Diversity of bacterial symbionts associated with the tropical plant bug Monalonion velezangeli (Hemiptera: Miridae) revealed by high-throughput 16S-rRNA sequencing.

Diversity of bacterial symbionts associated with the tropical plant bug Monalonion velezangeli (Hemiptera: Miridae) revealed by high-throughput 16S-rRNA sequencing. | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Diversity of bacterial symbionts associated with the tropical plant bug Monalonion velezangeli (Hemiptera: Miridae) revealed by high-throughput 16S-rRNA sequencing. Lucio Navarro-Escalante, Pablo Benavides, Flor Edith Acevedo This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2022560/v7 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 28 Jan, 2024 Read the published version in Peer Community Journal → Version 7 posted You are reading this latest preprint version Show more versions Abstract Insects and microbes have developed complex symbiotic relationships that evolutionarily and ecologically play beneficial roles for both, the symbiont and the host. In most Hemiptera insects, bacterial symbionts offer mainly nutritional, defensive, and reproductive roles in addition to promoting the adaptive radiation of several hemipteran phytophagous lineages. The tropical plant bug Monalonion velezangeli (Hemiptera: Miridae) is a polyphagous herbivore considered an important insect pest for several economically relevant tropical crops, but information about the composition of its bacterial microbiota was missing. In this study, we describe the diversity and structure of the bacterial microbiota in the nymph and adult life stages of M . velezangeli using Illumina high-throughput sequencing of 16S ribosomal RNA gene amplicons (meta-barcoding). We found that both insect life stages share a similar microbiota in terms of bacterial diversity and community structure. The intracellular symbiont Wolbachia dominated the overall microbiome composition (~92%) in these life stages. Members of the core microbiota include Wolbachia, Romboutsia , Ignavibacterium , Clostridium , Allobaculum , Paracoccus , Methylobacterium , Faecalibacterium , Collinsella , Rothia , Sphingomonas and 4 other undetermined bacterial genera. Based on PCR screening and DNA sequencing of the wsp gene, Wolbachia infection was confirmed in almost 80% of samples, and represented by two different isolates or strains within the supergroup B. This data offer opportunities for studying the contribution of symbiotic bacteria in the biological performance of this insect pest, and provides a base to explore other insect control methods. Entomology bacterial microbiota endosymbiont plant bug Wolbachia Monalonion Coffea Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Full Text Additional Declarations The authors declare no competing interests. Supplementary Files Supplementarydata.pdf Supplementary Information Cite Share Download PDF Status: Published Journal Publication published 28 Jan, 2024 Read the published version in Peer Community Journal → Version 7 posted You are reading this latest preprint version Show more versions Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-2022560","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":249386730,"identity":"1f2fc4ce-c5b3-4409-810f-ea22a9ffe4b1","order_by":0,"name":"Lucio Navarro-Escalante","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA4UlEQVRIiWNgGAWjYBACPiA+gOBWADEzcwNeLWyoWs6AtDAS1oIAjG1gkoAWieSHBz7usEvsn3324YOP82qj+duBWn5UbMOjJc3g4MwzyYkzzqUbG87cdjx3xmHGBsaeM7fxaMlhOMzbxmzMcIaNTZp327HcBqAWZsY2glrqjeVBWv7OOZY7n0gth+UMQFoYG2pyNxDUwvMM6Je243KGZ9iYDXuOHcjdCNRyEJ9f+NmTH3/42FbNI3eGjfHBj5q63HnnDx988KMCtxZ0cBhMHiBaPRDUkaJ4FIyCUTAKRggAAFFyVsNbxYDHAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0001-7315-5669","institution":"CENICAFE: Centro Nacional de Investigaciones de Cafe","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Lucio","middleName":"","lastName":"Navarro-Escalante","suffix":""},{"id":249386731,"identity":"ac7d9d4f-ed60-4ffe-94e5-9e8a34367fe8","order_by":1,"name":"Pablo Benavides","email":"","orcid":"https://orcid.org/0000-0003-2227-4232","institution":"CENICAFE: Centro Nacional de Investigaciones de Cafe","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Pablo","middleName":"","lastName":"Benavides","suffix":""},{"id":249386732,"identity":"73575fa5-c5ef-47d5-98ce-2e61d8c450f6","order_by":2,"name":"Flor Edith Acevedo","email":"","orcid":"https://orcid.org/0000-0002-0946-9951","institution":"Department of Entomology, Pennsylvania State University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Flor","middleName":"Edith","lastName":"Acevedo","suffix":""}],"badges":[],"createdAt":"2022-09-01 15:30:53","currentVersionCode":7,"declarations":{"humanSubjects":false,"vertebrateSubjects":false,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false,"coiExplicitlySet":false},"doi":"10.21203/rs.3.rs-2022560/v7","doiUrl":"https://doi.org/10.21203/rs.3.rs-2022560/v7","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.24072/pcjournal.362","type":"published","date":"2024-01-29T00:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":48934703,"identity":"b3f248b7-f6e5-4651-8e21-f15e9fc68448","added_by":"auto","created_at":"2023-12-28 22:40:02","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":309720,"visible":true,"origin":"","legend":"\u003cp\u003eDiversity and community structure of the bacterial microbiota in adult and nymph life stages of \u003cem\u003eMonalonion\u003c/em\u003e \u003cem\u003evelezangeli\u003c/em\u003e. (A) Rarefaction curves. (B) Alpha diversity indices and their corresponding p-value of the Mann-Whitney U test. (C) Principal coordinate analysis (PCoA) plot based on Bray-Curtis dissimilarity of bacterial communities in nymphs and adults. (D) Non-metric multidimensional scaling (NMDS) ordination analysis plot based on Bray-Curtis dissimilarity of bacterial communities in nymphs and adults. Stress value represents the goodness-of-fit for the NMDS analysis.\u003c/p\u003e","description":"","filename":"Figure1.png","url":"https://assets-eu.researchsquare.com/files/rs-2022560/v7/abd745e7c37e4418b29803cc.png"},{"id":48934701,"identity":"8dc54a3f-f059-4e34-9511-f5a0ca555028","added_by":"auto","created_at":"2023-12-28 22:40:02","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":390257,"visible":true,"origin":"","legend":"\u003cp\u003eTaxonomic composition of the bacterial microbiota in nymphs and adults of \u003cem\u003eMonalonion\u003c/em\u003e \u003cem\u003evelezangeli\u003c/em\u003e. (A) Relative abundance at Phylum level. (B) Relative abundance at Order level. (C) Heatmap for relative abundances at genus level. (D) Number of ASV sequences consistently detected on either adults of nymphs and number of shared ASVs (circle intersection) as members of the core microbiota.\u003c/p\u003e","description":"","filename":"Figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-2022560/v7/8d699160d27726b0754d16b1.png"},{"id":48935176,"identity":"2d1aa6d9-946a-4c72-b42e-1811325bbba3","added_by":"auto","created_at":"2023-12-28 22:48:03","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":281305,"visible":true,"origin":"","legend":"\u003cp\u003eRelative abundance of the bacterial microbiota, with the exclusion of \u003cem\u003eWolbachia\u003c/em\u003e-associated sequences, for nymphs and adults of \u003cem\u003eMonalonion\u003c/em\u003e \u003cem\u003evelezangeli\u003c/em\u003e. (A) Relative abundances at Phylum level. (B) Heatmap of relative abundances at genus level.\u003c/p\u003e","description":"","filename":"Figure3.png","url":"https://assets-eu.researchsquare.com/files/rs-2022560/v7/714e307d41fcf3b06cc3ce07.png"},{"id":48934704,"identity":"b58bb2fa-d115-4f99-ad26-1ab31b4522c5","added_by":"auto","created_at":"2023-12-28 22:40:02","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":318877,"visible":true,"origin":"","legend":"\u003cp\u003eMolecular screening for presence of \u003cem\u003eWolbachia\u003c/em\u003eendosymbiont in \u003cem\u003eMonalonion velezangeli\u003c/em\u003e samples. DNA samples from single insects (HU15.1 to HU20.6) were tested for PCR amplification of the \u003cem\u003eWolbachia\u003c/em\u003e \u003cem\u003ewsp\u003c/em\u003egene using wsp81F and wsp691R primers. Quality of DNA was tested by amplification of the 28S rRNA (28S) gene fragment (~700 bp). DNA from a \u003cem\u003eDrosophila\u003c/em\u003e \u003cem\u003emelanogaster \u003c/em\u003e(Dm) population was used as positive control for \u003cem\u003eWolbachia\u003c/em\u003einfection, and water (-) as negative control.\u003c/p\u003e","description":"","filename":"Figure4.png","url":"https://assets-eu.researchsquare.com/files/rs-2022560/v7/95d89b44c5c893395936f3e5.png"},{"id":48934699,"identity":"f3c0e3a1-fd41-4b92-9a6d-43e721f0a50c","added_by":"auto","created_at":"2023-12-28 22:40:02","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":1129682,"visible":true,"origin":"","legend":"\u003cp\u003eMaximum Likelihood phylogenetic tree of \u003cem\u003eWolbachia\u003c/em\u003e \u003cem\u003ewsp\u003c/em\u003esequences from \u003cem\u003eMonalonion\u003c/em\u003e \u003cem\u003evelezangeli \u003c/em\u003eand representative \u003cem\u003eWolbachia\u003c/em\u003estrains from other host insects at the GenBank database. \u003cem\u003eWolbachia\u003c/em\u003esupergroups A (green branch) and B (blue branch) clusters based on \u003cem\u003ewsp\u003c/em\u003esequences are shown. Sequence haplotypes clustering of the \u003cem\u003eM. velezangeli \u003c/em\u003ewsp isolates, wMvel1 and wMvel2, are shown in purple and pink colors respectively. Hemiptera species are highlighted in bold letters. The aLRT branch supports are indicated as red numbers. Genbank accession numbers precede each sequence name.\u003c/p\u003e","description":"","filename":"Figure5.png","url":"https://assets-eu.researchsquare.com/files/rs-2022560/v7/8518f6dfc78d445e6f326487.png"},{"id":50916702,"identity":"c3e39d3a-0239-4831-a7ee-efe56719305d","added_by":"auto","created_at":"2024-02-09 14:21:59","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1847171,"visible":true,"origin":"","legend":"","description":"","filename":"Monalonionmicrobiomemanuscript122023v7.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2022560/v7_covered_f56841c9-e152-402f-9512-97f763bbe33b.pdf"},{"id":48934700,"identity":"4d47e73d-f11e-4c20-83c2-f5a5dda7bf48","added_by":"auto","created_at":"2023-12-28 22:40:02","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":1203588,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSupplementary Information\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Supplementarydata.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2022560/v7/07e74f18af688b856d7db310.pdf"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003eDiversity of bacterial symbionts associated with the tropical plant bug \u003cem\u003eMonalonion velezangeli\u003c/em\u003e (Hemiptera: Miridae) revealed by high-throughput 16S-rRNA sequencing.\u003c/p\u003e","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"bacterial microbiota, endosymbiont, plant bug, Wolbachia, Monalonion, Coffea","lastPublishedDoi":"10.21203/rs.3.rs-2022560/v7","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2022560/v7","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eInsects and microbes have developed complex symbiotic relationships that evolutionarily and ecologically play beneficial roles for both, the symbiont and the host. In most Hemiptera insects, bacterial symbionts offer mainly nutritional, defensive, and reproductive roles in addition to promoting the adaptive radiation of several hemipteran phytophagous lineages. The tropical plant bug \u003cem\u003eMonalonion\u003c/em\u003e \u003cem\u003evelezangeli\u003c/em\u003e(Hemiptera: Miridae) is a polyphagous herbivore considered an important insect pest for several economically relevant tropical crops, but information about the composition of its bacterial microbiota was missing. In this study, we describe the diversity and structure of the bacterial microbiota in the nymph and adult life stages of \u003cem\u003eM\u003c/em\u003e. \u003cem\u003evelezangeli\u003c/em\u003eusing Illumina high-throughput sequencing of 16S ribosomal RNA gene amplicons (meta-barcoding). We found that both insect life stages share a similar microbiota in terms of bacterial diversity and community structure. The intracellular symbiont \u003cem\u003eWolbachia\u003c/em\u003edominated the overall microbiome composition (~92%) in these life stages. Members of the core microbiota include \u003cem\u003eWolbachia, Romboutsia\u003c/em\u003e, \u003cem\u003eIgnavibacterium\u003c/em\u003e, \u003cem\u003eClostridium\u003c/em\u003e, \u003cem\u003eAllobaculum\u003c/em\u003e, \u003cem\u003eParacoccus\u003c/em\u003e, \u003cem\u003eMethylobacterium\u003c/em\u003e, \u003cem\u003eFaecalibacterium\u003c/em\u003e, \u003cem\u003eCollinsella\u003c/em\u003e, \u003cem\u003eRothia\u003c/em\u003e, \u003cem\u003eSphingomonas\u003c/em\u003eand 4 other undetermined bacterial genera. Based on PCR screening and DNA sequencing of the \u003cem\u003ewsp\u003c/em\u003egene, \u003cem\u003eWolbachia\u003c/em\u003einfection was confirmed in almost 80% of samples, and represented by two different isolates or strains within the supergroup B. This data offer opportunities for studying the contribution of symbiotic bacteria in the biological performance of this insect pest, and provides a base to explore other insect control methods.\u003c/p\u003e","manuscriptTitle":"Diversity of bacterial symbionts associated with the tropical plant bug Monalonion velezangeli (Hemiptera: Miridae) revealed by high-throughput 16S-rRNA sequencing.","msid":"","msnumber":"","nonDraftVersions":[{"code":7,"date":"2023-12-28 22:39:58","doi":"10.21203/rs.3.rs-2022560/v7","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}},{"code":6,"date":"2023-11-01 20:04:53","doi":"10.21203/rs.3.rs-2022560/v6","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}},{"code":5,"date":"2023-09-05 17:46:40","doi":"10.21203/rs.3.rs-2022560/v5","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}},{"code":4,"date":"2023-07-14 18:04:36","doi":"10.21203/rs.3.rs-2022560/v4","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}},{"code":3,"date":"2023-07-14 16:41:44","doi":"10.21203/rs.3.rs-2022560/v3","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}},{"code":2,"date":"2022-11-28 16:30:07","doi":"10.21203/rs.3.rs-2022560/v2","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}},{"code":1,"date":"2022-09-13 21:23:00","doi":"10.21203/rs.3.rs-2022560/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"0c25f6b1-eea5-4bfc-a13a-087935bf3666","owner":[],"postedDate":"December 28th, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":27422714,"name":"Entomology"}],"tags":[],"updatedAt":"2024-02-09T14:13:14+00:00","versionOfRecord":{"articleIdentity":"rs-2022560","link":"https://doi.org/10.24072/pcjournal.362","journal":{"identity":"peer-community-journal","isVorOnly":true,"title":"Peer Community Journal"},"publishedOn":"2024-01-29 00:00:00","publishedOnDateReadable":"January 29th, 2024"},"versionCreatedAt":"2023-12-28 22:39:58","video":"","vorDoi":"10.24072/pcjournal.362","vorDoiUrl":"https://doi.org/10.24072/pcjournal.362","workflowStages":[]},"version":"v7","identity":"rs-2022560","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2022560","identity":"rs-2022560","version":["v7"]},"buildId":"WrCJVZZCHTDjtuVLN7oU0","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}