Development and evaluation of polyclonal antibodies for detection of Pythium aphanidermatum and Fusarium oxysporum in ginger

Polyclonal antibodies against Pythium aphanidermatum and Fusarium oxysporum proteins were developed for the detection of rhizome rot in ginger using serological assays. Under optimal experimental conditions, the detection limit of P. aphanidermatum by indirect ELISA was 10 µg/ml with a linear workin...

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Autores principales:	Monalisa Ray, Swagatika Dash, K. Gopinath Achary, Sanghamitra Nayak, Shikha Singh
Formato:	article
Lenguaje:	EN
Publicado:	Taylor & Francis Group 2018
Materias:	ginger rhizome rot pythium aphanidermatum fusarium oxysporum polyclonal antibodies sds-page elisa western blot Agriculture (General) S1-972 Immunologic diseases. Allergy RC581-607
Acceso en línea:	https://doaj.org/article/29babe2b6a93433783e649cffe52231f
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spelling	oai:doaj.org-article:29babe2b6a93433783e649cffe52231f2021-11-26T11:19:46ZDevelopment and evaluation of polyclonal antibodies for detection of Pythium aphanidermatum and Fusarium oxysporum in ginger0954-01051465-344310.1080/09540105.2017.1365820https://doaj.org/article/29babe2b6a93433783e649cffe52231f2018-01-01T00:00:00Zhttp://dx.doi.org/10.1080/09540105.2017.1365820https://doaj.org/toc/0954-0105https://doaj.org/toc/1465-3443Polyclonal antibodies against Pythium aphanidermatum and Fusarium oxysporum proteins were developed for the detection of rhizome rot in ginger using serological assays. Under optimal experimental conditions, the detection limit of P. aphanidermatum by indirect ELISA was 10 µg/ml with a linear working range from 5 to 100 µg/ml (R2 = 0.994). In case of F. oxysporum, the linear working range was 5–100 µg/ml (R2 = 0.991) and the limit of detection was 25 µg/ml. The developed antibodies showed the highest titer in ELISA at 1:2000 dilutions. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis identified proteins ranging from molecular weights 15–97 kDa and 14–116 kDa of P. aphanidermatum and F. oxysporum isolates, respectively. In Western blot analysis, the developed antisera gave positive reactions against the isolated antigens of the fungi. The antibodies revealed immune-reactive bands of molecular weights 59 and 65 kDa in P. aphanidermatum and 44 and 75 kDa in F. oxysporum. The results suggest that the developed antibodies could be successfully applied for the specific immunodetection of P. aphanidermatum and F. oxysporum at an early stage of rhizome rot disease.Monalisa RaySwagatika DashK. Gopinath AcharySanghamitra NayakShikha SinghTaylor & Francis Grouparticlegingerrhizome rotpythium aphanidermatumfusarium oxysporumpolyclonal antibodiessds-pageelisawestern blotAgriculture (General)S1-972Immunologic diseases. AllergyRC581-607ENFood and Agricultural Immunology, Vol 29, Iss 1, Pp 204-215 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	ginger rhizome rot pythium aphanidermatum fusarium oxysporum polyclonal antibodies sds-page elisa western blot Agriculture (General) S1-972 Immunologic diseases. Allergy RC581-607
spellingShingle	ginger rhizome rot pythium aphanidermatum fusarium oxysporum polyclonal antibodies sds-page elisa western blot Agriculture (General) S1-972 Immunologic diseases. Allergy RC581-607 Monalisa Ray Swagatika Dash K. Gopinath Achary Sanghamitra Nayak Shikha Singh Development and evaluation of polyclonal antibodies for detection of Pythium aphanidermatum and Fusarium oxysporum in ginger
description	Polyclonal antibodies against Pythium aphanidermatum and Fusarium oxysporum proteins were developed for the detection of rhizome rot in ginger using serological assays. Under optimal experimental conditions, the detection limit of P. aphanidermatum by indirect ELISA was 10 µg/ml with a linear working range from 5 to 100 µg/ml (R2 = 0.994). In case of F. oxysporum, the linear working range was 5–100 µg/ml (R2 = 0.991) and the limit of detection was 25 µg/ml. The developed antibodies showed the highest titer in ELISA at 1:2000 dilutions. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis identified proteins ranging from molecular weights 15–97 kDa and 14–116 kDa of P. aphanidermatum and F. oxysporum isolates, respectively. In Western blot analysis, the developed antisera gave positive reactions against the isolated antigens of the fungi. The antibodies revealed immune-reactive bands of molecular weights 59 and 65 kDa in P. aphanidermatum and 44 and 75 kDa in F. oxysporum. The results suggest that the developed antibodies could be successfully applied for the specific immunodetection of P. aphanidermatum and F. oxysporum at an early stage of rhizome rot disease.
format	article
author	Monalisa Ray Swagatika Dash K. Gopinath Achary Sanghamitra Nayak Shikha Singh
author_facet	Monalisa Ray Swagatika Dash K. Gopinath Achary Sanghamitra Nayak Shikha Singh
author_sort	Monalisa Ray
title	Development and evaluation of polyclonal antibodies for detection of Pythium aphanidermatum and Fusarium oxysporum in ginger
title_short	Development and evaluation of polyclonal antibodies for detection of Pythium aphanidermatum and Fusarium oxysporum in ginger
title_full	Development and evaluation of polyclonal antibodies for detection of Pythium aphanidermatum and Fusarium oxysporum in ginger
title_fullStr	Development and evaluation of polyclonal antibodies for detection of Pythium aphanidermatum and Fusarium oxysporum in ginger
title_full_unstemmed	Development and evaluation of polyclonal antibodies for detection of Pythium aphanidermatum and Fusarium oxysporum in ginger
title_sort	development and evaluation of polyclonal antibodies for detection of pythium aphanidermatum and fusarium oxysporum in ginger
publisher	Taylor & Francis Group
publishDate	2018
url	https://doaj.org/article/29babe2b6a93433783e649cffe52231f
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Development and evaluation of polyclonal antibodies for detection of Pythium aphanidermatum and Fusarium oxysporum in ginger

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