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Research on Crops

Evaluating the efficiency of Chelex 100 for DNA extraction in orange (Citrus sinensis)


DOI: 10.31830/2348-7542.2024.ROC-1018    | Article Id: ROC-1018 | Page : 292-295
Citation :- Evaluating the efficiency of Chelex 100 for DNA extraction in orange (Citrus sinensis). Res. Crop. 25: 292-295
GAVIN GELDENHUYS gavin.geldenhuys@ul.ac.za
Address : Aquaculture Research Unit, School of Agricultural and Environmental Sciences, University of Limpopo (Turfloop Campus), Private Bag X1106, Sovenga, 0727, South Africa
Submitted Date : 19-09-2023
Accepted Date : 2-05-2024
Online Published:

Abstract

Plant research has recently concentrated on the molecular level, which requires reliable, inexpensive, and fast protocols for extracting DNA. Many researchers have identified Chelex-based DNA extraction methods as having the potential to reduce costs. However, Chelex can also prevent DNA degradation by chelating metal ions. Citrus is one of the top three crops grown globally, with oranges being a well-known species. Therefore, study on the molecular of C. sinensis is essential. This study was conducted in 2022 at the genetic lab of the University of Limpopo- to compare two different extraction methods for DNA from C. sinensis. The DNA from the C. sinensis leaves was extracted using Chelex "method 1" (overnight incubation at 56°C) and "method 2" (Ten-minute incubation at 95°C). A spectrophotometer was employed to measure the extracted DNA's qualitative and quantitative parameters accurately. Furthermore, all samples were subjected to PCR reactions with primers for 28S. The results indicated that "method 2" (1.59-1.84) proved to be the most effective method to obtain high-quality extracted DNA. Additionally, the quantitative and qualitative tests for PCR reaction demonstrated that "method 2" produced better results. In conclusion, although both methods yielded favourable outcomes, "method 2" was ultimately the more successful approach.

Keywords

DNA extraction orange PCR spectrophotometer Citrus sinensis

References

Aminisarteshnizi, M. (2021). Evaluating the efficiency of Chelex 100 for DNA extraction in pomegranate (Punica granatum). Res. Crop. 23: 206-10. doi:10.31830/2348-7542. 2022.029.
Aminisarteshnizi, M. (2022). Comparison of different parts of lemon (Citrus limon) for DNA extraction. Trop. Agric. 99: 383-87.
Di Matteo, A., Di Rauso Simeone, G., Cirillo, A., Rao, M. A. and Di Vaio, C. (2022). Morphological characteristics, ascorbic acid and antioxidant activity during fruit ripening of four lemon (Citrus limon (L.) Burm. F.) cultivars. Sci. Hortic. 276: doi:10. 1016/j.scienta.2020.109741.
Geldenhuys, G. (2022). Comparison of two methods for DNA extraction from avocado (Persea americana). Res. Crop. 23: 870-73. doi:10.31830/2348-7542.2022.
Geldenhuys, G. (2023). Comparison of two methods for DNA extraction from Guava (Psidium guajava). Res. Crop. 24: 337-40. doi:10.31830/2348-7542.2023.
HwangBo, K., Son, S. H., Lee, J. S., Min, S. R., Ko, S. M., Liu, J. R., Choi, D. and Jeong, W. J. (2010). Rapid and simple method for DNA extraction from plant and algal species suitable for PCR amplification using a chelating resin Chelex 100. Plant Biotechnol. Rep. 4: 49-52. doi:10.1007/s11816-009-0117-4.
Liu, L., Wang, C. L., Peng, W. Y., Yang, J., Lan, M. Q., Zhang, B., Li, J. B., Zhu, Y. Y. and Li, C.Y. (2015). Direct DNA extraction method of an obligate parasitic fungus from infected plant tissue. Genet. Mol. Res. 14: 18546-551. doi:10.4238/2015.December.28.1.
Sajiba, A. A., Ashraful, M., Bhuiyab, I. and Huqueb, R. (2017). A simple, efficient and rapid method for good quality DNA extraction from rice grains. Rice Sci. 24: 119-22.  
Schlink, K. and Reski, R. (2002). Preparing high-quality DNA from Moss (Physcomitrella patens). Plant Mol. Biol. Rep. 20: 423a-23f. doi:10.1007/BF02772133.
Shokoohi, E. (2021). Morphological and molecular characters of Scutellonema brachyurus (Steiner, 1938) Andrassy, 1958 from South Africa. J. Nematol. 53: 1-13. doi:10. 21307/jofnem-2021-027.
Turan, C., Nanni, I. M., Brunelli, A. and Collina, M. (2015). New rapid DNA extraction method with Chelex from Venturia inaequalis spores. J. Microbiol. Methods 115: 139-43.             doi:10.1016/j.mimet.2015.06.005.
Turci, M., Sardaro, M. L. S., Visioli, G., Maestri, E., Marmiroli, M. and Marmiroli, N. (2010). Evaluation of DNA extraction procedures for traceability of various tomato products. Food Control 21: 143-49. doi:10.1016/J.FOODCONT.2009.04.012.
Vashisth, T. and Kadyampakeni, D. (2020). Diagnosis and management of nutrient constraints in citrus. In: Srivastava, A. K., Hu, C. (Eds.), Fruit Crops: Diagnosis and management of nutrient constraints. Elsevier, Kidlington, UK, pp. 723–37. doi:10.1016/B978-0-12-818732-6.00049-6.
Zahr. S., Zahr, R., El Hajj, L. and Khalil, M. (2023). Phytochemistry and biological activities of Citrus sinensis and Citrus limon: an update. J. Herbal Medicine 41:  doi:10.1016/j. hermed.2023.100737.

 
 

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