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ITS2 secondary structure data improves authentication of Moringa oleifera tea products when using with DNA barcoding

Adulteration is a severe issue affecting the herbal industry. Therefore, a robust tool is needed to address this problem. In the current study, Moringa-based products (tea) authentication was conducted using DNA barcoding. Two different barcode regions, rbcL and ITS2, were investigated in terms of t...

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Main Authors: Mohd. Nasarodin, Nur Syazwani, Akbar, Anisah, Wagiran, Alina
Format: Article
Language:English
Published: Horizon e-Publishing Group (HePG) 2023
Subjects:
Q Science (General)
Online Access:http://eprints.utm.my/106366/1/AlinaWagiran2023_ITS2SecondaryStructureDataImproves.pdf
http://eprints.utm.my/106366/
http://dx.doi.org/10.14719/pst.2094
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spelling my.utm.1063662024-06-29T07:04:35Z http://eprints.utm.my/106366/ ITS2 secondary structure data improves authentication of Moringa oleifera tea products when using with DNA barcoding Mohd. Nasarodin, Nur Syazwani Akbar, Anisah Wagiran, Alina Q Science (General) Adulteration is a severe issue affecting the herbal industry. Therefore, a robust tool is needed to address this problem. In the current study, Moringa-based products (tea) authentication was conducted using DNA barcoding. Two different barcode regions, rbcL and ITS2, were investigated in terms of their effectiveness in authenticating the herbal products. To proceed with the DNA barcoding, a good quality of gDNA is a prerequisite for PCR amplification. Hence, two lysis buffers, PL1 and PL2, were compared to obtain good quality gDNA. Results revealed that a higher gDNA yield was obtained from the fresh plants using PL2, but a lower gDNA yield was obtained for the tea products except for sample P3. The PCR reaction was successfully conducted by amplifying two different barcodes, rbcL and ITS2. The amplicon size for the fresh plant was 643 bp for rbcL and 404 bp for ITS2. In contrast, the generated rbcL amplicon sizes for herbal tea products were 672 bp for P1, 679 bp for P2, 679 bp for P3, and 674 bp for P4. For rbcL and ITS2 amplicon sizes were 395 bp for P1, 406 bp for P2, 398 bp for P3, and 413 bp for P4. The amplified PCR products were analyzed using bioinformatic tools. The neighbour-joining (NJ) analysis for the rbcL barcode indicated that the P2, P3, and P4 tea products were categorized in the same clade with the M. oleifera sequence obtained from GenBank. Simultaneously, P1 was clustered individually with other closely related species. The analysis for the ITS2 barcode showed that all samples were grouped in the same clade. Incorporating secondary structure prediction after NJ analysis improved the discrimination between species. Horizon e-Publishing Group (HePG) 2023-07-28 Article PeerReviewed application/pdf en http://eprints.utm.my/106366/1/AlinaWagiran2023_ITS2SecondaryStructureDataImproves.pdf Mohd. Nasarodin, Nur Syazwani and Akbar, Anisah and Wagiran, Alina (2023) ITS2 secondary structure data improves authentication of Moringa oleifera tea products when using with DNA barcoding. Plant Science Today, 10 (4). pp. 1-11. ISSN 2348-1900 http://dx.doi.org/10.14719/pst.2094 DOI:10.14719/pst.2094
institution Universiti Teknologi Malaysia
building UTM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Teknologi Malaysia
content_source UTM Institutional Repository
url_provider http://eprints.utm.my/
language English
topic Q Science (General)
spellingShingle Q Science (General)
Mohd. Nasarodin, Nur Syazwani
Akbar, Anisah
Wagiran, Alina
ITS2 secondary structure data improves authentication of Moringa oleifera tea products when using with DNA barcoding
description Adulteration is a severe issue affecting the herbal industry. Therefore, a robust tool is needed to address this problem. In the current study, Moringa-based products (tea) authentication was conducted using DNA barcoding. Two different barcode regions, rbcL and ITS2, were investigated in terms of their effectiveness in authenticating the herbal products. To proceed with the DNA barcoding, a good quality of gDNA is a prerequisite for PCR amplification. Hence, two lysis buffers, PL1 and PL2, were compared to obtain good quality gDNA. Results revealed that a higher gDNA yield was obtained from the fresh plants using PL2, but a lower gDNA yield was obtained for the tea products except for sample P3. The PCR reaction was successfully conducted by amplifying two different barcodes, rbcL and ITS2. The amplicon size for the fresh plant was 643 bp for rbcL and 404 bp for ITS2. In contrast, the generated rbcL amplicon sizes for herbal tea products were 672 bp for P1, 679 bp for P2, 679 bp for P3, and 674 bp for P4. For rbcL and ITS2 amplicon sizes were 395 bp for P1, 406 bp for P2, 398 bp for P3, and 413 bp for P4. The amplified PCR products were analyzed using bioinformatic tools. The neighbour-joining (NJ) analysis for the rbcL barcode indicated that the P2, P3, and P4 tea products were categorized in the same clade with the M. oleifera sequence obtained from GenBank. Simultaneously, P1 was clustered individually with other closely related species. The analysis for the ITS2 barcode showed that all samples were grouped in the same clade. Incorporating secondary structure prediction after NJ analysis improved the discrimination between species.
format Article
author Mohd. Nasarodin, Nur Syazwani
Akbar, Anisah
Wagiran, Alina
author_facet Mohd. Nasarodin, Nur Syazwani
Akbar, Anisah
Wagiran, Alina
author_sort Mohd. Nasarodin, Nur Syazwani
title ITS2 secondary structure data improves authentication of Moringa oleifera tea products when using with DNA barcoding
title_short ITS2 secondary structure data improves authentication of Moringa oleifera tea products when using with DNA barcoding
title_full ITS2 secondary structure data improves authentication of Moringa oleifera tea products when using with DNA barcoding
title_fullStr ITS2 secondary structure data improves authentication of Moringa oleifera tea products when using with DNA barcoding
title_full_unstemmed ITS2 secondary structure data improves authentication of Moringa oleifera tea products when using with DNA barcoding
title_sort its2 secondary structure data improves authentication of moringa oleifera tea products when using with dna barcoding
publisher Horizon e-Publishing Group (HePG)
publishDate 2023
url http://eprints.utm.my/106366/1/AlinaWagiran2023_ITS2SecondaryStructureDataImproves.pdf
http://eprints.utm.my/106366/
http://dx.doi.org/10.14719/pst.2094
_version_ 1803334997933817856
score 13.211869

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