DNA barcoding, phytochemical screening and antimicrobial activity of Rhododendron arboreum, a high altitudinal medicinal plant from Nepal

Yıl 2020, , 140 - 151, 15.06.2020

Deepak Sharma , Tribikram Bhandari Anaviggha Pradhan Navin Ghimire Sagar Basnet Saroj Pandey Janardan Lamichhane

https://doi.org/10.31195/ejejfs.696768

Cited By: 1

Öz

DNA barcoding is been proposed as a powerful taxonomic tool for species identification genetically. The Consortium for the Barcode of Life (CBOL) Plant Working Group has recommended the combination of rbcL & matK as the core plant barcode. Rhododendron of Nepal shows genetic diversity due to which 31 different species were reported till date, found in Nepal based on morphological character. We collected different samples from the central and western parts of Nepal (Province 3 & 5). For the phytochemical study, cold extraction was performed using methanol as solvents. The crude extracts were tested for the presence of phytochemicals such as alkaloids, saponins, glycosides, tannins, flavonoids, and coumarins. The same extracts were used for antimicrobial assay five different pathogenic micro-organisms. MIC was done for calculating the minimum effective concentration of the extract. Then TLC was done to separate the compound. The methanol extract was analyzed for core secondary metabolites by HPLC-QTOF and found more than 20 medicinal important compounds. DPPH assay was also done with crude extracts for identifying antioxidant potency of the extracts. Among samples compared, Gulmi and Palpa showed the best result for antioxidant assay with minimum IC50. Similarly, phenolic content was also checked and samples from Palpa showed high phenolic content. The rbcL and MatK gene loci show the 99.98% similarly for the R.arboreum in NCBI database and were submitted to the BOLD database for the final authentication and get success for the first time to get published DNA barcode of Nepalese plants.

Anahtar Kelimeler

DNA Barcode, rbcL+matK, Antimicrobial, Antioxidant, DPPH

Destekleyen Kurum

Kathmandu University, University Grant Commission and NAST Nepal

Teşekkür

The research part of this project has been supported by UGC-Nepal Faculty Grant 2017 and also support by NAST Ph.D. Grant 2014.

Kaynakça

• 1. Liu, Y., Zhang, L., Liu, Z., Luo, K., Chen, S., and Chen, K. (2012). Species identification of <i>Rhododendron</i> (Ericaceae) using the chloroplast deoxyribonucleic acid <i>PsbA-trnH</i> genetic marker. Pharmacognosy Magazine 8, 29-36. 2. COX PA, C.K. (1997). Encyclopedia of Rhododendron species. In Perth: Glendoick Publishing;. 3. Kress, W.J., Wurdack, K.J., Zimmer, E.A., Weigt, L.A., and Janzen, D.H. (2005). Use of DNA barcodes to identify flowering plants. Proc Natl Acad Sci U S A 102, 8369-8374. 4. Hebert, P.D.N., Cywinska, A., Ball, S.L., and deWaard, J.R. (2003). Biological identifications through DNA barcodes. Proc Biol Sci 270, 313-321. 5. Li, Y., Tong, Y., and Xing, F. (2016). DNA Barcoding Evaluation and Its Taxonomic Implications in the Recently Evolved Genus Oberonia Lindl. (Orchidaceae) in China. Frontiers in Plant Science 7. 6. Wong, K.-L., But, P., and Shaw, P.-C. (2013). Evaluation of seven DNA barcodes for differentiating closely related medicinal Gentiana species and their adulterants. Chinese medicine 8, 16. 7. Newmaster, S.G., Fazekas, A.J., Steeves, R.A.D., and Janovec, J. (2008). Testing candidate plant barcode regions in the Myristicaceae. Mol Ecol Resour 8, 480-490. 8. Garg, N. (2017). Chapter 8 - Technology for the Production of Agricultural Wines. In Science and Technology of Fruit Wine Production, M.R. Kosseva, V.K. Joshi and P.S. Panesar, eds. (San Diego: Academic Press), pp. 463-486. 9. Scott (2010). NEPAL'S MAGNIFICENT RHODODENDRON. In ECS Nepal. 10. Mainra, A., Badola, H., and Mohanty, B. (2010). RHODODENDRONS: CONSERVATION AND SUSTAINABLE USE International Conference RHODODENDRONS: CONSERVATION AND SUSTAINABLE USE. 11. pokhrel (1999). <pokhrel 1999.pdf>. 12. Paul, A., Khan, M., and Arunachalam, A. (2005). Biodiversity and conservation of rhododendrons in Arunachal Pradesh in the Indo-Burma biodiversity hotspot. Current Science 89. 13. Shrivastav, P. (2012). <Pallavi shrivastav.pdf>. Journal of Applied Pharmaceutical Science. 14. Andriani, Y., Ramli, N.M., Syamsumir, D.F., Kassim, M.N.I., Jaafar, J., Aziz, N.A., Marlina, L., Musa, N.S., and Mohamad, H. (2019). Phytochemical analysis, antioxidant, antibacterial and cytotoxicity properties of keys and cores part of Pandanus tectorius fruits. Arabian Journal of Chemistry 12, 3555-3564. 15. Deepak, S., Meena maiy, S., and Hitesh, B. (2019). The Effectiveness of Anti-Cancer Compounds Taxol Derived from Natural Products from Nepal. Acta Scientific Pharmaceutical Sciences 3, 116-123. 16. Venugopal, C., McFarlane, N.M., Nolte, S., Manoranjan, B., and Singh, S.K. (2012). Processing of primary brain tumor tissue for stem cell assays and flow sorting. J Vis Exp, 4111. 17. Sharma, D., Shrestha, T., Rapal, N., Poudel, S.C., Shrestha, S., Sharma, S., Poudel, K.P., and Lamichane, J. (2016). Screening and bioactivity measurement of high altitude medicinal plants of Nepal. Vegetos- An International Journal of Plant Research 29. 18. Deepak, S., Lamichhane J (2019). <qualitative-analysis-of-rhododendron-arboreum-leaves-extracts-using-hplcesiqtofms. Biotechnology an Indian Journal 15. 19. Kathirvel, A., and Sujatha, V. (2016). Phytochemical studies, antioxidant activities and identification of active compounds using GC–MS of Dryopteris cochleata leaves. Arabian Journal of Chemistry 9, S1435-S1442. 20. Zhang, Y.-J., Gan, R.-Y., Li, S., Zhou, Y., Li, A.-N., Xu, D.-P., and Li, H.-B. (2015). Antioxidant Phytochemicals for the Prevention and Treatment of Chronic Diseases. Molecules 20, 21138-21156. 21. Prajapati, K., Thapaliya, S., Dhakal, D., Acharya, S., Shrestha, S., Hora, R., and Sakha, H. (2018). Antimicrobial Activity of Ethanolic Extract of Medicinal Plants against Human Pathogenic Bacteria. Tribhuvan University Journal of Microbiology 5, 1-6. 22. Contreras-Lynch, S., Smith, P., Olmos, P., Loy, M.E., Finnegan, W., and Miranda, C.D. (2017). A Novel and Validated Protocol for Performing MIC Tests to Determine the Susceptibility of Piscirickettsia salmonis Isolates to Florfenicol and Oxytetracycline. Frontiers in Microbiology 8. 23. kit, Z.p.d. <quick-dna_plant-seed_miniprep_kit.pdf>. 24. Kress, W.J. (2017). Plant DNA barcodes: Applications today and in the future. Journal of Systematics and Evolution 55, 291-307. 25. Hollingsworth, P.M., Forrest, L.L., Spouge, J.L., Hajibabaei, M., Ratnasingham, S., van der Bank, M., Chase, M.W., Cowan, R.S., Erickson, D.L., Fazekas, A.J., et al. (2009). A DNA barcode for land plants. Proceedings of the National Academy of Sciences 106, 12794-12797. 26. Stecher, G., Tamura, K., and Kumar, S. (2020). Molecular Evolutionary Genetics Analysis (MEGA) for macOS. Mol Biol Evol. 27. Bold (2013). <BOLD_Handbook_Oct2013.pdf>.