Isolation and PCR Validation of DNA from Chicory (<i>Cichorium intybus</i> L.) Leaves Using an Optimized CTAB Method to Propel Medicinal Insights

Shadab Ahamad; Mamta Arya; Gohar Taj

doi:doi:10.11648/j.jdmp.20241004.12

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Isolation and PCR Validation of DNA from Chicory (Cichorium intybus L.) Leaves Using an Optimized CTAB Method to Propel Medicinal Insights

Shadab Ahamad^*

, Mamta Arya

, Gohar Taj

Published in Journal of Diseases and Medicinal Plants (Volume 10, Issue 4)

Received: 2 December 2024 Accepted: 12 December 2024 Published: 27 December 2024

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Abstract

Isolating and purifying DNA from plants, particularly from species like Cichorium intybus (chicory), poses significant challenges due to the presence of rigid cell walls and high levels of polyphenols and polysaccharides. These compounds can severely hinder the efficiency of traditional DNA extraction protocols, often resulting in poor-quality DNA. Conventional methods, such as the Cetyl-trimethyl-ammonium bromide (CTAB) protocol, are often found inadequate due to these biochemical barriers. In this study, we formulated an optimized DNA isolation protocol for chicory that eliminates the need of liquid nitrogen and phenol during processing. The refined approach resulted in the extraction of high-quality DNA suitable for downstream applications. The effectiveness of the optimized method was validated by polymerase chain reaction (PCR) amplification of the sucrose: sucrose 1-fructosyl transferase (1-SST) gene, a key gene involved in the biosynthesis of inulin- a compound with notable medicinal and nutritional value. The PCR amplification yielded the expected 2.01 kb product, confirming the high quality and integrity of the extracted DNA. Overall, the quantity and quality of the DNA were well-suited for molecular analysis, demonstrating the success of our method. This optimized protocol provides a valuable tool for molecular research on chicory and other plants with similar biochemical challenges, facilitating more efficient genetic studies and holding potential for advancing biotechnological applications.

Published in	Journal of Diseases and Medicinal Plants (Volume 10, Issue 4)
DOI	10.11648/j.jdmp.20241004.12
Page(s)	69-72
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), 2024. Published by Science Publishing Group

Keywords

Chicory, CTAB, DNA, Polyphenols, Polysaccharides, 1-SST Gene

References

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[2]	Qadir, I., Bazaz, M. R., Dar, R. M., Ovais, S., Mir, S. R., Zarkar, M. I., Rehman, M. U. Cichorium intybus: A comprehensive review on its pharmacological activity and phytochemistry. Edible plants in health and diseases. Singapore: Springer; 2022. https://doi.org/10.1007/978-981-16-4959-2_12
[3]	Ahamad, S., Upadhayay, V. K., Tiwari, A., Taj, G. In silico exploration of interaction profile of fructan 6G- fructosyltransferase (6G-FTT) enzyme with 1-kestose from Cichorium intybus in order to determine active site of this conjugation. Letters in Organic Chemistry. 2023, 20(7), 612-617. http://dx.doi.org/10.2174/1570178620666230123101752
[4]	Duda, Ł., Kłosiński, K. K., Budryn, G., Jaśkiewicz, A., Kołat, D., Kałuzińska-Kołat, Z., Pasieka, Z. W. Medicinal use of chicory (Cichorium intybus l.). Scientia Pharmaceutica. 2024, 92(2), 31. https://doi.org/10.3390/scipharm92020031
[5]	Shoorideh, H., Peighambari, S. A., Omidi, M., Naghavi, M. R., Maroufi, A. Spatial expression of genes in inulin biosynthesis pathway in wild and root type chicory. Journal of Agricultural Science and Technology. 2018, 20, 1049-1058. https://dorl.net/dor/20.1001.1.16807073.2018.20.5.12.2
[6]	Pouille, C. L., Ouaza, S., Roels, E., Behra, J., Tourret, M., Molinié, R., Fontaine, J. X., Mathiron, D., Gagneul, D., Taminiau, B., Daube, G., Ravallec, R., Rambaud, C., Hilbert, J. L., Cudennec, B., & Lucau-Danila, A. Chicory: Understanding the Effects and Effectors of This Functional Food. Nutrients. 2022, 14(5), 957. https://doi.org/10.3390/nu14050957
[7]	Jabeen, R., Habiba, U., Khalid, S., Shirazi, J. H. Extraction and comparison of quality and purity of DNA of medicinal plants by CTAB and kit method. Journal of Contemporary Pharmacy. 2022, 6(1). https://doi.org/10.56770/jcp2022613
[8]	Aboul-Ftooh, N., & Oraby, H. A. Extraction of high-quality genomic DNA from different plant orders applying a modified CTAB-based method. Bulletin of the National Research Centre. 2019, 43(1), 1-10. https://doi.org/10.1186/s42269-019-0066-1
[9]	Milala, J., Grzelak, K., Król, B., Juśkiewicz, J., Zduńczyk, Z. COMPOSITION AND PROPERTIES OF CHICORY EXTRACTS RICH IN FRUCTANS AND POLYPHENOLS. Polish Journal of Food and Nutrition Sciences. 2009, 59(1), 35-43.
[10]	Aggarwal, G., Edhigalla, P., Walia, P. A comprehensive review of high-quality plant DNA isolation. The Pharma Innovation Journal. 2022, 11(6), 171-176.
[11]	Gawel, N. J., Jarret, R. L. A modified CTAB DNA extraction procedure for Musa and Ipomoea. Plant Molecular Biology Reporter. 1991, 9, 262–266. https://doi.org/10.1007/bf02672076
[12]	Barnwell, P., Blanchard, A. N., Bryant, J. A. Smirnoff N and Weir AF: Isolation of DNA from the highly mucilaginous succulent plant Sedum telephium. Plant Molecular Biology Reporter. 1998, 16, 133–138. https://doi.org/10.1023/a:1007473302551
[13]	Michiels, A., Van Den Ende, Tucker, W. M., Van Riet, L. Van Laere, A. Extraction of high-quality genomic DNA from latex-containing plants. Analytical Biochemistry. 2003, 315(1), 85-89. https://doi.org/10.1016/s0003-2697(02)00665-6
[14]	Yadav, S., Ram, C., Singh, S., Rana, M. K. Plant genomic DNA isolation-the past and the present, A review. Indian Journal of Plant Genetic Resources. 2018, 31(3), 315-327. http://dx.doi.org/10.5958/0976-1926.2018.00037.2
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APA Style

Ahamad, S., Arya, M., Taj, G. (2024). Isolation and PCR Validation of DNA from Chicory (Cichorium intybus L.) Leaves Using an Optimized CTAB Method to Propel Medicinal Insights. Journal of Diseases and Medicinal Plants, 10(4), 69-72. https://doi.org/10.11648/j.jdmp.20241004.12

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ACS Style

Ahamad, S.; Arya, M.; Taj, G. Isolation and PCR Validation of DNA from Chicory (Cichorium intybus L.) Leaves Using an Optimized CTAB Method to Propel Medicinal Insights. J. Dis. Med. Plants 2024, 10(4), 69-72. doi: 10.11648/j.jdmp.20241004.12

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AMA Style

Ahamad S, Arya M, Taj G. Isolation and PCR Validation of DNA from Chicory (Cichorium intybus L.) Leaves Using an Optimized CTAB Method to Propel Medicinal Insights. J Dis Med Plants. 2024;10(4):69-72. doi: 10.11648/j.jdmp.20241004.12

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@article{10.11648/j.jdmp.20241004.12,
  author = {Shadab Ahamad and Mamta Arya and Gohar Taj},
  title = {Isolation and PCR Validation of DNA from Chicory (Cichorium intybus L.) Leaves Using an Optimized CTAB Method to Propel Medicinal Insights
},
  journal = {Journal of Diseases and Medicinal Plants},
  volume = {10},
  number = {4},
  pages = {69-72},
  doi = {10.11648/j.jdmp.20241004.12},
  url = {https://doi.org/10.11648/j.jdmp.20241004.12},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.jdmp.20241004.12},
  abstract = {Isolating and purifying DNA from plants, particularly from species like Cichorium intybus (chicory), poses significant challenges due to the presence of rigid cell walls and high levels of polyphenols and polysaccharides. These compounds can severely hinder the efficiency of traditional DNA extraction protocols, often resulting in poor-quality DNA. Conventional methods, such as the Cetyl-trimethyl-ammonium bromide (CTAB) protocol, are often found inadequate due to these biochemical barriers. In this study, we formulated an optimized DNA isolation protocol for chicory that eliminates the need of liquid nitrogen and phenol during processing. The refined approach resulted in the extraction of high-quality DNA suitable for downstream applications. The effectiveness of the optimized method was validated by polymerase chain reaction (PCR) amplification of the sucrose: sucrose 1-fructosyl transferase (1-SST) gene, a key gene involved in the biosynthesis of inulin- a compound with notable medicinal and nutritional value. The PCR amplification yielded the expected 2.01 kb product, confirming the high quality and integrity of the extracted DNA. Overall, the quantity and quality of the DNA were well-suited for molecular analysis, demonstrating the success of our method. This optimized protocol provides a valuable tool for molecular research on chicory and other plants with similar biochemical challenges, facilitating more efficient genetic studies and holding potential for advancing biotechnological applications.
},
 year = {2024}
}

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TY  - JOUR
T1  - Isolation and PCR Validation of DNA from Chicory (Cichorium intybus L.) Leaves Using an Optimized CTAB Method to Propel Medicinal Insights

AU  - Shadab Ahamad
AU  - Mamta Arya
AU  - Gohar Taj
Y1  - 2024/12/27
PY  - 2024
N1  - https://doi.org/10.11648/j.jdmp.20241004.12
DO  - 10.11648/j.jdmp.20241004.12
T2  - Journal of Diseases and Medicinal Plants
JF  - Journal of Diseases and Medicinal Plants
JO  - Journal of Diseases and Medicinal Plants
SP  - 69
EP  - 72
PB  - Science Publishing Group
SN  - 2469-8210
UR  - https://doi.org/10.11648/j.jdmp.20241004.12
AB  - Isolating and purifying DNA from plants, particularly from species like Cichorium intybus (chicory), poses significant challenges due to the presence of rigid cell walls and high levels of polyphenols and polysaccharides. These compounds can severely hinder the efficiency of traditional DNA extraction protocols, often resulting in poor-quality DNA. Conventional methods, such as the Cetyl-trimethyl-ammonium bromide (CTAB) protocol, are often found inadequate due to these biochemical barriers. In this study, we formulated an optimized DNA isolation protocol for chicory that eliminates the need of liquid nitrogen and phenol during processing. The refined approach resulted in the extraction of high-quality DNA suitable for downstream applications. The effectiveness of the optimized method was validated by polymerase chain reaction (PCR) amplification of the sucrose: sucrose 1-fructosyl transferase (1-SST) gene, a key gene involved in the biosynthesis of inulin- a compound with notable medicinal and nutritional value. The PCR amplification yielded the expected 2.01 kb product, confirming the high quality and integrity of the extracted DNA. Overall, the quantity and quality of the DNA were well-suited for molecular analysis, demonstrating the success of our method. This optimized protocol provides a valuable tool for molecular research on chicory and other plants with similar biochemical challenges, facilitating more efficient genetic studies and holding potential for advancing biotechnological applications.

VL  - 10
IS  - 4
ER  -

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Author Information

Shadab Ahamad

Department of Molecular Biology and Genetic Engineering, College of Basic Sciences and Humanities, G. B. Pant University of Agriculture and Technology, Pantnagar, India

Contact Email

http://orcid.org/0000-0003-0788-4817
Mamta Arya

Department of Molecular Biology and Genetic Engineering, College of Basic Sciences and Humanities, G. B. Pant University of Agriculture and Technology, Pantnagar, India

http://orcid.org/0009-0005-7216-6370
Gohar Taj

Department of Molecular Biology and Genetic Engineering, College of Basic Sciences and Humanities, G. B. Pant University of Agriculture and Technology, Pantnagar, India

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APA Style

Ahamad, S., Arya, M., Taj, G. (2024). Isolation and PCR Validation of DNA from Chicory (Cichorium intybus L.) Leaves Using an Optimized CTAB Method to Propel Medicinal Insights. Journal of Diseases and Medicinal Plants, 10(4), 69-72. https://doi.org/10.11648/j.jdmp.20241004.12

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ACS Style

Ahamad, S.; Arya, M.; Taj, G. Isolation and PCR Validation of DNA from Chicory (Cichorium intybus L.) Leaves Using an Optimized CTAB Method to Propel Medicinal Insights. J. Dis. Med. Plants 2024, 10(4), 69-72. doi: 10.11648/j.jdmp.20241004.12

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AMA Style

Ahamad S, Arya M, Taj G. Isolation and PCR Validation of DNA from Chicory (Cichorium intybus L.) Leaves Using an Optimized CTAB Method to Propel Medicinal Insights. J Dis Med Plants. 2024;10(4):69-72. doi: 10.11648/j.jdmp.20241004.12

Copy | Download

@article{10.11648/j.jdmp.20241004.12,
  author = {Shadab Ahamad and Mamta Arya and Gohar Taj},
  title = {Isolation and PCR Validation of DNA from Chicory (Cichorium intybus L.) Leaves Using an Optimized CTAB Method to Propel Medicinal Insights
},
  journal = {Journal of Diseases and Medicinal Plants},
  volume = {10},
  number = {4},
  pages = {69-72},
  doi = {10.11648/j.jdmp.20241004.12},
  url = {https://doi.org/10.11648/j.jdmp.20241004.12},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.jdmp.20241004.12},
  abstract = {Isolating and purifying DNA from plants, particularly from species like Cichorium intybus (chicory), poses significant challenges due to the presence of rigid cell walls and high levels of polyphenols and polysaccharides. These compounds can severely hinder the efficiency of traditional DNA extraction protocols, often resulting in poor-quality DNA. Conventional methods, such as the Cetyl-trimethyl-ammonium bromide (CTAB) protocol, are often found inadequate due to these biochemical barriers. In this study, we formulated an optimized DNA isolation protocol for chicory that eliminates the need of liquid nitrogen and phenol during processing. The refined approach resulted in the extraction of high-quality DNA suitable for downstream applications. The effectiveness of the optimized method was validated by polymerase chain reaction (PCR) amplification of the sucrose: sucrose 1-fructosyl transferase (1-SST) gene, a key gene involved in the biosynthesis of inulin- a compound with notable medicinal and nutritional value. The PCR amplification yielded the expected 2.01 kb product, confirming the high quality and integrity of the extracted DNA. Overall, the quantity and quality of the DNA were well-suited for molecular analysis, demonstrating the success of our method. This optimized protocol provides a valuable tool for molecular research on chicory and other plants with similar biochemical challenges, facilitating more efficient genetic studies and holding potential for advancing biotechnological applications.
},
 year = {2024}
}

Copy | Download

TY  - JOUR
T1  - Isolation and PCR Validation of DNA from Chicory (Cichorium intybus L.) Leaves Using an Optimized CTAB Method to Propel Medicinal Insights

AU  - Shadab Ahamad
AU  - Mamta Arya
AU  - Gohar Taj
Y1  - 2024/12/27
PY  - 2024
N1  - https://doi.org/10.11648/j.jdmp.20241004.12
DO  - 10.11648/j.jdmp.20241004.12
T2  - Journal of Diseases and Medicinal Plants
JF  - Journal of Diseases and Medicinal Plants
JO  - Journal of Diseases and Medicinal Plants
SP  - 69
EP  - 72
PB  - Science Publishing Group
SN  - 2469-8210
UR  - https://doi.org/10.11648/j.jdmp.20241004.12
AB  - Isolating and purifying DNA from plants, particularly from species like Cichorium intybus (chicory), poses significant challenges due to the presence of rigid cell walls and high levels of polyphenols and polysaccharides. These compounds can severely hinder the efficiency of traditional DNA extraction protocols, often resulting in poor-quality DNA. Conventional methods, such as the Cetyl-trimethyl-ammonium bromide (CTAB) protocol, are often found inadequate due to these biochemical barriers. In this study, we formulated an optimized DNA isolation protocol for chicory that eliminates the need of liquid nitrogen and phenol during processing. The refined approach resulted in the extraction of high-quality DNA suitable for downstream applications. The effectiveness of the optimized method was validated by polymerase chain reaction (PCR) amplification of the sucrose: sucrose 1-fructosyl transferase (1-SST) gene, a key gene involved in the biosynthesis of inulin- a compound with notable medicinal and nutritional value. The PCR amplification yielded the expected 2.01 kb product, confirming the high quality and integrity of the extracted DNA. Overall, the quantity and quality of the DNA were well-suited for molecular analysis, demonstrating the success of our method. This optimized protocol provides a valuable tool for molecular research on chicory and other plants with similar biochemical challenges, facilitating more efficient genetic studies and holding potential for advancing biotechnological applications.

VL  - 10
IS  - 4
ER  -

Copy | Download