Silencing in Apolygus lucorum of the olfactory coreceptor Orco gene by RNA interference induces EAG response declining to two putative semiochemicals

[Display omitted] •The AlucOrco gene were successfully cloned and characterized from Apolygus lucorum.•The AlucOrco gene expression level was the highest in the antennae.•We knocked down the AlucOrco gene by injecting siRNA in vivo.•The EAG responses of the siRNA-treated bugs to two chemicals were d...

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Veröffentlicht in:Journal of insect physiology 2014-01, Vol.60, p.31-39
Hauptverfasser: Zhou, Yan-Le, Zhu, Xiao-Qiang, Gu, Shao-Hua, Cui, Huan-huan, Guo, Yu-Yuan, Zhou, Jing-Jiang, Zhang, Yong-Jun
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container_title Journal of insect physiology
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creator Zhou, Yan-Le
Zhu, Xiao-Qiang
Gu, Shao-Hua
Cui, Huan-huan
Guo, Yu-Yuan
Zhou, Jing-Jiang
Zhang, Yong-Jun
description [Display omitted] •The AlucOrco gene were successfully cloned and characterized from Apolygus lucorum.•The AlucOrco gene expression level was the highest in the antennae.•We knocked down the AlucOrco gene by injecting siRNA in vivo.•The EAG responses of the siRNA-treated bugs to two chemicals were declined. Apolygus lucorum (Meyer-Dür) (Hemiptera: Miridae) is an agronomically important pest that causes severe economic damage to the cotton, fruit, and vegetable industries. Similar to other insects, A. lucorum can perceive and discriminate olfactory cues. A highly conserved and broadly expressed olfactory coreceptor (Orco) is crucial for insect olfaction, and Orco orthologs have been identified in several insect species. In this study, a homology-based polymerase chain reaction (PCR) method was utilized to identify AlucOrco, an Orco ortholog essential for olfaction in A. lucorum. AlucOrco shares significant sequence homology with known Orco proteins in other insects. Quantitative real-time PCR (qRT-PCR) analysis revealed that AlucOrco was abundantly expressed in adult A. lucorum. AlucOrco expression level was the highest in the antennae; by contrast, AlucOrco showed negligible expression level in other tissues. We injected AlucOrco siRNA into the conjunctivum between the prothorax and mesothorax of A. lucorum and evaluated its expression 36h after RNA interference. The results of qRT-PCR demonstrated that the level of mRNA expression was significantly reduced (>90%) in AlucOrco siRNA-treated A. lucorum than in water-injected and non-injected controls. The electroantennogram responses of A. lucorum to two putative semiochemicals, trans-2-hexenal and trans-2-hexenyl butyrate, were also reduced significantly (∼80%) in RNAi-treated A. lucorum than in the controls. These results suggest that AlucOrco is crucial in mediating odorant perception of A. lucorum, especially in perceiving trans-2-hexenal and trans-2-hexenyl butyrate.
doi_str_mv 10.1016/j.jinsphys.2013.10.006
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Apolygus lucorum (Meyer-Dür) (Hemiptera: Miridae) is an agronomically important pest that causes severe economic damage to the cotton, fruit, and vegetable industries. Similar to other insects, A. lucorum can perceive and discriminate olfactory cues. A highly conserved and broadly expressed olfactory coreceptor (Orco) is crucial for insect olfaction, and Orco orthologs have been identified in several insect species. In this study, a homology-based polymerase chain reaction (PCR) method was utilized to identify AlucOrco, an Orco ortholog essential for olfaction in A. lucorum. AlucOrco shares significant sequence homology with known Orco proteins in other insects. Quantitative real-time PCR (qRT-PCR) analysis revealed that AlucOrco was abundantly expressed in adult A. lucorum. AlucOrco expression level was the highest in the antennae; by contrast, AlucOrco showed negligible expression level in other tissues. We injected AlucOrco siRNA into the conjunctivum between the prothorax and mesothorax of A. lucorum and evaluated its expression 36h after RNA interference. The results of qRT-PCR demonstrated that the level of mRNA expression was significantly reduced (&gt;90%) in AlucOrco siRNA-treated A. lucorum than in water-injected and non-injected controls. The electroantennogram responses of A. lucorum to two putative semiochemicals, trans-2-hexenal and trans-2-hexenyl butyrate, were also reduced significantly (∼80%) in RNAi-treated A. lucorum than in the controls. 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All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c467t-63f99f80835bf953f1626e17d5946886bc0fce2270059a579a25b99c4b8b652b3</citedby><cites>FETCH-LOGICAL-c467t-63f99f80835bf953f1626e17d5946886bc0fce2270059a579a25b99c4b8b652b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.jinsphys.2013.10.006$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,777,781,3537,27905,27906,45976</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24216470$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhou, Yan-Le</creatorcontrib><creatorcontrib>Zhu, Xiao-Qiang</creatorcontrib><creatorcontrib>Gu, Shao-Hua</creatorcontrib><creatorcontrib>Cui, Huan-huan</creatorcontrib><creatorcontrib>Guo, Yu-Yuan</creatorcontrib><creatorcontrib>Zhou, Jing-Jiang</creatorcontrib><creatorcontrib>Zhang, Yong-Jun</creatorcontrib><title>Silencing in Apolygus lucorum of the olfactory coreceptor Orco gene by RNA interference induces EAG response declining to two putative semiochemicals</title><title>Journal of insect physiology</title><addtitle>J Insect Physiol</addtitle><description>[Display omitted] •The AlucOrco gene were successfully cloned and characterized from Apolygus lucorum.•The AlucOrco gene expression level was the highest in the antennae.•We knocked down the AlucOrco gene by injecting siRNA in vivo.•The EAG responses of the siRNA-treated bugs to two chemicals were declined. Apolygus lucorum (Meyer-Dür) (Hemiptera: Miridae) is an agronomically important pest that causes severe economic damage to the cotton, fruit, and vegetable industries. Similar to other insects, A. lucorum can perceive and discriminate olfactory cues. A highly conserved and broadly expressed olfactory coreceptor (Orco) is crucial for insect olfaction, and Orco orthologs have been identified in several insect species. In this study, a homology-based polymerase chain reaction (PCR) method was utilized to identify AlucOrco, an Orco ortholog essential for olfaction in A. lucorum. AlucOrco shares significant sequence homology with known Orco proteins in other insects. Quantitative real-time PCR (qRT-PCR) analysis revealed that AlucOrco was abundantly expressed in adult A. lucorum. AlucOrco expression level was the highest in the antennae; by contrast, AlucOrco showed negligible expression level in other tissues. We injected AlucOrco siRNA into the conjunctivum between the prothorax and mesothorax of A. lucorum and evaluated its expression 36h after RNA interference. The results of qRT-PCR demonstrated that the level of mRNA expression was significantly reduced (&gt;90%) in AlucOrco siRNA-treated A. lucorum than in water-injected and non-injected controls. The electroantennogram responses of A. lucorum to two putative semiochemicals, trans-2-hexenal and trans-2-hexenyl butyrate, were also reduced significantly (∼80%) in RNAi-treated A. lucorum than in the controls. 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Apolygus lucorum (Meyer-Dür) (Hemiptera: Miridae) is an agronomically important pest that causes severe economic damage to the cotton, fruit, and vegetable industries. Similar to other insects, A. lucorum can perceive and discriminate olfactory cues. A highly conserved and broadly expressed olfactory coreceptor (Orco) is crucial for insect olfaction, and Orco orthologs have been identified in several insect species. In this study, a homology-based polymerase chain reaction (PCR) method was utilized to identify AlucOrco, an Orco ortholog essential for olfaction in A. lucorum. AlucOrco shares significant sequence homology with known Orco proteins in other insects. Quantitative real-time PCR (qRT-PCR) analysis revealed that AlucOrco was abundantly expressed in adult A. lucorum. AlucOrco expression level was the highest in the antennae; by contrast, AlucOrco showed negligible expression level in other tissues. We injected AlucOrco siRNA into the conjunctivum between the prothorax and mesothorax of A. lucorum and evaluated its expression 36h after RNA interference. The results of qRT-PCR demonstrated that the level of mRNA expression was significantly reduced (&gt;90%) in AlucOrco siRNA-treated A. lucorum than in water-injected and non-injected controls. The electroantennogram responses of A. lucorum to two putative semiochemicals, trans-2-hexenal and trans-2-hexenyl butyrate, were also reduced significantly (∼80%) in RNAi-treated A. lucorum than in the controls. These results suggest that AlucOrco is crucial in mediating odorant perception of A. lucorum, especially in perceiving trans-2-hexenal and trans-2-hexenyl butyrate.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>24216470</pmid><doi>10.1016/j.jinsphys.2013.10.006</doi><tpages>9</tpages></addata></record>
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subjects Amino Acid Sequence
Animals
Apolygus lucorum
Arthropod Antennae - physiology
Base Sequence
Electroantennogram
Electrophysiological Phenomena
Expression profile
Female
Hemiptera
Hemiptera - genetics
Insect Proteins - metabolism
Male
Miridae
Molecular Sequence Data
Nymph - metabolism
Olfactory coreceptor
Ovum - metabolism
Pheromones - genetics
Phylogeny
Real-Time Polymerase Chain Reaction
Receptors, Odorant - metabolism
RNA Interference
Sequence Homology, Amino Acid
title Silencing in Apolygus lucorum of the olfactory coreceptor Orco gene by RNA interference induces EAG response declining to two putative semiochemicals
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