“Shining a LAMP” (Loop-Mediated Isothermal Amplification) on the Molecular Detection of Phytopathogens Phytophthora spp. and Phytophthora cactorum in Strawberry Fields

Phytopathogenic microorganisms belonging to the genus Phytophthora have been recognized many times as causal agents of diseases that lower the yield of many plants important for agriculture. Meanwhile, Phytophthora cactorum causes crown rot and leather rot of be...

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Autores principales:	Dominika G. Siegieda, Jacek Panek, Magdalena Frąc
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	Loop-Mediated Isothermal Amplification <i>Phytophthora cactorum</i> <i>Phytophthora</i> spp. rhizosphere soil shoots Medicine R
Acceso en línea:	https://doaj.org/article/80d84d71705c41f49803b40dbf3a873c
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spelling	oai:doaj.org-article:80d84d71705c41f49803b40dbf3a873c2021-11-25T18:38:22Z“Shining a LAMP” (Loop-Mediated Isothermal Amplification) on the Molecular Detection of Phytopathogens <i>Phytophthora</i> spp. and <i>Phytophthora cactorum</i> in Strawberry Fields10.3390/pathogens101114532076-0817https://doaj.org/article/80d84d71705c41f49803b40dbf3a873c2021-11-01T00:00:00Zhttps://www.mdpi.com/2076-0817/10/11/1453https://doaj.org/toc/2076-0817Phytopathogenic microorganisms belonging to the genus <i>Phytophthora</i> have been recognized many times as causal agents of diseases that lower the yield of many plants important for agriculture. Meanwhile, <i>Phytophthora cactorum</i> causes crown rot and leather rot of berry fruits, mainly strawberries. However, widely-applied culture-based methods used for the detection of pathogens are time-consuming and often inaccurate. What is more, molecular techniques require costly equipment. Here we show a rapid and effective detection method for the aforementioned targets, deploying a simple molecular biology technique, Loop-Mediated Isothermal Amplification (LAMP). We optimized assays to amplify the translation elongation factor 1-α (EF1a) gene for two targets: <i>Phytophthora</i> spp. And <i>Phytophthora cactorum</i>. We optimized the LAMP on pure strains of the pathogens, isolated from organic plantations of strawberry, and successfully validated the assay on biological material from the environment including soil samples, rhizosphere, shoots and roots of strawberry, and with SYBR Green. Our results demonstrate that a simple and reliable molecular detection method, that requires only a thermoblock and simple DNA isolation kit, can be successfully applied to detect pathogens that are difficult to separate from the field. We anticipate our findings to be a starting point for developing easier and faster modifications of the isothermal detection methods and which can be applied directly in the plantation, in particular with the use of freeze-dried reagents and chemistry, allowing observation of the results with the naked eye.Dominika G. SiegiedaJacek PanekMagdalena FrącMDPI AGarticleLoop-Mediated Isothermal Amplification<i>Phytophthora cactorum</i><i>Phytophthora</i> spp.rhizospheresoilshootsMedicineRENPathogens, Vol 10, Iss 1453, p 1453 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Loop-Mediated Isothermal Amplification <i>Phytophthora cactorum</i> <i>Phytophthora</i> spp. rhizosphere soil shoots Medicine R
spellingShingle	Loop-Mediated Isothermal Amplification <i>Phytophthora cactorum</i> <i>Phytophthora</i> spp. rhizosphere soil shoots Medicine R Dominika G. Siegieda Jacek Panek Magdalena Frąc “Shining a LAMP” (Loop-Mediated Isothermal Amplification) on the Molecular Detection of Phytopathogens <i>Phytophthora</i> spp. and <i>Phytophthora cactorum</i> in Strawberry Fields
description	Phytopathogenic microorganisms belonging to the genus <i>Phytophthora</i> have been recognized many times as causal agents of diseases that lower the yield of many plants important for agriculture. Meanwhile, <i>Phytophthora cactorum</i> causes crown rot and leather rot of berry fruits, mainly strawberries. However, widely-applied culture-based methods used for the detection of pathogens are time-consuming and often inaccurate. What is more, molecular techniques require costly equipment. Here we show a rapid and effective detection method for the aforementioned targets, deploying a simple molecular biology technique, Loop-Mediated Isothermal Amplification (LAMP). We optimized assays to amplify the translation elongation factor 1-α (EF1a) gene for two targets: <i>Phytophthora</i> spp. And <i>Phytophthora cactorum</i>. We optimized the LAMP on pure strains of the pathogens, isolated from organic plantations of strawberry, and successfully validated the assay on biological material from the environment including soil samples, rhizosphere, shoots and roots of strawberry, and with SYBR Green. Our results demonstrate that a simple and reliable molecular detection method, that requires only a thermoblock and simple DNA isolation kit, can be successfully applied to detect pathogens that are difficult to separate from the field. We anticipate our findings to be a starting point for developing easier and faster modifications of the isothermal detection methods and which can be applied directly in the plantation, in particular with the use of freeze-dried reagents and chemistry, allowing observation of the results with the naked eye.
format	article
author	Dominika G. Siegieda Jacek Panek Magdalena Frąc
author_facet	Dominika G. Siegieda Jacek Panek Magdalena Frąc
author_sort	Dominika G. Siegieda
title	“Shining a LAMP” (Loop-Mediated Isothermal Amplification) on the Molecular Detection of Phytopathogens <i>Phytophthora</i> spp. and <i>Phytophthora cactorum</i> in Strawberry Fields
title_short	“Shining a LAMP” (Loop-Mediated Isothermal Amplification) on the Molecular Detection of Phytopathogens <i>Phytophthora</i> spp. and <i>Phytophthora cactorum</i> in Strawberry Fields
title_full	“Shining a LAMP” (Loop-Mediated Isothermal Amplification) on the Molecular Detection of Phytopathogens <i>Phytophthora</i> spp. and <i>Phytophthora cactorum</i> in Strawberry Fields
title_fullStr	“Shining a LAMP” (Loop-Mediated Isothermal Amplification) on the Molecular Detection of Phytopathogens <i>Phytophthora</i> spp. and <i>Phytophthora cactorum</i> in Strawberry Fields
title_full_unstemmed	“Shining a LAMP” (Loop-Mediated Isothermal Amplification) on the Molecular Detection of Phytopathogens <i>Phytophthora</i> spp. and <i>Phytophthora cactorum</i> in Strawberry Fields
title_sort	“shining a lamp” (loop-mediated isothermal amplification) on the molecular detection of phytopathogens <i>phytophthora</i> spp. and <i>phytophthora cactorum</i> in strawberry fields
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/80d84d71705c41f49803b40dbf3a873c
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_version_	1718410900682047488

“Shining a LAMP” (Loop-Mediated Isothermal Amplification) on the Molecular Detection of Phytopathogens <i>Phytophthora</i> spp. and <i>Phytophthora cactorum</i> in Strawberry Fields

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