Antennal transcriptome analysis and expression profiles of odorant binding proteins in Eogystia hippophaecolus (Lepidoptera: Cossidae)

et al. 2017

PLoS ONE

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Seabuckthorn carpenter moth, Eogystia hippophaecolus (Lepidoptera: Cossidae), is an important pest of sea buckthorn (Hippophae rhamnoides), which is a shrub that has significant ecological and economic value in China. E. hippophaecolus is highly cold tolerant, but limited studies have been conducted to elucidate the molecular mechanisms underlying its cold resistance. Here we sequenced the E. hippophaecolus transcriptome using RNA-Seq technology and performed de novo assembly from the short paired-end reads. We investigated the larval response to cold stress by comparing gene expression profiles between treatments. We obtained 118,034 unigenes, of which 22,161 were annotated with gene descriptions, conserved domains, gene ontology terms, and metabolic pathways. These resulted in 57 GO terms and 193 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. By comparing transcriptome profiles for differential gene expression, we identified many differentially expressed proteins and genes, including heat shock proteins and cuticular proteins which have previously been reported to be involved in cold resistance of insects. This study provides a global transcriptome analysis and an assessment of differential gene expression in E. hippophaecolus under cold stress. We found seven differential expressed genes in common between developmental stages, which were verified with qPCR. Our findings facilitate future genomic studies aimed at improving our understanding of the molecular mechanisms underlying the response of insects to low temperatures.

“…A t-test was performed for the qPCR data. β-Actin was used as the reference gene to normalize target gene expression [ 53 ].…”

Section: Methodsmentioning

confidence: 99%

Differential transcriptome analysis reveals genes related to cold tolerance in seabuckthorn carpenter moth, Eogystia hippophaecolus

Cui

et al. 2017

PLoS ONE

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“…Specific primers (supplementary material 1) for qRT-PCR were designed based on the sequences of ORFs using Primer Premier 5.0 [45]. The E. hippophaecola actin gene was used as a reference [40,41,46] and we used Bestkeeper to evaluate the std value (<1) of actin gene under different temperatures which indicated that actin gene was stable [47]. The qRT-PCR was carried out on a Bio-Rad CFX96 PCR machine (Hercules, CA) in a reaction mixture of 12. in three biological replicates, and each sample was technically repeated three times.…”

Section: Expression Of Ehhsp90 Under Different Treatmentsmentioning

confidence: 99%

Different Responses of Cytoplasmic and Endoplasmic Reticulum Hsp90 Genes from Eogystia hippophaecola (Lepidoptera: Cossidae) to Cold Stress

Tao

et al. 2019

Forests

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Eogystia hippophaecola Hua, Chou, Fang et Chen (Lepidoptera: Cossidae) is an important borer pest of the sea buckthorn forest (Hippophae rhamnoides L.) in China. Its larvae, which are highly cold tolerant, mainly overwinter in sea buckthorn roots. Heat shock proteins (Hsps) are important molecular chaperones that have been linked to cold tolerance in insects. In this study, we cloned the open reading frames (ORFs) of two Hsp90 genes from E. hippophaecola, EhHsp90-1 and EhHsp90-2, and analyzed their expression under cold stress by qRT-PCR. EhHsp90-1 and EhHsp90-2 are 2154 and 2346 bp in length, respectively, encoding 717 and 781 amino acids. The deduced amino acid sequences contain the conserved signature sequences of the Hsp90 family and the C-terminus characteristic sequence of cytoplasmic or endoplasmic reticulum Hsp90 protein. Phylogenetic analysis revealed the amino acid sequences of EhHsp90-1 and EhHsp90-2 were very similar to the corresponding proteins from Lepidoptera. Under various low-temperature treatments lasting 2 h, EhHsp90-1 and EhHsp90-2 exhibited similar expression patterns, increasing first and then decreasing. At −5 °C, EhHsp90-1 was significantly up-regulated after 12 h, whereas EhHsp90-2 was up-regulated after just 1 h and reached its highest level at 2 h; however, the overall degree of upregulation was greater for EhHsp90-1. Subsequently, the expression level of EhHsp90-2 fluctuated with time. Our results suggest that the two Hsp90s play important roles in E. hippophaecola larvae response to cold stress, but that their response times and the magnitudes of their responses to low-temperature stress differed significantly, providing a theoretical basis for further studying the molecular mechanism of cold tolerance in E. hippophaecola larvae.

“…OBPs are soluble proteins with low molecular weights and are mainly expressed in the lymph of antenna sensilla [28][29]. Some OBPs exhibit specific binding with hydrophobic odorant molecules and can deliver the molecules to membrane-binding receptors [30][31][32][33]. Typical OBPs have six conserved cysteine residues, which form three pairs of disulfide bridges [28][29][34][35].…”

Section: Introductionmentioning

confidence: 99%

Antennal transcriptome analysis and expression profiles of odorant binding proteins in two woodwasps, Sirex noctilio and S. nitobei (Hymenoptera: Siricidae)

Guo

Qiao

et al. 2019

Preprint

Background The olfactory system is the foundation of insect behavior. Odorant binding proteins (OBPs) are key components of the insect olfactory system. The woodwasp Sirex noctilio Fabricius is a major quarantine pest worldwide that was first discovered in China in 2013 and mainly harms members of the Pinus genus. S. nitobei Matsumura is a native species in China and is closely related to S. noctilio. To gain insights into the olfactory mechanisms of these two woodwasp species, olfactory genes were identified using antennal transcriptome analysis. We also analyzed the expression profiles of OBPs with RT-qPCR. Results From our transcriptome analysis, 16 OBPs, 7 chemosensory proteins (CSPs), 41 odorant receptors (ORs), 8 gustatory receptors (GRs), 13 ionotropic receptors (IRs), and one sensory neuron membrane protein (SNMP) were identified in S. noctilio, while 15 OBPs, 6 CSPs, 43 ORs, 10 GRs, 16 IRs, and 1 SNMP were identified in S. nitobei. Most of the olfactory genes identified in two species were homologous. However, some species-specific olfactory genes were identified in the antennal transcriptomes, including SnocOBP13, SnocCSP6, SnocOR26, SnitGR9, and SnitIR17. In total, 14 OBPs (7 in S. noctilio and 7 in S. nitobei) were expressed primarily in the antennae of the two woodwasps. SnocOBP11 and SnitOBP11 were highly expressed in antennae and were also clearly expressed in the external genitalia. SnocOBP3 is highly expressed in the genitalia of females, and SnocOBP7 and SnitOBP7 are highly expressed in the genitalia of males. Meanwhile, SnocOBP10 was specifically expressed in male heads. Conclusion In total, 86 olfactory proteins were identified in S. noctilio, and 91 were identified in S. nitobei. Most SnocOBPs and SnitOBPs displayed enriched expression in the antennae, which are involved in odor recognition. A few OBPs were mainly expressed in the external genitals or heads and exhibited an obvious sex bias, which may indicate that the external genitals and heads are able to recognize sex pheromones or plant volatile compounds as a part of normal behaviors such as feeding, mating, or spawning. Our study provides key insights regarding the mechanism of interactions between the insect olfactory system and specific odor molecules.