Structural and functional insights into the Diabrotica virgifera virgifera ATP-binding cassette transporter gene family

- virgifera genome, we developed transcriptomic resources to use in identifying candidate genes likely to be involved in the evolution of resistance, starting with members of the ATP-binding cassette (ABC) transporter family..
- Phylogenetic analysis placed the deduced amino-acid sequences of the DvvABC transporters into eight subfamilies (A to H).
- Moreover, we have identified 65 members of the ABC transporter family and provided the first insights into the developmental and physiological roles of ABC transporters in this pest species..
- Keywords: ATP-binding cassette (ABC) transporter, Phylogenetic, Transcriptome, RNA interference (RNAi), Corn rootworm.
- virgifera in the Midwest United States circumvent the cultural-control practice of corn-soybean rotation [3, 5–7].
- 2019 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0.
- 1 Department of Entomology and Plant Pathology, North Carolina State University, Box Thomas Hall, Raleigh, NC USA Full list of author information is available at the end of the article.
- ATP-binding cassette (ABC) proteins comprise one of the largest gene families, and are found across prokary- otic and eukaryotic domains [24].
- Due to the relatively conserved sequence of the NBD, it has been used for the phylogenetic classification of the ABC transporter superfamily into eight subfamilies designated A to H (ABCA to ABCH) [29]..
- Changes in the expression level or structure of some ABCA, ABCC and ABCG subfamily members have been associated with Bt toxin resistance in species of Lepidoptera [37], while paralogs of an ABCB transporter were linked to Bt Cry3Aa resistance in the coleopteran species, Chrysomela tremula [38], and were found to be in proximity to a quantitative trait locus (QTL) for Cry3Bb1 resistance in D.
- Similar investigations of ABC transporters in D.
- Complicating the development of genomic tools is the 2.58 GB size and complex repeti- tive structure of the D.
- In the following, a de novo transcriptome as- sembly approach was used for the first prediction, annotation, and functional analysis of the ABC transporter gene family in D.
- Specifically, eight ABC trans- porters were identified as putative orthologs to those pre- viously reported to have a defining RNAi phenotype in the model coleopteran species, T.
- castaneum was the most frequent, representing 65% of the matches (Additional file 3: Figure S2).
- Among ontol- ogies assigned via mapping at GO level 2, a majority of the associated terms were assigned to cell struc- tural component, metabolic process, and catalytic ac- tivity respectively for GO Cellular Component, Biological Process and Molecular Function (Fig.
- The DNASTAR assembly showed a high degree of completeness based on a BUSCO score of 928, or 89.6%, of the 1066 genes in the arthropod reference set (v.
- Bioinformatic analysis of the D.
- virgifera ABC transporter family.
- castaneum orthologs from ABC transporter subfamilies A through H (Table 2.
- castaneum ABC transporters.
- The DvvABCH subfamily contained four members, which were similar to those of the ABCG sub- family in being HTs with a reverse domain organization..
- virgifera ABC transporter protein.
- 1 Gene ontology classification of the D.
- We were unable to identify a direct ortholog for these genes, but the closest homolog we found in the D.
- Finally, while the ABCG genes TcABCG-9A and TcABCG- 9B represent the orthologs of the T.
- virgifera ABC transcripts showed that all are expressed across all of the developmental stages examined (Fig.
- Moreover, injection of each of the eight dsRNAs resulted in defined phenotypes among dsRNA treated co- horts (Table 3.
- virgifera ABC transporters.
- Table 3 Results of RNAi knockdown of selected ABC transporters.
- Injection of dsRNA targeting DvvABCG_3712, DvvABCG_14042, and Dvvw resulted in phenotypes similar to those seen with RNAi knockdown of the corresponding T.
- Indeed, expression of the P.
- RNA isolated from Eggs (E), Larvae (L), Pupae (P), and adult Males (M) and Females (F) for each of the eight genes.
- virgifera as a major pest of cultivated maize (see Introduction) and current fragmented state of the unpublished draft genome assembly of this species (GenBank accession PXMJ the Illumina-based transcriptome assemblies reported here represent a particularly valu- able genetic tool for gene discovery, characterization,.
- Regardless, BLASTx analyses of the 65 putative D.
- This is probably a consequence of the extensive publicly available genomic data for both T.
- Since subfamily A transporter members are implicated in mammals with lipid transport, which can impact cell physiology [67], it is conceivable that the effects of the knockdown of DvvABCA_50718, and of its homologs, TcABCA-9A/9B, in T.
- castaneum orthologs, RNAi knockdown of the DvvABCG_14042 homolog TcABCG- 8A in T.
- In other species, orthologs of DvvABCH_5118 are known to transport cuticular lipids that are deposited in the outer epicuticle layer to form a waterproof barrier [30, 62]..
- Among insects, some of the first ABCGs to be characterized were the pigment transporters (white, scarlet and brown) in D..
- Studies have revealed that some ABCG proteins perform other crucial physiological roles in the transport of lipids, sterols, and drugs [73].
- In the current study, RNAi-mediated knockdown of Dvvw resulted in a white-eyed phenotype consistent with prior observations in T.
- Our findings support a prediction that Dvvw is part of the ommochrome path- way, where it is likely acting within a heterodimeric com- plex to import ommochrome pigments into the pigment granules of the compound eye.
- However, RNAi-mediated knockdown of the correspond- ing gene, ABCG-9A (scarlet), in T.
- This finding was not surpris- ing, since a previous report of RNAi targeting vermilion, a pivotal gene in the ommochrome pathway, also generates a white-eyed phenotype in T.
- Unfortunately, our initial survey of the D.
- So, it will be interesting to investigate in future studies if pigmentation of the D.
- melanogaster a third ABCG transporter, brown, is required for wild-type pigmentation of the eye..
- Although an ortho- log of brown has been identified in the T.
- This study provides a relatively large transcriptomic re- source comprising genes expressed across several life stages of the arthropod pest species, D.
- Regardless, our work represents the initial description of the ABC transporter gene family in D.
- virgifera ABC sequences were then used as BLASTx queries of the non-redundant NCBI protein database using the web blast interface (https://blast.ncbi.nlm.nih.gov/Blast.cgi) to confirm their identity as insect ABC genes.
- The number and positions of transmem- brane domains were assessed via query of the NCBI Con- served Domain Database [87].
- virgifera ABC transporter contigs were generated (Additional file 7).
- The unrooted Maximum Likeli- hood phylogenetic trees were constructed in the MEGAX program using default parameters in all cat- egories except: LG model of amino-acid substitution with Gamma distributed substitution rates (based on Best Model determination within the MEGA program), Partial Deletion treatment of gaps/missing data, and 1000 bootstrap replicates [89].
- ABC transporter subfam- ilies were assigned to D.
- virgifera ABC transporter transcript-specific primer pairs (Additional file 9: Table S3).
- Each of the synthesized dsRNAs were purified using the MEGAclear Kit (Ambion) and concentration determined using a Nanodrop 1000 (Thermo Scientific, Waltham, MA, USA) using the single- stranded RNA setting..
- RNAi assays were conducted by injecting dsRNA corre- sponding to each of the 8 specific D.
- virgifera ABC transporters..
- The fund- ing bodies had no role in the design of the study, collection, analysis, or in- terpretation of data, or in writing the manuscript..
- Adaptation of the western corn rootworm to crop rotation: evolution of a new strain in response to a management practice.
- Corn-rootworm (Coleoptera.
- Role of a gamma-aminobutryic acid (GABA) receptor mutation in the evolution and spread of Diabrotica virgifera virgifera resistance to cyclodiene insecticides.
- Field-Evolved Resistance to Bt Maize by Western Corn Rootworm.
- Multiple assays indicate varying levels of cross resistance in Cry3Bb1-selected field populations of the Western corn rootworm to mCry3A, eCry3.1Ab, and Cry34/35Ab1.
- Evidence of resistance to Cry34/35Ab1 corn by Western corn rootworm (Coleoptera:.
- Chrysomelidae): root injury in the field and larval survival in plant-based bioassays.
- Minnesota field population of western corn rootworm (Coleoptera:.
- Arthropod genomics research in the United States Department of Agriculture-Agricultural Research Service: applications of RNA interference and CRISPR gene editing technologies in pest control..
- Validation of RNA interference in western corn rootworm Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) adults.
- RNAi as a management tool for the western corn rootworm, Diabrotica virgifera virgifera.
- ABC transporters - from microorganisms to man.
- ABC transporters: the power to change..
- Evolution of the ATP-binding cassette (ABC) transporter superfamily in vertebrates.
- Functional analysis of the ATP-binding cassette (ABC) transporter gene family of Tribolium castaneum.
- Mutational analysis of the traffic Atpase (ABC) transporters involved in uptake of eye pigment precursors in Drosophila melanogaster - implications for structure-function-relationships.
- The ABCs of Eye Color in Tribolium castaneum: Orthologs of the Drosophila white, scarlet, and brown Genes.
- Genome-wide identification and characterization of ATP-binding cassette transporters in the silkworm, Bombyx mori.
- Mutations in the white gene of Drosophila melanogaster affecting ABC transporters that determine eye colouration.
- Interaction of pesticides with p-glycoprotein and other ABC proteins: a survey of the possible importance to insecticide, herbicide and fungicide resistance.
- Bacillus thuringiensis toxin resistance mechanisms among Lepidoptera: progress on genomic approaches to uncover causal mutations in the European corn borer, Ostrinia nubilalis.
- Genetic Markers for Western Corn Rootworm Resistance to Bt Toxin.
- Molecular Evolution of Glycoside Hydrolase Genes in the Western Corn Rootworm (Diabrotica virgifera virgifera).
- Distribution of Genes and Repetitive Elements in the Diabrotica virgifera virgifera Genome Estimated Using BAC Sequencing.
- Genetic analysis of the xenobiotic resistance-associated ABC gene subfamilies of the Lepidoptera.
- Genome-wide analysis of ATP-binding cassette (ABC) transporters in the sweetpotato whitefly, Bemisia tabaci.
- Roles of ABC proteins in the mechanism and Management of Bt Resistance.
- An ABC Transporter Mutation Is Correlated with Insect Resistance to Bacillus thuringiensis Cry1Ac Toxin..
- Mis-splicing of the ABCC2 gene linked with Bt toxin resistance in Helicoverpa armigera.
- Single amino acid mutation in an ATP-binding cassette transporter gene causes resistance to Bt toxin Cry1Ab in the silkworm, Bombyx mori.
- Mutational disruption of the ABCC2 gene in fall armyworm, Spodoptera frugiperda, confers resistance to the Cry1Fa and Cry1A.105 insecticidal proteins.
- Down- regulation of a novel ABC transporter gene (Pxwhite) is associated with Cry1Ac resistance in the diamondback moth, Plutella xylostella (L.
- Genome of the small hive beetle (Aethina tumida, Coleoptera: Nitidulidae), a worldwide parasite of social bee colonies, provides insights into detoxification and herbivory.
- The ABC transporter gene family of Daphnia pulex.
- A burst of ABC genes in the genome of the polyphagous spider mite Tetranychus urticae.
- Ribosome recycling depends on a mechanistic link between the FeS cluster domain and a conformational switch of the twin-ATPase ABCE1.
- Subcellular localization of the first three enzymes of the ommochrome synthetic pathway in Drosophila melanogaster.
- Cloning and characterization of the Tribolium castaneum eye-color genes encoding tryptophan oxygenase and kynurenine 3-monooxygenase.
- Germline transformation of the western corn rootworm, Diabrotica virgifera virgifera

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