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Three New Pierce's Disease Pathogenicity Effectors Identified Using Xylella fastidiosa Biocontrol Strain EB92-1

Identifieur interne : 000406 ( Main/Exploration ); précédent : 000405; suivant : 000407

Three New Pierce's Disease Pathogenicity Effectors Identified Using Xylella fastidiosa Biocontrol Strain EB92-1

Auteurs : Shujian Zhang [États-Unis] ; Pranjib K. Chakrabarty [États-Unis] ; Laura A. Fleites [États-Unis] ; Patricia A. Rayside [États-Unis] ; Donald L. Hopkins [États-Unis] ; Dean W. Gabriel [États-Unis]

Source :

RBID : PMC:4517913

Abstract

Xylella fastidiosa (X. fastidiosa) infects a wide range of plant hosts and causes economically serious diseases, including Pierce's Disease (PD) of grapevines. X. fastidiosa biocontrol strain EB92-1 was isolated from elderberry and is infectious and persistent in grapevines but causes only very slight symptoms under ideal conditions. The draft genome of EB92-1 revealed that it appeared to be missing genes encoding 10 potential PD pathogenicity effectors found in Temecula1. Subsequent PCR and sequencing analyses confirmed that EB92-1 was missing the following predicted effectors found in Temecula1: two type II secreted enzymes, including a lipase (LipA; PD1703) and a serine protease (PD0956); two identical genes encoding proteins similar to Zonula occludens toxins (Zot; PD0915 and PD0928), and at least one relatively short, hemagglutinin-like protein (PD0986). Leaves of tobacco and citrus inoculated with cell-free, crude protein extracts of E. coli BL21(DE3) overexpressing PD1703 exhibited a hypersensitive response (HR) in less than 24 hours. When cloned into shuttle vector pBBR1MCS-5, PD1703 conferred strong secreted lipase activity to Xanthomonas citri, E. coli and X. fastidiosa EB92-1 in plate assays. EB92-1/PD1703 transformants also showed significantly increased disease symptoms on grapevines, characteristic of PD. Genes predicted to encode PD0928 (Zot) and a PD0986 (hemagglutinin) were also cloned into pBBR1MCS-5 and moved into EB92-1; both transformants also showed significantly increased symptoms on V. vinifera vines, characteristic of PD. Together, these results reveal that PD effectors include at least a lipase, two Zot-like toxins and a possibly redundant hemagglutinin, none of which are necessary for parasitic survival of X. fastidiosa populations in grapevines or elderberry.


Url:
DOI: 10.1371/journal.pone.0133796
PubMed: 26218423
PubMed Central: 4517913


Affiliations:


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Le document en format XML

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<title xml:lang="en" level="a" type="main">Three New Pierce's Disease Pathogenicity Effectors Identified Using
<italic>Xylella fastidiosa</italic>
Biocontrol Strain EB92-1</title>
<author>
<name sortKey="Zhang, Shujian" sort="Zhang, Shujian" uniqKey="Zhang S" first="Shujian" last="Zhang">Shujian Zhang</name>
<affiliation wicri:level="2">
<nlm:aff id="aff001">
<addr-line>Plant Pathology Department, University of Florida, Gainesville, Florida, United States of America</addr-line>
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<name sortKey="Chakrabarty, Pranjib K" sort="Chakrabarty, Pranjib K" uniqKey="Chakrabarty P" first="Pranjib K." last="Chakrabarty">Pranjib K. Chakrabarty</name>
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<addr-line>Plant Pathology Department, University of Florida, Gainesville, Florida, United States of America</addr-line>
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<wicri:regionArea>Plant Pathology Department, University of Florida, Gainesville, Florida</wicri:regionArea>
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<name sortKey="Fleites, Laura A" sort="Fleites, Laura A" uniqKey="Fleites L" first="Laura A." last="Fleites">Laura A. Fleites</name>
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<nlm:aff id="aff001">
<addr-line>Plant Pathology Department, University of Florida, Gainesville, Florida, United States of America</addr-line>
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<wicri:regionArea>Plant Pathology Department, University of Florida, Gainesville, Florida</wicri:regionArea>
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<name sortKey="Rayside, Patricia A" sort="Rayside, Patricia A" uniqKey="Rayside P" first="Patricia A." last="Rayside">Patricia A. Rayside</name>
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<addr-line>Plant Pathology Department, University of Florida, Gainesville, Florida, United States of America</addr-line>
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<name sortKey="Hopkins, Donald L" sort="Hopkins, Donald L" uniqKey="Hopkins D" first="Donald L." last="Hopkins">Donald L. Hopkins</name>
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<italic>Xylella fastidiosa</italic>
(
<italic>X</italic>
.
<italic>fastidiosa</italic>
) infects a wide range of plant hosts and causes economically serious diseases, including Pierce's Disease (PD) of grapevines.
<italic>X</italic>
.
<italic>fastidiosa</italic>
biocontrol strain EB92-1 was isolated from elderberry and is infectious and persistent in grapevines but causes only very slight symptoms under ideal conditions. The draft genome of EB92-1 revealed that it appeared to be missing genes encoding 10 potential PD pathogenicity effectors found in Temecula1. Subsequent PCR and sequencing analyses confirmed that EB92-1 was missing the following predicted effectors found in Temecula1: two type II secreted enzymes, including a lipase (LipA; PD1703) and a serine protease (PD0956); two identical genes encoding proteins similar to Zonula occludens toxins (Zot; PD0915 and PD0928), and at least one relatively short, hemagglutinin-like protein (PD0986). Leaves of tobacco and citrus inoculated with cell-free, crude protein extracts of
<italic>E</italic>
.
<italic>coli</italic>
BL21(DE3) overexpressing PD1703 exhibited a hypersensitive response (HR) in less than 24 hours. When cloned into shuttle vector pBBR1MCS-5, PD1703 conferred strong secreted lipase activity to
<italic>Xanthomonas citri</italic>
,
<italic>E</italic>
.
<italic>coli</italic>
and
<italic>X</italic>
.
<italic>fastidiosa</italic>
EB92-1 in plate assays. EB92-1/PD1703 transformants also showed significantly increased disease symptoms on grapevines, characteristic of PD. Genes predicted to encode PD0928 (Zot) and a PD0986 (hemagglutinin) were also cloned into pBBR1MCS-5 and moved into EB92-1; both transformants also showed significantly increased symptoms on
<italic>V</italic>
.
<italic>vinifera</italic>
vines, characteristic of PD. Together, these results reveal that PD effectors include at least a lipase, two Zot-like toxins and a possibly redundant hemagglutinin, none of which are necessary for parasitic survival of
<italic>X</italic>
.
<italic>fastidiosa</italic>
populations in grapevines or elderberry.</p>
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