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Development of a TaqMan real-time PCR for sensitive detection of the novel phytoplasma associated with coconut yellow decline in Malaysia.

Coconut yellow decline (CYD) disease in Malaysia has been shown to be associated with two different phytoplasmas, one from the 16SrXIV group and one from a novel taxonomic group. Here we report the development of a real-time PCR assay using a 16S rDNA-based TaqMan primer–probe set, for sensitive, qu...

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Main Authors: Nejat, Naghmeh, Sijam, Kamaruzaman, Abdullah, Siti Nor Akmar, Vadamalai, Ganesan, Sidek, Zakaria, Dickinson, Matthew
Format: Article
Language:English
English
Published: EDIZIONI ETS 2010
Online Access:http://psasir.upm.edu.my/id/eprint/14002/1/Development%20of%20a%20TaqMan%20real.pdf
http://psasir.upm.edu.my/id/eprint/14002/
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spelling my.upm.eprints.140022015-12-16T07:51:16Z http://psasir.upm.edu.my/id/eprint/14002/ Development of a TaqMan real-time PCR for sensitive detection of the novel phytoplasma associated with coconut yellow decline in Malaysia. Nejat, Naghmeh Sijam, Kamaruzaman Abdullah, Siti Nor Akmar Vadamalai, Ganesan Sidek, Zakaria Dickinson, Matthew Coconut yellow decline (CYD) disease in Malaysia has been shown to be associated with two different phytoplasmas, one from the 16SrXIV group and one from a novel taxonomic group. Here we report the development of a real-time PCR assay using a 16S rDNA-based TaqMan primer–probe set, for sensitive, quantitative and rapid detection of the novel strain of CYD phytoplasmas. The TaqMan PCR assay was successfully used on genomic DNA extracts from symptomatic Malayan yellow dwarf (MYD) and Malayan red dwarf (MRD) palms, and the analysis of field-collected coconut samples showed that the real-time PCR was better for phytoplasma detection than conventional nested PCR using primers P1/P7 followed by R16F2n/R16R2. Due to lack of specificity, the designed primer probe set also amplified DNA from a range of other phytoplasma strains, including 16SrII chickpea phyllody, 16SrIV Cape St Paul wilt, 16SrVI brinjal little leaf and 16SrXI napier grass stunt, but not from the 16SrXIV phytoplasma associated with CYD, so it could be used to distinguish between infections caused by the two different CYD phytoplasmas present in Malaysia. EDIZIONI ETS 2010 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/14002/1/Development%20of%20a%20TaqMan%20real.pdf Nejat, Naghmeh and Sijam, Kamaruzaman and Abdullah, Siti Nor Akmar and Vadamalai, Ganesan and Sidek, Zakaria and Dickinson, Matthew (2010) Development of a TaqMan real-time PCR for sensitive detection of the novel phytoplasma associated with coconut yellow decline in Malaysia. Journal of Plant Pathology, 92 (3). pp. 769-773. ISSN 1125-4653 English
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
English
description Coconut yellow decline (CYD) disease in Malaysia has been shown to be associated with two different phytoplasmas, one from the 16SrXIV group and one from a novel taxonomic group. Here we report the development of a real-time PCR assay using a 16S rDNA-based TaqMan primer–probe set, for sensitive, quantitative and rapid detection of the novel strain of CYD phytoplasmas. The TaqMan PCR assay was successfully used on genomic DNA extracts from symptomatic Malayan yellow dwarf (MYD) and Malayan red dwarf (MRD) palms, and the analysis of field-collected coconut samples showed that the real-time PCR was better for phytoplasma detection than conventional nested PCR using primers P1/P7 followed by R16F2n/R16R2. Due to lack of specificity, the designed primer probe set also amplified DNA from a range of other phytoplasma strains, including 16SrII chickpea phyllody, 16SrIV Cape St Paul wilt, 16SrVI brinjal little leaf and 16SrXI napier grass stunt, but not from the 16SrXIV phytoplasma associated with CYD, so it could be used to distinguish between infections caused by the two different CYD phytoplasmas present in Malaysia.
format Article
author Nejat, Naghmeh
Sijam, Kamaruzaman
Abdullah, Siti Nor Akmar
Vadamalai, Ganesan
Sidek, Zakaria
Dickinson, Matthew
spellingShingle Nejat, Naghmeh
Sijam, Kamaruzaman
Abdullah, Siti Nor Akmar
Vadamalai, Ganesan
Sidek, Zakaria
Dickinson, Matthew
Development of a TaqMan real-time PCR for sensitive detection of the novel phytoplasma associated with coconut yellow decline in Malaysia.
author_facet Nejat, Naghmeh
Sijam, Kamaruzaman
Abdullah, Siti Nor Akmar
Vadamalai, Ganesan
Sidek, Zakaria
Dickinson, Matthew
author_sort Nejat, Naghmeh
title Development of a TaqMan real-time PCR for sensitive detection of the novel phytoplasma associated with coconut yellow decline in Malaysia.
title_short Development of a TaqMan real-time PCR for sensitive detection of the novel phytoplasma associated with coconut yellow decline in Malaysia.
title_full Development of a TaqMan real-time PCR for sensitive detection of the novel phytoplasma associated with coconut yellow decline in Malaysia.
title_fullStr Development of a TaqMan real-time PCR for sensitive detection of the novel phytoplasma associated with coconut yellow decline in Malaysia.
title_full_unstemmed Development of a TaqMan real-time PCR for sensitive detection of the novel phytoplasma associated with coconut yellow decline in Malaysia.
title_sort development of a taqman real-time pcr for sensitive detection of the novel phytoplasma associated with coconut yellow decline in malaysia.
publisher EDIZIONI ETS
publishDate 2010
url http://psasir.upm.edu.my/id/eprint/14002/1/Development%20of%20a%20TaqMan%20real.pdf
http://psasir.upm.edu.my/id/eprint/14002/
_version_ 1643825501111844864
score 13.211869

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