Development of the automated primer design workflow uniqprimer and diagnostic primers for the broad-host-range plant pathogen Dickeya dianthicola

Uniqprimer, a software pipeline developed in Python, was deployed as a user-friendly internet tool in Rice Galaxy for comparative genome analyses to design primer sets for PCRassays capable of detecting target bacterial taxa. The pipeline was trialed with Dickeya dianthicola, a destructive broad-hos...

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Main Authors: Karim, Shaista, McNally, R. Ryan, Nasaruddin, Afnan S., DeReeper, Alexis, Mauleon, Ramil P., Charkowski, Amy O., Leach, Jan E., Ben-Hur, Asa, Triplett, Lindsay R.
Format: Journal Article
Language:Inglés
Published: Scientific Societies 2019
Subjects:
Online Access:https://hdl.handle.net/10568/164603
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author Karim, Shaista
McNally, R. Ryan
Nasaruddin, Afnan S.
DeReeper, Alexis
Mauleon, Ramil P.
Charkowski, Amy O.
Leach, Jan E.
Ben-Hur, Asa
Triplett, Lindsay R.
author_browse Ben-Hur, Asa
Charkowski, Amy O.
DeReeper, Alexis
Karim, Shaista
Leach, Jan E.
Mauleon, Ramil P.
McNally, R. Ryan
Nasaruddin, Afnan S.
Triplett, Lindsay R.
author_facet Karim, Shaista
McNally, R. Ryan
Nasaruddin, Afnan S.
DeReeper, Alexis
Mauleon, Ramil P.
Charkowski, Amy O.
Leach, Jan E.
Ben-Hur, Asa
Triplett, Lindsay R.
author_sort Karim, Shaista
collection Repository of Agricultural Research Outputs (CGSpace)
description Uniqprimer, a software pipeline developed in Python, was deployed as a user-friendly internet tool in Rice Galaxy for comparative genome analyses to design primer sets for PCRassays capable of detecting target bacterial taxa. The pipeline was trialed with Dickeya dianthicola, a destructive broad-host-range bacterial pathogen found in most potato-growing regions. Dickeya is a highly variable genus, and some primers available to detect this genus and species exhibit common diagnostic failures. Upon uploading a selection of target and nontarget genomes, six primer sets were rapidly identified with Uniqprimer, of which two were specific and sensitive when tested with D. dianthicola. The remaining four amplified a minority of the nontarget strains tested. The two promising candidate primer sets were trialed with DNA isolated from 116 field samples from across the United States that were previously submitted for testing. D. dianthicola was detected in 41 samples, demonstrating the applicability of our detection primers and suggesting widespread occurrence of D. dianthicola in North America.
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spelling CGSpace1646032025-12-08T09:54:28Z Development of the automated primer design workflow uniqprimer and diagnostic primers for the broad-host-range plant pathogen Dickeya dianthicola Karim, Shaista McNally, R. Ryan Nasaruddin, Afnan S. DeReeper, Alexis Mauleon, Ramil P. Charkowski, Amy O. Leach, Jan E. Ben-Hur, Asa Triplett, Lindsay R. agronomy crop science plant science Uniqprimer, a software pipeline developed in Python, was deployed as a user-friendly internet tool in Rice Galaxy for comparative genome analyses to design primer sets for PCRassays capable of detecting target bacterial taxa. The pipeline was trialed with Dickeya dianthicola, a destructive broad-host-range bacterial pathogen found in most potato-growing regions. Dickeya is a highly variable genus, and some primers available to detect this genus and species exhibit common diagnostic failures. Upon uploading a selection of target and nontarget genomes, six primer sets were rapidly identified with Uniqprimer, of which two were specific and sensitive when tested with D. dianthicola. The remaining four amplified a minority of the nontarget strains tested. The two promising candidate primer sets were trialed with DNA isolated from 116 field samples from across the United States that were previously submitted for testing. D. dianthicola was detected in 41 samples, demonstrating the applicability of our detection primers and suggesting widespread occurrence of D. dianthicola in North America. 2019-11 2024-12-19T12:54:06Z 2024-12-19T12:54:06Z Journal Article https://hdl.handle.net/10568/164603 en Scientific Societies Karim, Shaista; McNally, R. Ryan; Nasaruddin, Afnan S.; DeReeper, Alexis; Mauleon, Ramil P.; Charkowski, Amy O.; Leach, Jan E.; Ben-Hur, Asa and Triplett, Lindsay R. 2019. Development of the automated primer design workflow uniqprimer and diagnostic primers for the broad-host-range plant pathogen Dickeya dianthicola. Plant Disease, Volume 103 no. 11 p. 2893-2902
spellingShingle agronomy
crop science
plant science
Karim, Shaista
McNally, R. Ryan
Nasaruddin, Afnan S.
DeReeper, Alexis
Mauleon, Ramil P.
Charkowski, Amy O.
Leach, Jan E.
Ben-Hur, Asa
Triplett, Lindsay R.
Development of the automated primer design workflow uniqprimer and diagnostic primers for the broad-host-range plant pathogen Dickeya dianthicola
title Development of the automated primer design workflow uniqprimer and diagnostic primers for the broad-host-range plant pathogen Dickeya dianthicola
title_full Development of the automated primer design workflow uniqprimer and diagnostic primers for the broad-host-range plant pathogen Dickeya dianthicola
title_fullStr Development of the automated primer design workflow uniqprimer and diagnostic primers for the broad-host-range plant pathogen Dickeya dianthicola
title_full_unstemmed Development of the automated primer design workflow uniqprimer and diagnostic primers for the broad-host-range plant pathogen Dickeya dianthicola
title_short Development of the automated primer design workflow uniqprimer and diagnostic primers for the broad-host-range plant pathogen Dickeya dianthicola
title_sort development of the automated primer design workflow uniqprimer and diagnostic primers for the broad host range plant pathogen dickeya dianthicola
topic agronomy
crop science
plant science
url https://hdl.handle.net/10568/164603
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