Transcriptome Sequencing Analysis and Functional Identification of Sex Differentiation Genes from the Mosquito Parasitic Nematode, Romanomermis wuchangensis
Mosquito-transmitted diseases like malaria and dengue fever are global problem and an estimated 50-100 million of dengue or dengue hemorrhagic fever cases are reported worldwide every year. The mermithid nematode Romanomermis wuchangensis has been successfully used as an ecosystem-friendly biocontro...
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| description | Mosquito-transmitted diseases like malaria and dengue fever are global problem and an estimated 50-100 million of dengue or dengue hemorrhagic fever cases are reported worldwide every year. The mermithid nematode Romanomermis wuchangensis has been successfully used as an ecosystem-friendly biocontrol agent for mosquito prevention in laboratory studies. However, this nematode can not undergo sex differentiation in vitro culture, which has seriously affected their application of biocontrol in the field. In this study, based on transcriptome sequencing analysis of R. wuchangensis, Rwucmab-3, Rwuclaf-1 and Rwuctra-2 were cloned and used to investigate molecular regulatory function of sex differentiation. qRT-PCR results demonstrated that the expression level of Rwucmab-3 between male and female displayed obvious difference on the 3rd day of parasitic stage, which was earlier than Rwuclaf-1 and Rwuctra-2, highlighting sex differentiation process may start on the 3rd day of parasitic stage. Besides, FITC was used as a marker to test dsRNA uptake efficiency of R. wuchangensis, which fluorescence intensity increased with FITC concentration after 16 h incubation, indicating this nematode can successfully ingest soaking solution via its cuticle. RNAi results revealed the sex ratio of R. wuchangensis from RNAi treated groups soaked in dsRNA of Rwucmab-3 was significantly higher than gfp dsRNA treated groups and control groups, highlighting RNAi of Rwumab-3 may hinder the development of male nematodes. These results suggest that Rwucmab-3 mainly involves in the initiation of sex differentiation and the development of male sexual dimorphism. Rwuclaf-1 and Rwuctra-2 may play vital role in nematode reproductive and developmental system. In conclusion, transcript sequences presented in this study could provide more bioinformatics resources for future studies on gene cloning and other molecular regulatory mechanism in R. wuchangensis. Moreover, identification and functional analysis of sex differentiation genes may clarify the sex differentiation mechanism of R. wuchangensis, which are helpful to solve the uncompleted sex differentiation problem in vitro culture and the potential large-scale field application controlling the larvae of C. quinquefasciatus, A. aegypti and A. albopictus. |
| doi_str_mv | 10.1371/journal.pone.0163127 |
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The mermithid nematode Romanomermis wuchangensis has been successfully used as an ecosystem-friendly biocontrol agent for mosquito prevention in laboratory studies. However, this nematode can not undergo sex differentiation in vitro culture, which has seriously affected their application of biocontrol in the field. In this study, based on transcriptome sequencing analysis of R. wuchangensis, Rwucmab-3, Rwuclaf-1 and Rwuctra-2 were cloned and used to investigate molecular regulatory function of sex differentiation. qRT-PCR results demonstrated that the expression level of Rwucmab-3 between male and female displayed obvious difference on the 3rd day of parasitic stage, which was earlier than Rwuclaf-1 and Rwuctra-2, highlighting sex differentiation process may start on the 3rd day of parasitic stage. Besides, FITC was used as a marker to test dsRNA uptake efficiency of R. wuchangensis, which fluorescence intensity increased with FITC concentration after 16 h incubation, indicating this nematode can successfully ingest soaking solution via its cuticle. RNAi results revealed the sex ratio of R. wuchangensis from RNAi treated groups soaked in dsRNA of Rwucmab-3 was significantly higher than gfp dsRNA treated groups and control groups, highlighting RNAi of Rwumab-3 may hinder the development of male nematodes. These results suggest that Rwucmab-3 mainly involves in the initiation of sex differentiation and the development of male sexual dimorphism. Rwuclaf-1 and Rwuctra-2 may play vital role in nematode reproductive and developmental system. In conclusion, transcript sequences presented in this study could provide more bioinformatics resources for future studies on gene cloning and other molecular regulatory mechanism in R. wuchangensis. Moreover, identification and functional analysis of sex differentiation genes may clarify the sex differentiation mechanism of R. wuchangensis, which are helpful to solve the uncompleted sex differentiation problem in vitro culture and the potential large-scale field application controlling the larvae of C. quinquefasciatus, A. aegypti and A. albopictus.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0163127</identifier><identifier>PMID: 27662191</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Analysis ; Aquatic insects ; Bioinformatics ; Biological control ; Biological pest control ; Biology ; Biology and Life Sciences ; Caenorhabditis ; Caenorhabditis elegans ; Cloning ; Culicidae ; Cuticle ; Dengue fever ; Dengue hemorrhagic fever ; Differentiation ; Disease transmission ; Double-stranded RNA ; Females ; Fever ; Fluorescence ; Functional analysis ; Gene expression ; Gene sequencing ; Genes ; Genetic aspects ; Genetic research ; Genomes ; Incubation ; Infections ; Insect control ; Insecticides ; Kinases ; Laboratories ; Larvae ; Life sciences ; Malaria ; Medicine and Health Sciences ; Mosquitoes ; Nematoda ; Nematodes ; Parasites ; Parasitic diseases ; Proteins ; Regulatory mechanisms (biology) ; Research and Analysis Methods ; RNA-mediated interference ; Romanomermis ; Roundworms ; Sex ; Sex differentiation ; Sex ratio ; Sexual dimorphism ; Spermatogenesis ; Transcription ; Tropical diseases ; Vector-borne diseases ; Viral diseases ; West Nile virus ; Worms</subject><ispartof>PloS one, 2016-09, Vol.11 (9), p.e0163127-e0163127</ispartof><rights>COPYRIGHT 2016 Public Library of Science</rights><rights>2016 Duan et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2016 Duan et al 2016 Duan et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c791t-ccee9cd61f248d0d624f7915d35b5a90f70b09bbe802a58282aa85a9bb52beb53</citedby><cites>FETCH-LOGICAL-c791t-ccee9cd61f248d0d624f7915d35b5a90f70b09bbe802a58282aa85a9bb52beb53</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5035087/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5035087/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2096,2915,23845,27901,27902,53766,53768,79342,79343</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27662191$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Duan, Mingyue</creatorcontrib><creatorcontrib>Xiong, Jinfeng</creatorcontrib><creatorcontrib>Lu, Dandan</creatorcontrib><creatorcontrib>Wang, Guoxiu</creatorcontrib><creatorcontrib>Ai, Hui</creatorcontrib><title>Transcriptome Sequencing Analysis and Functional Identification of Sex Differentiation Genes from the Mosquito Parasitic Nematode, Romanomermis wuchangensis</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Mosquito-transmitted diseases like malaria and dengue fever are global problem and an estimated 50-100 million of dengue or dengue hemorrhagic fever cases are reported worldwide every year. The mermithid nematode Romanomermis wuchangensis has been successfully used as an ecosystem-friendly biocontrol agent for mosquito prevention in laboratory studies. However, this nematode can not undergo sex differentiation in vitro culture, which has seriously affected their application of biocontrol in the field. In this study, based on transcriptome sequencing analysis of R. wuchangensis, Rwucmab-3, Rwuclaf-1 and Rwuctra-2 were cloned and used to investigate molecular regulatory function of sex differentiation. qRT-PCR results demonstrated that the expression level of Rwucmab-3 between male and female displayed obvious difference on the 3rd day of parasitic stage, which was earlier than Rwuclaf-1 and Rwuctra-2, highlighting sex differentiation process may start on the 3rd day of parasitic stage. Besides, FITC was used as a marker to test dsRNA uptake efficiency of R. wuchangensis, which fluorescence intensity increased with FITC concentration after 16 h incubation, indicating this nematode can successfully ingest soaking solution via its cuticle. RNAi results revealed the sex ratio of R. wuchangensis from RNAi treated groups soaked in dsRNA of Rwucmab-3 was significantly higher than gfp dsRNA treated groups and control groups, highlighting RNAi of Rwumab-3 may hinder the development of male nematodes. These results suggest that Rwucmab-3 mainly involves in the initiation of sex differentiation and the development of male sexual dimorphism. Rwuclaf-1 and Rwuctra-2 may play vital role in nematode reproductive and developmental system. In conclusion, transcript sequences presented in this study could provide more bioinformatics resources for future studies on gene cloning and other molecular regulatory mechanism in R. wuchangensis. Moreover, identification and functional analysis of sex differentiation genes may clarify the sex differentiation mechanism of R. wuchangensis, which are helpful to solve the uncompleted sex differentiation problem in vitro culture and the potential large-scale field application controlling the larvae of C. quinquefasciatus, A. aegypti and A. albopictus.</description><subject>Analysis</subject><subject>Aquatic insects</subject><subject>Bioinformatics</subject><subject>Biological control</subject><subject>Biological pest control</subject><subject>Biology</subject><subject>Biology and Life Sciences</subject><subject>Caenorhabditis</subject><subject>Caenorhabditis elegans</subject><subject>Cloning</subject><subject>Culicidae</subject><subject>Cuticle</subject><subject>Dengue fever</subject><subject>Dengue hemorrhagic fever</subject><subject>Differentiation</subject><subject>Disease transmission</subject><subject>Double-stranded 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interference</subject><subject>Romanomermis</subject><subject>Roundworms</subject><subject>Sex</subject><subject>Sex differentiation</subject><subject>Sex ratio</subject><subject>Sexual dimorphism</subject><subject>Spermatogenesis</subject><subject>Transcription</subject><subject>Tropical diseases</subject><subject>Vector-borne diseases</subject><subject>Viral diseases</subject><subject>West Nile 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Sequencing Analysis and Functional Identification of Sex Differentiation Genes from the Mosquito Parasitic Nematode, Romanomermis wuchangensis</title><author>Duan, Mingyue ; Xiong, Jinfeng ; Lu, Dandan ; Wang, Guoxiu ; Ai, Hui</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c791t-ccee9cd61f248d0d624f7915d35b5a90f70b09bbe802a58282aa85a9bb52beb53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Analysis</topic><topic>Aquatic insects</topic><topic>Bioinformatics</topic><topic>Biological control</topic><topic>Biological pest control</topic><topic>Biology</topic><topic>Biology and Life Sciences</topic><topic>Caenorhabditis</topic><topic>Caenorhabditis elegans</topic><topic>Cloning</topic><topic>Culicidae</topic><topic>Cuticle</topic><topic>Dengue fever</topic><topic>Dengue hemorrhagic fever</topic><topic>Differentiation</topic><topic>Disease 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One</addtitle><date>2016-09-23</date><risdate>2016</risdate><volume>11</volume><issue>9</issue><spage>e0163127</spage><epage>e0163127</epage><pages>e0163127-e0163127</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Mosquito-transmitted diseases like malaria and dengue fever are global problem and an estimated 50-100 million of dengue or dengue hemorrhagic fever cases are reported worldwide every year. The mermithid nematode Romanomermis wuchangensis has been successfully used as an ecosystem-friendly biocontrol agent for mosquito prevention in laboratory studies. However, this nematode can not undergo sex differentiation in vitro culture, which has seriously affected their application of biocontrol in the field. In this study, based on transcriptome sequencing analysis of R. wuchangensis, Rwucmab-3, Rwuclaf-1 and Rwuctra-2 were cloned and used to investigate molecular regulatory function of sex differentiation. qRT-PCR results demonstrated that the expression level of Rwucmab-3 between male and female displayed obvious difference on the 3rd day of parasitic stage, which was earlier than Rwuclaf-1 and Rwuctra-2, highlighting sex differentiation process may start on the 3rd day of parasitic stage. Besides, FITC was used as a marker to test dsRNA uptake efficiency of R. wuchangensis, which fluorescence intensity increased with FITC concentration after 16 h incubation, indicating this nematode can successfully ingest soaking solution via its cuticle. RNAi results revealed the sex ratio of R. wuchangensis from RNAi treated groups soaked in dsRNA of Rwucmab-3 was significantly higher than gfp dsRNA treated groups and control groups, highlighting RNAi of Rwumab-3 may hinder the development of male nematodes. These results suggest that Rwucmab-3 mainly involves in the initiation of sex differentiation and the development of male sexual dimorphism. Rwuclaf-1 and Rwuctra-2 may play vital role in nematode reproductive and developmental system. In conclusion, transcript sequences presented in this study could provide more bioinformatics resources for future studies on gene cloning and other molecular regulatory mechanism in R. wuchangensis. Moreover, identification and functional analysis of sex differentiation genes may clarify the sex differentiation mechanism of R. wuchangensis, which are helpful to solve the uncompleted sex differentiation problem in vitro culture and the potential large-scale field application controlling the larvae of C. quinquefasciatus, A. aegypti and A. albopictus.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>27662191</pmid><doi>10.1371/journal.pone.0163127</doi><tpages>e0163127</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2016-09, Vol.11 (9), p.e0163127-e0163127 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_1822644520 |
| source | Public Library of Science (PLoS) Journals Open Access; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Analysis Aquatic insects Bioinformatics Biological control Biological pest control Biology Biology and Life Sciences Caenorhabditis Caenorhabditis elegans Cloning Culicidae Cuticle Dengue fever Dengue hemorrhagic fever Differentiation Disease transmission Double-stranded RNA Females Fever Fluorescence Functional analysis Gene expression Gene sequencing Genes Genetic aspects Genetic research Genomes Incubation Infections Insect control Insecticides Kinases Laboratories Larvae Life sciences Malaria Medicine and Health Sciences Mosquitoes Nematoda Nematodes Parasites Parasitic diseases Proteins Regulatory mechanisms (biology) Research and Analysis Methods RNA-mediated interference Romanomermis Roundworms Sex Sex differentiation Sex ratio Sexual dimorphism Spermatogenesis Transcription Tropical diseases Vector-borne diseases Viral diseases West Nile virus Worms |
| title | Transcriptome Sequencing Analysis and Functional Identification of Sex Differentiation Genes from the Mosquito Parasitic Nematode, Romanomermis wuchangensis |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-11T14%3A10%3A11IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Transcriptome%20Sequencing%20Analysis%20and%20Functional%20Identification%20of%20Sex%20Differentiation%20Genes%20from%20the%20Mosquito%20Parasitic%20Nematode,%20Romanomermis%20wuchangensis&rft.jtitle=PloS%20one&rft.au=Duan,%20Mingyue&rft.date=2016-09-23&rft.volume=11&rft.issue=9&rft.spage=e0163127&rft.epage=e0163127&rft.pages=e0163127-e0163127&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0163127&rft_dat=%3Cgale_plos_%3EA471901743%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1822644520&rft_id=info:pmid/27662191&rft_galeid=A471901743&rft_doaj_id=oai_doaj_org_article_2e228869b7304f9184f6de164ece0384&rfr_iscdi=true |