Loop-Mediated Isothermal Amplification (LAMP) Method for Rapid Detection of Rf1 Event in Transgenic Rapeseed (<i>Brassica napus</i> L.)

Zhang Jihong; Wang Jiaying; Zhao Lei; Huang Suwen; Chen Xianfeng

doi:doi:10.11648/j.jps.20221003.11

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Loop-Mediated Isothermal Amplification (LAMP) Method for Rapid Detection of Rf1 Event in Transgenic Rapeseed (Brassica napus L.)

Zhang Jihong, Wang Jiaying, Zhao Lei, Huang Suwen, Chen Xianfeng

Published in Journal of Plant Sciences (Volume 10, Issue 3)

Received: 1 April 2022 Accepted: 18 April 2022 Published: 7 May 2022

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Abstract

Rapeseed (Brassica napus L.) is one of the most studied crops for genetic engineering. Up to now, at least 11 GM rapeseed varieties have been approved for commercial production. During this study, a simple and rapid LAMP method was established for detection of Rf1 event in transgenic rapeseed. Position 143-353 in the right border junction sequence of B. napus transgenic line Rf1 (accession number: EU090199.1) was chosen as the target region for primer design and LAMP detection. Reaction mixture at the volume of 25 μL contained 12.5 μL 2×buffer mix, 1.28 μM FIP, 1.28 μM BIP, 0.16 μM F3, 0.16 μM B3, 0.64 μM LF, 480 U/mL Bst DNA polymerase, 2.0 μL DNA template and 4.6 μL ddH₂O. SYBR Green I was added to the tube lid. After incubation at 60°C for 60 min, the reaction was terminated by heating at 80°C for 10 min. Then the tube was centrifuged for 60 s at 8,000 r/min to mix dye with reaction mixture. Results indicate that our LAMP assay is highly specific and sensitive (9.2×10³ copies/μL, 0.001%). This LAMP assay for Rf1 event in rapeseed, which is simple, time effective, specific, sensitive and result visible without expensive instruments, is suitable for application in wide fields.

Published in	Journal of Plant Sciences (Volume 10, Issue 3)
DOI	10.11648/j.jps.20221003.11
Page(s)	86-90
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), 2022. Published by Science Publishing Group

Keywords

Rf1 Event, Loop-Mediated Isothermal Amplification, Genetically Modified Organism, Sensitivity, Specificity

References

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[5]	Zhou XY, Wu YY, Peng Q, et al. Indoor Propagation of Transgenic Glyphosate-tolerant Rapeseed (Brassica napus L.) and Transfer Breeding for Herbicide Tolerance. Agricultural Biotechnology (2164-4993), 2021, 10 (1): 1-5. 5p.
[6]	Wu Y, Wu G, Xiao L, et al. Event-specific qualitative and quantitative PCR detection methods for transgenic rapeseed hybrids MS1×RF1 and MS1×RF2. Journal of agricultural and food chemistry, 2007, 55 (21): 8380-8389.
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[9]	Li Q, Fang J, Liu X, et al. Loop-mediated isothermal amplification (LAMP) method for rapid detection of cry1Ab gene in transgenic rice (Oryza sativa L.). European Food Research and Technology, 2013, 236 (4): 589-598.
[10]	Misawa Y, Saito R, Moriya K, et al. Application of loop-mediated isothermal amplification technique to rapid and direct detection of methicillin-resistant Staphylococcus aureus (MRSA) in blood cultures. Journal of Infection and Chemotherapy, 2007, 13 (3): 134-140.
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[15]	Huang X, Zhu SF, Gao HW, et al. Protocol of the real-time PCR method for detecting genetically modified plants and their derived products. SN/T 1204-2016. General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China. 2016-6-28.

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APA Style

Zhang Jihong, Wang Jiaying, Zhao Lei, Huang Suwen, Chen Xianfeng. (2022). Loop-Mediated Isothermal Amplification (LAMP) Method for Rapid Detection of Rf1 Event in Transgenic Rapeseed (Brassica napus L.). Journal of Plant Sciences, 10(3), 86-90. https://doi.org/10.11648/j.jps.20221003.11

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ACS Style

Zhang Jihong; Wang Jiaying; Zhao Lei; Huang Suwen; Chen Xianfeng. Loop-Mediated Isothermal Amplification (LAMP) Method for Rapid Detection of Rf1 Event in Transgenic Rapeseed (Brassica napus L.). J. Plant Sci. 2022, 10(3), 86-90. doi: 10.11648/j.jps.20221003.11

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AMA Style

Zhang Jihong, Wang Jiaying, Zhao Lei, Huang Suwen, Chen Xianfeng. Loop-Mediated Isothermal Amplification (LAMP) Method for Rapid Detection of Rf1 Event in Transgenic Rapeseed (Brassica napus L.). J Plant Sci. 2022;10(3):86-90. doi: 10.11648/j.jps.20221003.11

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@article{10.11648/j.jps.20221003.11,
  author = {Zhang Jihong and Wang Jiaying and Zhao Lei and Huang Suwen and Chen Xianfeng},
  title = {Loop-Mediated Isothermal Amplification (LAMP) Method for Rapid Detection of Rf1 Event in Transgenic Rapeseed (Brassica napus L.)},
  journal = {Journal of Plant Sciences},
  volume = {10},
  number = {3},
  pages = {86-90},
  doi = {10.11648/j.jps.20221003.11},
  url = {https://doi.org/10.11648/j.jps.20221003.11},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.jps.20221003.11},
  abstract = {Rapeseed (Brassica napus L.) is one of the most studied crops for genetic engineering. Up to now, at least 11 GM rapeseed varieties have been approved for commercial production. During this study, a simple and rapid LAMP method was established for detection of Rf1 event in transgenic rapeseed. Position 143-353 in the right border junction sequence of B. napus transgenic line Rf1 (accession number: EU090199.1) was chosen as the target region for primer design and LAMP detection. Reaction mixture at the volume of 25 μL contained 12.5 μL 2×buffer mix, 1.28 μM FIP, 1.28 μM BIP, 0.16 μM F3, 0.16 μM B3, 0.64 μM LF, 480 U/mL Bst DNA polymerase, 2.0 μL DNA template and 4.6 μL ddH2O. SYBR Green I was added to the tube lid. After incubation at 60°C for 60 min, the reaction was terminated by heating at 80°C for 10 min. Then the tube was centrifuged for 60 s at 8,000 r/min to mix dye with reaction mixture. Results indicate that our LAMP assay is highly specific and sensitive (9.2×103 copies/μL, 0.001%). This LAMP assay for Rf1 event in rapeseed, which is simple, time effective, specific, sensitive and result visible without expensive instruments, is suitable for application in wide fields.},
 year = {2022}
}

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TY - JOUR
T1 - Loop-Mediated Isothermal Amplification (LAMP) Method for Rapid Detection of Rf1 Event in Transgenic Rapeseed (Brassica napus L.)
AU - Zhang Jihong
AU - Wang Jiaying
AU - Zhao Lei
AU - Huang Suwen
AU - Chen Xianfeng
Y1 - 2022/05/07
PY - 2022
N1 - https://doi.org/10.11648/j.jps.20221003.11
DO - 10.11648/j.jps.20221003.11
T2 - Journal of Plant Sciences
JF - Journal of Plant Sciences
JO - Journal of Plant Sciences
SP - 86
EP - 90
PB - Science Publishing Group
SN - 2331-0731
UR - https://doi.org/10.11648/j.jps.20221003.11
AB - Rapeseed (Brassica napus L.) is one of the most studied crops for genetic engineering. Up to now, at least 11 GM rapeseed varieties have been approved for commercial production. During this study, a simple and rapid LAMP method was established for detection of Rf1 event in transgenic rapeseed. Position 143-353 in the right border junction sequence of B. napus transgenic line Rf1 (accession number: EU090199.1) was chosen as the target region for primer design and LAMP detection. Reaction mixture at the volume of 25 μL contained 12.5 μL 2×buffer mix, 1.28 μM FIP, 1.28 μM BIP, 0.16 μM F3, 0.16 μM B3, 0.64 μM LF, 480 U/mL Bst DNA polymerase, 2.0 μL DNA template and 4.6 μL ddH2O. SYBR Green I was added to the tube lid. After incubation at 60°C for 60 min, the reaction was terminated by heating at 80°C for 10 min. Then the tube was centrifuged for 60 s at 8,000 r/min to mix dye with reaction mixture. Results indicate that our LAMP assay is highly specific and sensitive (9.2×103 copies/μL, 0.001%). This LAMP assay for Rf1 event in rapeseed, which is simple, time effective, specific, sensitive and result visible without expensive instruments, is suitable for application in wide fields.
VL - 10
IS - 3
ER -

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Author Information

Zhang Jihong

Technical Center, Ningbo Customs, Ningbo, China
Wang Jiaying

Technical Center, Ningbo Customs, Ningbo, China
Zhao Lei

Ningbo Institute of Inspection and Quarantine Science and Technology, Ningbo, China
Huang Suwen

Technical Center, Ningbo Customs, Ningbo, China
Chen Xianfeng

Technical Center, Ningbo Customs, Ningbo, China

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Plain Text BibTeX RIS

APA Style

Zhang Jihong, Wang Jiaying, Zhao Lei, Huang Suwen, Chen Xianfeng. (2022). Loop-Mediated Isothermal Amplification (LAMP) Method for Rapid Detection of Rf1 Event in Transgenic Rapeseed (Brassica napus L.). Journal of Plant Sciences, 10(3), 86-90. https://doi.org/10.11648/j.jps.20221003.11

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ACS Style

Zhang Jihong; Wang Jiaying; Zhao Lei; Huang Suwen; Chen Xianfeng. Loop-Mediated Isothermal Amplification (LAMP) Method for Rapid Detection of Rf1 Event in Transgenic Rapeseed (Brassica napus L.). J. Plant Sci. 2022, 10(3), 86-90. doi: 10.11648/j.jps.20221003.11

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AMA Style

Zhang Jihong, Wang Jiaying, Zhao Lei, Huang Suwen, Chen Xianfeng. Loop-Mediated Isothermal Amplification (LAMP) Method for Rapid Detection of Rf1 Event in Transgenic Rapeseed (Brassica napus L.). J Plant Sci. 2022;10(3):86-90. doi: 10.11648/j.jps.20221003.11

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@article{10.11648/j.jps.20221003.11,
  author = {Zhang Jihong and Wang Jiaying and Zhao Lei and Huang Suwen and Chen Xianfeng},
  title = {Loop-Mediated Isothermal Amplification (LAMP) Method for Rapid Detection of Rf1 Event in Transgenic Rapeseed (Brassica napus L.)},
  journal = {Journal of Plant Sciences},
  volume = {10},
  number = {3},
  pages = {86-90},
  doi = {10.11648/j.jps.20221003.11},
  url = {https://doi.org/10.11648/j.jps.20221003.11},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.jps.20221003.11},
  abstract = {Rapeseed (Brassica napus L.) is one of the most studied crops for genetic engineering. Up to now, at least 11 GM rapeseed varieties have been approved for commercial production. During this study, a simple and rapid LAMP method was established for detection of Rf1 event in transgenic rapeseed. Position 143-353 in the right border junction sequence of B. napus transgenic line Rf1 (accession number: EU090199.1) was chosen as the target region for primer design and LAMP detection. Reaction mixture at the volume of 25 μL contained 12.5 μL 2×buffer mix, 1.28 μM FIP, 1.28 μM BIP, 0.16 μM F3, 0.16 μM B3, 0.64 μM LF, 480 U/mL Bst DNA polymerase, 2.0 μL DNA template and 4.6 μL ddH2O. SYBR Green I was added to the tube lid. After incubation at 60°C for 60 min, the reaction was terminated by heating at 80°C for 10 min. Then the tube was centrifuged for 60 s at 8,000 r/min to mix dye with reaction mixture. Results indicate that our LAMP assay is highly specific and sensitive (9.2×103 copies/μL, 0.001%). This LAMP assay for Rf1 event in rapeseed, which is simple, time effective, specific, sensitive and result visible without expensive instruments, is suitable for application in wide fields.},
 year = {2022}
}

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