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<input type="email" aria-label="Sender Email Address" placeholder="email@example.com" id="email-from" class="email-from" pattern="^[a-zA-Z0-9_.+-]+@[a-zA-Z0-9-]+\.[a-zA-Z0-9-.]+$" maxlength="256">
</div>
<div class="action-panel-control-wrap">
<label for="email-citation-format" class="action-panel-label">
Format:
</label>
<select id="email-citation-format" name="citation-format" class="action-panel-selector email-citation-format">
<option selected="selected" value="summary">Summary</option>
<option value="summary-text">Summary (text)</option>
<option value="abstract">Abstract</option>
<option value="abstract-text">Abstract (text)</option>
</select>
</div>
<div class="include-supplemental-container">
<input type="checkbox" aria-label="Include MeSH and other data" name="include-supplemental" id="email-include-supplemental" class="email-include-supplemental">
<label for="email-include-supplemental" class="email-include-supplemental-label">MeSH and other data</label>
</div>
<div class="form-field recaptcha ">
<div class="g-recaptcha" id="id-recaptcha" data-sitekey="6LfsWHMdAAAAAClKbtOpjQ2pMjgsGxvv7NdZW9uI"></div>
</div>
<div id="captcha-error-message" class="usa-input-error-message captcha-validation-message" role="alert"></div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Sending..."
data-ga-category="save_share"
data-ga-action="email"
data-ga-label="send">
Send email
</button>
<button class="action-panel-cancel"
aria-label="Close 'Email citations' panel"
ref="linksrc=close_email_panel"
aria-controls="email-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="email"
data-ga-label="cancel">
Cancel
</button>
</div>
<input type="hidden" name="email-search-details" value="" />
<input type="hidden" name="email-search-details-hash" value="0e42663a6c3bd85498fcb88798998fed7bfdc45d457db35281e41afe13cc0524" />
</form>
</div>
</div>
<div id="collections-action-panel"
class="collections-action-panel action-panel in-progress-dots-panel"
aria-hidden="true"
data-collections-open-panel-enabled="false"
data-collections-open-panel-url-hash="#open-collections-panel">
<div class="inner-wrap">
<h3 class="action-panel-heading">
Add to Collections
</h3>
<form id="collections-action-panel-form"
class="collections-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
data-existing-collections-url="/list-existing-collections/"
data-add-to-existing-collection-url="/add-to-existing-collection/"
data-create-and-add-to-new-collection-url="/create-and-add-to-new-collection/"
data-get-article-ids-by-search-url="/get-article-ids-by-search/"
data-myncbi-max-collection-name-length="100"
data-add-to-collection-max-amount="1000"
data-collections-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/">
<input type="hidden" name="csrfmiddlewaretoken" value="c8vuT60h4vQKdz9INAfXkyCnBJiBscY7tVfzwWtDE6De70fyqTSZOG819N0NlNwr">
<div class="choice-group" role="radiogroup">
<ul class="radio-group-items">
<li>
<input type="radio"
id="collections-action-panel-new"
class="collections-new"
name="collections"
value="new"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="collections_radio_new">
<label for="collections-action-panel-new">Create a new collection</label>
</li>
<li>
<input type="radio"
id="collections-action-panel-existing"
class="collections-existing"
name="collections"
value="existing"
checked="true"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="collections_radio_existing">
<label for="collections-action-panel-existing">Add to an existing collection</label>
</li>
</ul>
</div>
<div class="controls-wrapper">
<div class="action-panel-control-wrap new-collections-controls">
<label for="collections-action-panel-add-to-new" class="action-panel-label required-field-asterisk">
Name your collection:
</label>
<input
type="text"
name="add-to-new-collection"
id="collections-action-panel-add-to-new"
class="collections-action-add-to-new"
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/"
maxlength="100"
data-ga-category="save_share"
data-ga-action="create_collection"
data-ga-label="non_favorties_collection">
<div class="collections-new-name-too-long usa-input-error-message selection-validation-message">
Name must be less than 100 characters
</div>
</div>
<div class="action-panel-control-wrap existing-collections-controls">
<label for="collections-action-panel-add-to-existing" class="action-panel-label">
Choose a collection:
</label>
<select id="collections-action-panel-add-to-existing"
class="action-panel-selector collections-action-add-to-existing"
data-ga-category="save_share"
data-ga-action="select_collection"
data-ga-label="($('#collections-action-add-to-existing').val() === 'Favorites') ? 'Favorites' : 'non_favorites_collection'">
</select>
<div class="collections-retry-load-on-error usa-input-error-message selection-validation-message">
Unable to load your collection due to an error<br>
<a href="#">Please try again</a>
</div>
</div>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Adding..."
data-pinger-ignore
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="add">
Add
</button>
<button class="action-panel-cancel"
aria-label="Close 'Add to Collections' panel"
ref="linksrc=close_collections_panel"
aria-controls="collections-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="bibliography-action-panel"
class="bibliography-action-panel action-panel in-progress-dots-panel"
aria-hidden="true"
data-bibliography-open-panel-enabled="false"
data-bibliography-open-panel-url-hash="#open-bibliography-panel">
<div class="inner-wrap">
<h3 class="action-panel-heading">
Add to My Bibliography
</h3>
<form id="bibliography-action-panel-form"
class="bibliography-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
data-add-to-bibliography-max-amount="100"
data-add-to-bibliography-batch-size="10"
data-bibliography-delegates-url="/list-bibliography-delegates/"
data-add-to-bibliography-url="/add-to-bibliography/"
data-get-article-ids-by-search-url="/get-article-ids-by-search/"
data-mybib-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/mybibliography/">
<input type="hidden" name="csrfmiddlewaretoken" value="c8vuT60h4vQKdz9INAfXkyCnBJiBscY7tVfzwWtDE6De70fyqTSZOG819N0NlNwr">
<div class="action-panel-control-wrap bibliographies-controls">
<div class="choice-group">
<ul class="bibliographies-action-add radio-group-items">
<li>
<input name="bibliography" id="my-bibliography" class="my-bibliography" type="radio" checked/>
<label for="my-bibliography">My Bibliography</label>
</li>
</ul>
</div>
</div>
<div class="bibliographies-retry-load-on-error usa-input-error-message selection-validation-message">
Unable to load your delegates due to an error<br>
<a href="#">Please try again</a>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Adding..."
data-pinger-ignore>
Add
</button>
<button class="action-panel-cancel"
aria-label="Close 'Add to bibliography' panel"
ref="linksrc=close_bibliography_panel"
aria-controls="bibliography-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="mybib"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="saved-search-action-panel" class="saved-search-action-panel action-panel " aria-hidden="true"
data-saved-search-open-panel-enabled="false"
data-saved-search-open-panel-url-hash="#open-saved-search-panel">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Your saved search
</h2>
<form id="saved-search-action-panel-form"
class="saved-search-action-panel-form action-panel-content action-form"
data-create-saved-search-url="/create-saved-search/"
data-try-search-terms-url="/try-search-term/"
data-saved-search-root-url="https://www.ncbi.nlm.nih.gov/myncbi/searches/">
<input type="hidden" name="csrfmiddlewaretoken" value="c8vuT60h4vQKdz9INAfXkyCnBJiBscY7tVfzwWtDE6De70fyqTSZOG819N0NlNwr">
<div class="action-panel-control-wrap">
<label for="saved-search-name" class="action-panel-label saved-search-name-label required-field-asterisk">
Name of saved search:
</label>
<input maxlength="200"
type="text"
name="saved-search-name"
id="saved-search-name"
class="saved-search-name"
value=""
required
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/">
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-term" class="action-panel-label required-field-asterisk">
Search terms:
</label>
<textarea name="saved-search-term" id="saved-search-term" class="saved-search-term" required></textarea>
</div>
<div class="test-search-term-wrap">
<a href="#" class="try-search-term">Test search terms</a>
</div>
<div class="choice-group action-panel-extra-margin-top">
<span class="action-panel-label" id="fieldset-label">
Would you like email updates of new search results?
</span>
<fieldset id="saved-search-alert" aria-describedby="fieldset-label">
<legend class="usa-sr-only">Saved Search Alert Radio Buttons</legend>
<ul class="radio-group-items">
<li>
<input type="radio" id="saved-search-alert-yes" class="saved-search-alert-yes" name="saved-search-alert" value="yes" checked>
<label for="saved-search-alert-yes" class="action-panel-label">Yes</label>
</li>
<li>
<input aria-label="No radio input" type="radio" id="saved-search-alert-no" class="saved-search-alert-no" name="saved-search-alert" value="no">
<label for="saved-search-alert-no" class="action-panel-label">No</label>
</li>
</ul>
</fieldset>
</div>
<div class="alert-schedule-wrap">
<div class="action-panel-control-wrap">
<label class="action-panel-label">
Email:
</label>
<span aria-label="Email address" id="saved-search-email" class="action-panel-label"><span class="action-panel-label-bold"></span> (<a class="myncbi-account-settings" href="https://www.ncbi.nlm.nih.gov/account/settings/">change</a>)</span>
</div>
<div class="action-panel-control-wrap action-panel-extra-margin-top">
<label for="saved-search-frequency" class="action-panel-label">
Frequency:
</label>
<select id="saved-search-frequency" class="no-border-panel-selector saved-search-frequency">
<option value="monthly">Monthly</option>
<option value="weekly">Weekly</option>
<option value="daily">Daily</option>
</select>
</div>
<div class="action-panel-control-wrap saved-search-monthly-additional">
<label for="saved-search-monthly-on-day" class="action-panel-label">
Which day?
</label>
<select id="saved-search-monthly-on-day" class="no-border-panel-selector">
<option value="Sunday">The first Sunday</option>
<option value="Monday">The first Monday</option>
<option value="Tuesday">The first Tuesday</option>
<option value="Wednesday">The first Wednesday</option>
<option value="Thursday">The first Thursday</option>
<option value="Friday">The first Friday</option>
<option value="Saturday">The first Saturday</option>
<option value="day">The first day</option>
<option value="weekday">The first weekday</option>
</select>
</div>
<div class="action-panel-control-wrap saved-search-weekly-additional">
<label for="saved-search-weekly-on-day" class="action-panel-label">
Which day?
</label>
<select id="saved-search-weekly-on-day" class="no-border-panel-selector saved-search-weekly-on-day">
<option value="Sunday">Sunday</option>
<option value="Monday">Monday</option>
<option value="Tuesday">Tuesday</option>
<option value="Wednesday">Wednesday</option>
<option value="Thursday">Thursday</option>
<option value="Friday">Friday</option>
<option value="Saturday">Saturday</option>
</select>
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-report" class="action-panel-label">
Report format:
</label>
<select id="saved-search-report" class="no-border-panel-selector saved-search-report">
<option value="DocSum">Summary</option>
<option value="DocSumText">Summary (text)</option>
<option value="Abstract">Abstract</option>
<option value="AbstractText">Abstract (text)</option>
<option value="MEDLINE">PubMed</option>
</select>
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-amount" class="action-panel-label">
Send at most:
</label>
<select id="saved-search-amount" class="no-border-panel-selector saved-search-amount">
<option value="1">1 item</option>
<option value="5" selected>5 items</option>
<option value="10">10 items</option>
<option value="20">20 items</option>
<option value="50">50 items</option>
<option value="100">100 items</option>
<option value="200">200 items</option>
</select>
</div>
<div>
<input type="checkbox" id="saved-search-send-if-no-result" class="saved-search-send-if-no-result" name="saved-search-send-if-no-result">
<label for="saved-search-send-if-no-result" class="action-panel-label smaller-checkbox">
Send even when there aren't any new results
</label>
</div>
<div class="action-panel-control-wrap option-text-in-email-wrap">
<label for="saved-search-email-text" class="action-panel-label">
Optional text in email:
</label>
<textarea name="saved-search-email-text"
id="saved-search-email-text"
class="saved-search-email-text"></textarea>
</div>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Saving..."
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="save">
Save
</button>
<button class="action-panel-cancel"
aria-label="Close 'Your saved search' panel"
ref="linksrc=close_saved_search_panel"
aria-controls="saved-search-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="citation-manager-action-panel" class="citation-manager-action-panel action-panel" aria-hidden="true">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Create a file for external citation management software
</h2>
<form id="citation-manager-action-panel-form"
class="action-panel-content action-form"
action="/results-export-ids/"
data-by-search-action="/results-export-search-data/"
data-by-ids-action="/results-export-ids/"
method="post"
data-by-search-method="post"
data-by-ids-method="post">
<input type="hidden" name="csrfmiddlewaretoken" value="c8vuT60h4vQKdz9INAfXkyCnBJiBscY7tVfzwWtDE6De70fyqTSZOG819N0NlNwr">
<input name="results-format" type="hidden" value="pubmed"/>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Sending..."
data-ga-category="save_share"
data-ga-action="citation_manager"
data-ga-label="save">
Create file
</button>
<button class="action-panel-cancel"
aria-label="Close 'Send citations to citation manager' panel"
ref="linksrc=close_citation_manager_panel"
aria-controls="citation-manager-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="citation_manager"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="rss-action-panel" class="rss-action-panel action-panel " aria-hidden="true">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Your RSS Feed
</h2>
<form id="rss-action-panel-form"
class="rss-action-panel-form action-panel-content action-form"
data-create-rss-feed-url="/create-rss-feed-url/"
data-search-form-term-value="">
<input type="hidden" name="csrfmiddlewaretoken" value="c8vuT60h4vQKdz9INAfXkyCnBJiBscY7tVfzwWtDE6De70fyqTSZOG819N0NlNwr">
<div class="action-panel-control-wrap">
<label for="rss-name" class="action-panel-label required-field-asterisk">
Name of RSS Feed:
</label>
<input maxlength="200"
placeholder="Name"
type="text"
name="rss-name"
id="rss-name"
class="rss-name"
value=''
required
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/">
</div>
<div class="rss-limit-wrap">
<div class="action-panel-control-wrap action-panel-extra-margin-top">
<label for="rss-limit" class="action-panel-label">
Number of items displayed:
</label>
<select id="rss-limit" class="no-border-panel-selector rss-limit">
<option value="5">5</option>
<option value="10">10</option>
<option value="15" selected="selected">15</option>
<option value="20">20</option>
<option value="50">50</option>
<option value="100">100</option>
</select>
</div>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Creating..."
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="save">
Create RSS
</button>
<button class="action-panel-cancel"
aria-label="Close 'Your RSS' panel"
ref="linksrc=close_rss_panel"
aria-controls="rss-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="cancel">
Cancel
</button>
</div>
<div class="action-panel-control-wrap rss-link-copy-wrap">
<label for="rss-link" class="usa-sr-only">RSS Link</label>
<input placeholder="Your RSS Feed Link" type="text" name="rss-link" id="rss-link" class="rss-link" title="RSS Link">
<button
type="button"
disabled
class="rss-link-copy-button disabled"
data-ga-category="save_share"
data-ga-action="rss"
data-ga-label="copy">
Copy
</button>
</div>
</form>
</div>
</div>
</div>
</div>
<div class="article-page" id="article-page" data-article-pmid="33608298">
<aside class="page-sidebar">
<div class="inner-wrap">
<div class="full-text-links">
<div class="full-view">
<h3 class="title">
Full text links
</h3>
<div class="full-text-links-list">
<a class="link-item
dialog-focus"
href="https://journals.asm.org/doi/10.1128/AEM.00091-21?url_ver=Z39.88-2003&amp;rfr_id=ori:rid:crossref.org&amp;rfr_dat=cr_pub 0pubmed"
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ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=False&amp;PrId=4487&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=33608298&amp;db=pubmed&amp;nlmid=7605801"
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pmc
"
href="https://pmc.ncbi.nlm.nih.gov/articles/pmid/33608298/"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=True&amp;PrId=3494&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=33608298&amp;db=pubmed&amp;nlmid=7605801"
title="Free full text at PubMed Central"
data-ga-category="full_text"
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Free PMC article
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</div>
<div class="short-view">
<a href="#" class="full-text-links-button full-text-links-dialog-trigger">
Full text links
</a>
</div>
</div>
<div class="actions-buttons sidebar"><h3 class="title">Actions</h3><div class="inner-wrap"><button class="citation-button citation-dialog-trigger"
aria-label="Open dialog with citation text in different styles"
data-ga-category="save_share"
data-ga-action="cite"
data-ga-label="open"
data-all-citations-url="/33608298/citations/"
data-citation-style="nlm"
data-pubmed-format-link="/33608298/export/"><span class="button-label">Cite</span></button><link type="text/css" href="ncbi-overlay-block/src/overlay-block.css"><div class="collections-button-container" data-article-id="33608298" data-article-db="pubmed"><button class="collections-button collections-dialog-trigger"
aria-label="Save article in MyNCBI collections."
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_button"
data-collections-open-dialog-enabled="false"
data-collections-open-dialog-url="https://account.ncbi.nlm.nih.gov/?back_url=https%3A%2F%2Fpubmed.ncbi.nlm.nih.gov%2F33608298%2F%23open-collections-dialog"
data-in-collections="false"><span class="button-label">Collections</span></button><div class="overlay" role="dialog"><div id="collections-action-dialog"
class="dialog collections-dialog"
aria-hidden="true"><div class="title">Add to Collections</div><div class="collections-action-panel action-panel"><form id="collections-action-dialog-form"
class="collections-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
data-existing-collections-url="/list-existing-collections/"
data-add-to-existing-collection-url="/add-to-existing-collection/"
data-create-and-add-to-new-collection-url="/create-and-add-to-new-collection/"
data-myncbi-max-collection-name-length="100"
data-collections-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/"><input type="hidden" name="csrfmiddlewaretoken" value="c8vuT60h4vQKdz9INAfXkyCnBJiBscY7tVfzwWtDE6De70fyqTSZOG819N0NlNwr"><div class="choice-group" role="radiogroup"><ul class="radio-group-items"><li><input type="radio"
id="collections-action-dialog-new"
class="collections-new"
name="collections"
value="new"
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_radio_new"><label for="collections-action-dialog-new">Create a new collection</label></li><li><input type="radio"
id="collections-action-dialog-existing"
class="collections-existing"
name="collections"
value="existing"
checked="true"
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_radio_existing"><label for="collections-action-dialog-existing">Add to an existing collection</label></li></ul></div><div class="controls-wrapper"><div class="action-panel-control-wrap new-collections-controls"><label for="collections-action-dialog-add-to-new" class="action-panel-label required-field-asterisk">
Name your collection:
</label><input
type="text"
name="add-to-new-collection"
id="collections-action-dialog-add-to-new"
class="collections-action-add-to-new"
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/"
maxlength="100"
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<h1 class="heading-title">
Biosurfactants Produced by Phyllosphere-Colonizing Pseudomonads Impact Diesel Degradation but Not Colonization of Leaves of Gnotobiotic Arabidopsis thaliana
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Biosurfactants Produced by Phyllosphere-Colonizing Pseudomonads Impact Diesel Degradation but Not Colonization of Leaves of Gnotobiotic Arabidopsis thaliana
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<span class="authors-list-item "><a class="full-name"
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data-ga-label="S Oso">S Oso</a><sup class="affiliation-links"><span class="author-sup-separator">&nbsp;</span><a class="affiliation-link" title="School of Biological Sciences, University of Canterbury, Christchurch, New Zealand." href="#short-view-affiliation-1" ref="linksrc=author_aff">
1
</a></sup><span class="comma">,&nbsp;</span></span><span class="authors-list-item "><a class="full-name"
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data-ga-label="F Fuchs">F Fuchs</a><sup class="affiliation-links"><span class="author-sup-separator">&nbsp;</span><a class="affiliation-link" title="Institute for Cellular and Molecular Botany, University of Bonn, Bonn, Germany." href="#short-view-affiliation-2" ref="linksrc=author_aff">
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data-ga-label="C Übermuth">C Übermuth</a><sup class="affiliation-links"><span class="author-sup-separator">&nbsp;</span><a class="affiliation-link" title="Institute for Cellular and Molecular Botany, University of Bonn, Bonn, Germany." href="#short-view-affiliation-2" ref="linksrc=author_aff">
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data-ga-label="L Zander">L Zander</a><sup class="affiliation-links"><span class="author-sup-separator">&nbsp;</span><a class="affiliation-link" title="Institute for Cellular and Molecular Botany, University of Bonn, Bonn, Germany." href="#short-view-affiliation-2" ref="linksrc=author_aff">
2
</a></sup><span class="comma">,&nbsp;</span></span><span class="authors-list-item "><a class="full-name"
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data-ga-label="S Daunaraviciute">S Daunaraviciute</a><sup class="affiliation-links"><span class="author-sup-separator">&nbsp;</span><a class="affiliation-link" title="Institute for Cellular and Molecular Botany, University of Bonn, Bonn, Germany." href="#short-view-affiliation-2" ref="linksrc=author_aff">
2
</a></sup><span class="comma">,&nbsp;</span></span><span class="authors-list-item "><a class="full-name"
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data-ga-label="D M Remus">D M Remus</a><sup class="affiliation-links"><span class="author-sup-separator">&nbsp;</span><a class="affiliation-link" title="School of Biological Sciences, University of Canterbury, Christchurch, New Zealand." href="#short-view-affiliation-1" ref="linksrc=author_aff">
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</a><span class="author-sup-separator">&nbsp;</span><a class="affiliation-link" title="Protein Science &amp; Engineering, Callaghan Innovation, Christchurch, New Zealand." href="#short-view-affiliation-3" ref="linksrc=author_aff">
3
</a></sup><span class="comma">,&nbsp;</span></span><span class="authors-list-item "><a class="full-name"
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data-ga-label="I Stötzel">I Stötzel</a><sup class="affiliation-links"><span class="author-sup-separator">&nbsp;</span><a class="affiliation-link" title="Institute of Nutritional and Food Sciences, University of Bonn, Bonn, Germany." href="#short-view-affiliation-4" ref="linksrc=author_aff">
4
</a></sup><span class="comma">,&nbsp;</span></span><span class="authors-list-item "><a class="full-name"
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data-ga-label="M Wüst">M Wüst</a><sup class="affiliation-links"><span class="author-sup-separator">&nbsp;</span><a class="affiliation-link" title="Institute of Nutritional and Food Sciences, University of Bonn, Bonn, Germany." href="#short-view-affiliation-4" ref="linksrc=author_aff">
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</a></sup><span class="comma">,&nbsp;</span></span><span class="authors-list-item "><a class="full-name"
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data-ga-label="L Schreiber">L Schreiber</a><sup class="affiliation-links"><span class="author-sup-separator">&nbsp;</span><a class="affiliation-link" title="Institute for Cellular and Molecular Botany, University of Bonn, Bonn, Germany lukas.schreiber@uni-bonn.de mitja.remus-emsermann@canterbury.ac.nz." href="#short-view-affiliation-5" ref="linksrc=author_aff">
5
</a></sup><span class="comma">,&nbsp;</span></span><span class="authors-list-item "><a class="full-name"
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data-ga-label="M N P Remus-Emsermann">M N P Remus-Emsermann</a><sup class="affiliation-links"><span class="author-sup-separator">&nbsp;</span><a class="affiliation-link" title="School of Biological Sciences, University of Canterbury, Christchurch, New Zealand lukas.schreiber@uni-bonn.de mitja.remus-emsermann@canterbury.ac.nz." href="#short-view-affiliation-6" ref="linksrc=author_aff">
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</a><span class="author-sup-separator">&nbsp;</span><a class="affiliation-link" title="Biomolecular Interaction Centre, Christchurch, New Zealand." href="#short-view-affiliation-7" ref="linksrc=author_aff">
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</a><span class="author-sup-separator">&nbsp;</span><a class="affiliation-link" title="Bio-Protection Research Centre, School of Biological Sciences, University of Canterbury, Christchurch, New Zealand." href="#short-view-affiliation-8" ref="linksrc=author_aff">
8
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Affiliations
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<li data-affiliation-id="short-view-affiliation-1"
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><sup class="key">1</sup> School of Biological Sciences, University of Canterbury, Christchurch, New Zealand.</li>
<li data-affiliation-id="short-view-affiliation-2"
id="short-view-affiliation-2"
><sup class="key">2</sup> Institute for Cellular and Molecular Botany, University of Bonn, Bonn, Germany.</li>
<li data-affiliation-id="short-view-affiliation-3"
id="short-view-affiliation-3"
><sup class="key">3</sup> Protein Science &amp; Engineering, Callaghan Innovation, Christchurch, New Zealand.</li>
<li data-affiliation-id="short-view-affiliation-4"
id="short-view-affiliation-4"
><sup class="key">4</sup> Institute of Nutritional and Food Sciences, University of Bonn, Bonn, Germany.</li>
<li data-affiliation-id="short-view-affiliation-5"
id="short-view-affiliation-5"
><sup class="key">5</sup> Institute for Cellular and Molecular Botany, University of Bonn, Bonn, Germany lukas.schreiber@uni-bonn.de mitja.remus-emsermann@canterbury.ac.nz.</li>
<li data-affiliation-id="short-view-affiliation-6"
id="short-view-affiliation-6"
><sup class="key">6</sup> School of Biological Sciences, University of Canterbury, Christchurch, New Zealand lukas.schreiber@uni-bonn.de mitja.remus-emsermann@canterbury.ac.nz.</li>
<li data-affiliation-id="short-view-affiliation-7"
id="short-view-affiliation-7"
><sup class="key">7</sup> Biomolecular Interaction Centre, Christchurch, New Zealand.</li>
<li data-affiliation-id="short-view-affiliation-8"
id="short-view-affiliation-8"
><sup class="key">8</sup> Bio-Protection Research Centre, School of Biological Sciences, University of Canterbury, Christchurch, New Zealand.</li>
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Abstract
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<p>
Biosurfactant production is a common trait in leaf surface-colonizing bacteria that has been associated with increased survival and movement on leaves. At the same time, the ability to degrade aliphatics is common in biosurfactant-producing leaf colonizers. Pseudomonads are common leaf colonizers and have been recognized for their ability to produce biosurfactants and degrade aliphatic compounds. In this study, we investigated the role of biosurfactants in four non-plant-pathogenic <i>Pseudomonas</i> strains by performing a series of experiments to characterize their surfactant properties and their role during leaf colonization and diesel degradation. The biosurfactants produced were identified using mass spectrometry. Two strains produced viscosin-like biosurfactants, and the other two produced massetolide A-like biosurfactants, which aligned with the phylogenetic relatedness between the strains. To further investigate the role of surfactant production, random Tn<i>5</i> transposon mutagenesis was performed to generate knockout mutants. The knockout mutants were compared to their respective wild types with regard to their ability to colonize gnotobiotic <i>Arabidopsis thaliana</i> and to degrade diesel or dodecane. It was not possible to detect negative effects during plant colonization in direct competition or individual colonization experiments. When grown on diesel, knockout mutants grew significantly slower than their respective wild types. When grown on dodecane, knockout mutants were less impacted than during growth on diesel. By adding isolated wild-type biosurfactants, it was possible to complement the growth of the knockout mutants.<b>IMPORTANCE</b> Many leaf-colonizing bacteria produce surfactants and are able to degrade aliphatic compounds; however, whether surfactant production provides a competitive advantage during leaf colonization is unclear. Furthermore, it is unclear if leaf colonizers take advantage of the aliphatic compounds that constitute the leaf cuticle and cuticular waxes. Here, we tested the effect of surfactant production on leaf colonization, and we demonstrate that the lack of surfactant production decreases the ability to degrade aliphatic compounds. This indicates that leaf surface-dwelling, surfactant-producing bacteria contribute to degradation of environmental hydrocarbons and may be able to utilize leaf surface waxes. This has implications for plant-microbe interactions and future studies.
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<p>
<strong class="sub-title">
Keywords:
</strong>
aliphatics; leaf cuticle; leaf wax; massetolide A; phyllosphere-inhabiting microbes; plant-microbe interactions; viscosin.
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Copyright © 2021 American Society for Microbiology.
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data-image-width="760"
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data-pmc-id="PMC8091026"
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<img class="figure-thumb" itemprop="thumbnail"
id="article-image-0"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/327c/8091026/2df1a9fb9aec/AEM.00091-21-f0001.gif"
alt="FIG 1" />
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<p> FIG 1 </p>
</strong>
<div class="figure-caption-contents"><p> Phylogenetic placement of the four… </p></div>
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</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG 1 </p>
</strong>
<div class="figure-caption-contents"><p> Phylogenetic placement of the four isolated pseudomonads. The newly sequenced isolates are highlighted… </p></div>
</div>
<figcaption id="figure-caption-0" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG 1
</strong>
<div class="figure-caption-contents">Phylogenetic placement of the four isolated pseudomonads. The newly sequenced isolates are highlighted in bold. NCBI accession numbers of the respective sequences are given after the species names. Azotobacter chroococcum was used as an outgroup.</div>
</figcaption>
</figure>
<figure class="figure-item "
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itemprop="contentUrl"
aria-describedby="figure-caption-1"
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data-image-width="760"
data-image-height="243"
data-image-alt="FIG 2"
data-pmc-id="PMC8091026"
data-figure-id="F2">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-1"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/327c/8091026/f990fb966091/AEM.00091-21-f0002.gif"
alt="FIG 2" />
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<strong class="figure-label">
<p> FIG 2 </p>
</strong>
<div class="figure-caption-contents"><p> (A to F) Atomized-oil assays… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG 2 </p>
</strong>
<div class="figure-caption-contents"><p> (A to F) Atomized-oil assays to demonstrate the production of surfactants. (A to… </p></div>
</div>
<figcaption id="figure-caption-1" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG 2
</strong>
<div class="figure-caption-contents">(A to F) Atomized-oil assays to demonstrate the production of surfactants. (A to D) Wild-type colonies of <i>P</i>FF1, <i>P</i>FF2, <i>P</i>FF3, and <i>P</i>FF4, respectively, exhibiting a halo indicative of surfactant production. (E) Tween 20. (F) E. coli DH5α. (G to L) Drop collapse assays to demonstrate the production of surfactants. (G to J) Culture supernatants of wild-type <i>P</i>FF1, <i>P</i>FF2, <i>P</i>FF3, and <i>P</i>FF4, respectively, collapsed into oil, indicative of surfactant production. (K) Collapsed drop containing Tween 20. (J) Noncollapsed drop of E. coli culture supernatant (arrow).</div>
</figcaption>
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<figure class="figure-item "
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itemprop="contentUrl"
aria-describedby="figure-caption-2"
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data-image-width="771"
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<img class="figure-thumb" itemprop="thumbnail"
id="article-image-2"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/327c/8091026/8b075f5b36ce/AEM.00091-21-f0003.gif"
alt="FIG 3" />
</a>
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<div class="caption-wrap">
<strong class="figure-label">
<p> FIG 3 </p>
</strong>
<div class="figure-caption-contents"><p> MS/MS spectra of extracted surfactants… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG 3 </p>
</strong>
<div class="figure-caption-contents"><p> MS/MS spectra of extracted surfactants of <i> P </i> FF1, <i> P </i> FF2, <i> P </i> FF3, and… </p></div>
</div>
<figcaption id="figure-caption-2" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG 3
</strong>
<div class="figure-caption-contents">MS/MS spectra of extracted surfactants of <i>P</i>FF1, <i>P</i>FF2, <i>P</i>FF3, and <i>P</i>FF4 (A to D, respectively). <i>P</i>FF1 and <i>P</i>FF2 produce viscosin-like surfactants; <i>P</i>FF3 and <i>P</i>FF4 produce massetolide A-like surfactants. Spectra were normalized against the maximal intensity.</div>
</figcaption>
</figure>
<figure class="figure-item "
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itemprop="contentUrl"
aria-describedby="figure-caption-3"
role="button"
data-image-width="771"
data-image-height="378"
data-image-alt="FIG 4"
data-pmc-id="PMC8091026"
data-figure-id="F4">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-3"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/327c/8091026/638d8f284988/AEM.00091-21-f0004.gif"
alt="FIG 4" />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="771">
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<div class="caption-wrap">
<strong class="figure-label">
<p> FIG 4 </p>
</strong>
<div class="figure-caption-contents"><p> (A to D) Atomized-oil assay… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG 4 </p>
</strong>
<div class="figure-caption-contents"><p> (A to D) Atomized-oil assay to demonstrate the production of surfactants. Tn <i> 5… </i> </p></div>
</div>
<figcaption id="figure-caption-3" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG 4
</strong>
<div class="figure-caption-contents">(A to D) Atomized-oil assay to demonstrate the production of surfactants. Tn<i>5</i> insertion mutant colonies of <i>P</i>FF1::ezTn5-visB, <i>P</i>FF2::ezTn5-visB, <i>P</i>FF3::ezTn5-massB, and <i>P</i>FF4::ezTn5-massB, respectively, lacking a halo indicative of surfactant production. (E to H) Drop collapse assays to demonstrate the production of surfactants. Culture supernatants of Tn<i>5</i> insertion mutants <i>P</i>FF1::ezTn5-visB, <i>P</i>FF2::ezTn5-visB, <i>P</i>FF3::ezTn5-massB, and <i>P</i>FF4::ezTn5-massB, respectively, showing a beaded bubble swimming on top of the oil, indicative of the lack of surfactants. The noncollapsed droplets are indicated by arrows.</div>
</figcaption>
</figure>
<figure class="figure-item "
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data-slide-index="4"
data-label-slug="fig-5">
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href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/327c/8091026/d18632d41620/AEM.00091-21-f0005.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-4"
role="button"
data-image-width="771"
data-image-height="520"
data-image-alt="FIG 5"
data-pmc-id="PMC8091026"
data-figure-id="F5">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-4"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/327c/8091026/f9dbc5a40c0d/AEM.00091-21-f0005.gif"
alt="FIG 5" />
</a>
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<strong class="figure-label">
<p> FIG 5 </p>
</strong>
<div class="figure-caption-contents"><p> Knockout mutants show no sign… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG 5 </p>
</strong>
<div class="figure-caption-contents"><p> Knockout mutants show no sign of surfactant production. MS/MS spectra of extracts of… </p></div>
</div>
<figcaption id="figure-caption-4" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG 5
</strong>
<div class="figure-caption-contents">Knockout mutants show no sign of surfactant production. MS/MS spectra of extracts of <i>P</i>FF1::ezTn5-viscB, <i>P</i>FF2::ezTn5-viscB, <i>P</i>FF3::ezTn5-massB, and <i>P</i>FF4::ezTn5-massB (A to D, respectively), are shown. None of the random knockout mutants produced detectable surfactant peaks at the corresponding wild-type <i>m/z</i> values. Spectra were normalized against the maximal intensity.</div>
</figcaption>
</figure>
<figure class="figure-item "
itemscope itemtype="http://schema.org/ImageObject"
itemprop="associatedMedia"
data-slide-index="5"
data-label-slug="fig-6">
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href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/327c/8091026/eef4cfa917bf/AEM.00091-21-f0006.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-5"
role="button"
data-image-width="800"
data-image-height="496"
data-image-alt="FIG 6"
data-pmc-id="PMC8091026"
data-figure-id="F6">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-5"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/327c/8091026/ae336fc0d2e1/AEM.00091-21-f0006.gif"
alt="FIG 6" />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="800">
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="496">
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> FIG 6 </p>
</strong>
<div class="figure-caption-contents"><p> Utilization of diesel by biosurfactant… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG 6 </p>
</strong>
<div class="figure-caption-contents"><p> Utilization of diesel by biosurfactant knockout mutants and wild types. (A) <i> P </i> FF1;… </p></div>
</div>
<figcaption id="figure-caption-5" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG 6
</strong>
<div class="figure-caption-contents">Utilization of diesel by biosurfactant knockout mutants and wild types. (A) <i>P</i>FF1; (B) <i>P</i>FF2; (C) <i>P</i>FF3; (D) <i>P</i>FF4. Each wild type and knockout mutant was grown in Bushnell-Haas broth supplemented with diesel as the sole source of carbon (circles and squares, respectively). Knockout mutants were complemented with either wild-type surfactant (triangles) or Tween 20 (inverted triangles) or were incubated with no additional carbon source (diamonds). The statistical analysis can be found in Table S3. Error bars depict the standard deviations of the means. Experiments were performed in triplicate.</div>
</figcaption>
</figure>
<figure class="figure-item tail"
itemscope itemtype="http://schema.org/ImageObject"
itemprop="associatedMedia"
data-slide-index="6"
data-label-slug="fig-7">
<a class="figure-link"
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/327c/8091026/44b0b3361f5a/AEM.00091-21-f0007.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-6"
role="button"
data-image-width="720"
data-image-height="390"
data-image-alt="FIG 7"
data-pmc-id="PMC8091026"
data-figure-id="F7">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-6"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/327c/8091026/b2f85e5276fc/AEM.00091-21-f0007.gif"
alt="FIG 7" />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="720">
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="390">
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> FIG 7 </p>
</strong>
<div class="figure-caption-contents"><p> Growth of <i> Pseudomonas </i> knockout strains… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG 7 </p>
</strong>
<div class="figure-caption-contents"><p> Growth of <i> Pseudomonas </i> knockout strains on dodecane as the sole carbon source. Bushnell-Haas… </p></div>
</div>
<figcaption id="figure-caption-6" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG 7
</strong>
<div class="figure-caption-contents">Growth of <i>Pseudomonas</i> knockout strains on dodecane as the sole carbon source. Bushnell-Haas broth (BHB) supplemented with dodecane was inoculated with either wild-type strains (circles) or surfactant knockout mutant (squares) or was left noninoculated (diamonds). (A) Growth of <i>P</i>FF1 and the respective surfactant mutant. The growth of the surfactant mutant was significantly lower than that of the wild type on days 5 and 9. (B) Growth of <i>P</i>FF2 and the respective surfactant mutant. From day 7 to day 13, the growth of the surfactant mutant was significantly lower than that of the wild type. (C) Growth of <i>P</i>FF3 and the respective surfactant mutant. The growth of the surfactant mutant was significantly lower than that of the wild type on days 7, 11, and 15. (D) Growth of <i>P</i>FF4 and the respective surfactant mutant. From day 9 to day 19, the growth of the surfactant mutant was significantly lower than that of the wild type. The statistical analysis can be found in Table S3. Error bars depict the standard deviations of the means from three replicates.</div>
</figcaption>
</figure>
<figure class="figure-item tail"
itemscope itemtype="http://schema.org/ImageObject"
itemprop="associatedMedia"
data-slide-index="7"
data-label-slug="fig-8">
<a class="figure-link"
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/327c/8091026/60b460c76d2f/AEM.00091-21-f0008.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-7"
role="button"
data-image-width="771"
data-image-height="593"
data-image-alt="FIG 8"
data-pmc-id="PMC8091026"
data-figure-id="F8">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-7"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/327c/8091026/338bbd06ea19/AEM.00091-21-f0008.gif"
alt="FIG 8" />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="771">
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="593">
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> FIG 8 </p>
</strong>
<div class="figure-caption-contents"><i> In planta </i> competition of wild…</div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG 8 </p>
</strong>
<div class="figure-caption-contents"><i> In planta </i> competition of wild types (circles) and mutants (squares). (A) <i> P </i> FF1…</div>
</div>
<figcaption id="figure-caption-7" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG 8
</strong>
<div class="figure-caption-contents"><i>In planta</i> competition of wild types (circles) and mutants (squares). (A) <i>P</i>FF1 versus <i>P</i>FF1::ezTn5-visB; (B) <i>P</i>FF2 versus <i>P</i>FF2::ezTn5-visB; (C) <i>P</i>FF3 versus <i>P</i>FF3::ezTn5-massB; (D) <i>P</i>FF4 versus <i>P</i>FF2::ezTn5-massB. Symbols represent the mean number of CFU on five plants per measurement. The statistical analysis can be found in Table S3. Error bars depict the standard deviations of the means. Experiments were performed in quintuplicate.</div>
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Serrano M, Coluccia F, Torres M, LHaridon F, Métraux J-P. 2014. The cuticle and plant defense to pathogens. Front Plant Sci 5:274. doi:10.3389/fpls.2014.00274.
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