Dataset Information

Gene expression in Tribolium castaneum life stages: Identifying a species-specific target for pest control applications.

ABSTRACT: The red flour beetle, Tribolium castaneum, is a major agricultural pest of post-harvest products and stored grain. Control of T. castaneum in stored products and grain is primarily by fumigants and sprays, but insecticide resistance is a major problem, and new control strategies are needed. T. castaneum is a genetic model for coleopterans, and the reference genome can be used for discovery of candidate gene targets for molecular-based control, such as RNA interference. Gene targets need to be pest specific, and ideally, they are expressed at low levels for successful control. Therefore, we sequenced the transcriptome of four major life stages of T. castaneum, sorted data into groups based on high or low expression levels, and compared relative gene expression among all life stages. We narrowed our candidate gene list to a cuticle protein gene (CPG) for further analysis. We found that the CPG sequence was unique to T. castaneum and expressed only in the larval stage. RNA interference targeting CPG in newly-emerged larvae caused a significant (p < 0.05) decrease in CPG expression (1,491-fold) compared to control larvae and 64% mortality over 18 d. RNA-Seq of survivors after 18 d identified changes in the expression of other genes as well, including 52 long noncoding RNAs. Expression of three additional cuticle protein genes were increased and two chitinase genes were decreased in response to injection of CPG dsRNA. The data demonstrate that RNA-Seq can identify genes important for insect survival and thus may be used to develop novel biologically-based insect control products.

SUBMITTER: Perkin LC

PROVIDER: S-EPMC6535216 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Gene expression in Tribolium castaneum life stages: Identifying a species-specific target for pest control applications.

Perkin Lindsey C LC Oppert Brenda B

PeerJ 20190523

The red flour beetle, Tribolium castaneum, is a major agricultural pest of post-harvest products and stored grain. Control of T. castaneum in stored products and grain is primarily by fumigants and sprays, but insecticide resistance is a major problem, and new control strategies are needed. T. castaneum is a genetic model for coleopterans, and the reference genome can be used for discovery of candidate gene targets for molecular-based control, such as RNA interference. Gene ...[more]

PMID: 31198628

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Project description:In spite of great developments in the agricultural field and plant productivity in the last decades, the concern about the control of agricultural pests is still continuous. However, pest management is expected to have more effects from nanomaterials by providing innovative solutions. The current study confirms the biotransformation of copper nanoparticles (CuNPs) using a cell-free culture extract of metal copper-resistant bacteria Pseudomonas fluorescens MAL2, which was isolated from heavy metal-contaminated soils collected from Sharqia Governorate, Egypt. The local screened bacterial isolate, Pseudomonas fluorescens MAL2, is similar to Pseudomonas fluorescens DSM 12442T DSM. After optimization of growth conditions, F-Base medium was found to be the best medium and pH 7, temperature 35 °C, concentration of CuSO4·5H2O 300 ppm, 10 mL supernatant: 40 mL CuSO4·5H2O (300 ppm), and reaction time 90 min were recorded as the best growth conditions to the fabrication of CuNPs. The formed CuNPs were characterized using initially visual observation of the change in the color of the reaction mixture from blue color to the dark green as an indication of CuNPs biotransformation. Then, UV-Vis spectroscopy showed a maximum absorption at 610 nm under the optimum conditions performed. Transmission Electron Microscopy (TEM) revealed the formation of spherical aspect with size ranges from 10:70 nm; moreover, Energy Dispersive X-ray spectroscopy (EDX) indicated the presence of CuNPs and other elements. In addition, the presence of alcohols, phenols, alkenes, and amines is confirmed by Fourier-Transform Infrared spectroscopy (FTIR) spectroscopy analysis. Dynamic Light Scattering (DLS) supported that the Zeta-average size of nanoparticle was 48.07 with 0.227 PdI value. The Zeta potential showed -26.00mV with a single peak. The biosynthesized CuNPs (Bio CuNPs) showed toxicity against the stored grain pest (Tribolium castaneum), where LC50 value was 37 ppm after 5 days of treatment. However, the negligible effect was observed with chemical synthesis of CuNPs (Ch CuNPs) at the same concentration. The results suggest that Bio CuNPs could be used not only as a biocontrol agent, but also as an ecofriendly and inexpensive approach for controlling the stored grain pests.

Project description:BackgroundA complex life cycle, such as complete metamorphosis, is a key innovation that can promote diversification of species. The evolution of a morphologically distinct larval stage is thought to have enabled insects to occupy broader ecological niches and become the most diverse metazoan taxon, yet the extent to which larval and adult morphologies can evolve independently remains unknown. Perturbation of larval limb regeneration allows us to generate larval legs and antennae with altered limb morphologies, which may be used to explore the developmental continuity that might exist between larval and adult appendages. In this study, we determined the roles of several appendage patterning transcription factors, abrupt (ab), dachshund (dac), Distal-less (Dll), and spineless (ss), in the red flour beetle, Tribolium castaneum, during larval appendage regeneration. The functions of these genes in regenerating and non-regenerating limbs were compared using RNA interference.ResultsDuring limb regeneration, dac and ss were necessary to re-pattern the same larval structures as those patterned during embryogenesis. Removal of these two genes led to larval appendage patterning defects that were carried over to the adult legs. Surprisingly, even though maternal knockdown of ab had minimal effects on limb allocation and patterning in the embryo, it was necessary for blastema growth, an earlier phase of regeneration. Finally, knockdown of Dll prevented the blastema-like bumps from re-differentiating into appendages.ConclusionsOur results suggest that, similar to vertebrates, the re-patterning phase of Tribolium larval limb regeneration relies on the same genes that are used during embryonic limb patterning. Thus, the re-patterning phase of regeneration is likely to be regulated by taxon-specific patterning mechanisms. Furthermore, Ab and Dll appear to play important roles during blastema proliferation and re-differentiation, respectively. Finally, our results show that continuity exists between larval and adult limb patterning, and that larval and adult leg morphologies may be developmentally coupled. Thus, the evolution of imaginal discs may have been a key step towards completely removing any developmental constraints that existed between larval and adult phenotypes.

Dataset Information

Gene expression in Tribolium castaneum life stages: Identifying a species-specific target for pest control applications.

Publications

Gene expression in <i>Tribolium castaneum</i> life stages: Identifying a species-specific target for pest control applications.

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