@article{han_klobasa_oliveira_rotenberg_whitfield_lorenzen_2024, title={CRISPR/Cas9-mediated genome editing of <i>Frankliniella occidentalis</i>, the western flower thrips, via embryonic microinjection}, volume={4}, ISSN={["1365-2583"]}, DOI={10.1111/imb.12913}, abstractNote={Abstract The western flower thrips, Frankliniella occidentalis , poses a significant challenge in global agriculture as a notorious pest and a vector of economically significant orthotospoviruses. However, the limited availability of genetic tools for F. occidentalis hampers the advancement of functional genomics and the development of innovative pest control strategies. In this study, we present a robust methodology for generating heritable mutations in F. occidentalis using the CRISPR/Cas9 genome editing system. Two eye‐colour genes, white ( Fo‐w ) and cinnabar ( Fo‐cn ), frequently used to assess Cas9 function in insects were identified in the F. occidentalis genome and targeted for knockout through embryonic microinjection of Cas9 complexed with Fo‐w or Fo‐cn specific guide RNAs. Homozygous Fo‐w and Fo‐cn knockout lines were established by crossing mutant females and males. The Fo‐w knockout line revealed an age‐dependent modification of eye‐colour phenotype. Specifically, while young larvae exhibit orange‐coloured eyes, the colour transitions to bright red as they age. Unexpectedly, loss of Fo‐w function also altered body colour, with Fo‐w mutants having a lighter coloured body than wild type, suggesting a dual role for Fo‐w in thrips. In contrast, individuals from the Fo‐cn knockout line consistently displayed bright red eyes throughout all life stages. Molecular analyses validated precise editing of both target genes. This study offers a powerful tool to investigate thrips gene function and paves the way for the development of genetic technologies for population suppression and/or population replacement as a means of mitigating virus transmission by this vector.}, journal={INSECT MOLECULAR BIOLOGY}, author={Han, Jinlong and Klobasa, William and Oliveira, Lucas and Rotenberg, Dorith and Whitfield, Anna E. and Lorenzen, Marce D.}, year={2024}, month={Apr} }
 @article{almeida_maurastoni_sa-antunes_ventura_whitfield_fernandes_2024, title={Efforts to understand transmission of the papaya meleira virus complex by insects}, ISSN={["1983-2052"]}, DOI={10.1007/s40858-024-00661-5}, journal={TROPICAL PLANT PATHOLOGY}, author={Almeida, Joellington M. and Maurastoni, Marlonni and Sa-Antunes, Tathiana F. and Ventura, Jose A. and Whitfield, Anna E. and Fernandes, Patricia M. B.}, year={2024}, month={May} }
 @article{xavier_tyson_kerner_whitfield_2024, title={RNAi-mediated knockdown of exportin 1 negatively affected ovary development, survival and maize mosaic virus accumulation in its insect vector <i>Peregrinus maidis</i>}, ISSN={["1365-2583"]}, DOI={10.1111/imb.12910}, abstractNote={AbstractExportin 1 (XPO1) is the major karyopherin‐β nuclear receptor mediating the nuclear export of hundreds of proteins and some classes of RNA and regulates several critical processes in the cell, including cell‐cycle progression, transcription and translation. Viruses have co‐opted XPO1 to promote nucleocytoplasmic transport of viral proteins and RNA. Maize mosaic virus (MMV) is a plant‐infecting rhabdovirus transmitted in a circulative propagative manner by the corn planthopper, Peregrinus maidis. MMV replicates in the nucleus of plant and insect hosts, and it remains unknown whether MMV co‐opts P. maidis XPO1 (PmXPO1) to complete its life cycle. Because XPO1 plays multiple regulatory roles in cell functions and virus infection, we hypothesized that RNAi‐mediated silencing of XPO1 would negatively affect MMV accumulation and insect physiology. Although PmXPO1 expression was not modulated during MMV infection, PmXPO1 knockdown negatively affected MMV accumulation in P. maidis at 12 and 15 days after microinjection. Likewise, PmXPO1 knockdown negatively affected P. maidis survival and reproduction. PmXPO1 exhibited tissue‐specific expression patterns with higher expression in the ovaries compared with the guts of adult females. Survival rate was significantly lower for PmXPO1 knockdown females, compared with controls, but no effect was observed for males. PmXPO1 knockdown experiments revealed a role for PmXPO1 in ovary function and egg production. Oviposition and egg hatch on plants were dramatically reduced in females treated with dsRNA PmXPO1. These results suggest that PmXPO1 is a positive regulator of P. maidis reproduction and that it plays a proviral role in the insect vector supporting MMV infection.}, journal={INSECT MOLECULAR BIOLOGY}, author={Xavier, Cesar A. D. and Tyson, Clara and Kerner, Leo M. and Whitfield, Anna E.}, year={2024}, month={Mar} }
 @article{maurastoni_antunes_abreu_ribeiro_mehta_sanches_fontes_kitajima_cruz_santos_et al._2023, title={A Capsid Protein Fragment of a Fusagra-like Virus Found in Carica papaya Latex Interacts with the 50S Ribosomal Protein L17}, volume={15}, ISSN={["1999-4915"]}, DOI={10.3390/v15020541}, abstractNote={Papaya sticky disease is caused by the association of a fusagra-like and an umbra-like virus, named papaya meleira virus (PMeV) and papaya meleira virus 2 (PMeV2), respectively. Both viral genomes are encapsidated in particles formed by the PMeV ORF1 product, which has the potential to encode a protein with 1563 amino acids (aa). However, the structural components of the viral capsid are unknown. To characterize the structural proteins of PMeV and PMeV2, virions were purified from Carica papaya latex. SDS-PAGE analysis of purified virus revealed two major proteins of ~40 kDa and ~55 kDa. Amino-terminal sequencing of the ~55 kDa protein and LC-MS/MS of purified virions indicated that this protein starts at aa 263 of the deduced ORF1 product as a result of either degradation or proteolytic processing. A yeast two-hybrid assay was used to identify Arabidopsis proteins interacting with two PMeV ORF1 product fragments (aa 321–670 and 961–1200). The 50S ribosomal protein L17 (AtRPL17) was identified as potentially associated with modulated translation-related proteins. In plant cells, AtRPL17 co-localized and interacted with the PMeV ORF1 fragments. These findings support the hypothesis that the interaction between PMeV/PMeV2 structural proteins and RPL17 is important for virus–host interactions.}, number={2}, journal={VIRUSES-BASEL}, author={Maurastoni, Marlonni and Antunes, Tathiana Sa F. and Abreu, Emanuel F. M. and Ribeiro, Simone G. and Mehta, Angela and Sanches, Marcio M. and Fontes, Wagner and Kitajima, Elliot W. and Cruz, Fabiano T. and Santos, Alexandre M. C. and et al.}, year={2023}, month={Feb} }
 @article{maurastoni_han_whitfield_rotenberg_2023, title={A call to arms: novel strategies for thrips and tospovirus control}, volume={57}, ISSN={["2214-5753"]}, DOI={10.1016/j.cois.2023.101033}, abstractNote={Thrips and the tospoviruses they transmit are some of the most significant threats to food and ornamental crop production globally. Control of the insect and virus is challenging and new strategies are needed. Characterizing the thrips-virus interactome provides new targets for disrupting the transmission cycle. Viral and insect determinants of vector competence are being defined, including the viral attachment protein and its structure as well as thrips proteins that interact with and respond to tospovirus infection. Additional thrips control strategies such as RNA interference need further refinement and field-applicable delivery systems, but they show promise for the knockdown of essential genes for thrips survival and virus transmission. The identification of a toxin that acts to deter thrips oviposition on cotton also presents new opportunities for control of this important pest.}, journal={CURRENT OPINION IN INSECT SCIENCE}, author={Maurastoni, Marlonni and Han, Jinlong and Whitfield, Anna E. and Rotenberg, Dorith}, year={2023}, month={Jun} }
 @article{lee_hossain_jamalzadegan_liu_wang_saville_shymanovich_paul_rotenberg_whitfield_et al._2023, title={Abaxial leaf surface-mounted multimodal wearable sensor for continuous plant physiology monitoring}, volume={9}, ISSN={["2375-2548"]}, DOI={10.1126/sciadv.ade2232}, abstractNote={Wearable plant sensors hold tremendous potential for smart agriculture. We report a lower leaf surface-attached multimodal wearable sensor for continuous monitoring of plant physiology by tracking both biochemical and biophysical signals of the plant and its microenvironment. Sensors for detecting volatile organic compounds (VOCs), temperature, and humidity are integrated into a single platform. The abaxial leaf attachment position is selected on the basis of the stomata density to improve the sensor signal strength. This versatile platform enables various stress monitoring applications, ranging from tracking plant water loss to early detection of plant pathogens. A machine learning model was also developed to analyze multichannel sensor data for quantitative detection of tomato spotted wilt virus as early as 4 days after inoculation. The model also evaluates different sensor combinations for early disease detection and predicts that minimally three sensors are required including the VOC sensors.}, number={15}, journal={SCIENCE ADVANCES}, author={Lee, Giwon and Hossain, Oindrila and Jamalzadegan, Sina and Liu, Yuxuan and Wang, Hongyu and Saville, Amanda C. and Shymanovich, Tatsiana and Paul, Rajesh and Rotenberg, Dorith and Whitfield, Anna E. and et al.}, year={2023}, month={Apr} }
 @article{lahre_shekasteband_meadows_whitfield_rotenberg_2023, title={First Report of Resistance-Breaking Variants of Tomato Spotted Wilt Virus (TSWV) Infecting Tomatoes with the Sw-5 Resistance Gene in North Carolina}, volume={1}, ISSN={["1943-7692"]}, url={https://doi.org/10.1094/PDIS-11-22-2637-PDN}, DOI={10.1094/PDIS-11-22-2637-PDN}, abstractNote={Widespread use of tomato cultivars with the Sw-5 resistance gene has led to the emergence of resistance-breaking (RB) strains of tomato spotted wilt virus across the globe. In June of 2022, tomato spotted wilt (TSW) symptoms were observed at two farms (A and B, within 15 miles of each other) in Rowan County, NC on several commercial TSW resistant tomato cultivars (all heterozygous for the Sw-5 gene). At farm A, ~10% of plants had symptomatic foliage with ~30% of fruit with symptoms, while at farm B, up to 50% of plants had symptomatic foliage with ~80% of fruit with symptoms. Visual symptoms included stunting, severe leaf curling and bronzing, necrotic lesions on leaves, petioles and stems, and concentric ring spots on fruit (Supplementary Fig. 1). TSWV ImmunoStrips (AgDia, Elkhart, IN) and reverse-transcription (RT)-PCR with NSm primers (di Rienzo et al 2018) confirmed the presence of TSWV in 12 symptomatic plants sampled across the two farms. Primers designed to detect Impatiens necrotic spot virus, groundnut ringspot virus, tomato chlorotic spot virus, tomato chlorosis virus, alfalfa mosaic virus, and tomato necrotic streak virus (ilarvirus, Badillo et al., 2016) failed to generate amplicons of the expected size from cDNA generated from these field samples. The amplicons from full-length NSm cDNA were sequenced from independent, single-leaflet isolates from the TSWV-positive plants (three from farm A, nine from farm B) with the expectation of finding an amino acid (aa) substitution associated with the Sw-5 RB phenotype identified previously in CA (C118Y, Batuman et al. 2017) or Spain (C118Y and T120N, Lopez et al. 2011). All three nucleotide sequences from farm A contained the NSm C118Y substitution reported in CA. All three sequences were 99% identical (including the C118Y mutation) to NCBI GenBank accession KU179600.1, a TSWV isolate collected from GA in 2014 with no cultivar information reported. The nine nucleotide sequences from farm B contained neither of the two previously reported aa substitutions associated with the RB phenotype. Instead, all contained a D122G substitution within a conserved region of the TSWV NSm protein reported to be involved in direct interaction with the Sw-5 protein (Zhu et al 2017). Likewise, Huang et al (2021) generated a D122A mutation in TSWV-NSm, resulting in failure to elicit a Sw-5 mediated hypersensitive response. Three NSm sequences retrieved from GenBank contained the D122G substitution (AY848921.1, HM015516.1, KU179582.1), however, this mutation was not implicated directly with RB phenotypes (Ciuffo et al., 2005; Lopez et al., 2011; Marshall, 2016). The RB phenotype was confirmed with the NC variants on 'Mountain Merit' (Sw-5) by two means of virus inoculation: mechanical, rub-inoculation with extracted sap from infected plants, and thrips transmission assays with lab colony-maintained, Frankliniella occidentalis, the western flower thrips. Symptomatic leaf tissue obtained from these inoculation assays tested positive for TSWV by DAS-ELISA (AgDia, Elkhart, IN) and RT-PCR with NSm primers, providing definitive evidence of the occurrence of RB-TSWV at both farms, and subsequent sequencing confirmed the C118Y and D122G substitutions. This report warrants further investigation of the putative origins, prevalence and epidemiological implications of RB-TSWV variants in NC tomato production, and the development of new sources of resistance to TSWV.}, number={7}, journal={PLANT DISEASE}, author={Lahre, K. and Shekasteband, R. and Meadows, I. and Whitfield, A. E. and Rotenberg, D.}, year={2023}, month={Jan} }
 @article{oliver_rotenberg_agosto-shaw_mcinnes_lahre_mulot_adkins_whitfield_2024, title={Multigenic Hairpin Transgenes in Tomato Confer Resistance to Multiple Orthotospoviruses Including Sw-5 Resistance-Breaking Tomato Spotted Wilt Virus}, volume={4}, ISSN={["1943-7684"]}, DOI={10.1094/PHYTO-07-23-0256-KC}, abstractNote={Tomato spotted wilt virus (TSWV) and related thrips-borne orthotospoviruses are a threat to food and ornamental crops. Orthotospoviruses have the capacity for rapid genetic change by genome segment reassortment and mutation. Genetic resistance is one of the most effective strategies for managing orthotospoviruses, but there are multiple examples of resistance gene breakdown. Our goal was to develop effective multigenic, broad-spectrum resistance to TSWV and other orthotospoviruses. The most conserved sequences for each open reading frame (ORF) of the TSWV genome were identified, and comparison with other orthotospoviruses revealed sequence conservation within virus clades; some overlapped with domains with well-documented biological functions. We made six hairpin constructs, each of which incorporated sequences matching portions of all five ORFs. Tomato plants expressing the hairpin transgene were challenged with TSWV by thrips and leaf-rub inoculation, and four constructs provided strong protection against TSWV in foliage and fruit. To determine if the hairpin constructs provided protection against other emerging orthotospoviruses, we challenged the plants with tomato chlorotic spot virus and resistance-breaking TSWV and found that the same constructs also provided resistance to these related viruses. Antiviral hairpin constructs are an effective way to protect plants from multiple orthotospoviruses and are an important strategy in the fight against resistance-breaking TSWV and emerging viruses. Targeting of all five viral ORFs is expected to increase the durability of resistance, and combining them with other resistance genes could further extend the utility of this disease control strategy. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.}, journal={PHYTOPATHOLOGY}, author={Oliver, Jonathan E. and Rotenberg, Dorith and Agosto-Shaw, Karolyn and McInnes, Holly A. and Lahre, Kirsten A. and Mulot, Michael and Adkins, Scott and Whitfield, Anna E.}, year={2024}, month={Apr} }
 @article{whitfield_rotenberg_2023, title={Pests and resistance: The biology and control of supervectors and superpests}, volume={58}, ISSN={["2214-5753"]}, DOI={10.1016/j.cois.2023.101060}, journal={CURRENT OPINION IN INSECT SCIENCE}, author={Whitfield, Anna E. and Rotenberg, Dorith}, year={2023}, month={Aug} }
 @article{wang_klobasa_chu_huot_whitfield_lorenzen_2023, title={Structural and functional insights into the ATP-binding cassette transporter family in the corn planthopper, Peregrinus maidis}, volume={32}, ISSN={0962-1075 1365-2583}, url={http://dx.doi.org/10.1111/imb.12840}, DOI={10.1111/imb.12840}, abstractNote={AbstractThe corn planthopper, Peregrinus maidis, is an economically important pest of maize and sorghum. Its feeding behaviour and the viruses it transmits can significantly reduce crop yield. The control of P. maidis and its associated viruses relies heavily on insecticides. However, control has proven difficult due to limited direct exposure of P. maidis to insecticides and rapid development of resistance. As such, alternative control methods are needed. In the absence of a genome assembly for this species, we first developed transcriptomic resources. Then, with the goal of finding targets for RNAi‐based control, we identified members of the ATP‐binding cassette transporter family and targeted specific members via RNAi. PmABCB_160306_3, PmABCE_118332_5 and PmABCF_24241_1, whose orthologs in other insects have proven important in development, were selected for knockdown. We found that RNAi‐mediated silencing of PmABCB_160306_3 impeded ovary development; disruption of PmABCE_118332_5 resulted in localized melanization; and knockdown of PmABCE_118332_5 or PmABCF_24241_1 each led to high mortality within five days. Each phenotype is similar to that found when targeting the orthologous gene in other species and it demonstrates their potential for use in RNAi‐based P. maidis control. The transcriptomic data and RNAi results presented here will no doubt assist with the development of new control methods for this pest.}, number={4}, journal={Insect Molecular Biology}, publisher={Wiley}, author={Wang, Yu‐Hui and Klobasa, William and Chu, Fu‐Chyun and Huot, Ordom and Whitfield, Anna E. and Lorenzen, Marcé}, year={2023}, month={Apr}, pages={412–423} }
 @article{walker_freitas-astua_bejerman_blasdell_breyta_dietzgen_fooks_kondo_kurath_kuzmin_et al._2022, title={ICTV Virus Taxonomy Profile: Rhabdoviridae 2022}, volume={103}, ISSN={["1465-2099"]}, DOI={10.1099/jgv.0.001689}, abstractNote={The family Rhabdoviridae comprises viruses with negative-sense (−) RNA genomes of 10–16 kb. Virions are typically enveloped with bullet-shaped or bacilliform morphology but can also be non-enveloped filaments. Rhabdoviruses infect plants or animals, including mammals, birds, reptiles, amphibians or fish, as well as arthropods, which serve as single hosts or act as biological vectors for transmission to animals or plants. Rhabdoviruses include important pathogens of humans, livestock, fish or agricultural crops. This is a summary of the International Committee on Taxonomy of Viruses (ICTV) Report on the family Rhabdoviridae, which is available at ictv.global/report/rhabdoviridae.}, number={6}, journal={JOURNAL OF GENERAL VIROLOGY}, author={Walker, Peter J. and Freitas-Astua, Juliana and Bejerman, Nicolas and Blasdell, Kim R. and Breyta, Rachel and Dietzgen, Ralf G. and Fooks, Anthony R. and Kondo, Hideki and Kurath, Gael and Kuzmin, Ivan V and et al.}, year={2022} }
 @article{castrosanto_clemente_whitfield_alviar_2022, title={In silico analysis of the predicted protein-protein interaction of syntaxin-18, a putative receptor of Peregrinus maidis Ashmead (Hemiptera: Delphacidae) with Maize mosaic virus glycoprotein}, ISSN={["1538-0254"]}, DOI={10.1080/07391102.2022.2059569}, abstractNote={Abstract The corn planthopper, Peregrinus maidis Ashmead (Hemiptera:Delphacidae), is a widely distributed insect pest which serves as a vector of two phytopathogenic viruses, Maize mosaic virus (MMV) and Maize stripe virus (MStV). It transmits the viruses in a persistent and propagative manner. MMV is an alphanucleorhabdovirus with a negative-sense, single-stranded RNA unsegmented genome. One identified insect vector protein that may serve as receptor to MMV is Syntaxin-18 (PmStx18) which belongs to the SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) proteins. SNAREs play major roles in the final stage of docking and subsequent fusion of diverse vesicle-mediated transport events. In this work, in silico analysis of the interaction of MMV glycoprotein (MMV G) and PmStx18 was performed. Various freely available protein-protein docking web servers were used to predict the 3 D complex of MMV G and PmStx18. Analysis and protein-protein interaction (PPI) count showed that the complex predicted by the ZDOCK server has the highest number of interaction and highest affinity, as suggested by the calculated solvation free energy gain upon formation of the interface (ΔiG = −31 kcal/mol). Molecular dynamics simulation of the complex revealed important interactions at the interface over the course of 25 ns. This is the first in silico analysis performed for the interaction on a putative receptor of P. maidis and MMV G. The results of the PPI prediction provide novel information for studying the role of Stx18 in the transport, docking and fusion events involved in virus particle transport in the insect vector cells and its release. Communicated by Ramaswamy H. Sarma}, journal={JOURNAL OF BIOMOLECULAR STRUCTURE & DYNAMICS}, author={Castrosanto, Melvin A. and Clemente, Apel Jae N. and Whitfield, Anna E. and Alviar, Karen B.}, year={2022}, month={Mar} }
 @article{kanakala_xavier_martin_tran_redinbaugh_whitfield_2022, title={Rescue of the first alphanucleorhabdovirus entirely from cloned complementary DNA: An efficient vector for systemic expression of foreign genes in maize and insect vectors}, ISSN={["1364-3703"]}, DOI={10.1111/mpp.13273}, abstractNote={AbstractRecent reverse genetics technologies have enabled genetic manipulation of plant negative‐strand RNA virus (NSR) genomes. Here, we report construction of an infectious clone for the maize‐infecting Alphanucleorhabdovirus maydis, the first efficient NSR vector for maize. The full‐length infectious clone was established using agrobacterium‐mediated delivery of full‐length maize mosaic virus (MMV) antigenomic RNA and the viral core proteins (nucleoprotein N, phosphoprotein P, and RNA‐directed RNA polymerase L) required for viral transcription and replication into Nicotiana benthamiana. Insertion of intron 2 ST‐LS1 into the viral L gene increased stability of the infectious clone in Escherichia coli and Agrobacterium tumefaciens. To monitor virus infection in vivo, a green fluorescent protein (GFP) gene was inserted in between the N and P gene junctions to generate recombinant MMV‐GFP. Complementary DNA (cDNA) clones of MMV‐wild type (WT) and MMV‐GFP replicated in single cells of agroinfiltrated N. benthamiana. Uniform systemic infection and high GFP expression were observed in maize inoculated with extracts of the infiltrated N. benthamiana leaves. Insect vectors supported virus infection when inoculated via feeding on infected maize or microinjection. Both MMV‐WT and MMV‐GFP were efficiently transmitted to maize by planthopper vectors. The GFP reporter gene was stable in the virus genome and expression remained high over three cycles of transmission in plants and insects. The MMV infectious clone will be a versatile tool for expression of proteins of interest in maize and cross‐kingdom studies of virus replication in plant and insect hosts.}, journal={MOLECULAR PLANT PATHOLOGY}, author={Kanakala, Surapathrudu and Xavier, Cesar A. D. and Martin, Kathleen M. and Tran, Hong Hanh and Redinbaugh, Margaret G. and Whitfield, Anna E.}, year={2022}, month={Oct} }
 @article{rajarapu_ben-mahmoud_benoit_ullman_whitfield_rotenberg_2022, title={Sex-biased proteomic response to tomato spotted wilt virus infection of the salivary glands of Frankliniella occidentalis, the western flower thrips}, volume={149}, ISSN={["1879-0240"]}, DOI={10.1016/j.ibmb.2022.103843}, abstractNote={Successful transmission of tomato spotted wilt virus (TSWV) by Frankliniella occidentalis requires robust infection of the salivary glands (SGs) and virus delivery to plants during salivation. Feeding behavior and transmission efficiency are sexually-dimorphic traits of this thrips vector species. Proteins secreted from male and female SG tissues, and the effect of TSWV infection on the thrips SG proteome are unknown. To begin to discern thrips factors that facilitate virus infection of SGs and transmission by F. occidentalis, we used gel- and label-free quantitative and qualitative proteomics to address two hypotheses: (i) TSWV infection modifies the composition and/or abundance of SG-expressed proteins in adults; and (ii) TSWV has a differential effect on the male and female SG proteome and secreted saliva. Our study revealed a sex-biased SG proteome for F. occidentalis, and TSWV infection modulated the SG proteome in a sex-dependent manner as evident by the number, differential abundance, identities and generalized roles of the proteins. Male SGs exhibited a larger proteomic response to the virus than female SGs. Intracellular processes modulated by TSWV in males indicated perturbation of SG cytoskeletal networks and cell-cell interactions, i.e., basement membrane (BM) and extracellular matrix (ECM) proteins, and subcellular processes consistent with a metabolic slow-down under infection. Several differentially-abundant proteins in infected male SGs play critical roles in viral life cycles of other host-virus pathosystems. In females, TSWV modulated processes consistent with tissue integrity and active translational and transcriptional regulation. A core set of proteins known for their roles in plant cell-wall degradation and protein metabolism were identified in saliva of both sexes, regardless of virus infection status. Saliva proteins secreted by TSWV-infected adults indicated energy generation, consumption and protein turnover, with an enrichment of cytoskeletal/BM/ECM proteins and tricarboxylic acid cycle proteins in male and female saliva, respectively. The nonstructural TSWV protein NSs - a multifunctional viral effector protein reported to target plant defenses against TSWV and thrips - was identified in female saliva. This study represents the first description of the SG proteome and secretome of a thysanopteran and provides many candidate proteins to further unravel the complex interplay between the virus, insect vector, and plant host.}, journal={INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY}, author={Rajarapu, Swapna Priya and Ben-Mahmoud, Sulley and Benoit, Joshua B. and Ullman, Diane E. and Whitfield, Anna E. and Rotenberg, Dorith}, year={2022}, month={Oct} }
 @article{alviar_rotenberg_martin_whitfield_2022, title={The physical interactome between Peregrinus maidis proteins and the maize mosaic virus glycoprotein provides insights into the cellular biology of a rhabdovirus in the insect vector.}, volume={577}, ISSN={["1089-862X"]}, DOI={10.1016/j.virol.2022.10.002}, abstractNote={Rhabdovirus glycoproteins (G) serve multifunctional roles in virus entry, assembly, and exit from animal cells. We hypothesize that maize mosaic virus (MMV) G is required for invasion, infection, and spread in Peregrinus maidis, the planthopper vector. Using a membrane-based yeast two-hybrid assay, we identified 107 P. maidis proteins that physically interacted with MMV G, of which approximately 53% matched proteins with known functions including endocytosis, vesicle-mediated transport, protein synthesis and turnover, nuclear export, metabolism and host defense. Physical interaction networks among conserved proteins indicated a possible cellular coordination of processes associated with MMV G translation, protein folding and trafficking. Non-annotated proteins contained predicted functional sites, including a diverse array of ligand binding sites. Cyclophilin A and apolipophorin III co-immunoprecipitated with MMV G, and each showed different patterns of localization with G in insect cells. This study describes the first protein interactome for a rhabdovirus spike protein and insect vector.}, journal={VIROLOGY}, author={Alviar, Karen B. and Rotenberg, Dorith and Martin, Kathleen M. and Whitfield, Anna E.}, year={2022}, month={Dec}, pages={163–173} }
 @article{kuhn_adkins_agwanda_al kubrusli_alkhovsky_amarasinghe_avsic-zupanc_ayllon_bahl_balkema-buschmann_et al._2021, title={2021 Taxonomic update of phylum Negarnaviricota (Riboviria: Orthornavirae), including the large orders Bunyavirales and Mononegavirales}, ISSN={["1432-8798"]}, DOI={10.1007/s00705-021-05143-6}, abstractNote={In March 2021, following the annual International Committee on Taxonomy of Viruses (ICTV) ratification vote on newly proposed taxa, the phylum Negarnaviricota was amended and emended. The phylum was expanded by four families (Aliusviridae, Crepuscuviridae, Myriaviridae, and Natareviridae), three subfamilies (Alpharhabdovirinae, Betarhabdovirinae, and Gammarhabdovirinae), 42 genera, and 200 species. Thirty-nine species were renamed and/or moved and seven species were abolished. This article presents the updated taxonomy of Negarnaviricota as now accepted by the ICTV.}, journal={ARCHIVES OF VIROLOGY}, author={Kuhn, Jens H. and Adkins, Scott and Agwanda, Bernard R. and Al Kubrusli, Rim and Alkhovsky, Sergey V. and Amarasinghe, Gaya K. and Avsic-Zupanc, Tatjana and Ayllon, Maria A. and Bahl, Justin and Balkema-Buschmann, Anne and et al.}, year={2021}, month={Aug} }
 @article{kuhn_adkins_agwanda_al kubrusli_alkhovsky_amarasinghe_avsic-zupanc_ayllon_bahl_balkema-buschmann_et al._2021, title={2021 Taxonomic update of phylum Negarnaviricota (Riboviria: Orthornavirae), including the large orders Bunyavirales and Mononegavirales (Aug, 10.1007/s00705-021-05143-6, 2021)}, volume={166}, ISSN={["1432-8798"]}, DOI={10.1007/s00705-021-05266-w}, number={12}, journal={ARCHIVES OF VIROLOGY}, author={Kuhn, Jens H. and Adkins, Scott and Agwanda, Bernard R. and Al Kubrusli, Rim and Alkhovsky, Sergey V. and Amarasinghe, Gaya K. and Avsic-Zupanc, Tatjana and Ayllon, Maria A. and Bahl, Justin and Balkema-Buschmann, Anne and et al.}, year={2021}, month={Dec}, pages={3567–3579} }
 @article{xavier_allen_whitfield_2021, title={Ever-increasing viral diversity associated with the red imported fire ant Solenopsis invicta (Formicidae: Hymenoptera)}, volume={18}, ISSN={["1743-422X"]}, DOI={10.1186/s12985-020-01469-w}, abstractNote={Abstract
                Background
                Advances in sequencing and analysis tools have facilitated discovery of many new viruses from invertebrates, including ants. Solenopsis invicta is an invasive ant that has quickly spread worldwide causing significant ecological and economic impacts. Its virome has begun to be characterized pertaining to potential use of viruses as natural enemies. Although the S. invicta virome is the best characterized among ants, most studies have been performed in its native range, with less information from invaded areas.
              
                Methods
                Using a metatranscriptome approach, we further identified and molecularly characterized virus sequences associated with S. invicta, in two introduced areas, U.S and Taiwan. The data set used here was obtained from different stages (larvae, pupa, and adults) of S. invicta life cycle. Publicly available RNA sequences from GenBank’s Sequence Read Archive were downloaded and de novo assembled using CLC Genomics Workbench 20.0.1. Contigs were compared against the non-redundant protein sequences and those showing similarity to viral sequences were further analyzed.
              
                Results
                We characterized five putative new viruses associated with S. invicta transcriptomes. Sequence comparisons revealed extensive divergence across ORFs and genomic regions with most of them sharing less than 40% amino acid identity with those closest homologous sequences previously characterized. The first negative-sense single-stranded RNA virus genomic sequences included in the orders Bunyavirales and Mononegavirales are reported. In addition, two positive single-strand virus genome sequences and one single strand DNA virus genome sequence were also identified. While the presence of a putative tenuivirus associated with S. invicta was previously suggested to be a contamination, here we characterized and present strong evidence that Solenopsis invicta virus 14 (SINV-14) is a tenui-like virus that has a long-term association with the ant. Furthermore, based on virus sequence abundance compared to housekeeping genes, phylogenetic relationships, and completeness of viral coding sequences, our results suggest that four of five virus sequences reported, those being SINV-14, SINV-15, SINV-16 and SINV-17, may be associated to viruses actively replicating in the ant S. invicta.
              
                Conclusions
                The present study expands our knowledge about viral diversity associated with S. invicta in introduced areas with potential to be used as biological control agents, which will require further biological characterization.
              }, number={1}, journal={VIROLOGY JOURNAL}, author={Xavier, Cesar Augusto Diniz and Allen, Margaret Louise and Whitfield, Anna Elizabeth}, year={2021}, month={Jan} }
 @article{paul_ostermann_chen_saville_yang_gu_whitfield_ristaino_wei_2021, title={Integrated microneedle-smartphone nucleic acid amplification platform for in-field diagnosis of plant diseases}, volume={187}, ISSN={["1873-4235"]}, url={https://doi.org/10.1016/j.bios.2021.113312}, DOI={10.1016/j.bios.2021.113312}, abstractNote={We demonstrate an integrated microneedle (MN)-smartphone nucleic acid amplification platform for "sample-to-answer" diagnosis of multiplexed plant pathogens within 30 min. This portable system consists of a polymeric MN patch for rapid nucleic acid extraction within a minute and a 3D-printed smartphone imaging device for loop-mediated isothermal amplification (LAMP) reaction and detection. We expanded the extraction of the MN technology for DNA targets as in the previous study (ACS Nano, 2019, 13, 6540–6549) to more fragile RNA biomarkers, evaluated the storability of the extracted nucleic acid samples on MN surfaces, and developed a smartphone-based LAMP amplification and fluorescent reader device that can quantify four LAMP reactions on the same chip. In addition, we have found that the MN patch containing as few as a single needle tip successfully extracted enough RNA for RT-PCR or RT-LAMP analysis. Moreover, MN-extracted RNA samples remained stable on MN surfaces for up to three days. The MN-smartphone platform has been used to detect both Phytophthora infestans DNA and tomato spotted wilt virus (TSWV) RNA down to 1 pg, comparable to the results from a benchtop thermal cycler. Finally, multiplexed detection of P. infestans and TSWV through a single extraction from infected tomato leaves and amplification on the smartphone without benchtop equipment was demonstrated.}, journal={BIOSENSORS & BIOELECTRONICS}, publisher={Elsevier BV}, author={Paul, Rajesh and Ostermann, Emily and Chen, Yuting and Saville, Amanda C. and Yang, Yuming and Gu, Zhen and Whitfield, Anna E. and Ristaino, Jean B. and Wei, Qingshan}, year={2021}, month={Sep} }
 @article{klobasa_chu_huot_grubbs_rotenberg_whitfield_lorenzen_2021, title={Microinjection of Corn Planthopper, <em>Peregrinus maidis</em>, Embryos for CRISPR/Cas9 Genome Editing}, volume={3}, ISSN={1940-087X}, url={http://dx.doi.org/10.3791/62417}, DOI={10.3791/62417}, abstractNote={The corn planthopper, Peregrinus maidis, is a pest of maize and a vector of several maize viruses. Previously published methods describe the triggering of RNA interference (RNAi) in P. maidis through microinjection of double-stranded RNAs (dsRNAs) into nymphs and adults. Despite the power of RNAi, phenotypes generated via this technique are transient and lack long-term Mendelian inheritance. Therefore, the P. maidis toolbox needs to be expanded to include functional genomic tools that would enable the production of stable mutant strains, opening the door for researchers to bring new control methods to bear on this economically important pest. However, unlike the dsRNAs used for RNAi, the components used in CRISPR/Cas9-based genome editing and germline transformation do not easily cross cell membranes. As a result, plasmid DNAs, RNAs, and/or proteins must be microinjected into embryos before the embryo cellularizes, making the timing of injection a critical factor for success. To that end, an agarose-based egg-lay method was developed to allow embryos to be harvested from P. maidis females at relatively short intervals. Herein are provided detailed protocols for collecting and microinjecting precellular P. maidis embryos with CRISPR components (Cas9 nuclease that has been complexed with guide RNAs), and results of Cas9-based gene knockout of a P. maidis eye-color gene, white, are presented. Although these protocols describe CRISPR/Cas9-genome editing in P. maidis, they can also be used for producing transgenic P. maidis via germline transformation by simply changing the composition of the injection solution.}, number={169}, journal={Journal of Visualized Experiments}, publisher={MyJove Corporation}, author={Klobasa, William and Chu, Fu-Chyun and Huot, Ordom and Grubbs, Nathaniel and Rotenberg, Dorith and Whitfield, Anna E. and Lorenzen, Marcé D.}, year={2021}, month={Mar} }
 @article{kuhn_adkins_alioto_alkhovsky_amarasinghe_anthony_avsic-zupanc_ayllon_bahl_balkema-buschmann_et al._2020, title={2020 taxonomic update for phylumNegarnaviricota(Riboviria:Orthornavirae), including the large ordersBunyaviralesandMononegavirales}, volume={165}, ISSN={["1432-8798"]}, DOI={10.1007/s00705-020-04731-2}, abstractNote={In March 2020, following the annual International Committee on Taxonomy of Viruses (ICTV) ratification vote on newly proposed taxa, the phylum Negarnaviricota was amended and emended. At the genus rank, 20 new genera were added, two were deleted, one was moved, and three were renamed. At the species rank, 160 species were added, four were deleted, ten were moved and renamed, and 30 species were renamed. This article presents the updated taxonomy of Negarnaviricota as now accepted by the ICTV.}, number={12}, journal={ARCHIVES OF VIROLOGY}, author={Kuhn, Jens H. and Adkins, Scott and Alioto, Daniela and Alkhovsky, Sergey V. and Amarasinghe, Gaya K. and Anthony, Simon J. and Avsic-Zupanc, Tatjana and Ayllon, Maria A. and Bahl, Justin and Balkema-Buschmann, Anne and et al.}, year={2020}, month={Dec}, pages={3023–3072} }
 @article{rotenberg_baumann_ben-mahmoud_christiaens_dermauw_ioannidis_jacobs_vargas jentzsch_oliver_poelchau_et al._2020, title={Genome-enabled insights into the biology of thrips as crop pests}, volume={18}, ISSN={["1741-7007"]}, DOI={10.1186/s12915-020-00862-9}, abstractNote={AbstractBackgroundThe western flower thrips,Frankliniella occidentalis(Pergande), is a globally invasive pest and plant virus vector on a wide array of food, fiber, and ornamental crops. The underlying genetic mechanisms of the processes governing thrips pest and vector biology, feeding behaviors, ecology, and insecticide resistance are largely unknown. To address this gap, we present theF. occidentalisdraft genome assembly and official gene set.ResultsWe report on the first genome sequence for any member of the insect order Thysanoptera. Benchmarking Universal Single-Copy Ortholog (BUSCO) assessments of the genome assembly (size = 415.8 Mb, scaffold N50 = 948.9 kb) revealed a relatively complete and well-annotated assembly in comparison to other insect genomes. The genome is unusually GC-rich (50%) compared to other insect genomes to date. The official gene set (OGS v1.0) contains 16,859 genes, of which ~ 10% were manually verified and corrected by our consortium. We focused on manual annotation, phylogenetic, and expression evidence analyses for gene sets centered on primary themes in the life histories and activities of plant-colonizing insects. Highlights include the following: (1) divergent clades and large expansions in genes associated with environmental sensing (chemosensory receptors) and detoxification (CYP4, CYP6, and CCE enzymes) of substances encountered in agricultural environments; (2) a comprehensive set of salivary gland genes supported by enriched expression; (3) apparent absence of members of the IMD innate immune defense pathway; and (4) developmental- and sex-specific expression analyses of genes associated with progression from larvae to adulthood through neometaboly, a distinct form of maturation differing from either incomplete or complete metamorphosis in the Insecta.ConclusionsAnalysis of theF. occidentalisgenome offers insights into the polyphagous behavior of this insect pest that finds, colonizes, and survives on a widely diverse array of plants. The genomic resources presented here enable a more complete analysis of insect evolution and biology, providing a missing taxon for contemporary insect genomics-based analyses. Our study also offers a genomic benchmark for molecular and evolutionary investigations of other Thysanoptera species.}, number={1}, journal={BMC BIOLOGY}, author={Rotenberg, Dorith and Baumann, Aaron A. and Ben-Mahmoud, Sulley and Christiaens, Olivier and Dermauw, Wannes and Ioannidis, Panagiotis and Jacobs, Chris G. C. and Vargas Jentzsch, Iris M. and Oliver, Jonathan E. and Poelchau, Monica F. and et al.}, year={2020}, month={Oct} }
 @article{rotenberg_baumann_ben-mahmoud_christiaens_dermauw_ioannidis_jacobs_jentzsch_oliver_poelchau_et al._2020, title={Genome-enabled insights into the biology of thrips as crop pests (vol 18, 142, 2020)}, volume={18}, ISSN={["1741-7007"]}, DOI={10.1186/s12915-020-00915-z}, abstractNote={An amendment to this paper has been published and can be accessed via the original article.}, number={1}, journal={BMC BIOLOGY}, author={Rotenberg, Dorith and Baumann, Aaron A. and Ben-Mahmoud, Sulley and Christiaens, Olivier and Dermauw, Wannes and Ioannidis, Panagiotis and Jacobs, Chris G. C. and Jentzsch, Iris M. Vargas and Oliver, Jonathan E. and Poelchau, Monica F. and et al.}, year={2020}, month={Nov} }
 @article{german_lorenzen_grubbs_whitfield_2020, title={New Technologies for Studying Negative-Strand RNA Viruses in Plant and Arthropod Hosts}, volume={33}, ISSN={0894-0282 1943-7706}, url={http://dx.doi.org/10.1094/MPMI-10-19-0281-FI}, DOI={10.1094/MPMI-10-19-0281-FI}, abstractNote={ The plant viruses in the phylum Negarnaviricota, orders Bunyavirales and Mononegavirales, have common features of single-stranded, negative-sense RNA genomes and replication in the biological vector. Due to the similarities in biology, comparative functional analysis in plant and vector hosts is helpful for understanding host–virus interactions for negative-strand RNA viruses. In this review, we will highlight recent technological advances that are breaking new ground in the study of these recalcitrant virus systems. The development of infectious clones for plant rhabdoviruses and bunyaviruses is enabling unprecedented examination of gene function in plants and these advances are also being transferred to study virus biology in the vector. In addition, genome and transcriptome projects for critical nonmodel arthropods has enabled characterization of insect response to viruses and identification of interacting proteins. Functional analysis of genes using genome editing will provide future pathways for further study of the transmission cycle and new control strategies for these viruses and their vectors. }, number={3}, journal={Molecular Plant-Microbe Interactions®}, publisher={Scientific Societies}, author={German, Thomas L. and Lorenzen, Marcé D. and Grubbs, Nathaniel and Whitfield, Anna E.}, year={2020}, month={Mar}, pages={382–393} }
 @article{nachappa_challacombe_margolies_nechols_whitfield_rotenberg_2020, title={Tomato Spotted Wilt Virus Benefits Its Thrips Vector by Modulating Metabolic and Plant Defense Pathways in Tomato}, volume={11}, ISSN={["1664-462X"]}, DOI={10.3389/fpls.2020.575564}, abstractNote={Several plant viruses modulate vector fitness and behavior in ways that may enhance virus transmission. Previous studies have documented indirect, plant-mediated effects of tomato spotted wilt virus (TSWV) infection on the fecundity, growth and survival of its principal thrips vector, Frankliniella occidentalis, the western flower thrips. We conducted thrips performance and preference experiments combined with plant gene expression, phytohormone and total free amino acid analyses to determine if systemically-infected tomato plants modulate primary metabolic and defense-related pathways to culminate into a more favorable environment for the vector. In a greenhouse setting, we documented a significant increase in the number of offspring produced by F. occidentalis on TSWV-infected tomato plants compared to mock-inoculated plants, and in choice test assays, females exhibited enhanced settling on TSWV-infected leaves. Microarray analysis combined with phytohormone signaling pathway analysis revealed reciprocal modulation of key phytohormone pathways under dual attack, possibly indicating a coordinated and dampening defense against the vector on infected plants. TSWV infection, alone or in combination with thrips, suppressed genes associated with photosynthesis and chloroplast function thereby significantly impacting primary metabolism of the host plant, and hierarchical cluster and network analyses revealed that many of these genes were co-regulated with phytohormone defense signaling genes. TSWV infection increased expression of genes related to protein synthesis and degradation which was reflected in the increased total free amino acid content in virus-infected plants that harbored higher thrips populations. These results suggest coordinated gene networks that regulate plant primary metabolism and defense responses rendering virus-infected plants more conducive for vector colonization, an outcome that is potentially beneficial to the vector and the virus when considered within the context of the complex transmission biology of TSWV. To our knowledge this is the first study to identify global transcriptional networks that underlie the TSWV-thrips interaction as compared to a single mechanistic approach. Findings of this study increase our fundamental knowledge of host plant-virus-vector interactions and identifies underlying mechanisms of induced host susceptibility to the insect vector.}, journal={FRONTIERS IN PLANT SCIENCE}, author={Nachappa, Punya and Challacombe, Jean and Margolies, David C. and Nechols, James R. and Whitfield, Anna E. and Rotenberg, Dorith}, year={2020}, month={Dec} }
 @article{martin_whitfield_2019, title={Complete Genome Sequence of Maize Mosaic Nucleorhabdovirus}, volume={8}, ISSN={["2576-098X"]}, DOI={10.1128/MRA.00637-19}, abstractNote={
            The complete genome sequence of maize mosaic virus (MMV) was obtained using next-generation sequencing from infected
            Peregrinus maidis
            and rapid amplification of cDNA ends from infected
            Zea mays
            . The genome of MMV is 12,170 bases, and this project completed the 5′ and 3′ ends and amended the polymerase sequence.
          }, number={29}, journal={MICROBIOLOGY RESOURCE ANNOUNCEMENTS}, author={Martin, Kathleen M. and Whitfield, Anna E.}, year={2019}, month={Jul} }
 @article{yao_rotenberg_whitfield_2019, title={Delivery of maize mosaic virus to planthopper vectors by microinjection increases infection efficiency and facilitates functional genomics experiments in the vector}, volume={270}, ISBN={1879-0984}, DOI={10.1016/j.jviromet.2019.05.010}, abstractNote={The corn planthopper, Peregrinus maidis, not only causes direct damage to plants by feeding, but also transmits maize mosaic virus (MMV) to the plant hosts. The virus is transmitted in a propagative manner but the acquisition of MMV by the vector feeding on infected plants can result in low acquisition and inoculation efficiency. Here, we increased the acquisition efficiency by delivering the virus directly into the hemocoel through microinjection, which resulted in efficient virus infection of the insect and transmission to maize. We found that delivery of virus by injection of 10 ng MMV (50 nl, 200 μg/ml virions) into P. maidis resulted in 93% transmission efficiency. In dose-response experiments, MMV abundance in insects and transmission efficiency decreased as the amount of virus inoculum delivered into the hemocoel was reduced. Examination of virus distribution in the vector using immunolabeling and confocal microscopy revealed similar tissue distributions in the injected insects when compared to those of previous studies using feeding on plants for virus acquisition. The utility of virus inoculation by microinjection for functional analysis in virus-vector interaction was explored. Co-microinjection of MMV virions and the dsRNA of PI3Kδ (a transcript that is less abundant in MMV-infected insects), resulted in a reduction in PI3Kδ expression and higher virus titers in P. maidis. These findings demonstrated that virus microinjection is a robust method for obtaining large numbers of infected planthoppers that are competent in transmitting MMV and, in combination with RNAi, could significantly facilitate the functional analysis of P. maidis-MMV interactions.}, journal={JOURNAL OF VIROLOGICAL METHODS}, author={Yao, Jianxiu and Rotenberg, Dorith and Whitfield, Anna E.}, year={2019}, month={Aug}, pages={153–162} }
 @article{badillo-vargas_chen_martin_rotenberg_whitfield_2019, title={Discovery of Novel Thrips Vector Proteins That Bind to the Viral Attachment Protein of the Plant Bunyavirus Tomato Spotted Wilt Virus}, volume={93}, ISSN={["1098-5514"]}, url={https://doi.org/10.1101/416560}, DOI={10.1128/JVI.00699-19}, abstractNote={
            Thrips-transmitted viruses cause devastating losses to numerous food crops worldwide. For negative-sense RNA viruses that infect plants, the arthropod serves as a host as well by supporting virus replication in specific tissues and organs of the vector. The goal of this work was to identify thrips proteins that bind directly to the viral attachment protein and thus may play a role in the infection cycle in the insect. Using the model plant bunyavirus tomato spotted wilt virus (TSWV), and the most efficient thrips vector, we identified and validated six TSWV-interacting proteins from
            Frankliniella occidentalis
            first-instar larvae. Two proteins, an endocuticle structural glycoprotein and cyclophilin, were able to interact directly with the TSWV attachment protein, G
            N
            , in insect cells. The TSWV G
            N
            -interacting proteins provide new targets for disrupting the viral disease cycle in the arthropod vector and could be putative determinants of vector competence.
          }, number={21}, journal={Journal of Virology}, author={Badillo-Vargas, I.E. and Chen, Y. and Martin, K.M. and Rotenberg, D. and Whitfield, A.E.}, year={2019}, pages={e00699–19} }
 @article{chen_dessau_rotenberg_rasmussen_whitfield_2019, title={Entry of bunyaviruses into plants and vectors}, volume={104}, ISBN={["978-0-12-818394-6"]}, ISSN={["1557-8399"]}, DOI={10.1016/bs.aivir.2019.07.001}, abstractNote={The majority of plant-infecting viruses are transmitted by arthropod vectors that deliver them directly into a living plant cell. There are diverse mechanisms of transmission ranging from direct binding to the insect stylet (non-persistent transmission) to persistent-propagative transmission in which the virus replicates in the insect vector. Despite this diversity in interactions, most arthropods that serve as efficient vectors have feeding strategies that enable them to deliver the virus into the plant cell without extensive damage to the plant and thus effectively inoculate the plant. As such, the primary virus entry mechanism for plant viruses is mediated by the biological vector. Remarkably, viruses that are transmitted in a propagative manner (bunyaviruses, rhabdoviruses, and reoviruses) have developed an ability to replicate in hosts from two kingdoms. Viruses in the order Bunyavirales are of emerging importance and with the advent of new sequencing technologies, we are getting unprecedented glimpses into the diversity of these viruses. Plant-infecting bunyaviruses are transmitted in a persistent, propagative manner must enter two unique types of host cells, plant and insect. In the insect phase of the virus life cycle, the propagative viruses likely use typical cellular entry strategies to traverse cell membranes. In this review, we highlight the transmission and entry strategies of three genera of plant-infecting bunyaviruses: orthotospoviruses, tenuiviruses, and emaraviruses.}, journal={VIRUS ENTRY}, author={Chen, Yuting and Dessau, Moshe and Rotenberg, Dorith and Rasmussen, David A. and Whitfield, Anna E.}, year={2019}, pages={65–96} }
 @article{maes_adkins_alkhovsky_avsic-zupanc_ballinger_bente_beer_bergeron_blair_briese_et al._2019, title={Taxonomy of the order Bunyavirales: second update 2018}, volume={164}, ISSN={["1432-8798"]}, DOI={10.1007/s00705-018-04127-3}, abstractNote={In October 2018, the order Bunyavirales was amended by inclusion of the family Arenaviridae, abolishment of three families, creation of three new families, 19 new genera, and 14 new species, and renaming of three genera and 22 species. This article presents the updated taxonomy of the order Bunyavirales as now accepted by the International Committee on Taxonomy of Viruses (ICTV).}, number={3}, journal={ARCHIVES OF VIROLOGY}, author={Maes, Piet and Adkins, Scott and Alkhovsky, Sergey V. and Avsic-Zupanc, Tatjana and Ballinger, Matthew J. and Bente, Dennis A. and Beer, Martin and Bergeron, Eric and Blair, Carol D. and Briese, Thomas and et al.}, year={2019}, month={Mar}, pages={927–941} }
 @article{abudurexiti_adkins_alioto_alkhovsky_avsic-zupanc_ballinger_bente_beer_bergeron_blair_et al._2019, title={Taxonomy of the order Bunyavirales: update 2019}, volume={164}, ISSN={["1432-8798"]}, DOI={10.1007/s00705-019-04253-6}, abstractNote={In February 2019, following the annual taxon ratification vote, the order Bunyavirales was amended by creation of two new families, four new subfamilies, 11 new genera and 77 new species, merging of two species, and deletion of one species. This article presents the updated taxonomy of the order Bunyavirales now accepted by the International Committee on Taxonomy of Viruses (ICTV).}, number={7}, journal={ARCHIVES OF VIROLOGY}, author={Abudurexiti, Abulikemu and Adkins, Scott and Alioto, Daniela and Alkhovsky, Sergey V. and Avsic-Zupanc, Tatjana and Ballinger, Matthew J. and Bente, Dennis A. and Beer, Martin and Bergeron, Eric and Blair, Carol D. and et al.}, year={2019}, month={Jul}, pages={1949–1965} }
 @article{maes_amarasinghe_ayllon_basler_bavari_blasdell_briese_brown_bukreyev_balkema-buschmann_et al._2019, title={Taxonomy of the order Mononegavirales: second update 2018}, volume={164}, ISSN={["1432-8798"]}, DOI={10.1007/s00705-018-04126-4}, abstractNote={In October 2018, the order Mononegavirales was amended by the establishment of three new families and three new genera, abolishment of two genera, and creation of 28 novel species. This article presents the updated taxonomy of the order Mononegavirales as now accepted by the International Committee on Taxonomy of Viruses (ICTV).}, number={4}, journal={ARCHIVES OF VIROLOGY}, author={Maes, Piet and Amarasinghe, Gaya K. and Ayllon, Maria A. and Basler, Christopher F. and Bavari, Sina and Blasdell, Kim R. and Briese, Thomas and Brown, Paul A. and Bukreyev, Alexander and Balkema-Buschmann, Anne and et al.}, year={2019}, month={Apr}, pages={1233–1244} }
 @article{amarasinghe_ayllon_bao_basler_bavari_blasdell_briese_brown_bukreyev_balkema-buschmann_et al._2019, title={Taxonomy of the order Mononegavirales: update 2019}, volume={164}, ISSN={["1432-8798"]}, DOI={10.1007/s00705-019-04247-4}, abstractNote={In February 2019, following the annual taxon ratification vote, the order Mononegavirales was amended by the addition of four new subfamilies and 12 new genera and the creation of 28 novel species. This article presents the updated taxonomy of the order Mononegavirales as now accepted by the International Committee on Taxonomy of Viruses (ICTV).}, number={7}, journal={ARCHIVES OF VIROLOGY}, author={Amarasinghe, Gaya K. and Ayllon, Maria A. and Bao, Yiming and Basler, Christopher F. and Bavari, Sina and Blasdell, Kim R. and Briese, Thomas and Brown, Paul A. and Bukreyev, Alexander and Balkema-Buschmann, Anne and et al.}, year={2019}, month={Jul}, pages={1967–1980} }
 @article{martin_whitfield_2018, title={Cellular localization and interactions of nucleorhabdovirus proteins are conserved between insect and plant cells}, volume={523}, ISSN={["0042-6822"]}, DOI={10.1016/j.virol.2018.06.019}, abstractNote={Maize mosaic virus (MMV), similar to other nucleorhabdoviruses, replicates in divergent hosts: plants and insects. To compare MMV protein localization and interactions, we visualized autofluorescent protein fusions in both cell types. Nucleoprotein (N) and glycoprotein (G) localized to the nucleus and cytoplasm, phosphoprotein (P) was only found in the nucleus, and 3 (movement) and matrix (M) were present in the cytoplasm. This localization pattern is consistent with the model of nucleorhabdoviral replication of N, P, L and viral RNA forming a complex in the nucleus and the subvirion associating with M and then G during budding into perinuclear space. The comparable localization patterns in both organisms indicates a similar replication cycle. Changes in localization when proteins were co-expressed suggested viral proteins interact thus altering organelle targeting. We documented a limited number of direct protein interactions indicating host factors play a role in the virus protein interactions during the infection cycle.}, journal={VIROLOGY}, author={Martin, Kathleen M. and Whitfield, Anna E.}, year={2018}, month={Oct}, pages={6–14} }
 @article{rotenberg_whitfield_2018, title={Molecular interactions between tospoviruses and thrips vectors}, volume={33}, ISSN={["1879-6265"]}, DOI={10.1016/j.coviro.2018.11.007}, abstractNote={Thrips-transmitted tospoviruses are an emerging and re-emerging threat to crop production worldwide. Tospoviruses are transstadially transmitted from larval to pupal stages of development, with adults serving as the primary inoculators of plants. A unique feature of the transmission cycle is that adults-while they can acquire virus from plants directly-are competent as vectors only if they acquire virus as larvae. Thrips vectors also serve as hosts for the virus, supporting its replication in midgut tissues and salivary glands. There is a tight link between thrips development and virus dissemination in the insect, and recent transcriptome studies point to stage-specific responses that coincide with localization of the virus in the insect body. Transcriptome sequencing of thrips vectors is leading to identification of virus-responsive thrips genes and possibly new targets to disrupt the virus transmission cycle. Accumulation of thrips-omics resources and advancements in functional biology tools will propel new and exciting molecular studies of thrips-tospoviruses interactions.}, journal={CURRENT OPINION IN VIROLOGY}, author={Rotenberg, Dorith and Whitfield, Anna E.}, year={2018}, month={Dec}, pages={191–197} }
 @article{whitfield_huot_martin_kondo_dietzgen_2018, title={Plant rhabdoviruses-their origins and vector interactions}, volume={33}, ISSN={["1879-6265"]}, DOI={10.1016/j.coviro.2018.11.002}, abstractNote={Classical plant rhabdoviruses infect monocot and dicot plants, have unsegmented negative-sense RNA genomes and have been taxonomically classified in the genera Cytorhabdovirus and Nucleorhabdovirus. These viruses replicate in their hemipteran vectors and are transmitted in a circulative-propagative mode and virus infection persists for the life of the insect. Based on the discovery of numerous novel rhabdoviruses in arthropods during metagenomic studies and extensive phylogenetic analyses of the family Rhabdoviridae, it is hypothesized that plant-infecting rhabdoviruses are derived from insect viruses. Analyses of viral gene function in plants and insects is beginning to reveal conserved and unique biology for these plant viruses in the two diverse hosts. New tools for insect molecular biology and infectious clones for plant rhabdoviruses are increasing our understanding of the lifestyles of these viruses.}, journal={CURRENT OPINION IN VIROLOGY}, author={Whitfield, Anna E. and Huot, Ordom Brian and Martin, Kathleen M. and Kondo, Hideki and Dietzgen, Ralf G.}, year={2018}, month={Dec}, pages={198–207} }
 @article{whitfield_dietzgen_2018, title={Plant virus-vector interactions}, volume={33}, ISSN={["1879-6265"]}, DOI={10.1016/j.coviro.2018.11.008}, journal={CURRENT OPINION IN VIROLOGY}, author={Whitfield, Anna E. and Dietzgen, Ralf G.}, year={2018}, month={Dec}, pages={III-V} }
 @article{amarasinghe_aréchiga ceballos_banyard_basler_bavari_bennett_blasdell_briese_bukreyev_caì_et al._2018, title={Taxonomy of the order Mononegavirales: update 2018}, volume={163}, ISSN={0304-8608 1432-8798}, url={http://dx.doi.org/10.1007/S00705-018-3814-X}, DOI={10.1007/S00705-018-3814-X}, abstractNote={In 2018, the order Mononegavirales was expanded by inclusion of 1 new genus and 12 novel species. This article presents the updated taxonomy of the order Mononegavirales as now accepted by the International Committee on Taxonomy of Viruses (ICTV) and summarizes additional taxonomic proposals that may affect the order in the near future.}, number={8}, journal={Archives of Virology}, publisher={Springer Science and Business Media LLC}, author={Amarasinghe, Gaya K. and Aréchiga Ceballos, Nidia G. and Banyard, Ashley C. and Basler, Christopher F. and Bavari, Sina and Bennett, Andrew J. and Blasdell, Kim R. and Briese, Thomas and Bukreyev, Alexander and Caì, Yíngyún and et al.}, year={2018}, month={Apr}, pages={2283–2294} }
 @article{amarasinghe_bào_basler_bavari_beer_bejerman_blasdell_bochnowski_briese_bukreyev_et al._2017, title={Taxonomy of the order Mononegavirales: update 2017}, volume={162}, ISSN={0304-8608 1432-8798}, url={http://dx.doi.org/10.1007/S00705-017-3311-7}, DOI={10.1007/S00705-017-3311-7}, abstractNote={In 2017, the order Mononegavirales was expanded by the inclusion of a total of 69 novel species. Five new rhabdovirus genera and one new nyamivirus genus were established to harbor 41 of these species, whereas the remaining new species were assigned to already established genera. Furthermore, non-Latinized binomial species names replaced all paramyxovirus and pneumovirus species names, thereby accomplishing application of binomial species names throughout the entire order. This article presents the updated taxonomy of the order Mononegavirales as now accepted by the International Committee on Taxonomy of Viruses (ICTV).}, number={8}, journal={Archives of Virology}, publisher={Springer Science and Business Media LLC}, author={Amarasinghe, Gaya K. and Bào, Yīmíng and Basler, Christopher F. and Bavari, Sina and Beer, Martin and Bejerman, Nicolás and Blasdell, Kim R. and Bochnowski, Alisa and Briese, Thomas and Bukreyev, Alexander and et al.}, year={2017}, month={Apr}, pages={2493–2504} }
 @article{martin_barandoc-alviar_schneweis_stewart_rotenberg_whitfield_2017, title={Transcriptomic response of the insect vector, Peregrinus maidis, to Maize mosaic rhabdovirus and identification of conserved responses to propagative viruses in hopper vectors}, volume={509}, ISSN={0042-6822}, url={http://dx.doi.org/10.1016/j.virol.2017.05.019}, DOI={10.1016/j.virol.2017.05.019}, abstractNote={Maize mosaic virus (MMV) is a plant-pathogenic rhabdovirus that is transmitted by the corn planthopper, Peregrinus maidis, in a propagative manner. P. maidis supports long-term MMV infections with no negative effects on insect performance. To elucidate whole-body transcriptome responses to virus infection, RNA-Seq was used to examine differential gene expression of virus-infected adult insects, and libraries were prepared from replicated groups of virus-exposed insects and non-exposed insects. From the 68,003 de novo-assembled transcripts, 144 were differentially-expressed (DE) during viral infection with comparable numbers up- and down-regulated. DE transcripts with similarity to genes associated with transposable elements (i.e., RNA-directed DNA polymerases) were enriched and may represent a mechanisim for modulating virus infection. Comparison of the P. maidis DE transcripts to published propagative virus-responsive transcript databases for two other hopper vectors revealed that 16% of the DE transcripts were shared across the three systems and may represent conserved responses to propagative viruses.}, journal={Virology}, publisher={Elsevier BV}, author={Martin, Kathleen M. and Barandoc-Alviar, Karen and Schneweis, Derek J. and Stewart, Catherine L. and Rotenberg, Dorith and Whitfield, Anna E.}, year={2017}, month={Sep}, pages={71–81} }
 @article{montero-astúa_ullman_whitfield_2016, title={Salivary gland morphology, tissue tropism and the progression of tospovirus infection in Frankliniella occidentalis}, volume={493}, ISSN={0042-6822}, url={http://dx.doi.org/10.1016/J.VIROL.2016.03.003}, DOI={10.1016/J.VIROL.2016.03.003}, abstractNote={Tomato spotted wilt virus (TSWV) is transmitted by thrips in a propagative manner; however, progression of virus infection in the insect is not fully understood. The goal of this work was to study the morphology and infection of thrips salivary glands. The primary salivary glands (PSG) are complex, with three distinct regions that may have unique functions. Analysis of TSWV progression in thrips revealed the presence of viral proteins in the foregut, midgut, ligaments, tubular salivary glands (TSG), and efferent duct and filament structures connecting the TSG and PSG of first and second instar larvae. The primary site of virus infection shifted from the midgut and TSG in the larvae to the PSG in adults, suggesting that tissue tropism changes with insect development. TSG infection was detected in advance of PSG infection. These findings support the hypothesis that the TSG are involved in trafficking of TSWV to the PSG.}, journal={Virology}, publisher={Elsevier BV}, author={Montero-Astúa, Mauricio and Ullman, Diane E. and Whitfield, Anna E.}, year={2016}, month={Jun}, pages={39–51} }
 @article{afonso_amarasinghe_bányai_bào_basler_bavari_bejerman_blasdell_briand_briese_et al._2016, title={Taxonomy of the order Mononegavirales: update 2016}, volume={161}, ISSN={0304-8608 1432-8798}, url={http://dx.doi.org/10.1007/S00705-016-2880-1}, DOI={10.1007/S00705-016-2880-1}, abstractNote={In 2016, the order Mononegavirales was emended through the addition of two new families (Mymonaviridae and Sunviridae), the elevation of the paramyxoviral subfamily Pneumovirinae to family status (Pneumoviridae), the addition of five free-floating genera (Anphevirus, Arlivirus, Chengtivirus, Crustavirus, and Wastrivirus), and several other changes at the genus and species levels. This article presents the updated taxonomy of the order Mononegavirales as now accepted by the International Committee on Taxonomy of Viruses (ICTV).}, number={8}, journal={Archives of Virology}, publisher={Springer Science and Business Media LLC}, author={Afonso, Claudio L. and Amarasinghe, Gaya K. and Bányai, Krisztián and Bào, Yīmíng and Basler, Christopher F. and Bavari, Sina and Bejerman, Nicolás and Blasdell, Kim R. and Briand, François-Xavier and Briese, Thomas and et al.}, year={2016}, month={May}, pages={2351–2360} }
 @article{schneweis_whitfield_rotenberg_2017, title={Thrips developmental stage-specific transcriptome response to tomato spotted wilt virus during the virus infection cycle in Frankliniella occidentalis, the primary vector}, volume={500}, ISSN={0042-6822}, url={http://dx.doi.org/10.1016/J.VIROL.2016.10.009}, DOI={10.1016/J.VIROL.2016.10.009}, abstractNote={Tomato spotted wilt virus (TSWV) is transmitted by Frankliniella occidentalis in a circulative-propagative manner. Little is known about thrips vector response to TSWV during the infection process from larval acquisition to adult inoculation of plants. Whole-body transcriptome response to virus infection was determined for first-instar larval, pre-pupal and adult thrips using RNA-Seq. TSWV responsive genes were identified using preliminary sequence of a draft genome of F. occidentalis as a reference and three developmental-stage transcriptomes were assembled. Processes and functions associated with host defense, insect cuticle structure and development, metabolism and transport were perturbed by TSWV infection as inferred by ontologies of responsive genes. The repertoire of genes responsive to TSWV varied between developmental stages, possibly reflecting the link between thrips development and the virus dissemination route in the vector. This study provides the foundation for exploration of tissue-specific expression in response to TSWV and functional analysis of thrips gene function.}, journal={Virology}, publisher={Elsevier BV}, author={Schneweis, Derek J. and Whitfield, Anna E. and Rotenberg, Dorith}, year={2017}, month={Jan}, pages={226–237} }
 @article{whitfield_rotenberg_2015, title={Disruption of insect transmission of plant viruses}, volume={8}, ISSN={2214-5745}, url={http://dx.doi.org/10.1016/J.COIS.2015.01.009}, DOI={10.1016/J.COIS.2015.01.009}, abstractNote={Plant-infecting viruses are transmitted by a diverse array of organisms including insects, mites, nematodes, fungi, and plasmodiophorids. Virus interactions with these vectors are diverse, but there are some commonalities. Generally the infection cycle begins with the vector encountering the virus in the plant and the virus is acquired by the vector. The virus must then persist in or on the vector long enough for the virus to be transported to a new host and delivered into the plant cell. Plant viruses rely on their vectors for breaching the plant cell wall to be delivered directly into the cytosol. In most cases, viral capsid or membrane glycoproteins are the specific viral proteins that are required for transmission and determinants of vector specificity. Specific molecules in vectors also interact with the virus and while there are few-identified to no-identified receptors, candidate recognition molecules are being further explored in these systems. Due to the specificity of virus transmission by vectors, there are defined steps that represent good targets for interdiction strategies to disrupt the disease cycle. This review focuses on new technologies that aim to disrupt the virus–vector interaction and focuses on a few of the well-characterized virus–vector interactions in the field. In closing, we discuss the importance of integration of these technologies with current methods for plant virus disease control.}, journal={Current Opinion in Insect Science}, publisher={Elsevier BV}, author={Whitfield, Anna E and Rotenberg, Dorith}, year={2015}, month={Apr}, pages={79–87} }
 @article{whitfield_falk_rotenberg_2015, title={Insect vector-mediated transmission of plant viruses}, volume={479-480}, ISSN={0042-6822}, url={http://dx.doi.org/10.1016/J.VIROL.2015.03.026}, DOI={10.1016/J.VIROL.2015.03.026}, abstractNote={The majority of plant-infecting viruses are transmitted to their host plants by vectors. The interactions between viruses and vector vary in duration and specificity but some common themes in vector transmission have emerged: 1) plant viruses encode structural proteins on the surface of the virion that are essential for transmission, and in some cases additional non-structural helper proteins that act to bridge the virion to the vector binding site; 2) viruses bind to specific sites in or on vectors and are retained there until they are transmitted to their plant hosts; and 3) viral determinants of vector transmission are promising candidates for translational research aimed at disrupting transmission or decreasing vector populations. In this review, we focus on well-characterized insect vector-transmitted viruses in the following genera: Caulimovirus, Crinivirus, Luteovirus, Geminiviridae, Reovirus, Tospovirus, and Tenuivirus. New discoveries regarding these genera have increased our understanding of the basic mechanisms of virus transmission by arthropods, which in turn have enabled the development of innovative strategies for breaking the transmission cycle.}, journal={Virology}, publisher={Elsevier BV}, author={Whitfield, Anna E. and Falk, Bryce W. and Rotenberg, Dorith}, year={2015}, month={May}, pages={278–289} }
 @article{badillo-vargas_rotenberg_schneweis_whitfield_2015, title={RNA interference tools for the western flower thrips, Frankliniella occidentalis}, volume={76}, ISSN={0022-1910}, url={http://dx.doi.org/10.1016/J.JINSPHYS.2015.03.009}, DOI={10.1016/J.JINSPHYS.2015.03.009}, abstractNote={The insect order Thysanoptera is exclusively comprised of small insects commonly known as thrips. The western flower thrips, Frankliniella occidentalis, is an economically important pest amongst thysanopterans due to extensive feeding damage and tospovirus transmission to hundreds of plant species worldwide. Geographically-distinct populations of F. occidentalis have developed resistance against many types of traditional chemical insecticides, and as such, management of thrips and tospoviruses are a persistent challenge in agriculture. Molecular methods for defining the role(s) of specific genes in thrips–tospovirus interactions and for assessing their potential as gene targets in thrips management strategies is currently lacking. The goal of this work was to develop an RNA interference (RNAi) tool that enables functional genomic assays and to evaluate RNAi for its potential as a biologically-based approach for controlling F. occidentalis. Using a microinjection system, we delivered double-stranded RNA (dsRNA) directly to the hemocoel of female thrips to target the vacuolar ATP synthase subunit B (V-ATPase-B) gene of F. occidentalis. Gene expression analysis using real-time quantitative reverse transcriptase-PCR (qRT-PCR) revealed significant reductions of V-ATPase-B transcripts at 2 and 3 days post-injection (dpi) with dsRNA of V-ATPase-B compared to injection with dsRNA of GFP. Furthermore, the effect of knockdown of the V-ATPase-B gene in females at these two time points was mirrored by the decreased abundance of V-ATPase-B protein as determined by quantitative analysis of Western blots. Reduction in V-ATPase-B expression in thrips resulted in increased female mortality and reduced fertility, i.e., number of viable offspring produced. Survivorship decreased significantly by six dpi compared to the dsRNA-GFP control group, which continued decreasing significantly until the end of the bioassay. Surviving female thrips injected with dsRNA-V-ATPase-B produced significantly fewer offspring compared to those in the dsRNA-GFP control group. Our findings indicate that an RNAi-based strategy to study gene function in thrips is feasible, can result in quantifiable phenotypes, and provides a much-needed tool for investigating the molecular mechanisms of thrips–tospovirus interactions. To our knowledge, this represents the first report of RNAi for any member of the insect order Thysanoptera and demonstrates the potential for translational research in the area of thrips pest control.}, journal={Journal of Insect Physiology}, publisher={Elsevier BV}, author={Badillo-Vargas, Ismael E. and Rotenberg, Dorith and Schneweis, Brandi A. and Whitfield, Anna E.}, year={2015}, month={May}, pages={36–46} }
 @article{rotenberg_jacobson_schneweis_whitfield_2015, title={Thrips transmission of tospoviruses}, volume={15}, ISSN={1879-6257}, url={http://dx.doi.org/10.1016/J.COVIRO.2015.08.003}, DOI={10.1016/J.COVIRO.2015.08.003}, abstractNote={One hundred years ago, the disease tomato spotted wilt was first described in Australia. Since that time, knowledge of this disease caused by Tomato spotted wilt virus (TSWV) and transmitted by thrips (insects in the order Thysanoptera) has revealed a complex relationship between the virus, vector, plant host, and environment. Numerous tospoviruses and thrips vectors have been described, revealing diversity in plant host range and geographical distributions. Advances in characterization of the tripartite interaction between the virus, vector, and plant host have provided insight into molecular and ecological relationships. Comparison to animal-infecting viruses in the family Bunyaviridae has enabled the identification of commonalities between tospoviruses and other bunyaviruses in transmission by arthropod vectors and molecular interactions with hosts. This review provides a special emphasis on TSWV and Frankliniella occidentalis, the model tospovirus–thrips pathosystem. However, other virus–vector combinations are also of importance and where possible, comparisons are made between different viruses and thrips vectors.}, journal={Current Opinion in Virology}, publisher={Elsevier BV}, author={Rotenberg, Dorith and Jacobson, Alana L and Schneweis, Derek J and Whitfield, Anna E}, year={2015}, month={Dec}, pages={80–89} }
 @article{alexander_mauck_whitfield_garrett_malmstrom_2013, title={Plant-virus interactions and the agro-ecological interface}, volume={138}, ISSN={0929-1873 1573-8469}, url={http://dx.doi.org/10.1007/S10658-013-0317-1}, DOI={10.1007/S10658-013-0317-1}, number={3}, journal={European Journal of Plant Pathology}, publisher={Springer Science and Business Media LLC}, author={Alexander, H. M. and Mauck, K. E. and Whitfield, A. E. and Garrett, K. A. and Malmstrom, C. M.}, year={2013}, month={Nov}, pages={529–547} }
 @article{elbeaino_whitfield_sharma_digiaro_2013, title={Emaravirus-specific degenerate PCR primers allowed the identification of partial RNA-dependent RNA polymerase sequences of Maize red stripe virus and Pigeonpea sterility mosaic virus}, volume={188}, ISSN={0166-0934}, url={http://dx.doi.org/10.1016/j.jviromet.2012.11.037}, DOI={10.1016/j.jviromet.2012.11.037}, abstractNote={Emaravirus is a recently established viral genus that includes two approved virus species: European mountain ash ringspot-associated virus (EMARaV) and Fig mosaic virus (FMV). Other described but unclassified viruses appear to share biological characteristics similar to emaraviruses, including segmented, negative-single stranded RNA genomes with enveloped virions approximately 80-200nm in diameter. Sequence analysis of emaravirus genomes revealed the presence of conserved amino acid sequences in the RNA-dependent RNA polymerase gene (RdRp) denoted as pre-motif A, motifs A and C. Degenerate oligonucleotide primers were developed to these conserved sequences and were shown to amplify in reverse transcription-polymerase chain reaction assay (RT-PCR) DNA fragments of 276bp and 360bp in size. These primers efficiently detected emaraviruses with known sequences available in the database (FMV and EMARaV); they also detected viruses with limited sequence information such as Pigeonpea sterility mosaic virus (PPSMV) and Maize red stripe virus (MRSV). The degenerate primers designed on pre-motif A and motif A sequences successfully amplified the four species used as positive controls (276bp), whereas those of motifs A and C failed to detect only MRSV. The amino acid sequences obtained from PPSMV and MRSV shared the highest identity with those of two other tentative species of the Emaravirus genus, Rose rosette virus (RRV) (69%) and Redbud yellow ringspot virus (RYRV) (60%), respectively. The phylogenetic tree constructed with 92 amino acid-long portions of polypeptide putatively encoded by RNA1 of definitive and tentative emaravirus species clustered PPSMV and MRSV in two separate clades close to RRV and Raspberry leaf blotch virus (RLBV), respectively. The newly developed degenerate primers have proved their efficacy in amplifying new emaravirus-specific sequences; accordingly, they could be useful in identifying new emaravirus-like species in nature.}, number={1-2}, journal={Journal of Virological Methods}, publisher={Elsevier BV}, author={Elbeaino, Toufic and Whitfield, Anna and Sharma, Mamta and Digiaro, Michele}, year={2013}, month={Mar}, pages={37–40} }
 @article{rotenberg_wells_chapman_whitfield_goodman_cooperband_2007, title={Soil properties associated with organic matter-mediated suppression of bean root rot in field soil amended with fresh and composted paper mill residuals}, volume={39}, ISSN={0038-0717}, url={http://dx.doi.org/10.1016/j.soilbio.2007.06.011}, DOI={10.1016/j.soilbio.2007.06.011}, abstractNote={The ability of an organic amendment to suppress soil-borne disease is mediated by the complex interactions between biotic and abiotic soil factors. Various microbiological and physicochemical soil properties were measured in field soils with histories of receiving 4 or 5 years of spring additions of paper mill residuals (PMR), PMR composted alone (PMRC), PMR composted with bark (PMRB), or no amendment under a conventionally managed vegetable crop rotation. The objectives of this study were to (i) determine the residual and re-amendment effects of the organic materials on root rot disease severity; (ii) determine the influence of amendment type on the structure of bacterial communities associated with snap bean roots grown in these soils; and (iii) quantify the relative contributions of microbiological and physicochemical properties to root rot suppression in the field and greenhouse. While all amendment types significantly suppressed root rot disease compared to non-amended soils in both environments, only soils amended with PMR or PMRB sustained suppressive conditions 1 year after the most recent amendment event. Disease severity was inversely related to microbial activity (fluorescein diacetate assay) in recently amended soils only. Terminal restriction fragment length polymorphism (T-RFLP) analysis of the 16s rRNA gene was performed to obtain bacterial profiles. Principal component analysis (PCA) of terminal restriction fragments (TRFs) revealed general differences in bacterial community composition (PC1) among amendment types, and specific TRFs contributed to these differences. Correlation and multiple regression analyses of the measured soil variables revealed that the composition of root-associated bacterial communities and the amount of particulate organic matter—carbon in bulk soils imparted independent and relatively equal contributions to the variation in disease severity documented in the field and greenhouse. Together, our findings provide evidence that disease suppression induced by annual PMR inputs was mediated by their differential effects on bacterial communities and the amount and quality of organic matter in these soils.}, number={11}, journal={Soil Biology and Biochemistry}, publisher={Elsevier BV}, author={Rotenberg, Dorith and Wells, Ana Jiménez and Chapman, Elisabeth J. and Whitfield, Anna E. and Goodman, Robert M. and Cooperband, Leslie R.}, year={2007}, month={Nov}, pages={2936–2948} }
 @article{ullman_whitfield_german_2005, title={Thrips and tospoviruses come of age: Mapping determinants of insect transmission}, volume={102}, ISSN={0027-8424 1091-6490}, url={http://dx.doi.org/10.1073/PNAS.0501341102}, DOI={10.1073/PNAS.0501341102}, abstractNote={Insect vectors play a key role in dissemination of viruses that cause important diseases in humans, animals, and plants. Specific understanding of insect–virus interactions leading to successful transmission is a central problem in vector biology and critical to developing effective control strategies. In this issue of PNAS, Sin et al. (1) provide evidence that the genetic determinants of insect transmissibility for tomato spotted wilt virus (TSWV) reside on the middle RNA (M RNA) segment encoding the viral membrane glycoproteins (GPs) [N-terminal GP (GN) and C-terminal GP(GC)]. This virus is transmitted between plants by insects called thrips (Thripidae, Thysanoptera) and is the type member of the genus Tospovirus. The tospoviruses are the only plant-infecting members in the family Bunyaviridae , which consists of many viruses that cause diseases in animals and humans (2). Thus, tospoviruses and their thrips vectors are ideal model systems for elucidating processes of virus infection in disparate hosts that can be extended to viruses of importance to human health.

The complex nature of the interplay between thrips, tospoviruses, and their shared plant hosts was first recognized with the discovery that TSWV multiplies in its insect vectors (3, 4). This discovery opened rich and exciting avenues of exploration, including understanding biological and molecular interactions underlying TSWV pathogenesis in plant and insect hosts and the role these processes play in virus evolution. Findings of the last decade show that insect inoculation of tospoviruses into a plant host cannot occur without viral passage across at least three insect organs (the midgut, visceral muscle cells, and salivary glands) that include six membrane barriers (2). Previous hypotheses that tospovirus GPs are essential determinants of thrips acquisition were based on several pieces of experimental evidence: ( i ) assembly-deficient TSWV isolates could be passed mechanically between plants but were not insect transmissible …}, number={14}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Ullman, D. E. and Whitfield, A. E. and German, T. L.}, year={2005}, month={Mar}, pages={4931–4932} }
 @article{whitfield_ullman_german_2005, title={Tomato spotted wilt virus glycoprotein GC is cleaved at acidic pH}, volume={110}, ISSN={0168-1702}, url={http://dx.doi.org/10.1016/j.virusres.2005.01.007}, DOI={10.1016/j.virusres.2005.01.007}, abstractNote={Tomato spotted wilt virus (TSWV) is a plant-infecting member of the family Bunyaviridae. TSWV encodes two envelope glycoproteins, GN and GC, which are required for virus infection of the arthropod vector. Other members of the Bunyaviridae enter host cells by pH-dependent endocytosis. During this process, the glycoproteins are exposed to conditions of acidic pH within endocytic vesicles causing the GC protein to change conformation. This conformational change renders GC more sensitive to protease cleavage. We subjected TSWV virions to varying pH conditions and determined that TSWV GC, but not GN, was cleaved under acidic pH conditions, and that this phenomenon did not occur at neutral or alkaline pH. This data provides evidence that GC changes conformation at low pH which results in altered protease sensitivity. Furthermore, sequence analysis of GC predicts the presence of internal hydrophobic domains, regions that are characteristic of fusion proteins. Like studies with other members of the Bunyaviridae, this study is the first step towards characterizing the nature of cell entry by TSWV.}, number={1-2}, journal={Virus Research}, publisher={Elsevier BV}, author={Whitfield, Anna E. and Ullman, Diane E. and German, Thomas L.}, year={2005}, month={Jun}, pages={183–186} }