particularly the study of larval movement between plants is important to determine the feasibility of refuge configurations. Exposure to Bt maize, Zea mays L. (Poaceae), has been suggested to increase larval movement in lepidopteran species but few studies have examined the potential for resistance to interact with behavioral responses to Bt toxins. Choice and no-choice experiments were conducted with Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae) and Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) to determine whether Cry1F resistance influences neonate movement. Leaf discs …

Evidence Of Field-Evolved Resistance To Bifenthrin In Western Corn Rootworm (Diabrotica Virgifera Virgifera Leconte) Populations In Western Nebraska And Kansas, Adriano E. Pereira, Haichuan Wang, Sarah N. Zukoff, Lance Meinke, B. Wade French, Blair Siegfried

Department of Entomology: Faculty Publications

Pyrethroid insecticides have been used to control larvae or adults of the western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, a key pest of field corn in the United States. In response to reports of reduced efficacy of pyrethroids in WCR management programs in southwestern areas of Nebraska and Kansas the present research was designed to establish a baseline of susceptibility to the pyrethroid insecticide, bifenthrin, using susceptible laboratory populations and to compare this baseline with susceptibility of field populations. Concentration-response bioassays were performed to estimate the baseline susceptibility. From the baseline data, a diagnostic concentration (LC99) was determined and …

Transcriptional Analysis Of Susceptible And Resistant European Corn Borer Strains And Their Response To Cry1f Protoxin, Neetha Nanoth Vellichirammal, Haichuan Wang, Seong-Il Eyun, Estuko N. Moriyama, Brad Coates, Nicholas Miller, Blair Siegfried

Department of Entomology: Faculty Publications

Background: Despite a number of recent reports of insect resistance to transgenic crops expressing insecticidal toxins from Bacillus thuringiensis (Bt), little is known about the mechanism of resistance to these toxins. The purpose of this study is to identify genes associated with the mechanism of Cry1F toxin resistance in European corn borer (Ostrinia nubilalis Hübner). For this, we compared the global transcriptomic response of laboratory selected resistant and susceptible O. nubilalis strain to Cry1F toxin. We further identified constitutive transcriptional differences between the two strains.

Results: An O. nubilalis midgut transcriptome of 36,125 transcripts was assembled de novo from …

Parental Rna Interference Of Genes Involved In Embryonic Development Of The Western Corn Rootworm, Diabrotica Virgifera Virgifera Leconte, Chitvan Khajuria, Ana Maria Vélez, Murugesan Rangasamy, Haichuan Wang, Elane Fishilevich, Meghan L.F. Frey, Newton Portilho Carneiro, Premchand Gandra, Kenneth E. Narva, B. D. Siegfried

Department of Entomology: Faculty Publications

RNA interference (RNAi) is being developed as a potential tool for insect pest management and one of the most likely target pest species for transgenic plants that express double stranded RNA (dsRNA) is the western corn rootworm. Thus far, most genes proposed as targets for RNAi in rootworm cause lethality in the larval stage. In this study, we describe RNAi-mediated knockdown of two developmental genes, hunchback (hb) and brahma (brm), in the western corn rootworm delivered via dsRNA fed to adult females. dsRNA feeding caused a significant decrease in hb and brm transcripts in the adult …

Genetic Variation In Field And Laboratory Populations Of The Spined Soldier Bug, Podisus Maculiventris, Kathleen Kneeland, Thomas A. Coudron, Erica Lindroth, David Stanley, John E. Foster

Department of Entomology: Faculty Publications

The predatory spined soldier bug, Podisus maculiventris (Say) (Heteroptera: Pentatomidae), is an economically important and highly valued biological control agent. There is substantial information on the biology, ecology, behavior, and rearing of this stink bug. However, virtually nothing is known of its genetic variation, in natural or domesticated populations. To address this lacuna, we used amplified fragment length polymorphism (AFLP) to assess the genetic variability of field and laboratory populations. Four AFLP universal primer combinations yielded a total of 209 usable loci. The AFLP results showed greater genetic variability between populations from Missouri and Mississippi (both USA), and relatively low …

Spatial Genetic Variation Among Spodoptera Frugiperda (Lepidoptera: Noctuidae) Sampled From The United States, Puerto Rico, Panama, And Argentina, Difabachew K. Belay, Pete L. Clark, Steven R. Skoda, David J. Isenhour, Jaime Molina-Ochoa, Claudia Gianni, John E. Foster

Department of Entomology: Faculty Publications

Spatial genetic variability of the fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), was studied by collecting samples from 31 locations in the United States, Argentina, Panama, and Puerto Rico, and then using amplified fragment length polymorphism to detect genetic variation. Analysis of molecular variance showed significant genetic variation in fall armyworm among all (28%) sample locations and individuals within (71%) sample locations; genetic variation of fall armyworm was minimal between sample locations grouped into regions. The pairwise fixation index (F_ST) comparisons showed significant genetic differentiation (0.288) among the 31 locations. However, dendrograms of results from cluster …

Distribution Of Genes And Repetitive Elements In The Diabrotica Virgifera Virgifera Genome Estimated Using Bac Sequencing, Brad S. Coates, Analiza P. Alves, Haichuan Wang, Kimberly K. O. Walden, B. Wade French, Nicholas J. Miller, Craig A. Abel, Hugh M. Robertson, Thomas W. Sappington, Blair D. Siegfried

Department of Entomology: Faculty Publications

Feeding damage caused by the western corn rootworm, Diabrotica virgifera virgifera, is destructive to corn plants in North America and Europe where control remains challenging due to evolution of resistance to chemical and transgenic toxins. A BAC library, DvvBAC1, containing 109,486 clones with 104 ± 34.5 kb inserts was created, which has an ~4.56X genome coverage based upon a 2.58 Gb (2.80 pg) flow cytometry-estimated haploid genome size. Paired end sequencing of 1037 BAC inserts produced 1.17Mb of data (~0.05% genome coverage) and indicated ~9.4 and 16.0% of reads encode, respectively, endogenous genes and transposable elements (TEs). …

Inbreeding-Stress Interactions: Evolutionary And Conservation Consequences, David H. Reed, Charles W. Fox, Laramy S. Enders, Torsten N. Kristensen

Department of Entomology: Faculty Publications

The effect of environmental stress on the magnitude of inbreeding depression has a long history of intensive study. Inbreeding-stress interactions are of great importance to the viability of populations of conservation concern and have numerous evolutionary ramifications. However, such interactions are controversial. Several meta-analyses over the last decade, combined with omic studies, have provided considerable insight into the generality of inbreeding-stress interactions, its physiological basis, and have provided the foundation for future studies. In this review, we examine the genetic and physiological mechanisms proposed to explain why inbreeding-stress interactions occur. We specifically examine whether the increase in inbreeding depression with …

Morphological Characterization And Molecular Mediated Genetic Variation Of Thief Ants (Hymenoptera: Formicidae), Ralph B. Narain, Shripat T. Kamble, Thomas O. Powers

Department of Entomology: Faculty Publications

The morphological characterization and molecular genetic variation were determined in populations of thief ants, Solenopsis molesta (Say). The genetic variations were elucidated using mitochondrial deoxyribonucleic acid (mDNA) sequences of cytochrome oxidase I. DNA from thief ants was extracted with Qiagen’s Gentra PUREGENE DNA Isolation Kit using their solid tissue protocol. Polymerase chain reactions (PCR) were run on the extracted DNA using primers Lep-F1 (forward) and Lep-R1 (reverse). The DNA products were concentrated and purified by Microcon Centrifugal Filter Unit YM-100. Purified DNA samples were sequenced at the University of Arkansas Medical Sciences (UAMS) DNA Sequencing Core Facility. The sequences were …

Efficacy Of Genetically Modified Bt Toxins Against Insects With Different Genetic Mechanisms Of Resistance, Bruce E. Tabashnik, Fangneng P. Huang, Mukti N. Ghimire, B. Rogers Leonard, Blair D. Siegfried, Murugesan Rangasamy, Yajun Yang, Yidong Wu, Linda J. Gahan, David G. Heckel, Alejandro Bravo, Mario Soberón

Department of Entomology: Faculty Publications

Transgenic crops that produce Bacillus thuringiensis (Bt) toxins are grown widely for pest control, but insect adaptation can reduce their efficacy. The genetically modified Bt toxins Cry1AbMod and Cry1AcMod were designed to counter insect resistance to native Bt toxins Cry1Ab and Cry1Ac. Previous results suggested that the modified toxins would be effective only if resistance was linked with mutations in genes encoding toxin-binding cadherin proteins. Here we report evidence from five major crop pests refuting this hypothesis. Relative to native toxins, the potency of modified toxins was > 350-fold higher against resistant strains of Plutella xylostella and Ostrinia nubilalis …

Field Introgression Of Diabrotica Barberi And Diabrotica Longicornis (Coleoptera: Chrysomelidae) Based On Genetic And Morphological Characters, Laura A. Campbell, Thomas L. Clark, Pete L. Clark, Lance Meinke, John E. Foster

Department of Entomology: Faculty Publications

Diabrotica barberi Smith & Lawrence and Diabrotica longicornis (Say) (Coleoptera: Chrysomelidae) are considered to be sister species, and it has been proposed that the two species may hybridize under field conditions. The objective of this study was to examine genetic and morphological characters of D. barberi and D. longicornis for evidence of field introgression. Both species were collected from sympatric and allopatric areas. Amplified fragment length polymorphisms and morphological characters (color and head capsule width) were used to examine variation within and among populations of D. barberi and D. longicornis. Relatively little of the overall …

Coordinated Diabrotica Genetics Research: Accelerating Progress On An Urgent Insect Pest Problem, Thomas W. Sappington, Blair D. Siegfried, Thomas Guillemaud

Department of Entomology: Faculty Publications

Diabrotica spp. (western, northern, and Mexican corn rootworms) represent the main pest complex of continuous field corn, Zea mays (L.), in North America. The western corn rootworm, Diabrotica virgifera virgifera LeConte, also has become the main pest of continuous corn in Central and Southeastern Europe since its introduction near Belgrade 15–20 years ago, and it represents a major risk to Western Europe. It has already caused economic losses in Eastern Europe, and Western countries such as France have committed large expenditures for containment and/or eradication.

Rootworm larvae feed on corn roots, and damaged plants are more susceptible to drought and …

Registration Of N547 Maize Germplasm Line, Nora E. D'Croz-Mason, John E. Foster, Stephen C. Mason

Department of Entomology: Faculty Publications

N547 germplasm line (Reg. no. GP-367, PI 615526) was released by the Agriculture Research Division, Institute of Agriculture and Natural Resources, University of Nebraska, on 24 Aug. 1999. N547 is a yellow endosperm maize inbred germplasm with resistance to stalk and shank tunneling damage caused by the second generation of the European corn borer (ECB) Ostrinia nubilalis (Hubner).

N547 was derived by selfing a selected full-sib family from MBITA, a tropical population developed by CIMMYT, Mexico. MBITA was created to recombine sources of resistance to temperate and subtropical stalk borers. These insects included ECB, southwestern corn borer (SWCB) Diatraea grandiosella …

Molecular Phylogeny Of Diabrotica Beetles (Coleoptera: Chrysomelidae) Inferred From Analysis Of Combined Mitochondrial And Nuclear Dna Sequences, Thomas L. Clark, Lance J. Meinke, John E. Foster

Department of Entomology: Faculty Publications

The phylogenetic relationships of thirteen Diabrotica (representing virgifera and fucata species groups) and two outgroup Acalymma beetle species (Coleoptera: Chrysomelidae) were inferred from the phylogenetic analysis of a combined data set of 1323 bp of mitochondrial DNA (mtDNA) cytochrome oxidase subunit 1 (COI) and the entire second internal transcribed spacer region (ITS-2) of nuclear ribosomal DNA of 362 characters. Species investigated were D. adelpha, D. balteata, D. barberi, D. cristata, D. lemniscata, D. longicornis, D. porracea, D. speciosa, D. undecimpunctata howardi, D. u. undecimpunctata, D. virgifera virgifera, D. v. zeae, D. viridula, and outgroup A. blandulum and A. vittatum. Maximum …

Molecular Phylogeny And Typing Of Blackflies (Diptera: Simuliidae) That Serve As Vectors Of Human Or Bovine Onchocerciasis, Jianming Tang, Kenneth Pruess, Eddie W. Cupp, Thomas R. Unnasch

Department of Entomology: Faculty Publications

A subregion of the mitochondrial large subunit (16s) rRNA gene was amplified by polymerase chain reaction (PCR) from nine species of blackflies (Diptera: Simuliidae) which serve as natural or experimental vectors of human or bovine Onchocerca parasites. PCR products from each species of blackfly were tested by directed heteroduplex analysis (DHDA), and their genotypes established according to diagnostic banding patterns of the heteroduplex products. Three alleles of mitochondrial 16s rRNA were found to exist in members of the Simulium (Edwardsellum) damnosum sensu lato complex from West Africa, and two alleles were found in the Neotropical Simulium (Psilopelmia …

Molecular Differentiation Of Alfalfa Weevil Strains (Coleoptera: Curculionidae), S. J. Erney, K. P. Pruess, S. D. Danielson, T. O. Powers

Department of Entomology: Faculty Publications

Mitochondrial DNA was amplified and sequenced from eastern, western, and Egyptian strains of alfalfa weevil, Hypera postica (Gyllenhal). Eastern and Egyptian weevils differed at only 2 nucleotide sites in 1,031 base pairs sequenced; western weevils differed by 5% sequence divergence. Three restriction sites were identified which separated eastern and western haplotypes. No intrastrain polymorphism was detected in 150 weevils from Nebraska. Collections from Lincoln in eastern Nebraska and Scottsbluff in western Nebraska were fixed for the eastern and western haplotypes, respectively. Eastern and western haplotypes were found together in the same fields in a broad overlap region in central Nebraska.

Mitochondrial Transfer Rna Genes In A Black Fly, Simulium Vittatum (Diptera: Simuliidae), Indicate Long Divergence From Mosquito (Diptera: Culicidae) And Fruit Fly (Diptera: Drosophilidae), K. P. Pruess, X. Zhu, T. O. Powers

Department of Entomology: Faculty Publications

Sequences are given for nine complete genes and one partial mitochondrial tRNA gene of the black fly, Simulium vittatum (Zetterstedt). Sequenced tRNA genes were for alanine, arginine, asparagine, aspartic acid, glutamic acid, glycine, leucine(CUN), lysine, serine(AGN), and valine. Nucleotides were aligned with the same previously sequenced genes in Aedes albopictus Skuse and Drosophila yakuba Burla. A cluster of six tRNA genes, which differ in arrangement in Ae. albopictus and D. yakuba, was amplified by PCR and found to have the same position and orientation in S. vittatum as in D. yakuba. Overall, similarity with either D. yakuba or …