Optimization of nettle (Urtica dioica L.) callogenesis and the effect of methyl jasmonate elicitor on biochemical properties of callus | ||
| تحقیقات گیاهان دارویی و معطر ایران | ||
| Article 9, Volume 40, Issue 2 - Serial Number 120, June 2024, Pages 331-351 PDF (878.9 K) | ||
| Document Type: Research Paper | ||
| DOI: 10.22092/ijmapr.2023.363123.3353 | ||
| Authors | ||
| M. Sarrafi1; M. Mahmoodnia Meimand* 2; M. Dahajipour3; M.R. Dehghani3 | ||
| 1Genetics and Plant Production Dept, Agriculture College, Vali-eAsr University of Rafsanjan, Rafsanjan, Iran | ||
| 2Genetics and Plant Production Dept. Agriculture College, Vali-e-Asr University of Rafsanjan, Rafsanjan, Iran | ||
| 3Genetics and Plant Production Dept, Agriculture College, Vali-e-Asr University of Rafsanjan, Rafsanjan, Iran | ||
| Abstract | ||
| Background and objectives: In recent years, Urtica dioica L. has attracted the attention of researchers due to its significant effect on many diseases, especially diabetes. However, more research needs to be done in the field of tissue culture and the effect of elicitors on antioxidant properties and the production of its secondary metabolites. This research aimed to investigate the best hormonal combination and light conditions effective on nettle callogenesis and the effect of methyl jasmonate elicitor on its biochemical properties. Methodology: For this purpose, using stem explants, the effect of different levels of auxin hormones, naphthalene acetic acid (NAA) (1.5 and 3 mg/l) and 2,4-dichlorophenoxy acetic acid (2,4 D) (0.5 and 1.5 mg/l) and cytokinin hormone, benzylaminopurine (BAP) (0.5 and 1.5 mg/l) were evaluated in two light conditions (light and dark) by measuring fresh weight, dry weight and percentage of callogenesis. The nettle plant's callus growth curve was obtained using liquid culture. After starting the suspension culture, methyl jasmonate treatment was applied with three levels of 1, 0.1 and 0.01 mM, and sampling was done at 24, 48 and 96 hours. The enzyme activity of phenylalanine ammonialyase and polyphenol oxidase and total protein content by extracting in 50 mM Tris-HCl buffer with 8.8 acidity and total phenol, total flavonoid content by extracting in 80% methanol was measured using spectrophotometric and standard methods for control samples and those under different treatments of methyl jasmonate. Also, three metabolites of quercetin, kaempferol and rutin were measured using an HPLC device with a UV detector and 18C column stationary phase. Data from the callogenesis experiment were analyzed in a two-factor experiment in a completely randomized design. The data from the biochemical and HPLC experiments were analyzed in a split-plot in time design, and the means were compared with Duncan's test. Results: According to the results, the best hormonal combination to produce fresh and dry weight in light conditions includes 1.5 mg/l of NAA and 1.5 mg/l of BAP and in dark conditions, it includes 1.5 mg/l naphthalene acetic acid and 0.5 mg/l of benzyl aminopurine. Also, according to the results, naphthalene acetic acid (62.5%) performed better than 2,4-D (43.5%) in the nettle callogenesis percentage. Lighting was determined as the best light condition for callogenesis in nettle, so the amount of fresh and dry weight and percent of callogenesis in light compared to darkness increased by 4.79, 4.91 and 1.92 times, respectively. There was an increase in polyphenol oxidase enzyme activity, total phenol and total protein in some samples treated with methyl jasmonate compared to the control, and no significant increase was observed in the activity of phenylalanine ammonialyase enzyme and total flavonoid content. According to the HPLC results, the highest amount of rutin metabolite was obtained in 96 hours after the addition of the elicitor and at the concentration of 0.01 mM methyl jasmonate. Conclusion: The naphthalene acetic acid hormone was more effective than the 2,4-D hormone and light conditions than darkness in nettle callogenesis. Methyl jasmonate elicitor increased the antioxidant property of nettle (48 hours after treatment) by increasing the polyphenol oxidase enzyme activity. However, compared to the control, it had no significant effect on the phenylalanine ammonialyase enzyme activity and total flavonoid. It is suggested that other concentrations of methyl jasmonate be tested to reach optimal conditions for increasing the production of secondary metabolites in nettle in vitro culture conditions. | ||
| Keywords | ||
| Auxin; antioxidant; polyphenol oxidase; secondary metabolite; rutin; nettle (Urtica dioica L.) | ||
| References | ||
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- Adhikari, S.R. and Pant, B., 2013. Induction and proliferation of in vitro mass of callus of Withania somnifera (L.) Dunal. Research in Plant Sciences, 1(3): 58-61. - Agarwal, S. and Pandey, V., 2004. Antioxidant enzyme responses to NaCl stress in Cassia angustifolia. Biologia Plantarum, 48(4): 555-560. - Alirezalo, A., malekzadeh, M., Babaei, A. and Kosravi, H., 2011. The effect of different hormonal treatments on the callus formation of the leaves of the medicinal plant Catharanthus roseus L. and the evaluation of the amount of callus alkaloid. Medicinal Plant National Congress, Sari, Iran, 2-4 March. - Al-Rifai, A.A., Aqel, A., Awaad, A. and Alothman, Z.A., 2015. Analysis of Quercetin and Kaempferol in an Alcoholic Extract of Convolvulus pilosellifolius using HPLC. Communications in Soil Science and Plant Analysis, 46(11): 1411-1418. - Asgarpanah, J. and Mohajerani, R., 2012. Phytochemistry and pharmacologic properties of Urtica dioica L. Journal of Medicinal Plants Research, 6(46): 5714-5719. - Babashpour-Asl, M., Baleghi, M., Sajadi, P. and Golalipour, M.J., 2014. Different Aspects and Results of Modern Studies of Urtica Dioica: A Review. Journal of Babol University of Medical Sciences, 16(3): 47-54. - Bradford, M.M., 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72(1-2): 248-254. - Brouki Milan, E., Abdollahi Mandoulakani, B. and Kheradmand, F., 2014. The effect of methyl jasmonate on the activity of poly phenol oxidase and phenil alanin amonia lyase enzymes in basil (Ocimum basilicum L.). 13th Iranian Crop Sciences Congress & 3rd Iranian Seed Science and Technology Conference. Karaj, Iran, 24-26 August, Conference Hall Research institute for improvement and preparation of seedlings and seeds: 1-4. - Cao, S., Zheng, Y., Wang, K., Rui, H. and Tang, S., 2010. Effect of methyl jasmonate on cell wall modification of loquat fruit in relation to chilling injury after harvest. Food Chemistry, 118(3): 641-647. - Chaoui, A., Mazhoudi, S., Ghorbal, M.H. and El Ferjani, E., 1997. Cadmium and zinc induction of lipid peroxidation and effects on antioxidant enzyme activities in bean (Phaseolus vulgaris L.). Plant Science, 127(2): 139-147. - Colins, G.B., Vian, W.E. and Phillips, G.C., 1978. Use of 4-Amino-3, 5, 6-trichloropicolinic Acid as an Auxin Source in Plant Tissue Cultures. Crop Science, 18(2): 286-288. - Dar, S.A., Nawchoo, I.A., Tyub, S. and Kamili, A.N., 2021. Effect of plant growth regulators on in vitro induction and maintenance of callus from leaf and root explants of Atropa acuminata Royal ex Lindl. Biotechnology Report: 32: 1-5. - El Haouari, M. and Rosado, J.A., 2019. Phytochemical, anti-diabetic and cardiovascular properties of Urtica dioica L. (Urticaceae): A Review. Mini Reviews in Medicinal Chemistry, 19(1): 63-71. - Farooq, M.A., Gill, R.A., Islam, F., Ali, B., Liu, H., Xu, J., He, S. and Zhou, W., 2016. Methyl jasmonate regulates antioxidant defense and suppresses arsenic uptake in Brassica napus L. Frontiers in Plant Science, 7(468): 1-16. - Fazili, M.A., Bashir, I., Ahmad, M., Yaqoob, U. and Geelani, S.N., 2022. In vitro strategies for the enhancement of secondary metabolite production in plants: a review. Bulletin of the National Research Centre, 46(1): 1-12. - Friis, I., 1993. Urticaceae. 612-613, In: Kubitzki, K., Rohwer, J.G. and Bittrich, V. (eds.), The Families and Genera of Vascular Plants. Springer, Berlin, Heidelberg, 478p. - Ghanati, F., Bakhtiyarian, S. and Abdolmaleki, P., 2010. Effects of methyl jasmonate on the secondary metabolites of Calendula officinalis L. Modares Journal of Biotechnology, 1(1): 21-33. - Gopi, C. and Vatsala, T.M., 2006. In vitro studies on effects of plant growth regulators on callus and suspension culture biomass yield from Gymnema sylvestre R. Br. African Journal of Biotechnology, 5(12): 1215-1219. - Guan, Y., Hu, W., Jiang, A., Xu, Y., Sa, R., Feng, K., Zhao, M., Yu, J., Ji, Y., Hou, M. and Yang, X., 2019. Effect of Methyl Jasmonate on Phenolic Accumulation in Wounded Broccoli. Molecules, 24(19): 3537. - Hashemyan, M., Ganjeali, A. and Cheniany, M., 2020. Effect of Methyl Jasmonate and Salicylic Acid Elicitors on the Production of Secondary Metabolites and Antioxidant Capacity of Teucrium polium L. in-vitro. Iranian Journal of Plant Biology, 12(2): 61-76. - Jaimand, K., Rezaee, M.B., Asareh, M.H., Tabaei Aghdaei, S.R. and Meshkizadeh, S., 2010. Extraction and determination of Kaempferol and Quercetin in petals of 10 genotypes of Rosa damascena Mill. from western Iran. Iranian Journal of Medicinal and Aromatic Plants, 25(4): 547-555. - Ketabchi, S., Majzoob, Sh. and Charegani, H.A., 2015. Effect of salicylic acid and methyl jasmonate on phenylalanine ammonia-lyase activity and total phenol in wheat infected by Pratylenchus thornei. Archives of Phytopathology and Plant Protection, 48(1): 10-17. - Kim, H.J., Chen, F., Wang, X. and Choi, J.H., 2006. Effect of methyl jasmonate on phenolics, isothiocyanate, and metabolic enzymes in radish sprout (Raphanus sativus L.). Journal of agricultural and food chemistry, 54(19): 7263-7269. - Kintzios, S.E., Hiureas, G., Shortsianitis, E., Sereti, E., Blouhos, P., Manos, C., Makri, O., Taravira, N., Drossopoulos, J.B. and Holevas, C.D., 1998. The effect of light on the induction, development and maturation of somatic embryos from various horticultural and ornamental species. In International Symposium on Biotechnology of Tropical and Subtropical Species Part 2, 461: 427-432. - Kintzios, S., Papanastasiou, I., Tourgelis, P., Papastellatos, C., Georgopoulos, V. and Drossopoulos, J., 2002. The effects of light on callus growth and somatic embryogenesis from Lavandula vera and Teucrium chamaedrys: A preliminary study. Journal of Herbs, Spices & Medicinal Plants, 9(2-3): 223-227. - Liu, R., Wang, Z., Zheng, J., Xu, Z., Tang, X., Huang, Z., Zhang, N., Dong, Y. and Li, T., 2022. The effects of methyl jasmonate on growth, gene expression and metabolite accumulation in Isatis indigotica Fort. Industrial Crops and Products, 177: 1144-1182. - Mungole, A.J., Doifode, V.D., Kamble, R.B., Alka, C. and Prakash, Z., 2011. In-vitro callus induction and shoot regeneration in Physalis minima L. Annals of Biological Research, 2(2): 79-85. - Murashige, T. and Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiologia Planetarium, 15(1): 473-479. - Pinelli, P., Ieri, F., Vignolini, P., Bacci, L., Baronti, S. and Romani, A., 2008. Extraction and HPLC analysis of phenolic compounds in leaves, stalks, and textile fibers of Urtica dioica L. Journal of Agricultural and Food Chemistry, 56(19): 9127-9132. - Said, A.A.H., Otmani, I.S.E., Derfoufi, S. and Benmoussa, A., 2015. Highlights on nutritional and therapeutic value of stinging nettle (Urtica dioica). International Journal of Pharmacy and Pharmaceutical Sciences, 7(10): 8-14. - Samadi, S. and Ghasemnezhad, A. and Alizadeh, M., 2019. Influence of Salicylic acid and Methyl jasmonate on fresh weight, total phenol, total flavonoids, antioxidant and PAL enzyme activity of Stevia callus. Plant Process and Function, 8(32): 324-337. - Sarrafi, M., 2022. Optimization of nettle (Urtica diocia) callogenesis and the effect of methyl jasmonate elicitor on its biochemical properties. M.Sc. thesis, Department of Genetics and Plant Production, Vali-e-Asr University of Rafsanjan, Rafsanjan. - Sayed-Tabatabaei, B.E. and Omidi, M., 2021. Plant Cell and Tissue Culture. University of Tehran Press, Tehran, 388p. - Sidhu, Y., 2011. In vitro micropropagation of medicinal plants by tissue culture. The Plymouth Student Scientist, 4(1): 432-449. - Sood, H., 2020. Production of medicinal compounds from endangered and commercially important medicinal plants through cell and tissue culture technology for herbal industry. In: Sharma, K., Mishra, K., Kamal Senapati, K., Danciu, C., (Eds.). Bioactive Compounds in Nutraceutical and Functional Food for Good Human Health. IntechOpen, United Kingdom, 358p. - Šrůtek, M. and Teckelmann, M., 1998. Review of biology and ecology of Urtica dioica. Preslia Praha, 70: 1-19. - Tapas, D.A., Sakarkar, D.M. and Kakde, R.B., 2008. Flavonoids as Nutraceuticals: A Review. Tropical Journal of Pharmaceutical, 7(3): 1089-1099. - Thiem, B. and Krawczyk, A., 2010. Enhanced isoflavones accumulation in methyl jasmonate-treated in vitro cultures of kudzu [Pueraria lobata Ohwi]. Herba Polonica, 56(1): 48-56. - Tripathi, L. and Tripathi, J.N., 2003. Role of biotechnology in medicinal plants. Tropical Journal of Pharmaceutical Research, 2(2): 243-253. - Vanisree, M. and Tsay, H.S., 2007. Plant cell cultures: Production of biologically important secondary metabolites from medicinal plants of Taiwan. Medicinal plant biotechnology. From basic research to industrial application, 267-285. - Wang, J., Qian, J., Yao, L. and Lu, Y., 2015. Enhanced production of flavonoids by methyl jasmonate elicitation in cell suspension culture of Hypericum perforatum. Bioresources and Bioprocessing, 2(1): 1-9. - Yao, D., Zhang, Z., Chen, Y., Lin, Y., Xu, X. and Lai, Z., 2021. Transcriptome Analysis Reveals Differentially Expressed Genes That Regulate Biosynthesis of the Active Compounds with Methyl Jasmonate in Rosemary Suspension Cells. Genes, 13(1): 2-21. - Zamani, M., Moradi, H., Chalavi, V. and Kazemitabar, S.K., 2019. Effect of Salicylic Asid and Methyle Jasmonat Elicitors on Hypericin production in stJohn’s wort (Hypericum perforatum L.) cv. Topas Callus culture. Iranian Journal of Horticultural Science, 49(4): 915-923. - Zhao, P., Wu, F., Feng, F.S. and Wang, W.J., 2008. Protocom-like body (PLB) formation and plant regeneration from the callus culture of Dendrobium candidm Wall ex Lindl. In Vitro Cellular & Developmental Biology-Plant, 44(3): 178-185. - Zu, Y., Li, C., Fu, Y. and Zhao, C., 2006. Simultaneous determination of catechin, rutin, quercetin, kaempferol and isorhamnetin in the extract of sea buckthorn (Hippophae rhamnoides L.) leaves by RP-HPLC with DAD. Journal of Pharmaceutical and Biomedical Analysis, 41(3): 714-719. | ||
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