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. 2019 Nov 6;20(1):808.
doi: 10.1186/s12864-019-6208-x.

Sex- and tissue-specific transcriptome analyses and expression profiling of olfactory-related genes in Ceracris nigricornis Walker (Orthoptera: Acrididae)

Hao Yuan  1 , Huihui Chang  1 , Lina Zhao  1 , Chao Yang  1   2 , Yuan Huang  3
Affiliations

Sex- and tissue-specific transcriptome analyses and expression profiling of olfactory-related genes in Ceracris nigricornis Walker (Orthoptera: Acrididae)

Hao Yuan et al. BMC Genomics. .

Abstract

Background: The sophisticated insect olfactory system plays an important role in recognizing external odors and enabling insects to adapt to environment. Foraging, host seeking, mating, ovipositing and other forms of chemical communication are based on olfaction, which requires the participation of multiple olfactory genes. The exclusive evolutionary trend of the olfactory system in Orthoptera insects is an excellent model for studying olfactory evolution, but limited olfaction research is available for these species. The olfactory-related genes of Ceracris nigricornis Walker (Orthoptera: Acrididae), a severe pest of bamboos, have not yet been reported.

Results: We sequenced and analyzed the transcriptomes from different tissues of C. nigricornis and obtained 223.76 Gb clean data that were assembled into 43,603 unigenes with an N50 length of 2235 bp. Among the transcripts, 66.79% of unigenes were annotated. Based on annotation and tBLASTn results, 112 candidate olfactory-related genes were identified for the first time, including 20 odorant-binding proteins (OBPs), 10 chemosensory-binding proteins (CSPs), 71 odorant receptors (ORs), eight ionotropic receptors (IRs) and three sensory neuron membrane proteins (SNMPs). The fragments per kilobase per million mapped fragments (FPKM) values showed that most olfactory-related differentially expressed genes (DEGs) were enriched in the antennae, and these results were confirmed by detecting the expression of olfactory-related genes with quantitative real-time PCR (qRT-PCR). Among these antennae-enriched genes, some were sex-biased, indicating their different roles in the olfactory system of C. nigricornis.

Conclusions: This study provides the first comprehensive list and expression profiles of olfactory-related genes in C. nigricornis and a foundation for functional studies of these olfactory-related genes at the molecular level.

Keywords: Ceracris nigricornis; Chemosensory-binding protein; Expression profiles analysis; Ionotropic receptor; Odorant receptor; Odorant-binding protein; Sensory neuron membrane protein; Transcriptome.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Differentially expressed genes (DEGs) analysis of C. nigricornis. a Gene ontology (GO) classifications of DEGs in C. nigricornis. b GO enrichment analysis in the top 25 DEGs of biological process category. c K-means clustering of 111 olfactory-related DEGs. A: antennae; H: head (antennae were cut off); L: leg; W: wing; T: abdomen-thorax. Log10 (FPKM+ 1) values were used, and FPKM values were the average values of each tissues, including female three biological repeats and male three biological repeats
Fig. 2
Fig. 2
Hierarchical clustering of differentially expressed olfactory-related genes with difference tissues in C. nigricornis. a odorant receptors (ORs); b odorant binding proteins (OBPs); c chemosensory proteins (CSPs); d ionotropic receptors (IRs); e sensory neuron membrane proteins (SNMPs). A: antennae; H: head (antennae were cut off); L: leg; W: wing; T: abdomen-thorax. Log10 (FPKM+ 1) values were used, and FPKM values were the average values of each tissues including female three biological repeats and male three biological repeats
Fig. 3
Fig. 3
Quantitative real-time PCR analysis of relative expression levels of odorant binding proteins (OBPs) from C. nigricornis. A: antennae; H: head (antennae were cut off); L: leg; W: wing; T: abdomen-thorax. The β-actin was used as the reference gene and CnigOBP3 in female antennae as a positive control. The error bars represent the standard error of three independent experiments. Different small letters above bars indicate significant differences among different tissues (P < 0.05). * indicates significant difference between both sexes in the same tissue (P < 0.05), and ns indicates no significant difference
Fig. 4
Fig. 4
Quantitative real-time PCR analysis of relative expression levels of chemosensory proteins (CSPs) from C. nigricornis. A: antennae; H: head (antennae were cut off); L: leg; W: wing; T: abdomen-thorax. The β-actin was used as the reference gene and CnigCSP1 in female antennae as a positive control. The error bars represent the standard error of three independent experiments. Different small letters above bars indicate significant differences among different tissues (P < 0.05). * indicates significant difference between both sexes in the same tissue (P < 0.05), and ns indicates no significant difference
Fig. 5
Fig. 5
Quantitative real-time PCR analysis of relative expression levels of odorant receptors (ORs) from C. nigricornis. A: antennae; H: head (antennae were cut off); L: leg; W: wing; T: abdomen-thorax. The β-actin was used as the reference gene and CnigOR37 in female antennae as a positive control. The error bars represent the standard error of three independent experiments. Different small letters above bars indicate significant differences among different tissues (P < 0.05). * indicates significant difference between both sexes in the same tissue (P < 0.05), and ns indicates no significant difference
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