Detection and Quantification of Alternaria solani in Tomato by Real Time PCR

A conventional and real-time PCR assays using SYBR Green for the detection and quantification of A. solanihave been developed and validated. A primer set (ALP and ITS4) designed from the ITS region of A. linicola/ A. solani complex, yielded a 536 bp product when DNA from 38 isolates of A. solani wer...

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Autores principales:	P Chowdappa, B J Nirmal Kumar, B Reddi Bhargavi, K R Hema, S P Mohan Kumar
Formato:	article
Lenguaje:	EN
Publicado:	Society for Promotion of Horticulture - Indian Institute of Horticultural Research 2014
Materias:	alternaria solani molecular diagnosis real time pcr early blight Plant culture SB1-1110
Acceso en línea:	https://doaj.org/article/123932889fba476fa168dcc2dfe043bf
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spelling	oai:doaj.org-article:123932889fba476fa168dcc2dfe043bf2021-12-02T11:12:39ZDetection and Quantification of Alternaria solani in Tomato by Real Time PCR0973-354X2582-4899https://doaj.org/article/123932889fba476fa168dcc2dfe043bf2014-12-01T00:00:00Zhttps://jhs.iihr.res.in/index.php/jhs/article/view/197https://doaj.org/toc/0973-354Xhttps://doaj.org/toc/2582-4899A conventional and real-time PCR assays using SYBR Green for the detection and quantification of A. solanihave been developed and validated. A primer set (ALP and ITS4) designed from the ITS region of A. linicola/ A. solani complex, yielded a 536 bp product when DNA from 38 isolates of A. solani were amplified. No product was amplified from A. alternata, A. brassicae, A. brassicicola, A.helianthi, A. porri, A. sesami, A.carthami, A.ricini, Colletotrichum gloeosporioides, C. capsici, C. falcatum, Cercospora canescens, C. capsici, Phytophthora infestans, Sclerotium rolfsii, Fusarium equiseti, F. oxysporum, Rhizoctonia solani, Phoma exigua, Curvularia spp and Drechslera. In addition, ALP/ITS4 primers were successfully utilized in real-time PCR assays of A. solani. The efficiency of conventional and real-time PCR assays was compared. The conventional PCR was able to detect the pathogen on symptomatic artificially infected tomato plants 5 days after pathogen inoculation. The detection limit was 100 conidia and 10 pg of DNA in the case of conventional PCR. Real-time PCR exhibited a detection limit 10 times lower (10 conidia, 10fg of DNA). The application of real time PCR assay for rapid detection of A.solani in infected tomato plant material is discussed.P ChowdappaB J Nirmal KumarB Reddi BhargaviK R HemaS P Mohan KumarSociety for Promotion of Horticulture - Indian Institute of Horticultural Researcharticlealternaria solanimolecular diagnosisreal time pcrearly blightPlant cultureSB1-1110ENJournal of Horticultural Sciences, Vol 9, Iss 2, Pp 196-201 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	alternaria solani molecular diagnosis real time pcr early blight Plant culture SB1-1110
spellingShingle	alternaria solani molecular diagnosis real time pcr early blight Plant culture SB1-1110 P Chowdappa B J Nirmal Kumar B Reddi Bhargavi K R Hema S P Mohan Kumar Detection and Quantification of Alternaria solani in Tomato by Real Time PCR
description	A conventional and real-time PCR assays using SYBR Green for the detection and quantification of A. solanihave been developed and validated. A primer set (ALP and ITS4) designed from the ITS region of A. linicola/ A. solani complex, yielded a 536 bp product when DNA from 38 isolates of A. solani were amplified. No product was amplified from A. alternata, A. brassicae, A. brassicicola, A.helianthi, A. porri, A. sesami, A.carthami, A.ricini, Colletotrichum gloeosporioides, C. capsici, C. falcatum, Cercospora canescens, C. capsici, Phytophthora infestans, Sclerotium rolfsii, Fusarium equiseti, F. oxysporum, Rhizoctonia solani, Phoma exigua, Curvularia spp and Drechslera. In addition, ALP/ITS4 primers were successfully utilized in real-time PCR assays of A. solani. The efficiency of conventional and real-time PCR assays was compared. The conventional PCR was able to detect the pathogen on symptomatic artificially infected tomato plants 5 days after pathogen inoculation. The detection limit was 100 conidia and 10 pg of DNA in the case of conventional PCR. Real-time PCR exhibited a detection limit 10 times lower (10 conidia, 10fg of DNA). The application of real time PCR assay for rapid detection of A.solani in infected tomato plant material is discussed.
format	article
author	P Chowdappa B J Nirmal Kumar B Reddi Bhargavi K R Hema S P Mohan Kumar
author_facet	P Chowdappa B J Nirmal Kumar B Reddi Bhargavi K R Hema S P Mohan Kumar
author_sort	P Chowdappa
title	Detection and Quantification of Alternaria solani in Tomato by Real Time PCR
title_short	Detection and Quantification of Alternaria solani in Tomato by Real Time PCR
title_full	Detection and Quantification of Alternaria solani in Tomato by Real Time PCR
title_fullStr	Detection and Quantification of Alternaria solani in Tomato by Real Time PCR
title_full_unstemmed	Detection and Quantification of Alternaria solani in Tomato by Real Time PCR
title_sort	detection and quantification of alternaria solani in tomato by real time pcr
publisher	Society for Promotion of Horticulture - Indian Institute of Horticultural Research
publishDate	2014
url	https://doaj.org/article/123932889fba476fa168dcc2dfe043bf
work_keys_str_mv	AT pchowdappa detectionandquantificationofalternariasolaniintomatobyrealtimepcr AT bjnirmalkumar detectionandquantificationofalternariasolaniintomatobyrealtimepcr AT breddibhargavi detectionandquantificationofalternariasolaniintomatobyrealtimepcr AT krhema detectionandquantificationofalternariasolaniintomatobyrealtimepcr AT spmohankumar detectionandquantificationofalternariasolaniintomatobyrealtimepcr
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Detection and Quantification of Alternaria solani in Tomato by Real Time PCR

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