Detection and Control of Fusarium oxysporum from Soft Rot in Dendrobium officinale by Loop-Mediated Isothermal Amplification Assays

Soft rot causing Fusarium oxysporum is one of the most destructive diseases of Dendrobium officinale Kimura et Migo in China that reduces D. officinale yield and quality. A key challenge for an integrated management strategy for this disease i...

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Autores principales:	Caiyun Xiao, Rongyu Li
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	<i>Fusarium oxysporum</i> soft rot on <i>Dendrobium officinale</i> loop-mediated isothermal amplification translation elongation factor-1α Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/4191f7a03b824bfbae70aed79d894c92
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spelling	oai:doaj.org-article:4191f7a03b824bfbae70aed79d894c922021-11-25T16:47:22ZDetection and Control of <i>Fusarium oxysporum</i> from Soft Rot in <i>Dendrobium officinale</i> by Loop-Mediated Isothermal Amplification Assays10.3390/biology101111362079-7737https://doaj.org/article/4191f7a03b824bfbae70aed79d894c922021-11-01T00:00:00Zhttps://www.mdpi.com/2079-7737/10/11/1136https://doaj.org/toc/2079-7737Soft rot causing <i>Fusarium oxysporum</i> is one of the most destructive diseases of <i>Dendrobium officinale</i> Kimura et Migo in China that reduces <i>D. officinale</i> yield and quality. A key challenge for an integrated management strategy for this disease is the rapid and accurate detection of <i>F. oxysporum</i> on <i>D. officinale</i>. Therefore, a new loop-mediated isothermal amplification (LAMP) assay was developed for this purpose. In this study, the primers were selected and designed using the translation elongation factor-1α (<i>TEF-1α</i>) gene region as the target DNA sequence in order to screen the best system of reaction of LAMP to detect <i>F. oxysporum</i> through optimizing different conditions of the LAMP reaction, including time, temperature, concentrations of MgSO<sub>4</sub>, and concentrations of inner and outer primers. The optimized system was able to efficiently amplify the target gene at 62 °C for 60 min with 1.2 μM internal primers, 0.4 μM external primers, 7 mM Mg<sup>2+</sup>, and 5 fg/µL minimum detection concentration of DNA for <i>F. oxysporum</i>. The amplified products could be detected with the naked eye after completion of the reaction with SYBR green I. We were better able to control the effect of soft rot in <i>D. officinale</i> using fungicides following a positive test result. Additionally, the control effect of synergism combinations against soft rot was higher than 75%. Thus, LAMP assays could detect <i>F. oxysporum</i> in infected tissues of <i>D. officinale</i> and soils in field, allowing for early diagnosis of the disease.Caiyun XiaoRongyu LiMDPI AGarticle<i>Fusarium oxysporum</i>soft rot on <i>Dendrobium officinale</i>loop-mediated isothermal amplificationtranslation elongation factor-1αBiology (General)QH301-705.5ENBiology, Vol 10, Iss 1136, p 1136 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	<i>Fusarium oxysporum</i> soft rot on <i>Dendrobium officinale</i> loop-mediated isothermal amplification translation elongation factor-1α Biology (General) QH301-705.5
spellingShingle	<i>Fusarium oxysporum</i> soft rot on <i>Dendrobium officinale</i> loop-mediated isothermal amplification translation elongation factor-1α Biology (General) QH301-705.5 Caiyun Xiao Rongyu Li Detection and Control of <i>Fusarium oxysporum</i> from Soft Rot in <i>Dendrobium officinale</i> by Loop-Mediated Isothermal Amplification Assays
description	Soft rot causing <i>Fusarium oxysporum</i> is one of the most destructive diseases of <i>Dendrobium officinale</i> Kimura et Migo in China that reduces <i>D. officinale</i> yield and quality. A key challenge for an integrated management strategy for this disease is the rapid and accurate detection of <i>F. oxysporum</i> on <i>D. officinale</i>. Therefore, a new loop-mediated isothermal amplification (LAMP) assay was developed for this purpose. In this study, the primers were selected and designed using the translation elongation factor-1α (<i>TEF-1α</i>) gene region as the target DNA sequence in order to screen the best system of reaction of LAMP to detect <i>F. oxysporum</i> through optimizing different conditions of the LAMP reaction, including time, temperature, concentrations of MgSO<sub>4</sub>, and concentrations of inner and outer primers. The optimized system was able to efficiently amplify the target gene at 62 °C for 60 min with 1.2 μM internal primers, 0.4 μM external primers, 7 mM Mg<sup>2+</sup>, and 5 fg/µL minimum detection concentration of DNA for <i>F. oxysporum</i>. The amplified products could be detected with the naked eye after completion of the reaction with SYBR green I. We were better able to control the effect of soft rot in <i>D. officinale</i> using fungicides following a positive test result. Additionally, the control effect of synergism combinations against soft rot was higher than 75%. Thus, LAMP assays could detect <i>F. oxysporum</i> in infected tissues of <i>D. officinale</i> and soils in field, allowing for early diagnosis of the disease.
format	article
author	Caiyun Xiao Rongyu Li
author_facet	Caiyun Xiao Rongyu Li
author_sort	Caiyun Xiao
title	Detection and Control of <i>Fusarium oxysporum</i> from Soft Rot in <i>Dendrobium officinale</i> by Loop-Mediated Isothermal Amplification Assays
title_short	Detection and Control of <i>Fusarium oxysporum</i> from Soft Rot in <i>Dendrobium officinale</i> by Loop-Mediated Isothermal Amplification Assays
title_full	Detection and Control of <i>Fusarium oxysporum</i> from Soft Rot in <i>Dendrobium officinale</i> by Loop-Mediated Isothermal Amplification Assays
title_fullStr	Detection and Control of <i>Fusarium oxysporum</i> from Soft Rot in <i>Dendrobium officinale</i> by Loop-Mediated Isothermal Amplification Assays
title_full_unstemmed	Detection and Control of <i>Fusarium oxysporum</i> from Soft Rot in <i>Dendrobium officinale</i> by Loop-Mediated Isothermal Amplification Assays
title_sort	detection and control of <i>fusarium oxysporum</i> from soft rot in <i>dendrobium officinale</i> by loop-mediated isothermal amplification assays
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/4191f7a03b824bfbae70aed79d894c92
work_keys_str_mv	AT caiyunxiao detectionandcontrolofifusariumoxysporumifromsoftrotinidendrobiumofficinaleibyloopmediatedisothermalamplificationassays AT rongyuli detectionandcontrolofifusariumoxysporumifromsoftrotinidendrobiumofficinaleibyloopmediatedisothermalamplificationassays
_version_	1718412949726429184

Detection and Control of <i>Fusarium oxysporum</i> from Soft Rot in <i>Dendrobium officinale</i> by Loop-Mediated Isothermal Amplification Assays

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