Signatures of plant defense response specificity mediated by herbivore‐associated molecular patterns in legumes
SUMMARY Chewing herbivores activate plant defense responses through a combination of mechanical wounding and elicitation by herbivore‐associated molecular patterns (HAMPs). HAMPs are wound response amplifiers; however, specific defense outputs may also exist that strictly require HAMP‐mediated defen...
Gespeichert in:
Veröffentlicht in: | The Plant journal : for cell and molecular biology 2022-06, Vol.110 (5), p.1255-1270 |
---|---|
Hauptverfasser: | , , , , , |
Format: | Artikel |
Sprache: | eng |
Schlagworte: | |
Online-Zugang: | Volltext |
Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
container_end_page | 1270 |
---|---|
container_issue | 5 |
container_start_page | 1255 |
container_title | The Plant journal : for cell and molecular biology |
container_volume | 110 |
creator | Steinbrenner, Adam D. Saldivar, Evan Hodges, Nile Guayazán‐Palacios, Natalia Chaparro, Antonio F. Schmelz, Eric A. |
description | SUMMARY
Chewing herbivores activate plant defense responses through a combination of mechanical wounding and elicitation by herbivore‐associated molecular patterns (HAMPs). HAMPs are wound response amplifiers; however, specific defense outputs may also exist that strictly require HAMP‐mediated defense signaling. To investigate HAMP‐mediated signaling and defense responses, we characterized cowpea (Vigna unguiculata) transcriptome changes following elicitation by inceptin, a peptide HAMP common in Lepidoptera larvae oral secretions. Following inceptin treatment, we observed large‐scale reprogramming of the transcriptome consistent with three different response categories: (i) amplification of mechanical wound responses, (ii) temporal extension through accelerated or prolonged responses, and (iii) examples of inceptin‐specific elicitation and suppression. At both early and late timepoints, namely 1 and 6 h, large sets of transcripts specifically accumulated following inceptin elicitation. Further early inceptin‐regulated transcripts were classified as reversing changes induced by wounding alone. Within key signaling‐ and defense‐related gene families, inceptin‐elicited responses included target subsets of wound‐induced transcripts. Transcripts displaying the largest inceptin‐elicited fold changes included transcripts encoding terpene synthases (TPSs) and peroxidases (POXs) that correspond with induced volatile production and increased POX activity in cowpea. Characterization of inceptin‐elicited cowpea defenses via heterologous expression in Nicotiana benthamiana demonstrated that specific cowpea TPSs and POXs were able to confer terpene emission and the reduced growth of beet armyworm (Spodoptera exigua) herbivores, respectively. Collectively, our present findings in cowpea support a model where HAMP elicitation both amplifies concurrent wound responses and specifically contributes to the activation of selective outputs associated with direct and indirect antiherbivore defenses.
Significance Statement
Plants recognize herbivore‐associated molecular patterns (HAMPs) and increase defense production, but interactions with the more general wound response are not well understood. We examined an established HAMP–receptor interaction to characterize the transcriptomic modulation of wound responses by the HAMP peptide inceptin. Inceptin not only amplifies wound responses, but can also specifically induce or suppress transcripts with demonstrated functions in direct |
doi_str_mv | 10.1111/tpj.15732 |
format | Article |
fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_2641865179</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2672256112</sourcerecordid><originalsourceid>FETCH-LOGICAL-c3882-6558bd87cd7132b80c73e18f8dbb6e25b7e31ee84bdfbbc1ac7ede5ecd87494d3</originalsourceid><addsrcrecordid>eNp1kctu1TAQhi0EoofCghdAltiURdqMHV_OElW0UFUCiSKxi3yZFB8lcWonVGfHI_CMPAkuaVkg4c1YM9_8mpmfkJdQH0N5J_O0OwahOHtENsClqDjwr4_Jpt7KulINsAPyLOddXYPisnlKDrjgIISQG3LzOVyPZl4SZho7OvVmnKnHDseMtCSnePfJE7rQBRfmPR3QBzOjp3ZPv2Gy4XtM-OvHT5NzdGtliD26pTeJTmaeMY2ZhpH2eL0MmJ-TJ53pM764j4fky9m7q9P31eXH8w-nby8rx7VmlRRCW6-V8wo4s7p2iiPoTntrJTJhFXJA1I31nbUOjFPoUaArPc228fyQHK26U4o3C-a5HUJ22JcNMS65ZbIBLQWobUFf_4Pu4pLGMl2hFGNCArBCvVkpl2LOCbt2SmEwad9C3d750BYf2j8-FPbVveJiy8H-kg-HL8DJCtyGHvf_V2qvPl2skr8BkoCV3Q</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2672256112</pqid></control><display><type>article</type><title>Signatures of plant defense response specificity mediated by herbivore‐associated molecular patterns in legumes</title><source>Wiley Free Content</source><source>MEDLINE</source><source>IngentaConnect Free/Open Access Journals</source><source>Wiley Online Library Journals Frontfile Complete</source><source>EZB-FREE-00999 freely available EZB journals</source><creator>Steinbrenner, Adam D. ; Saldivar, Evan ; Hodges, Nile ; Guayazán‐Palacios, Natalia ; Chaparro, Antonio F. ; Schmelz, Eric A.</creator><creatorcontrib>Steinbrenner, Adam D. ; Saldivar, Evan ; Hodges, Nile ; Guayazán‐Palacios, Natalia ; Chaparro, Antonio F. ; Schmelz, Eric A.</creatorcontrib><description>SUMMARY
Chewing herbivores activate plant defense responses through a combination of mechanical wounding and elicitation by herbivore‐associated molecular patterns (HAMPs). HAMPs are wound response amplifiers; however, specific defense outputs may also exist that strictly require HAMP‐mediated defense signaling. To investigate HAMP‐mediated signaling and defense responses, we characterized cowpea (Vigna unguiculata) transcriptome changes following elicitation by inceptin, a peptide HAMP common in Lepidoptera larvae oral secretions. Following inceptin treatment, we observed large‐scale reprogramming of the transcriptome consistent with three different response categories: (i) amplification of mechanical wound responses, (ii) temporal extension through accelerated or prolonged responses, and (iii) examples of inceptin‐specific elicitation and suppression. At both early and late timepoints, namely 1 and 6 h, large sets of transcripts specifically accumulated following inceptin elicitation. Further early inceptin‐regulated transcripts were classified as reversing changes induced by wounding alone. Within key signaling‐ and defense‐related gene families, inceptin‐elicited responses included target subsets of wound‐induced transcripts. Transcripts displaying the largest inceptin‐elicited fold changes included transcripts encoding terpene synthases (TPSs) and peroxidases (POXs) that correspond with induced volatile production and increased POX activity in cowpea. Characterization of inceptin‐elicited cowpea defenses via heterologous expression in Nicotiana benthamiana demonstrated that specific cowpea TPSs and POXs were able to confer terpene emission and the reduced growth of beet armyworm (Spodoptera exigua) herbivores, respectively. Collectively, our present findings in cowpea support a model where HAMP elicitation both amplifies concurrent wound responses and specifically contributes to the activation of selective outputs associated with direct and indirect antiherbivore defenses.
Significance Statement
Plants recognize herbivore‐associated molecular patterns (HAMPs) and increase defense production, but interactions with the more general wound response are not well understood. We examined an established HAMP–receptor interaction to characterize the transcriptomic modulation of wound responses by the HAMP peptide inceptin. Inceptin not only amplifies wound responses, but can also specifically induce or suppress transcripts with demonstrated functions in direct and indirect defense against herbivores. The plant immune system thus recognizes HAMPs to fine‐tune wound responses against insect herbivores.</description><identifier>ISSN: 0960-7412</identifier><identifier>EISSN: 1365-313X</identifier><identifier>DOI: 10.1111/tpj.15732</identifier><identifier>PMID: 35315556</identifier><language>eng</language><publisher>England: Blackwell Publishing Ltd</publisher><subject>Animals ; cowpea ; Cowpeas ; Defense mechanisms ; Emissions control ; Fabaceae - genetics ; Gene Expression Regulation, Plant ; Gene families ; HAMP ; Herbivores ; Herbivory - physiology ; inceptin ; INR ; Larvae ; Legumes ; peroxidase ; Plants ; Secretions ; Signaling ; Spodoptera ; Spodoptera exigua ; terpene ; Terpenes - metabolism ; Transcriptomes ; Vigna - genetics ; Vigna unguiculata ; Wounding</subject><ispartof>The Plant journal : for cell and molecular biology, 2022-06, Vol.110 (5), p.1255-1270</ispartof><rights>2022 Society for Experimental Biology and John Wiley & Sons Ltd.</rights><rights>Copyright © 2022 John Wiley & Sons Ltd and the Society for Experimental Biology</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3882-6558bd87cd7132b80c73e18f8dbb6e25b7e31ee84bdfbbc1ac7ede5ecd87494d3</citedby><cites>FETCH-LOGICAL-c3882-6558bd87cd7132b80c73e18f8dbb6e25b7e31ee84bdfbbc1ac7ede5ecd87494d3</cites><orcidid>0000-0002-2837-734X ; 0000-0002-7493-678X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Ftpj.15732$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Ftpj.15732$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,1427,27901,27902,45550,45551,46384,46808</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/35315556$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Steinbrenner, Adam D.</creatorcontrib><creatorcontrib>Saldivar, Evan</creatorcontrib><creatorcontrib>Hodges, Nile</creatorcontrib><creatorcontrib>Guayazán‐Palacios, Natalia</creatorcontrib><creatorcontrib>Chaparro, Antonio F.</creatorcontrib><creatorcontrib>Schmelz, Eric A.</creatorcontrib><title>Signatures of plant defense response specificity mediated by herbivore‐associated molecular patterns in legumes</title><title>The Plant journal : for cell and molecular biology</title><addtitle>Plant J</addtitle><description>SUMMARY
Chewing herbivores activate plant defense responses through a combination of mechanical wounding and elicitation by herbivore‐associated molecular patterns (HAMPs). HAMPs are wound response amplifiers; however, specific defense outputs may also exist that strictly require HAMP‐mediated defense signaling. To investigate HAMP‐mediated signaling and defense responses, we characterized cowpea (Vigna unguiculata) transcriptome changes following elicitation by inceptin, a peptide HAMP common in Lepidoptera larvae oral secretions. Following inceptin treatment, we observed large‐scale reprogramming of the transcriptome consistent with three different response categories: (i) amplification of mechanical wound responses, (ii) temporal extension through accelerated or prolonged responses, and (iii) examples of inceptin‐specific elicitation and suppression. At both early and late timepoints, namely 1 and 6 h, large sets of transcripts specifically accumulated following inceptin elicitation. Further early inceptin‐regulated transcripts were classified as reversing changes induced by wounding alone. Within key signaling‐ and defense‐related gene families, inceptin‐elicited responses included target subsets of wound‐induced transcripts. Transcripts displaying the largest inceptin‐elicited fold changes included transcripts encoding terpene synthases (TPSs) and peroxidases (POXs) that correspond with induced volatile production and increased POX activity in cowpea. Characterization of inceptin‐elicited cowpea defenses via heterologous expression in Nicotiana benthamiana demonstrated that specific cowpea TPSs and POXs were able to confer terpene emission and the reduced growth of beet armyworm (Spodoptera exigua) herbivores, respectively. Collectively, our present findings in cowpea support a model where HAMP elicitation both amplifies concurrent wound responses and specifically contributes to the activation of selective outputs associated with direct and indirect antiherbivore defenses.
Significance Statement
Plants recognize herbivore‐associated molecular patterns (HAMPs) and increase defense production, but interactions with the more general wound response are not well understood. We examined an established HAMP–receptor interaction to characterize the transcriptomic modulation of wound responses by the HAMP peptide inceptin. Inceptin not only amplifies wound responses, but can also specifically induce or suppress transcripts with demonstrated functions in direct and indirect defense against herbivores. The plant immune system thus recognizes HAMPs to fine‐tune wound responses against insect herbivores.</description><subject>Animals</subject><subject>cowpea</subject><subject>Cowpeas</subject><subject>Defense mechanisms</subject><subject>Emissions control</subject><subject>Fabaceae - genetics</subject><subject>Gene Expression Regulation, Plant</subject><subject>Gene families</subject><subject>HAMP</subject><subject>Herbivores</subject><subject>Herbivory - physiology</subject><subject>inceptin</subject><subject>INR</subject><subject>Larvae</subject><subject>Legumes</subject><subject>peroxidase</subject><subject>Plants</subject><subject>Secretions</subject><subject>Signaling</subject><subject>Spodoptera</subject><subject>Spodoptera exigua</subject><subject>terpene</subject><subject>Terpenes - metabolism</subject><subject>Transcriptomes</subject><subject>Vigna - genetics</subject><subject>Vigna unguiculata</subject><subject>Wounding</subject><issn>0960-7412</issn><issn>1365-313X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kctu1TAQhi0EoofCghdAltiURdqMHV_OElW0UFUCiSKxi3yZFB8lcWonVGfHI_CMPAkuaVkg4c1YM9_8mpmfkJdQH0N5J_O0OwahOHtENsClqDjwr4_Jpt7KulINsAPyLOddXYPisnlKDrjgIISQG3LzOVyPZl4SZho7OvVmnKnHDseMtCSnePfJE7rQBRfmPR3QBzOjp3ZPv2Gy4XtM-OvHT5NzdGtliD26pTeJTmaeMY2ZhpH2eL0MmJ-TJ53pM764j4fky9m7q9P31eXH8w-nby8rx7VmlRRCW6-V8wo4s7p2iiPoTntrJTJhFXJA1I31nbUOjFPoUaArPc228fyQHK26U4o3C-a5HUJ22JcNMS65ZbIBLQWobUFf_4Pu4pLGMl2hFGNCArBCvVkpl2LOCbt2SmEwad9C3d750BYf2j8-FPbVveJiy8H-kg-HL8DJCtyGHvf_V2qvPl2skr8BkoCV3Q</recordid><startdate>202206</startdate><enddate>202206</enddate><creator>Steinbrenner, Adam D.</creator><creator>Saldivar, Evan</creator><creator>Hodges, Nile</creator><creator>Guayazán‐Palacios, Natalia</creator><creator>Chaparro, Antonio F.</creator><creator>Schmelz, Eric A.</creator><general>Blackwell Publishing Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>7QP</scope><scope>7QR</scope><scope>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-2837-734X</orcidid><orcidid>https://orcid.org/0000-0002-7493-678X</orcidid></search><sort><creationdate>202206</creationdate><title>Signatures of plant defense response specificity mediated by herbivore‐associated molecular patterns in legumes</title><author>Steinbrenner, Adam D. ; Saldivar, Evan ; Hodges, Nile ; Guayazán‐Palacios, Natalia ; Chaparro, Antonio F. ; Schmelz, Eric A.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3882-6558bd87cd7132b80c73e18f8dbb6e25b7e31ee84bdfbbc1ac7ede5ecd87494d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Animals</topic><topic>cowpea</topic><topic>Cowpeas</topic><topic>Defense mechanisms</topic><topic>Emissions control</topic><topic>Fabaceae - genetics</topic><topic>Gene Expression Regulation, Plant</topic><topic>Gene families</topic><topic>HAMP</topic><topic>Herbivores</topic><topic>Herbivory - physiology</topic><topic>inceptin</topic><topic>INR</topic><topic>Larvae</topic><topic>Legumes</topic><topic>peroxidase</topic><topic>Plants</topic><topic>Secretions</topic><topic>Signaling</topic><topic>Spodoptera</topic><topic>Spodoptera exigua</topic><topic>terpene</topic><topic>Terpenes - metabolism</topic><topic>Transcriptomes</topic><topic>Vigna - genetics</topic><topic>Vigna unguiculata</topic><topic>Wounding</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Steinbrenner, Adam D.</creatorcontrib><creatorcontrib>Saldivar, Evan</creatorcontrib><creatorcontrib>Hodges, Nile</creatorcontrib><creatorcontrib>Guayazán‐Palacios, Natalia</creatorcontrib><creatorcontrib>Chaparro, Antonio F.</creatorcontrib><creatorcontrib>Schmelz, Eric A.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>The Plant journal : for cell and molecular biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Steinbrenner, Adam D.</au><au>Saldivar, Evan</au><au>Hodges, Nile</au><au>Guayazán‐Palacios, Natalia</au><au>Chaparro, Antonio F.</au><au>Schmelz, Eric A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Signatures of plant defense response specificity mediated by herbivore‐associated molecular patterns in legumes</atitle><jtitle>The Plant journal : for cell and molecular biology</jtitle><addtitle>Plant J</addtitle><date>2022-06</date><risdate>2022</risdate><volume>110</volume><issue>5</issue><spage>1255</spage><epage>1270</epage><pages>1255-1270</pages><issn>0960-7412</issn><eissn>1365-313X</eissn><abstract>SUMMARY
Chewing herbivores activate plant defense responses through a combination of mechanical wounding and elicitation by herbivore‐associated molecular patterns (HAMPs). HAMPs are wound response amplifiers; however, specific defense outputs may also exist that strictly require HAMP‐mediated defense signaling. To investigate HAMP‐mediated signaling and defense responses, we characterized cowpea (Vigna unguiculata) transcriptome changes following elicitation by inceptin, a peptide HAMP common in Lepidoptera larvae oral secretions. Following inceptin treatment, we observed large‐scale reprogramming of the transcriptome consistent with three different response categories: (i) amplification of mechanical wound responses, (ii) temporal extension through accelerated or prolonged responses, and (iii) examples of inceptin‐specific elicitation and suppression. At both early and late timepoints, namely 1 and 6 h, large sets of transcripts specifically accumulated following inceptin elicitation. Further early inceptin‐regulated transcripts were classified as reversing changes induced by wounding alone. Within key signaling‐ and defense‐related gene families, inceptin‐elicited responses included target subsets of wound‐induced transcripts. Transcripts displaying the largest inceptin‐elicited fold changes included transcripts encoding terpene synthases (TPSs) and peroxidases (POXs) that correspond with induced volatile production and increased POX activity in cowpea. Characterization of inceptin‐elicited cowpea defenses via heterologous expression in Nicotiana benthamiana demonstrated that specific cowpea TPSs and POXs were able to confer terpene emission and the reduced growth of beet armyworm (Spodoptera exigua) herbivores, respectively. Collectively, our present findings in cowpea support a model where HAMP elicitation both amplifies concurrent wound responses and specifically contributes to the activation of selective outputs associated with direct and indirect antiherbivore defenses.
Significance Statement
Plants recognize herbivore‐associated molecular patterns (HAMPs) and increase defense production, but interactions with the more general wound response are not well understood. We examined an established HAMP–receptor interaction to characterize the transcriptomic modulation of wound responses by the HAMP peptide inceptin. Inceptin not only amplifies wound responses, but can also specifically induce or suppress transcripts with demonstrated functions in direct and indirect defense against herbivores. The plant immune system thus recognizes HAMPs to fine‐tune wound responses against insect herbivores.</abstract><cop>England</cop><pub>Blackwell Publishing Ltd</pub><pmid>35315556</pmid><doi>10.1111/tpj.15732</doi><tpages>1270</tpages><orcidid>https://orcid.org/0000-0002-2837-734X</orcidid><orcidid>https://orcid.org/0000-0002-7493-678X</orcidid><oa>free_for_read</oa></addata></record> |
fulltext | fulltext |
identifier | ISSN: 0960-7412 |
ispartof | The Plant journal : for cell and molecular biology, 2022-06, Vol.110 (5), p.1255-1270 |
issn | 0960-7412 1365-313X |
language | eng |
recordid | cdi_proquest_miscellaneous_2641865179 |
source | Wiley Free Content; MEDLINE; IngentaConnect Free/Open Access Journals; Wiley Online Library Journals Frontfile Complete; EZB-FREE-00999 freely available EZB journals |
subjects | Animals cowpea Cowpeas Defense mechanisms Emissions control Fabaceae - genetics Gene Expression Regulation, Plant Gene families HAMP Herbivores Herbivory - physiology inceptin INR Larvae Legumes peroxidase Plants Secretions Signaling Spodoptera Spodoptera exigua terpene Terpenes - metabolism Transcriptomes Vigna - genetics Vigna unguiculata Wounding |
title | Signatures of plant defense response specificity mediated by herbivore‐associated molecular patterns in legumes |
url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-05T17%3A52%3A16IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Signatures%20of%20plant%20defense%20response%20specificity%20mediated%20by%20herbivore%E2%80%90associated%20molecular%20patterns%20in%20legumes&rft.jtitle=The%20Plant%20journal%20:%20for%20cell%20and%20molecular%20biology&rft.au=Steinbrenner,%20Adam%20D.&rft.date=2022-06&rft.volume=110&rft.issue=5&rft.spage=1255&rft.epage=1270&rft.pages=1255-1270&rft.issn=0960-7412&rft.eissn=1365-313X&rft_id=info:doi/10.1111/tpj.15732&rft_dat=%3Cproquest_cross%3E2672256112%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2672256112&rft_id=info:pmid/35315556&rfr_iscdi=true |