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Molecular Detection of Harmful Raphidophyte Chattonella subsalsa Biecheler by Whole-Cell Fluorescence in situ Hybridisation Assay

Species of the genus Chattonella (Raphidophyceae) are a group of marine protists that are commonly found in coastal waters. Some are known as harmful microalgae that form noxious blooms and cause massive fish mortality in finfish aquaculture. In Malaysia, blooms of Chattonella have been recorded si...

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Main Authors: Winnie Lau, Lik Sing, Teng, Sing Tung, Lim, Hong Chang, Hii, Kieng Soon, Sandric Leong, Chee Yew, Leaw, Chui Pin, Lim, Po Teen
Format: Article
Language:English
Published: Penerbit Universiti Sains Malaysia 2023
Subjects:
QH301 Biology
QK Botany
Online Access:http://ir.unimas.my/id/eprint/44769/1/Molecular%20Detection.pdf
http://ir.unimas.my/id/eprint/44769/
https://ejournal.usm.my/tlsr/article/view/tlsr_vol34-no-1-2023_7
https://doi.org/10.21315/tlsr2023.34.1.7
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spelling my.unimas.ir.447692024-05-16T06:38:40Z http://ir.unimas.my/id/eprint/44769/ Molecular Detection of Harmful Raphidophyte Chattonella subsalsa Biecheler by Whole-Cell Fluorescence in situ Hybridisation Assay Winnie Lau, Lik Sing Teng, Sing Tung Lim, Hong Chang Hii, Kieng Soon Sandric Leong, Chee Yew Leaw, Chui Pin Lim, Po Teen QH301 Biology QK Botany Species of the genus Chattonella (Raphidophyceae) are a group of marine protists that are commonly found in coastal waters. Some are known as harmful microalgae that form noxious blooms and cause massive fish mortality in finfish aquaculture. In Malaysia, blooms of Chattonella have been recorded since the 1980s in the Johor Strait. In this study, two strains of Chattonella were established from the strait, and morphological examination revealed characteristics resembling Chattonella subsalsa. The molecular characterization further confirmed the species’ identity as C. subsalsa. To precisely detect the cells of C. subsalsa in the environment, a whole-cell fluorescence in-situ hybridisation (FISH) assay was developed. The species-specific oligonucleotide probes were designed in silico based on the nucleotide sequences of the large subunit (LSU) and internal transcribed spacer 2 (ITS2) of the ribosomal DNA (rDNA). The best candidate signature regions in the LSU-rRNA and ITS2-rDNA were selected based on hybridisation efficiency and probe parameters. The probes were synthesised as biotinylated probes and tested by tyramide signal amplification with FISH (FISH-TSA). The results showed the specificity of the probes toward the target cells. FISH-TSA has been proven to be a potential tool in the detection of harmful algae in the environment and could be applied to the harmful algal monitoring program. Penerbit Universiti Sains Malaysia 2023 Article PeerReviewed text en http://ir.unimas.my/id/eprint/44769/1/Molecular%20Detection.pdf Winnie Lau, Lik Sing and Teng, Sing Tung and Lim, Hong Chang and Hii, Kieng Soon and Sandric Leong, Chee Yew and Leaw, Chui Pin and Lim, Po Teen (2023) Molecular Detection of Harmful Raphidophyte Chattonella subsalsa Biecheler by Whole-Cell Fluorescence in situ Hybridisation Assay. Tropical Life Sciences Research, 34 (1). pp. 99-120. ISSN 2180-4249 https://ejournal.usm.my/tlsr/article/view/tlsr_vol34-no-1-2023_7 https://doi.org/10.21315/tlsr2023.34.1.7
institution Universiti Malaysia Sarawak
building Centre for Academic Information Services (CAIS)
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaysia Sarawak
content_source UNIMAS Institutional Repository
url_provider http://ir.unimas.my/
language English
topic QH301 Biology
QK Botany
spellingShingle QH301 Biology
QK Botany
Winnie Lau, Lik Sing
Teng, Sing Tung
Lim, Hong Chang
Hii, Kieng Soon
Sandric Leong, Chee Yew
Leaw, Chui Pin
Lim, Po Teen
Molecular Detection of Harmful Raphidophyte Chattonella subsalsa Biecheler by Whole-Cell Fluorescence in situ Hybridisation Assay
description Species of the genus Chattonella (Raphidophyceae) are a group of marine protists that are commonly found in coastal waters. Some are known as harmful microalgae that form noxious blooms and cause massive fish mortality in finfish aquaculture. In Malaysia, blooms of Chattonella have been recorded since the 1980s in the Johor Strait. In this study, two strains of Chattonella were established from the strait, and morphological examination revealed characteristics resembling Chattonella subsalsa. The molecular characterization further confirmed the species’ identity as C. subsalsa. To precisely detect the cells of C. subsalsa in the environment, a whole-cell fluorescence in-situ hybridisation (FISH) assay was developed. The species-specific oligonucleotide probes were designed in silico based on the nucleotide sequences of the large subunit (LSU) and internal transcribed spacer 2 (ITS2) of the ribosomal DNA (rDNA). The best candidate signature regions in the LSU-rRNA and ITS2-rDNA were selected based on hybridisation efficiency and probe parameters. The probes were synthesised as biotinylated probes and tested by tyramide signal amplification with FISH (FISH-TSA). The results showed the specificity of the probes toward the target cells. FISH-TSA has been proven to be a potential tool in the detection of harmful algae in the environment and could be applied to the harmful algal monitoring program.
format Article
author Winnie Lau, Lik Sing
Teng, Sing Tung
Lim, Hong Chang
Hii, Kieng Soon
Sandric Leong, Chee Yew
Leaw, Chui Pin
Lim, Po Teen
author_facet Winnie Lau, Lik Sing
Teng, Sing Tung
Lim, Hong Chang
Hii, Kieng Soon
Sandric Leong, Chee Yew
Leaw, Chui Pin
Lim, Po Teen
author_sort Winnie Lau, Lik Sing
title Molecular Detection of Harmful Raphidophyte Chattonella subsalsa Biecheler by Whole-Cell Fluorescence in situ Hybridisation Assay
title_short Molecular Detection of Harmful Raphidophyte Chattonella subsalsa Biecheler by Whole-Cell Fluorescence in situ Hybridisation Assay
title_full Molecular Detection of Harmful Raphidophyte Chattonella subsalsa Biecheler by Whole-Cell Fluorescence in situ Hybridisation Assay
title_fullStr Molecular Detection of Harmful Raphidophyte Chattonella subsalsa Biecheler by Whole-Cell Fluorescence in situ Hybridisation Assay
title_full_unstemmed Molecular Detection of Harmful Raphidophyte Chattonella subsalsa Biecheler by Whole-Cell Fluorescence in situ Hybridisation Assay
title_sort molecular detection of harmful raphidophyte chattonella subsalsa biecheler by whole-cell fluorescence in situ hybridisation assay
publisher Penerbit Universiti Sains Malaysia
publishDate 2023
url http://ir.unimas.my/id/eprint/44769/1/Molecular%20Detection.pdf
http://ir.unimas.my/id/eprint/44769/
https://ejournal.usm.my/tlsr/article/view/tlsr_vol34-no-1-2023_7
https://doi.org/10.21315/tlsr2023.34.1.7
_version_ 1800728098904211456
score 13.211869

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