Effect of salicylic acid stimulant on the biomass content and some biochemical characteristics of deadly nightshade (Atropa belladonna L.) hairy roots

Document Type : Research Paper

Authors

1 M.Sc. Graduated. Dept. Horticultural Science, Faculty of Agriculture, Urmia University, Urmia, I.R. Iran.

2 Corresponding author, Assoc. Prof., Dept. Horticultural Science, Faculty of Agriculture, Urmia University, Urmia, I.R. Iran

10.22092/ijrfpbgr.2023.360647.1427

Abstract

The rapid and mass production of secondary metabolites through chemical methods is mostly expensive, difficult, or impossible. Considering the economic importance of these metabolites and their small production in medicinal plants, some methods such as the cultivation of hairy roots and the use of biotic and abiotic stimuli have been suggested to increase these metabolites. The accumulation of the tropane alkaloid atropine in Atropa belladonna L. is of great importance in the pharmaceutical industry. In this research, transgenic hairy roots of A. belladonna was produced using an A7 strain of Agrobacterium rhizogenes. The transformation of hairy roots was confirmed by PCR test. The results of the PCR test showed the correctness of the amplification of the bp780 fragment for the rolB gene in hairy roots. In addition, the effect of different concentrations (0, 125, 250, and 500 μM) of salicylic acid in two times of 24 and 48 hours on the amount of atropine content, the plant growth rate and the activity of antioxidant enzymes of A. belladonna hairy roots was investigated. The data statistically analyzed using factorial experiment based on a completely random design with three replications. The results of the analysis of variance showed significant effect of salicylic acid on all the traits (P<0.01). The effect of time was significant for the amount of atropine and ascorbate peroxidase (APX). The salicylic acid by time interaction effects were significant for the amount of atropine, catalase (CAT), and guaiacol peroxidase (GPX) (P<0.01, 0.05). The results showed that by increasing salicylic acid concentrations the growth of hairy roots significantly decrease compared to the control and the lowest values of fresh and dry weight of the hairy roots (2.79 and 0.148 g) were obtained in 500 μM of salicylic acid compared to the control (6.76 and 0.273 g). However, based on GC/MS analysis, the highest amount of atropine (11 times more than the control) was obtained in the 250 μM salicylic acid in a period of 24 hours. The highest catalase and guaiacol peroxidase activities were obtained at concentrations of 250 and 125µM salicylic acid for 24 hours, respectively. The highest level of ascorbate peroxidase enzyme activity was obtained at the concentration of 250 µM salicylic acid compared to the control. It was concluded that the salicylic acid stimulant at low concentrations is useful for cell signaling and stimulating the production of secondary metabolites, but at high concentrations, it disrupts the growth of hairy roots.
 

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References

  • Abdala, G., Miersch, O., Kramell, R., Vigliocco, A., Agostini, E., Forchetti, G. and Alemano, S., 2003. Jasmonate and octadecanoid occurrence in tomato hairy roots. Endogenous level changes in response to NaCl. Plant Growth Regulation, 40(1): 21-27.
  • Aebi, H., 1984. Catalase in vitro. Methods in enzymology, 105, 121-126.
  • Ahmadian Chashmi, N., Sharifi, M., Karimi, F. and Rahnama, H., 2010. Comparative study of tropane alkaloids production in hairy roots and plantlet cultures of Atropa belladonna by salicylic acid treatments. Iranian Journal of Plant Biology, 2(3): 63-76.
  • Ajungla, L., Patil, P. P., Barmukh, R. B. and Nikam, T. D. 2009. Influence of biotic and abiotic elicitors on accumulation of hyoscyamine and scopolamine in root cultures of Datura metel L. Indian Journal of Biotechnology, 8(3): 317-322.
  • Asghari, M., 2015. New (Non-Classic) Plant Hormones and Growth Regulators. Urmia University, Urmia, Iran, 352p. (In Persian).
  • Bavi, K., Khavari-Nejad, R.A., Najafi, F. and Ghanati, F., 2022. Stimulation of phenolic compounds production in Zataria multiflora Boiss. Cell suspension culture through salicylic acid elicitation. Journal of Plant Research, 34(4): 806-817. (In Persian).
  • Cai, Z., Kastell, A., Mewis, I., Knorr, D. and Smetanska, I., 2012. Polysaccharide elicitors enhance anthocyanin and phenolic acid accumulation in cell suspension cultures of Vitis vinifera. Plant Cell, Tissue and Organ Culture (PCTOC), 108(3): 401-409.
  • Chaichana, N. and Dheeranupattana, S., 2012. Effects of Methyl Jasmonate and Salicylic Acid on Alkaloid Production from in vitro Culture of Stemona sp. International Journal of Bioscience, Biochemistry and Bioinformatics, 2(3):
  • Chen, J. Y., Wen, P. F., Kong, W. F., Pan, Q. H., Zhan, J. C., Li, J. M. and Huang, W. D., 2006. Effect of salicylic acid on phenylpropanoids and phenylalanine ammonia-lyase in harvested grape berries. Postharvest Biology and Technology, 40(1): 64-72.
  • Cheruvathur, M. K. and Thomas, T. D. 2014. Effect of plant growth regulators and elicitors on rhinacanthin accumulation in hairy root cultures of Rhinacanthus nasutus (L.) Kurz. Plant Cell, Tissue and Organ Culture (PCTOC), 118(1): 169-177.
  • Chotikadachanarong, K., Dheeranupattana, S., Jatisatienr, C., Wangkarn, S., Mungkornasawakul, P., Pyne, S. G. and Sastraruji, T., 2011. Influence of Salicylic Acid on Alkaloid Production by Root Cultures of Stemona curtisii Hook. F. Journal of Biological Sciences, 3(4): 322-325.
  • Farag, M.A., Al-Mahdy, D.A., Meyer, A., Westphal, H. and Wessjohann, L.A., 2017. Metabolomics reveals biotic and abiotic elicitor effects on the soft coral Sarcophyton ehrenbergi terpenoid content. Scientific reports, 7(1): 1-11.
  • Hayat, S., Ali, B. and Ahmad, A., 2007. Salicylic acid: biosynthesis, metabolism and physiological role in plants. In Salicylic acid: A plant hormone (pp. 1-14). Springer Netherlands.
  • Horvath, E., Szalai, G. and Janda, T., 2007. Induction of abiotic stress tolerance by salicylic acid signaling. Journal of Plant Growth Regulation 26: 290-300.
  • Kai, G., Yang, S., Zhang, Y., Luo, X., Fu, X., Zhang, A. and Xiao, J., 2012. Effects of different elicitors on yield of tropane alkaloids in hairy roots of Anisodus acutangulus. Molecular biology reports, 39(2): 1721-1729.
  • Kamada, H., Okamura, N., Satake, M., Harada, H. and Shimomura, K., 1986. Alkaloid production by hairy root cultures in Atropa belladonna.Plant cell reports, 5(4): 239-242.
  • Kamalipourazad, M., Sharifi, M., Maivan, H. Z., Behmanesh, M., and Chashmi, N. A., 2016. Induction of aromatic amino acids and phenylpropanoid compounds in Scrophularia striata Boiss. Cell culture in response to chitosan-induced oxidative stress. Plant Physiology and Biochemistry, 107: 374-384.
  • Kang, S. M., Jung, H. Y., Kang, Y. M., Yun, D. J., Bahk, J. D., Yang, J. K. and Choi, M. S., 2004. Effects of methyl jasmonate and salicylic acid on the production of tropane alkaloids and the expression of PMT and H6H in adventitious root cultures of Scopolia parviflora. Plant Science, 166(3): 745-751.
  • Khalili, M., Hasanloo, T., Kazemi Tabar, S.K. and Rahnama, H., 2009. Influence of exogenous salicylic acid on flavonolignans and lipoxygenase activity in the hairy root cultures of Silybum marianum. Cell Biology International, 33: 988-994.
  • Khalili, M., Hasanloo, T., Kazemi Tabar, S.K. and Sepehrifar, R., 2010. Effect of salicylic acid on antioxidant activity in milk thistle hairy root cultures. Journal of Medicinal Plants, 35: 51-60
  • Khan, S., Qureshi, M. I., Alam, T. and Abdin, M. Z., 2007. Protocol for isolation of genomic DNA from dry and fresh roots of medicinal plants suitable for RAPD and restriction digestion. African Journal of Biotechnology, 6(3):
  • Laspina, N. V., Groppa, M. D., Tomaro, M. L. and Benavides, M. P., 2005. Nitric oxide protects sunflower leaves against Cd-induced oxidative stress. Plant Science, 169(2): 323-330.
  • Li, J., Wang, J., Li, J., Li, J., Liu, S. and Gao, W., 2016. Salicylic acid induces the change in the adventitious root of Glycyrrhiza uralensis Fisch: bioactive compounds and antioxidant enzymes. Research on Chemical Intermediates, 42(2): 1503-1519.
  • Liu, Z. B., Chen, J. G., Yin, Z. P., Shangguan, X. C., Peng, D. Y., Lu, T., and Lin, P., 2018. Methyl jasmonate and salicylic acid elicitation increase content and yield of chlorogenic acid and its derivatives in Gardenia jasminoides cell suspension cultures. Plant Cell, Tissue and Organ Culture (PCTOC), 134(1): 79-93.
  • Maqbool, F., Singh, S., Kaloo, Z. A. and Jan, M., 2014. Medicinal importance of Genus Atropa Royle-A review. International Journal of Advanced Research, 2(2): 48-54.
  • Naeini, M. S., Naghavi, M. R., Bihamta, M. R., Sabokdast, M. and Salehi, M., 2021. Production of some benzylisoquinoline alkaloids in Papaver armeniacum L. hairy root cultures elicited with salicylic acid and methyl jasmonate. In Vitro Cellular & Developmental Biology-Plant, 57(2), 261-271.
  • Nakano, Y. and Asada, K., 1981. Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant and Cell Physiology, 22(5): 867-880.
  • Parsa, M. and Zeinali, A., 2016. Effects of salicylic acid elicitor on the production of tropane alkaloids (atropine and scopolamine) in hairy roots and in vitro roots cultures of Hyoscyamus niger L. Iranian Journal of Medicinal and Aromatic Plants, 32(4): 655-666. (In Persian).
  • Pu, G. B., Ma, D. M., Chen, J. L., Ma, L. Q., Wang, H., Li, G. F. and Liu, B. Y., 2009. Salicylic acid activates artemisinin biosynthesis in Artemisia annua L. Plant Cell Reports, 28(7), 1127-1135.
  • Rani, A. and Prassad, M. P., 2013. Studies on the Organogenesis of Atropa Belladonna in In-vitro Conditions. International Journal of Biotechnology and Bioengineering Research, 4(5): 457-464.
  • Roy, S., Sen, C. K. and Hänninen, O., 1996. Monitoring of polycyclic aromatic hydrocarbons using ‘moss bags’: Bioaccumulation and responses of antioxidant enzymes in Fontinalis antipyretica Hedw. Chemosphere, 32(12): 2305-2315.
  • Samadi, S., Ghasemnajad, A. and Alizadeh, M., 2015. Investigation on phenylalanine ammonia-lyase activity of artichoke (Cynara scolymus L.) affected by methyl jasmonate and salicylic acid in in-vitro conditions. Journal of Plant Production Research, 21(4): 135-148.
  • Shuncang, Z., Hongyan, L., Xiao, L., Yan, Y., Pengguo, X., Yanyan, J. and Zongsuo, L., 2015. Enhanced production of phenolic acids in Salvia miltiorrhiza hairy root cultures by combing the RNAi-mediated silencing of chalcone synthase gene with salicylic acid treatment. Biochemical Engineering Journal, 103: 185–192.
  • Sivanandhan, G., Selvaraj, N., Ganapathi, A. and Manickavasagam, M., 2016. Elicitation Approaches for Withanolide Production in Hairy Root Culture of Withania somnifera (L.) Dunal. Biotechnology of Plant Secondary Metabolism: Methods and Protocols, 1-18.
  • Taghizadeh, M., Nasibi, F., Kalantari, K. M., and Ghanati, F., 2019. Evaluation of secondary metabolites and antioxidant activity in Dracocephalum polychaetum Bornm. Cell suspension culture under magnetite nanoparticles and static magnetic field elicitation. Plant Cell, Tissue and Organ Culture (PCTOC), 136(3), 489-498.
  • Teixeira da Silva, J.A., 2006. Floriculture, Ornamental and Plant Biotechnology: Advances and Topical Issues. Global Science Books, Ltd: London, 646p.
  • Vasconsuelo, A. and Boland, R., 2007. Molecular aspects of the early stages of elicitation of secondary metabolites in plants. Plant Science, 172(5): 861-875.
  • Wen, P. F., Chen, J. Y., Kong, W. F., Pan, Q. H., Wan, S. B. and Huang, W. D., 2005. Salicylic acid induced the expression of phenylalanine ammonia-lyase gene in grape berry. Plant Science, 169(5): 928-934.
  • Xo, H., Park, J., Kim, Y., Park, N., Lee, S., Park, S. 2009. Optimization of growth and pyranocoumarins production in hairy root culture of Angelica gigas Nakai. Journal of Medicinal Plants Research, 3: 978-981.
  • Zarei, B., Taghipour, Z. and Kahrizi, D., 2020. Effect of Growth Regulators on In Vitro Callogenesis and Regeneration of Atropa Belladonna. Iranian Journal of Rangelands and Forests Plant Breeding and Genetic Research, 27(2): 231-239. (In Persian).

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