DNA Extraction from Heartwood and Quick Species Authentication Using Real-Time PCR: A Case Study of the Rosewood (<i>Pterocarpus Indicus</i>)

Zhang Jihong; Wang Jiaying; Xu Ying; Lv Xiaoling; Wei Jianhua; Ma Ming; Cui Junxia

doi:doi:10.11648/j.jps.20231102.11

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DNA Extraction from Heartwood and Quick Species Authentication Using Real-Time PCR: A Case Study of the Rosewood (Pterocarpus Indicus)

Zhang Jihong, Wang Jiaying, Xu Ying, Lv Xiaoling, Wei Jianhua, Ma Ming, Cui Junxia

Published in Journal of Plant Sciences (Volume 11, Issue 2)

Received: 20 March 2023 Accepted: 4 April 2023 Published: 27 April 2023

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Abstract

Illegal logging, felling and timber trade have continued to increase over the past two decades, leading to a decline in forest biodiversity and the extinction of some wood species. Pterocarpus indicus Willd., listed in the National Standards of the People’s Republic of China for Hongmu (GB/T 18107-2017), is widely used in production of high-end furniture, decorative flooring and musical instruments due to its high-quality timber. For molecular species identification, the quality and quantity of DNA extracted from wood samples should first be ensured. However, extracting DNA from dried, aged timber heartwood is difficult, as heartwood contains little fragmented DNA, along with lots of phenolic compounds known to impede sequence amplification. In order to protect P. indicus from over-exploitation and to achieve accurate species-level identification, we established a particular extraction method for obtaining amplifiable DNA from heartwood samples and the real-time PCR assay for species discrimination of P. indicus in this study. The quantity and quality of DNA extracted from dry heartwood samples using the modified CTAB method were 2.40-37.70 ng/μL and 1.55-2.12 demonstrated by OD_260/280, respectively. Primer set P9, targeting P. indicus specific microsatellite Pin2-20 sequence, was amplifiable in newly established real-time PCR. Through analysis, this real time PCR was shown to be specific and sensitive with a detection limit around 0.17 ng/μL. Hopefully, this study will contribute to heartwood DNA extraction and species identification of timber logs for forensic discrimination, law enforcement and natural resource conservation.

Published in	Journal of Plant Sciences (Volume 11, Issue 2)
DOI	10.11648/j.jps.20231102.11
Page(s)	28-34
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2023. Published by Science Publishing Group

Keywords

DNA Extraction, Heartwood, Species Discrimination, Biodiversity

References

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APA Style

Zhang Jihong, Wang Jiaying, Xu Ying, Lv Xiaoling, Wei Jianhua, et al. (2023). DNA Extraction from Heartwood and Quick Species Authentication Using Real-Time PCR: A Case Study of the Rosewood (Pterocarpus Indicus). Journal of Plant Sciences, 11(2), 28-34. https://doi.org/10.11648/j.jps.20231102.11

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ACS Style

Zhang Jihong; Wang Jiaying; Xu Ying; Lv Xiaoling; Wei Jianhua, et al. DNA Extraction from Heartwood and Quick Species Authentication Using Real-Time PCR: A Case Study of the Rosewood (Pterocarpus Indicus). J. Plant Sci. 2023, 11(2), 28-34. doi: 10.11648/j.jps.20231102.11

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AMA Style

Zhang Jihong, Wang Jiaying, Xu Ying, Lv Xiaoling, Wei Jianhua, et al. DNA Extraction from Heartwood and Quick Species Authentication Using Real-Time PCR: A Case Study of the Rosewood (Pterocarpus Indicus). J Plant Sci. 2023;11(2):28-34. doi: 10.11648/j.jps.20231102.11

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@article{10.11648/j.jps.20231102.11,
  author = {Zhang Jihong and Wang Jiaying and Xu Ying and Lv Xiaoling and Wei Jianhua and Ma Ming and Cui Junxia},
  title = {DNA Extraction from Heartwood and Quick Species Authentication Using Real-Time PCR: A Case Study of the Rosewood (Pterocarpus Indicus)},
  journal = {Journal of Plant Sciences},
  volume = {11},
  number = {2},
  pages = {28-34},
  doi = {10.11648/j.jps.20231102.11},
  url = {https://doi.org/10.11648/j.jps.20231102.11},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.jps.20231102.11},
  abstract = {Illegal logging, felling and timber trade have continued to increase over the past two decades, leading to a decline in forest biodiversity and the extinction of some wood species. Pterocarpus indicus Willd., listed in the National Standards of the People’s Republic of China for Hongmu (GB/T 18107-2017), is widely used in production of high-end furniture, decorative flooring and musical instruments due to its high-quality timber. For molecular species identification, the quality and quantity of DNA extracted from wood samples should first be ensured. However, extracting DNA from dried, aged timber heartwood is difficult, as heartwood contains little fragmented DNA, along with lots of phenolic compounds known to impede sequence amplification. In order to protect P. indicus from over-exploitation and to achieve accurate species-level identification, we established a particular extraction method for obtaining amplifiable DNA from heartwood samples and the real-time PCR assay for species discrimination of P. indicus in this study. The quantity and quality of DNA extracted from dry heartwood samples using the modified CTAB method were 2.40-37.70 ng/μL and 1.55-2.12 demonstrated by OD260/280, respectively. Primer set P9, targeting P. indicus specific microsatellite Pin2-20 sequence, was amplifiable in newly established real-time PCR. Through analysis, this real time PCR was shown to be specific and sensitive with a detection limit around 0.17 ng/μL. Hopefully, this study will contribute to heartwood DNA extraction and species identification of timber logs for forensic discrimination, law enforcement and natural resource conservation.},
 year = {2023}
}

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TY - JOUR
T1 - DNA Extraction from Heartwood and Quick Species Authentication Using Real-Time PCR: A Case Study of the Rosewood (Pterocarpus Indicus)
AU - Zhang Jihong
AU - Wang Jiaying
AU - Xu Ying
AU - Lv Xiaoling
AU - Wei Jianhua
AU - Ma Ming
AU - Cui Junxia
Y1 - 2023/04/27
PY - 2023
N1 - https://doi.org/10.11648/j.jps.20231102.11
DO - 10.11648/j.jps.20231102.11
T2 - Journal of Plant Sciences
JF - Journal of Plant Sciences
JO - Journal of Plant Sciences
SP - 28
EP - 34
PB - Science Publishing Group
SN - 2331-0731
UR - https://doi.org/10.11648/j.jps.20231102.11
AB - Illegal logging, felling and timber trade have continued to increase over the past two decades, leading to a decline in forest biodiversity and the extinction of some wood species. Pterocarpus indicus Willd., listed in the National Standards of the People’s Republic of China for Hongmu (GB/T 18107-2017), is widely used in production of high-end furniture, decorative flooring and musical instruments due to its high-quality timber. For molecular species identification, the quality and quantity of DNA extracted from wood samples should first be ensured. However, extracting DNA from dried, aged timber heartwood is difficult, as heartwood contains little fragmented DNA, along with lots of phenolic compounds known to impede sequence amplification. In order to protect P. indicus from over-exploitation and to achieve accurate species-level identification, we established a particular extraction method for obtaining amplifiable DNA from heartwood samples and the real-time PCR assay for species discrimination of P. indicus in this study. The quantity and quality of DNA extracted from dry heartwood samples using the modified CTAB method were 2.40-37.70 ng/μL and 1.55-2.12 demonstrated by OD260/280, respectively. Primer set P9, targeting P. indicus specific microsatellite Pin2-20 sequence, was amplifiable in newly established real-time PCR. Through analysis, this real time PCR was shown to be specific and sensitive with a detection limit around 0.17 ng/μL. Hopefully, this study will contribute to heartwood DNA extraction and species identification of timber logs for forensic discrimination, law enforcement and natural resource conservation.
VL - 11
IS - 2
ER -

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Author Information

Zhang Jihong

Technical Center, Ningbo Customs, Ningbo, China
Wang Jiaying

Technical Center, Ningbo Customs, Ningbo, China
Xu Ying

Technical Center, Ningbo Customs, Ningbo, China
Lv Xiaoling

Technical Center, Ningbo Customs, Ningbo, China
Wei Jianhua

Technical Center, Shantou Customs, Guangdong, China
Ma Ming

Technical Center, Ningbo Customs, Ningbo, China
Cui Junxia

Technical Center, Ningbo Customs, Ningbo, China

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APA Style

Zhang Jihong, Wang Jiaying, Xu Ying, Lv Xiaoling, Wei Jianhua, et al. (2023). DNA Extraction from Heartwood and Quick Species Authentication Using Real-Time PCR: A Case Study of the Rosewood (Pterocarpus Indicus). Journal of Plant Sciences, 11(2), 28-34. https://doi.org/10.11648/j.jps.20231102.11

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ACS Style

Zhang Jihong; Wang Jiaying; Xu Ying; Lv Xiaoling; Wei Jianhua, et al. DNA Extraction from Heartwood and Quick Species Authentication Using Real-Time PCR: A Case Study of the Rosewood (Pterocarpus Indicus). J. Plant Sci. 2023, 11(2), 28-34. doi: 10.11648/j.jps.20231102.11

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AMA Style

Zhang Jihong, Wang Jiaying, Xu Ying, Lv Xiaoling, Wei Jianhua, et al. DNA Extraction from Heartwood and Quick Species Authentication Using Real-Time PCR: A Case Study of the Rosewood (Pterocarpus Indicus). J Plant Sci. 2023;11(2):28-34. doi: 10.11648/j.jps.20231102.11

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@article{10.11648/j.jps.20231102.11,
  author = {Zhang Jihong and Wang Jiaying and Xu Ying and Lv Xiaoling and Wei Jianhua and Ma Ming and Cui Junxia},
  title = {DNA Extraction from Heartwood and Quick Species Authentication Using Real-Time PCR: A Case Study of the Rosewood (Pterocarpus Indicus)},
  journal = {Journal of Plant Sciences},
  volume = {11},
  number = {2},
  pages = {28-34},
  doi = {10.11648/j.jps.20231102.11},
  url = {https://doi.org/10.11648/j.jps.20231102.11},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.jps.20231102.11},
  abstract = {Illegal logging, felling and timber trade have continued to increase over the past two decades, leading to a decline in forest biodiversity and the extinction of some wood species. Pterocarpus indicus Willd., listed in the National Standards of the People’s Republic of China for Hongmu (GB/T 18107-2017), is widely used in production of high-end furniture, decorative flooring and musical instruments due to its high-quality timber. For molecular species identification, the quality and quantity of DNA extracted from wood samples should first be ensured. However, extracting DNA from dried, aged timber heartwood is difficult, as heartwood contains little fragmented DNA, along with lots of phenolic compounds known to impede sequence amplification. In order to protect P. indicus from over-exploitation and to achieve accurate species-level identification, we established a particular extraction method for obtaining amplifiable DNA from heartwood samples and the real-time PCR assay for species discrimination of P. indicus in this study. The quantity and quality of DNA extracted from dry heartwood samples using the modified CTAB method were 2.40-37.70 ng/μL and 1.55-2.12 demonstrated by OD260/280, respectively. Primer set P9, targeting P. indicus specific microsatellite Pin2-20 sequence, was amplifiable in newly established real-time PCR. Through analysis, this real time PCR was shown to be specific and sensitive with a detection limit around 0.17 ng/μL. Hopefully, this study will contribute to heartwood DNA extraction and species identification of timber logs for forensic discrimination, law enforcement and natural resource conservation.},
 year = {2023}
}

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