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

Mostrar todas las versiones(3)

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:...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Llorente, B., Bravo-Almonacid, F., Cvitanich, C., Orlowska, E., Torres, H.N., Flawiá, M.M., Alonso, G.D.
Formato:	Artículo publishedVersion
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:	http://hdl.handle.net/20.500.12110/paper_02668254_v51_n6_p603_Llorente https://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=artiaex&d=paper_02668254_v51_n6_p603_Llorente_oai
Aporte de:	Repositorio Digital de la Universidad de Buenos Aires (UBA) de Universidad de Buenos Aires

id	I28-R145-paper_02668254_v51_n6_p603_Llorente_oai
record_format	dspace
spelling	I28-R145-paper_02668254_v51_n6_p603_Llorente_oai2024-08-16 Llorente, B. Bravo-Almonacid, F. Cvitanich, C. Orlowska, E. Torres, H.N. Flawiá, M.M. Alonso, G.D. 2010 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. application/pdf http://hdl.handle.net/20.500.12110/paper_02668254_v51_n6_p603_Llorente info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar Lett. Appl. Microbiol. 2010;51(6):603-610 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 A quantitative real-time PCR method for in planta monitoring of Phytophthora infestans growth info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion https://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=artiaex&d=paper_02668254_v51_n6_p603_Llorente_oai
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-145
collection	Repositorio Digital de la Universidad de Buenos Aires (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, B. Bravo-Almonacid, F. Cvitanich, C. Orlowska, E. Torres, H.N. Flawiá, M.M. Alonso, G.D. 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.
format	Artículo Artículo publishedVersion
author	Llorente, B. Bravo-Almonacid, F. Cvitanich, C. Orlowska, E. Torres, H.N. Flawiá, M.M. Alonso, G.D.
author_facet	Llorente, B. Bravo-Almonacid, F. Cvitanich, C. Orlowska, E. Torres, H.N. Flawiá, M.M. Alonso, G.D.
author_sort	Llorente, B.
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	http://hdl.handle.net/20.500.12110/paper_02668254_v51_n6_p603_Llorente https://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=artiaex&d=paper_02668254_v51_n6_p603_Llorente_oai
work_keys_str_mv	AT llorenteb aquantitativerealtimepcrmethodforinplantamonitoringofphytophthorainfestansgrowth AT bravoalmonacidf aquantitativerealtimepcrmethodforinplantamonitoringofphytophthorainfestansgrowth AT cvitanichc aquantitativerealtimepcrmethodforinplantamonitoringofphytophthorainfestansgrowth AT orlowskae aquantitativerealtimepcrmethodforinplantamonitoringofphytophthorainfestansgrowth AT torreshn aquantitativerealtimepcrmethodforinplantamonitoringofphytophthorainfestansgrowth AT flawiamm aquantitativerealtimepcrmethodforinplantamonitoringofphytophthorainfestansgrowth AT alonsogd aquantitativerealtimepcrmethodforinplantamonitoringofphytophthorainfestansgrowth AT llorenteb quantitativerealtimepcrmethodforinplantamonitoringofphytophthorainfestansgrowth AT bravoalmonacidf quantitativerealtimepcrmethodforinplantamonitoringofphytophthorainfestansgrowth AT cvitanichc quantitativerealtimepcrmethodforinplantamonitoringofphytophthorainfestansgrowth AT orlowskae quantitativerealtimepcrmethodforinplantamonitoringofphytophthorainfestansgrowth AT torreshn quantitativerealtimepcrmethodforinplantamonitoringofphytophthorainfestansgrowth AT flawiamm quantitativerealtimepcrmethodforinplantamonitoringofphytophthorainfestansgrowth AT alonsogd quantitativerealtimepcrmethodforinplantamonitoringofphytophthorainfestansgrowth
_version_	1809357092766613504

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

Ejemplares similares