Dataset Information

Analysis of the transcriptome of blowfly Chrysomya megacephala (Fabricius) larvae in responses to different edible oils.

ABSTRACT:

Background

Chrysomya megacephala (Fabricius), a prevalent necrophagous blowfly that is easily mass reared, is noted for being a mechanical vector of pathogenic microorganisms, a pollinator of numerous crops, and a resource insect in forensic investigation in the postmortem interval. In the present study, in order to comprehensively understand the physiological and biochemical functions of C. megacephala, we performed RNA-sequencing and digital gene expression (DGE) profiling using Solexa/Illumina sequencing technology.

Methodology/principal findings

A total of 39,098,662 clean reads were assembled into 27,588 unigenes with a mean length of 768 nt. All unigenes were searched against the Nt database, Nr database, Swiss-Prot, Cluster of Orthologous Groups (COG) and Kyoto Encyclopedia of Genes and Genome (KEGG) with the BLASTn or BLASTx algorithm (E-value<0.00001) for annotations. In total, 7,081 unigenes and 14,099 unigenes were functionally classified into 25 COG categories and 240 KEGG pathways, respectively. Furthermore, 20,216 unigenes were grouped into 48 sub-categories belonging to 3 main Gene Ontology (GO) categories (ontologies). Using the transcriptome data as references, we analyzed the differential gene expressions between a soybean oil-fed group (SOF) and a lard oil-fed group (LOF), compared to the negative control group (NC), using the DGE approach. We finally obtained 1,566 differentially expressed genes in SOF/NC, and 1,099 genes in LOF/NC. For further analysis, GO and KEGG functional enrichment were performed on all differentially expressed genes, and a group of differentially expressed candidate genes related to lipometabolism were identified.

Conclusions/significance

This study provides a global survey of C. megacephala and provides the basis for further research on the functional genomics of this insect.

SUBMITTER: Zhang M

PROVIDER: S-EPMC3653882 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Analysis of the transcriptome of blowfly Chrysomya megacephala (Fabricius) larvae in responses to different edible oils.

Zhang Min M Yu Hao H Yang Yanyan Y Song Chao C Hu Xinjun X Zhang Guren G

PloS one 20130514 5

<h4>Background</h4>Chrysomya megacephala (Fabricius), a prevalent necrophagous blowfly that is easily mass reared, is noted for being a mechanical vector of pathogenic microorganisms, a pollinator of numerous crops, and a resource insect in forensic investigation in the postmortem interval. In the present study, in order to comprehensively understand the physiological and biochemical functions of C. megacephala, we performed RNA-sequencing and digital gene expression (DGE) profiling using Solexa ...[more]

PMID: 23690992

Similar Datasets

Project description:BackgroundChrysomya megacephala (Fabricius) is a prevalent and synanthropic blowfly which has two sides, for being a pathogenic vector, an efficient pollinator, a promising resource of proteins, lipids, chitosan, biofuel et al., and an important forensic indicator. Moreover olfactory proteins are crucial component to function in related processes. However, the genomic platform of C. megacephala remains relatively unavailable. Developmental transcriptomes of eggs, larvae from 1st instar to before pupa stage and adults from emergence to egg laying period were built by RNA-sequencing to establish sequence background of C. megacephala with special lights on olfactory proteins.ResultsClean reads in eggs, larvae and adults were annotated into 59486 transcripts. Transcripts were assembled into 22286, 17180, 18934 and 35900 unigenes in eggs, larvae, adults and the combined datasets, respectively. Unigenes were annotated using Nr (NCBI non-redundant protein sequences), Nt (NCBI non-redundant nucleotide sequences), GO (Gene Ontology), PFAM (Protein family), KOG/COG (Clusters of Orthologous Groups of proteins), Swiss-Prot (A manually annotated and reviewed protein sequence database), and KO (KEGG Orthology). Totally 12196 unigenes were annotated into 51 sub-categories belonging to three main GO categories; 8462 unigenes were classified functionally into 26 categories to KOG classifications; 5160 unigenes were functionally classified into 5 KEGG categories. Moreover, according to RSEM, the number of differentially expressed genes between larvae and eggs, adults and eggs, adults and larvae, and the common differentially expressed genes were 2637, 1804, 2628 and 258, respectively. Among them, 17 odorant-binding proteins (OBPs), 7 chemosensory proteins (CSPs) and 8 ionotropic receptors (IRs) were differently expressed in adults and larvae. Ten were confirmed as significant differentially expressed genes. Furthermore, OBP Cmeg32081-c4 was highly expressed in the female head and Cmeg33593_c0 were up-regulated with the increase of larval age.ConclusionsA comprehensive sequence resource with desirable quality was built by comparative transcriptome of eggs, larvae and adults, enriching the genomic platform of C. megacephala. The identified differentially expressed genes would facilitate the understanding of metamorphosis, development and the fitness to environmental change of C. megacephala. OBP Cmeg32081-c4 and Cmeg33593_c0 might play a crucial role in the interactions between olfactory system and biological processes.

Project description:BackgroundStable reference genes are essential for accurate normalization in gene expression studies with reverse transcription quantitative polymerase chain reaction (qPCR). A synanthropic fly, Chrysomya megacephala, is a well known medical vector and forensic indicator. Unfortunately, previous studies did not look at the stability of reference genes used in C. megacephala.ResultsIn this study, the expression level of Actin, ribosomal protein L8 (Rpl8), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), elongation factor 1α (EF1), α-tubulin (α-TUB), β-tubulin (β-TUB), TATA binding box (TBP), 18S rRNA (18S) and ribosomal protein S7 (Rps7) were evaluated for their stability using online software RefFinder, which combines the normal software of the ΔCt method, BestKeeper, Normfinder, and geNorm. Moreover the number of suitable reference gene pairs was also suggested by Excel-based geNorm. The expression levels of these reference genes were evaluated under different experimental conditions with special perspectives of forensic applications: developmental stages (eggs, first, second and third instar larvae, pupae and adults); food sources of larvae (pork, fish and chicken); feeding larvae with drugs (untreated control, Estazolam and Marvelon); feeding larvae with heavy metals (untreated control, cadmium and zinc); tissues of adults (head, thorax, abdomen, legs and wings). According to RefFinder, EF1 was the most suitable reference gene of developmental stages, food and tissues; 18S and GAPDH were the most suitable reference genes for drugs and heavy metals, respectively, which could be widely used for quantification of target gene expression with qPCR in C. megacephala. Suitable reference gene pairs were also suggested by geNorm.ConclusionThis fundamental but vital work should facilitate the gene studies of related biological processes and deepen the understanding in physiology, toxicology, and especially medical and forensic entomology of C. megacephala.

Dataset Information

Analysis of the transcriptome of blowfly Chrysomya megacephala (Fabricius) larvae in responses to different edible oils.

Background

Methodology/principal findings

Conclusions/significance

Publications

Analysis of the transcriptome of blowfly Chrysomya megacephala (Fabricius) larvae in responses to different edible oils.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets