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Project description:Female reproductive potential is crucial for a population to multiply. MicroRNAs (miRNAs) have been demonstrated to be critical regulators in the post-transcriptional regulation of numerous biological processes, including reproduction. However, these studies mainly focus on model species. The oriental fruit fly, Bactrocera dorsalis, is a highly invasive pest in East Asia and the Pacific. Thus, it is of great significance to identify miRNAs involved in reproduction in this notorious pest. In this study, high-throughput sequencing was carried out to identify miRNAs potentially involved in reproductive development in female B. dorsalis. Eight miRNA libraries were established in the ovary and fat body from four developmental stages [1 days (d), 5 d, 9 d, and 13 d post-eclosion]. 116 and 84 miRNAs were found to be differentially expressed in ovary and fat body, respectively, during sexual maturation of female adults. The Gene Ontology analysis of the target genes involved in oogenesis and lipid particle accounted for 33% and 15% respectively . Among these differentially expressed miRNAs, we found five highly expressed miRNAs that are shared in the ovary and fat body that comprised four (miR-305-3p, miR-317-3p, miR-2779, and miR-305) conserved and one novel (Bdo-pc-3p-116) miRNA. Quantitative polymerase chain reaction results revealed that Bdo-pc-3p-116 was found to be specifically expressed in the ovary and down-regulated during sexual maturation. Injection of agomir-pc-3p-116 resulted in arrested ovarian development, and reduced egg deposition and progeny viability in B. dorsalis. CYP6a14 and endophilin B were confirmed to be the targets of pc-3p-116 via dual-luciferase assay and expression profiling. RNAi depletion of CYP6a14 led to reproductive defects similar to those caused by the injection of pc-3p-116 agomir. Thus, pc-3p-116 might play a critical role in female reproduction by targeting CYP6a14 and endophilin B and serve as a monitor to control CYP6a14 and endophilin B1 mRNA levels. Our data will not only provide abundant miRNAs as well as target genes for molecular research in female reproduction but also promote environment-friendly control strategies for this agricultural pest.
2025-03-03 | GSE146379 | GEO
Project description:Female reproductive potential is crucial for a population to multiply. MicroRNAs (miRNAs) have been demonstrated to be critical regulators in the post-transcriptional regulation of numerous biological processes, including reproduction. However, these studies mainly focus on model species. The oriental fruit fly, Bactrocera dorsalis, is a highly invasive pest in East Asia and the Pacific. Thus, it is of great significance to identify miRNAs involved in reproduction in this notorious pest. In this study, high-throughput sequencing was carried out to identify miRNAs potentially involved in reproductive development in female B. dorsalis. Eight miRNA libraries were established in the ovary and fat body from four developmental stages [1 days (d), 5 d, 9 d, and 13 d post-eclosion]. 116 and 84 miRNAs were found to be differentially expressed in ovary and fat body, respectively, during sexual maturation of female adults. The Gene Ontology analysis of the target genes involved in oogenesis and lipid particle accounted for 33% and 15% respectively . Among these differentially expressed miRNAs, we found five highly expressed miRNAs that are shared in the ovary and fat body that comprised four (miR-305-3p, miR-317-3p, miR-2779, and miR-305) conserved and one novel (Bdo-pc-3p-116) miRNA. Quantitative polymerase chain reaction results revealed that Bdo-pc-3p-116 was found to be specifically expressed in the ovary and down-regulated during sexual maturation. Injection of agomir-pc-3p-116 resulted in arrested ovarian development, and reduced egg deposition and progeny viability in B. dorsalis. CYP6a14 and endophilin B were confirmed to be the targets of pc-3p-116 via dual-luciferase assay and expression profiling. RNAi depletion of CYP6a14 led to reproductive defects similar to those caused by the injection of pc-3p-116 agomir. Thus, pc-3p-116 might play a critical role in female reproduction by targeting CYP6a14 and endophilin B and serve as a monitor to control CYP6a14 and endophilin B1 mRNA levels. Our data will not only provide abundant miRNAs as well as target genes for molecular research in female reproduction but also promote environment-friendly control strategies for this agricultural pest.
2025-03-03 | GSE146375 | GEO
Project description:Comprehensive RNA sequencing was performed on a laboratory colony of B. dorsalis with a focus on attempting to capture as many genes in the sequencing from throughout the entire developmental life history. De novo assembly and analysis of the resulting sequence One sample each for the egg, larvae, pupae, adult male, adult female and mated female life stages was sequenced.
2013-04-24 | E-GEOD-46310 | biostudies-arrayexpress
Project description:BACKGROUND:The gut bacteria of tephritid fruit flies play prominent roles in nutrition, reproduction, maintenance and ecological adaptations of the host. Here, we adopted an approach based on direct observation of symbiotic or axenic flies feeding on dishes seeded with drops of full diet (containing all amino acids) or full diet supplemented with bacteria at similar concentrations to explore the effects of intestinal bacteria on foraging decision and fitness of Bactrocera dorsalis. RESULTS:The results show that intestinal probiotics elicit beneficial foraging decision and enhance the female reproduction fitness and survival of B. dorsalis (symbiotic and axenic), yet preferences for probiotic diets were significantly higher in axenic flies to which they responded faster compared to full diet. Moreover, females fed diet supplemented with Pantoea dispersa and Enterobacter cloacae laid more eggs but had shorter lifespan while female fed Enterococcus faecalis and Klebsiella oxytoca enriched diets lived longer but had lower fecundity compared to the positive control. Conversely, flies fed sugar diet (negative control) were not able to produce eggs, but lived longer than those from the positive control. CONCLUSIONS:These results suggest that intestinal bacteria can drive the foraging decision in a way which promotes the reproduction and survival of B. dorsalis. Our data highlight the potentials of gut bacterial isolates to control the foraging behavior of the fly and empower the sterile insect technique (SIT) program through the mass rearing.
| S-EPMC6805663 | biostudies-literature
Project description:The oriental fruit fly, Bactrocera dorsalis, is a destructive polyphagous pest that causes damage to various fruit crops, and their distribution is currently expanding worldwide. Temperature is an important abiotic factor that influences insect population dynamics and distribution by affecting their survival, development, and reproduction. We examined the fecundity, pre-oviposition and oviposition periods, and longevity of adult B. dorsalis at various constant temperatures ranging from13°C to 35°C. The longevity of female B. dorsalis ranged from 116.8 days (18.8°C) to 22.4 days (34.9°C), and the maximum fecundity per female was 1,684 eggs at 28.1°C. Females were only able to lay eggs at 16.7°C to 34.9°C, and both the pre-oviposition and oviposition periods were different depending on the temperature. We modeled female reproduction in two oviposition models (OMs): 1) the current model developed by Kim and Lee, an OM composed of a fecundity model, age-specific survival model, and age-specific cumulative oviposition rate model, and 2) a two-phase OM modified the logic structure of the current model by separating pre-oviposition, so that oviposition was estimated with the female in oviposition phase who had complete pre-oviposition phase. The results of the two-phase OM provided more realistic outputs at lower and higher temperatures than those of the current model. We discussed the usefulness of the two-phase OM for the reproduction of insects with long pre-oviposition periods.
| S-EPMC7363081 | biostudies-literature
Project description:Comprehensive RNA sequencing was performed on a laboratory colony of B. dorsalis with a focus on attempting to capture as many genes in the sequencing from throughout the entire developmental life history. De novo assembly and analysis of the resulting sequence
2013-04-24 | GSE46310 | GEO
Project description:Genetics-enhanced sterile insect techniques (SIT) are promising novel approaches to control Bactrocera dorsalis, the most destructive horticultural pest in East Asia and the Pacific region. To identify novel genetic agents to alter male fertility of B. dorsalis, previous studies investigated miRNA expression in testes of B. dorsalis. One miRNA, miR-8-3p was predicted to bind the 3'UTR of putative B. dorsalis mitoferrin (bmfrn). The ortholog of bmfrn in D. melanogaster is essential for male fertility. Here we show that bmfrn has all conserved amino acid residues of known mitoferrins and is most abundantly expressed in B. dorsalis testes, making miR-8-3p and mitoferrin candidates for genetics-enhanced SIT. Furthermore, using a dual-luciferase reporter system, we show in HeLa cells that miR-8-3p interacts with the 3'UTR of bmfrn. Dietary treatments of adult male flies with miR-8-3p mimic, antagomiR, or bmfrn dsRNA, altered mitoferrin expression in the testes and resulted in reduced male reproductive capacity due to reduced numbers and viability of spermatozoa. We show for the first time that a mitoferrin is regulated by a miRNA and we demonstrate miR-8-3p as well as bmfrn dsRNA to be promising novel agents that could be used for genetics-enhanced SIT.
| S-EPMC4773865 | biostudies-literature
Project description:BackgroundNitrogen is considered the most limiting nutrient element for herbivorous insects. To alleviate nitrogen limitation, insects have evolved various symbiotically mediated strategies that enable them to colonize nitrogen-poor habitats or exploit nitrogen-poor diets. In frugivorous tephritid larvae developing in fruit pulp under nitrogen stress, it remains largely unknown how nitrogen is obtained and larval development is completed.ResultsIn this study, we used metagenomics and metatranscriptomics sequencing technologies as well as in vitro verification tests to uncover the mechanism underlying the nitrogen exploitation in the larvae of Bactrocera dorsalis. Our results showed that nitrogenous waste recycling (NWR) could be successfully driven by symbiotic bacteria, including Enterobacterales, Lactobacillales, Orbales, Pseudomonadales, Flavobacteriales, and Bacteroidales. In this process, urea hydrolysis in the larval gut was mainly mediated by Morganella morganii and Klebsiella oxytoca. In addition, core bacteria mediated essential amino acid (arginine excluded) biosynthesis by ammonium assimilation and transamination.ConclusionsSymbiotic bacteria contribute to nitrogen transformation in the larvae of B. dorsalis in fruit pulp. Our findings suggest that the pattern of NWR is more likely to be applied by B. dorsalis, and M. morganii, K. oxytoca, and other urease-positive strains play vital roles in hydrolysing nitrogenous waste and providing metabolizable nitrogen for B. dorsalis.
| S-EPMC9476588 | biostudies-literature
Project description:BACKGROUND: Tephritid fruit flies in the genus Bactrocera are of major economic significance in agriculture causing considerable loss to the fruit and vegetable industry. Currently, there is no ideal control program. Molecular means is an effective method for pest control at present, but genomic or transcriptomic data for members of this genus remains limited. To facilitate molecular research into reproduction and development mechanisms, and finally effective control on these pests, an extensive transcriptome for the oriental fruit fly Bactrocera dorsalis was produced using the Roche 454-FLX platform. RESULTS: We obtained over 350 million bases of cDNA derived from the whole body of B. dorsalis at different developmental stages. In a single run, 747,206 sequencing reads with a mean read length of 382 bp were obtained. These reads were assembled into 28,782 contigs and 169,966 singletons. The mean contig size was 750 bp and many nearly full-length transcripts were assembled. Additionally, we identified a great number of genes that are involved in reproduction and development as well as genes that represent nearly all major conserved metazoan signal transduction pathways, such as insulin signal transduction. Furthermore, transcriptome changes during development were analyzed. A total of 2,977 differentially expressed genes (DEGs) were detected between larvae and pupae libraries, while there were 1,621 DEGs between adults and larvae, and 2,002 between adults and pupae. These DEGs were functionally annotated with KEGG pathway annotation and 9 genes were validated by qRT-PCR. CONCLUSION: Our data represent the extensive sequence resources available for B. dorsalis and provide for the first time access to the genetic architecture of reproduction and development as well as major signal transduction pathways in the Tephritid fruit fly pests, allowing us to elucidate the molecular mechanisms underlying courtship, ovipositing, development and detailed analyses of the signal transduction pathways.
| S-EPMC3343016 | biostudies-literature
Project description:Regulation of male sexual differentiation by a Y chromosome-linked male determining factor (M-factor) is one of a diverse array of sex determination mechanisms found in insects. By deep sequencing of small RNAs from Bactrocera dorsalis early embryos, we identified an autosomal-derived microRNA, miR-1-3p, that has predicted target sites in the transformer gene (Bdtra) required for female sex determination. We further demonstrate by both in vitro and in vivo tests that miR-1-3p suppresses Bdtra expression. Injection of a miR-1-3p mimic in early embryos results in 87-92% phenotypic males, whereas knockdown of miR-1-3p by an inhibitor results in 67-77% phenotypic females. Finally, CRISPR/Cas9-mediated knockout of miR-1-3p results in the expression of female-specific splice variants of Bdtra and doublesex (Bddsx), and induced sex reversal of XY individuals into phenotypic females. These results indicate that miR-1-3p is required for male sex determination in early embryogenesis in B. dorsalis as an intermediate male determiner.
| S-EPMC7029022 | biostudies-literature