Serveur d'exploration Phytophthora

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Detection of the Genus Phytophthora and the Species Phytophthora nicotianae by LAMP with a QProbe.

Identifieur interne : 000247 ( Main/Exploration ); précédent : 000246; suivant : 000248

Detection of the Genus Phytophthora and the Species Phytophthora nicotianae by LAMP with a QProbe.

Auteurs : Ayaka Hieno [Japon] ; Mingzhu Li [République populaire de Chine] ; Auliana Afandi [Indonésie] ; Kayoko Otsubo [Japon] ; Haruhisa Suga [Japon] ; Koji Kageyama [Japon]

Source :

RBID : pubmed:32628090

Descripteurs français

English descriptors

Abstract

Phytophthora is an oomycete genus with worldwide distribution, and many of its species cause destructive diseases. In Japan, Phytophthora species are listed as quarantine organisms with the exception of Phytophthora nicotianae. For effective quarantine control, we designed a Phytophthora genus-specific loop-mediated isothermal amplification (LAMP) primer set and a P. nicotianae species-specific quenching probe (QProbe) to establish a simultaneous LAMP-based detection method. We confirmed the specificity of the genus-specific primers, and all 161 taxa were detected. No other species in the closely related genera Pythium and Phytopythium gave positive results with the exception of two species, Phytopythium delawarense and Phytopythium fagopyri. These two species gave inconsistent results. We used annealing curve analysis with the QProbe to demonstrate that P. nicotianae could be distinguished from other species. DNA from inoculated and naturally infected plants was extracted using a time-saving extraction kit and subjected to the simultaneous detection method. We confirmed that all Phytophthora DNAs in the plant samples were detected, and P. nicotianae was specifically identified. This simultaneous detection method will make quarantine inspections faster and easier.

DOI: 10.1094/PDIS-12-19-2523-RE
PubMed: 32628090


Affiliations:


Links toward previous steps (curation, corpus...)


Le document en format XML

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<i>Phytophthora nicotianae</i>
by LAMP with a QProbe.</title>
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<term>Phytophthora (genetics)</term>
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<div type="abstract" xml:lang="en">
<i>Phytophthora</i>
is an oomycete genus with worldwide distribution, and many of its species cause destructive diseases. In Japan,
<i>Phytophthora</i>
species are listed as quarantine organisms with the exception of
<i>Phytophthora nicotianae</i>
. For effective quarantine control, we designed a
<i>Phytophthora</i>
genus-specific loop-mediated isothermal amplification (LAMP) primer set and a
<i>P. nicotianae</i>
species-specific quenching probe (QProbe) to establish a simultaneous LAMP-based detection method. We confirmed the specificity of the genus-specific primers, and all 161 taxa were detected. No other species in the closely related genera
<i>Pythium</i>
and
<i>Phytopythium</i>
gave positive results with the exception of two species,
<i>Phytopythium delawarense</i>
and
<i>Phytopythium fagopyri</i>
. These two species gave inconsistent results. We used annealing curve analysis with the QProbe to demonstrate that
<i>P. nicotianae</i>
could be distinguished from other species. DNA from inoculated and naturally infected plants was extracted using a time-saving extraction kit and subjected to the simultaneous detection method. We confirmed that all
<i>Phytophthora</i>
DNAs in the plant samples were detected, and
<i>P. nicotianae</i>
was specifically identified. This simultaneous detection method will make quarantine inspections faster and easier.</div>
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<i>Phytophthora</i>
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<i>Phytophthora nicotianae</i>
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<i>Phytophthora</i>
is an oomycete genus with worldwide distribution, and many of its species cause destructive diseases. In Japan,
<i>Phytophthora</i>
species are listed as quarantine organisms with the exception of
<i>Phytophthora nicotianae</i>
. For effective quarantine control, we designed a
<i>Phytophthora</i>
genus-specific loop-mediated isothermal amplification (LAMP) primer set and a
<i>P. nicotianae</i>
species-specific quenching probe (QProbe) to establish a simultaneous LAMP-based detection method. We confirmed the specificity of the genus-specific primers, and all 161 taxa were detected. No other species in the closely related genera
<i>Pythium</i>
and
<i>Phytopythium</i>
gave positive results with the exception of two species,
<i>Phytopythium delawarense</i>
and
<i>Phytopythium fagopyri</i>
. These two species gave inconsistent results. We used annealing curve analysis with the QProbe to demonstrate that
<i>P. nicotianae</i>
could be distinguished from other species. DNA from inoculated and naturally infected plants was extracted using a time-saving extraction kit and subjected to the simultaneous detection method. We confirmed that all
<i>Phytophthora</i>
DNAs in the plant samples were detected, and
<i>P. nicotianae</i>
was specifically identified. This simultaneous detection method will make quarantine inspections faster and easier.</AbstractText>
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