A quantitative real-time PCR method for in planta monitoring of Phytophthora infestans growth

Aims: To establish a reliable and rapid protocol to simultaneously obtain high quality DNA from an infected host plant and the infecting pathogen. To develop an accurate and sensitive low-cost assay for the quantification and in planta monitoring of Phytophthora infestans growth.Methods and Results:...

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Autores principales:	Llorente, Briardo Ernesto, Torres, Héctor Norberto, Flawiá, Mirtha María, Alonso, Guillermo Daniel
Publicado:	2010
Materias:	Biomass monitoring Late blight Phytophthora infestans Potato crop Quantitative real-time PCR Solanum tuberosum adaptation bioassay cultivar disease resistance fungal disease growth rate host plant host-pathogen interaction monitoring system plant breeding polymerase chain reaction real time symptom tuber article DNA purification fungus growth monitoring nonhuman nucleotide sequence pathogenesis plant plant disease plant gene plant leaf potato quantitative analysis real time polymerase chain reaction DNA DNA Primers DNA, Plant Organic Chemicals Plant Diseases Plant Leaves Plant Tubers Polymerase Chain Reaction Species Specificity Lycopersicon esculentum Nicotiana benthamiana
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_02668254_v51_n6_p603_Llorente http://hdl.handle.net/20.500.12110/paper_02668254_v51_n6_p603_Llorente
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_02668254_v51_n6_p603_Llorente
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spelling	paper:paper_02668254_v51_n6_p603_Llorente2023-06-08T15:23:37Z A quantitative real-time PCR method for in planta monitoring of Phytophthora infestans growth Llorente, Briardo Ernesto Torres, Héctor Norberto Flawiá, Mirtha María Alonso, Guillermo Daniel Biomass monitoring Late blight Phytophthora infestans Potato crop Quantitative real-time PCR Solanum tuberosum adaptation bioassay cultivar disease resistance fungal disease growth rate host plant host-pathogen interaction monitoring system plant breeding polymerase chain reaction real time symptom tuber article DNA purification fungus growth monitoring nonhuman nucleotide sequence pathogenesis Phytophthora infestans plant plant disease plant gene plant leaf potato quantitative analysis real time polymerase chain reaction DNA DNA Primers DNA, Plant Organic Chemicals Phytophthora infestans Plant Diseases Plant Leaves Plant Tubers Polymerase Chain Reaction Solanum tuberosum Species Specificity Lycopersicon esculentum Nicotiana benthamiana Phytophthora infestans Solanum tuberosum Aims: To establish a reliable and rapid protocol to simultaneously obtain high quality DNA from an infected host plant and the infecting pathogen. To develop an accurate and sensitive low-cost assay for the quantification and in planta monitoring of Phytophthora infestans growth.Methods and Results: In this study, we describe a SYBR Green-based quantitative real-time PCR (qPCR) method for the quantification of P. infestans. The method is based on a simultaneous plant-pathogen DNA purification followed by a qPCR in which the relative quantification of pathogen and plant DNA is performed. Besides assuring an accurate quantification, the use of a plant gene provides a reliable indicator of sample quality, allowing the exclusion of inappropriate samples. By applying this methodology, we were able to detect P. infestans in potato leaf and tuber tissue before the first symptoms of the disease were observed and to monitor the in planta growth of the pathogen for 6 days.Conclusions: This is a reliable low-cost assay that provides rapid, accurate and sensitive quantification of the late blight pathogen, allowing the in planta monitoring of P. infestans growth.Significance and Impact of the Study: The quantitative nature of the assay described in this study may be useful in plant breeding programmes and basic research. The method is appropriate for the comparison of cultivars with different, and even subtle, degrees of pathogen resistance and in the screening of new anti-oomycete compounds. The method can be easily adapted to tomato and the model plant Nicotiana benthamiana. © No claim to Argentinean Government works. Letters in Applied Microbiology 51, 603-610 © 2010 The Society for Applied Microbiology. Fil:Llorente, B. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Torres, H.N. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Flawiá, M.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Alonso, G.D. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2010 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_02668254_v51_n6_p603_Llorente http://hdl.handle.net/20.500.12110/paper_02668254_v51_n6_p603_Llorente
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Biomass monitoring Late blight Phytophthora infestans Potato crop Quantitative real-time PCR Solanum tuberosum adaptation bioassay cultivar disease resistance fungal disease growth rate host plant host-pathogen interaction monitoring system plant breeding polymerase chain reaction real time symptom tuber article DNA purification fungus growth monitoring nonhuman nucleotide sequence pathogenesis Phytophthora infestans plant plant disease plant gene plant leaf potato quantitative analysis real time polymerase chain reaction DNA DNA Primers DNA, Plant Organic Chemicals Phytophthora infestans Plant Diseases Plant Leaves Plant Tubers Polymerase Chain Reaction Solanum tuberosum Species Specificity Lycopersicon esculentum Nicotiana benthamiana Phytophthora infestans Solanum tuberosum
spellingShingle	Biomass monitoring Late blight Phytophthora infestans Potato crop Quantitative real-time PCR Solanum tuberosum adaptation bioassay cultivar disease resistance fungal disease growth rate host plant host-pathogen interaction monitoring system plant breeding polymerase chain reaction real time symptom tuber article DNA purification fungus growth monitoring nonhuman nucleotide sequence pathogenesis Phytophthora infestans plant plant disease plant gene plant leaf potato quantitative analysis real time polymerase chain reaction DNA DNA Primers DNA, Plant Organic Chemicals Phytophthora infestans Plant Diseases Plant Leaves Plant Tubers Polymerase Chain Reaction Solanum tuberosum Species Specificity Lycopersicon esculentum Nicotiana benthamiana Phytophthora infestans Solanum tuberosum Llorente, Briardo Ernesto Torres, Héctor Norberto Flawiá, Mirtha María Alonso, Guillermo Daniel A quantitative real-time PCR method for in planta monitoring of Phytophthora infestans growth
topic_facet	Biomass monitoring Late blight Phytophthora infestans Potato crop Quantitative real-time PCR Solanum tuberosum adaptation bioassay cultivar disease resistance fungal disease growth rate host plant host-pathogen interaction monitoring system plant breeding polymerase chain reaction real time symptom tuber article DNA purification fungus growth monitoring nonhuman nucleotide sequence pathogenesis Phytophthora infestans plant plant disease plant gene plant leaf potato quantitative analysis real time polymerase chain reaction DNA DNA Primers DNA, Plant Organic Chemicals Phytophthora infestans Plant Diseases Plant Leaves Plant Tubers Polymerase Chain Reaction Solanum tuberosum Species Specificity Lycopersicon esculentum Nicotiana benthamiana Phytophthora infestans Solanum tuberosum
description	Aims: To establish a reliable and rapid protocol to simultaneously obtain high quality DNA from an infected host plant and the infecting pathogen. To develop an accurate and sensitive low-cost assay for the quantification and in planta monitoring of Phytophthora infestans growth.Methods and Results: In this study, we describe a SYBR Green-based quantitative real-time PCR (qPCR) method for the quantification of P. infestans. The method is based on a simultaneous plant-pathogen DNA purification followed by a qPCR in which the relative quantification of pathogen and plant DNA is performed. Besides assuring an accurate quantification, the use of a plant gene provides a reliable indicator of sample quality, allowing the exclusion of inappropriate samples. By applying this methodology, we were able to detect P. infestans in potato leaf and tuber tissue before the first symptoms of the disease were observed and to monitor the in planta growth of the pathogen for 6 days.Conclusions: This is a reliable low-cost assay that provides rapid, accurate and sensitive quantification of the late blight pathogen, allowing the in planta monitoring of P. infestans growth.Significance and Impact of the Study: The quantitative nature of the assay described in this study may be useful in plant breeding programmes and basic research. The method is appropriate for the comparison of cultivars with different, and even subtle, degrees of pathogen resistance and in the screening of new anti-oomycete compounds. The method can be easily adapted to tomato and the model plant Nicotiana benthamiana. © No claim to Argentinean Government works. Letters in Applied Microbiology 51, 603-610 © 2010 The Society for Applied Microbiology.
author	Llorente, Briardo Ernesto Torres, Héctor Norberto Flawiá, Mirtha María Alonso, Guillermo Daniel
author_facet	Llorente, Briardo Ernesto Torres, Héctor Norberto Flawiá, Mirtha María Alonso, Guillermo Daniel
author_sort	Llorente, Briardo Ernesto
title	A quantitative real-time PCR method for in planta monitoring of Phytophthora infestans growth
title_short	A quantitative real-time PCR method for in planta monitoring of Phytophthora infestans growth
title_full	A quantitative real-time PCR method for in planta monitoring of Phytophthora infestans growth
title_fullStr	A quantitative real-time PCR method for in planta monitoring of Phytophthora infestans growth
title_full_unstemmed	A quantitative real-time PCR method for in planta monitoring of Phytophthora infestans growth
title_sort	quantitative real-time pcr method for in planta monitoring of phytophthora infestans growth
publishDate	2010
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_02668254_v51_n6_p603_Llorente http://hdl.handle.net/20.500.12110/paper_02668254_v51_n6_p603_Llorente
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A quantitative real-time PCR method for in planta monitoring of Phytophthora infestans growth

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