Cargando…

Tobacco Mosaic Virus Infection of Chrysanthemums in Thailand: Development of Colorimetric Reverse-Transcription Loop-Mediated Isothermal Amplification (RT–LAMP) Technique for Sensitive and Rapid Detection

We detected tobacco mosaic virus (TMV), a member of the genus Tobamovirus and one of the most significant plant-infecting viruses, for the first time in a chrysanthemum in Thailand using reverse-transcription polymerase chain reaction (RT–PCR). The TMV-infected chrysanthemum leaves exhibited mosaic...

Descripción completa

Detalles Bibliográficos
Autores principales: Supakitthanakorn, Salit, Vichittragoontavorn, Kanjana, Sunpapao, Anurag, Kunasakdakul, Kaewalin, Thapanapongworakul, Pilunthana, Ruangwong, On-Uma
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9325109/
https://www.ncbi.nlm.nih.gov/pubmed/35890422
http://dx.doi.org/10.3390/plants11141788
_version_ 1784756966096109568
author Supakitthanakorn, Salit
Vichittragoontavorn, Kanjana
Sunpapao, Anurag
Kunasakdakul, Kaewalin
Thapanapongworakul, Pilunthana
Ruangwong, On-Uma
author_facet Supakitthanakorn, Salit
Vichittragoontavorn, Kanjana
Sunpapao, Anurag
Kunasakdakul, Kaewalin
Thapanapongworakul, Pilunthana
Ruangwong, On-Uma
author_sort Supakitthanakorn, Salit
collection PubMed
description We detected tobacco mosaic virus (TMV), a member of the genus Tobamovirus and one of the most significant plant-infecting viruses, for the first time in a chrysanthemum in Thailand using reverse-transcription polymerase chain reaction (RT–PCR). The TMV-infected chrysanthemum leaves exhibited mosaic symptoms. We conducted a sequence analysis of the coat protein (CP) gene and found that the TMV detected in the chrysanthemum had 98% identity with other TMV isolates in GenBank. We carried out bioassays and showed that TMV induced mosaic and stunting symptoms in inoculated chrysanthemums. We observed the rigid rod structure of TMV under a transmission electron microscope (TEM). To enhance the speed and sensitivity of detection, we developed a colorimetric RT loop-mediated isothermal amplification (LAMP) technique. We achieved LAMP detection after 30 min incubation in isothermal conditions at 65 °C, and distinguished the positive results according to the color change from pink to yellow. The sensitivity of the LAMP technique was 1000-fold greater than that of RT–PCR, and we found no cross-reactivity with other viruses or viroids. This is the first reported case of a TMV-infected chrysanthemum in Thailand, and our colorimetric RT–LAMP TMV detection method is the first of its kind.
format Online
Article
Text
id pubmed-9325109
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-93251092022-07-27 Tobacco Mosaic Virus Infection of Chrysanthemums in Thailand: Development of Colorimetric Reverse-Transcription Loop-Mediated Isothermal Amplification (RT–LAMP) Technique for Sensitive and Rapid Detection Supakitthanakorn, Salit Vichittragoontavorn, Kanjana Sunpapao, Anurag Kunasakdakul, Kaewalin Thapanapongworakul, Pilunthana Ruangwong, On-Uma Plants (Basel) Article We detected tobacco mosaic virus (TMV), a member of the genus Tobamovirus and one of the most significant plant-infecting viruses, for the first time in a chrysanthemum in Thailand using reverse-transcription polymerase chain reaction (RT–PCR). The TMV-infected chrysanthemum leaves exhibited mosaic symptoms. We conducted a sequence analysis of the coat protein (CP) gene and found that the TMV detected in the chrysanthemum had 98% identity with other TMV isolates in GenBank. We carried out bioassays and showed that TMV induced mosaic and stunting symptoms in inoculated chrysanthemums. We observed the rigid rod structure of TMV under a transmission electron microscope (TEM). To enhance the speed and sensitivity of detection, we developed a colorimetric RT loop-mediated isothermal amplification (LAMP) technique. We achieved LAMP detection after 30 min incubation in isothermal conditions at 65 °C, and distinguished the positive results according to the color change from pink to yellow. The sensitivity of the LAMP technique was 1000-fold greater than that of RT–PCR, and we found no cross-reactivity with other viruses or viroids. This is the first reported case of a TMV-infected chrysanthemum in Thailand, and our colorimetric RT–LAMP TMV detection method is the first of its kind. MDPI 2022-07-06 /pmc/articles/PMC9325109/ /pubmed/35890422 http://dx.doi.org/10.3390/plants11141788 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Supakitthanakorn, Salit
Vichittragoontavorn, Kanjana
Sunpapao, Anurag
Kunasakdakul, Kaewalin
Thapanapongworakul, Pilunthana
Ruangwong, On-Uma
Tobacco Mosaic Virus Infection of Chrysanthemums in Thailand: Development of Colorimetric Reverse-Transcription Loop-Mediated Isothermal Amplification (RT–LAMP) Technique for Sensitive and Rapid Detection
title Tobacco Mosaic Virus Infection of Chrysanthemums in Thailand: Development of Colorimetric Reverse-Transcription Loop-Mediated Isothermal Amplification (RT–LAMP) Technique for Sensitive and Rapid Detection
title_full Tobacco Mosaic Virus Infection of Chrysanthemums in Thailand: Development of Colorimetric Reverse-Transcription Loop-Mediated Isothermal Amplification (RT–LAMP) Technique for Sensitive and Rapid Detection
title_fullStr Tobacco Mosaic Virus Infection of Chrysanthemums in Thailand: Development of Colorimetric Reverse-Transcription Loop-Mediated Isothermal Amplification (RT–LAMP) Technique for Sensitive and Rapid Detection
title_full_unstemmed Tobacco Mosaic Virus Infection of Chrysanthemums in Thailand: Development of Colorimetric Reverse-Transcription Loop-Mediated Isothermal Amplification (RT–LAMP) Technique for Sensitive and Rapid Detection
title_short Tobacco Mosaic Virus Infection of Chrysanthemums in Thailand: Development of Colorimetric Reverse-Transcription Loop-Mediated Isothermal Amplification (RT–LAMP) Technique for Sensitive and Rapid Detection
title_sort tobacco mosaic virus infection of chrysanthemums in thailand: development of colorimetric reverse-transcription loop-mediated isothermal amplification (rt–lamp) technique for sensitive and rapid detection
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9325109/
https://www.ncbi.nlm.nih.gov/pubmed/35890422
http://dx.doi.org/10.3390/plants11141788
work_keys_str_mv AT supakitthanakornsalit tobaccomosaicvirusinfectionofchrysanthemumsinthailanddevelopmentofcolorimetricreversetranscriptionloopmediatedisothermalamplificationrtlamptechniqueforsensitiveandrapiddetection
AT vichittragoontavornkanjana tobaccomosaicvirusinfectionofchrysanthemumsinthailanddevelopmentofcolorimetricreversetranscriptionloopmediatedisothermalamplificationrtlamptechniqueforsensitiveandrapiddetection
AT sunpapaoanurag tobaccomosaicvirusinfectionofchrysanthemumsinthailanddevelopmentofcolorimetricreversetranscriptionloopmediatedisothermalamplificationrtlamptechniqueforsensitiveandrapiddetection
AT kunasakdakulkaewalin tobaccomosaicvirusinfectionofchrysanthemumsinthailanddevelopmentofcolorimetricreversetranscriptionloopmediatedisothermalamplificationrtlamptechniqueforsensitiveandrapiddetection
AT thapanapongworakulpilunthana tobaccomosaicvirusinfectionofchrysanthemumsinthailanddevelopmentofcolorimetricreversetranscriptionloopmediatedisothermalamplificationrtlamptechniqueforsensitiveandrapiddetection
AT ruangwongonuma tobaccomosaicvirusinfectionofchrysanthemumsinthailanddevelopmentofcolorimetricreversetranscriptionloopmediatedisothermalamplificationrtlamptechniqueforsensitiveandrapiddetection