Callus and hairy Root Induction in the Medicinal Plant of Withania coagulans (Stocks) Dunal

Document Type : Research Paper

Authors

1 Department of Production Engineering and Plant Genetics, Faculty of Science and Agricultural Engineering, Razi University, Kermanshah, Iran

2 Department of Agricultural Biotechnology, Tarbiat Modares University, Tehran. Iran

3 Nuclear Agriculture Research School, Nuclear Science and Technology Research Institute, AEOI, Karaj, Iran

Abstract

Callus induction from the leaf explants of Withania coagulans was assessed using the MS culture media containing co-application of BAP (0.1, 0.3, and 0.5 mg/L) and 2,4-D (0.1, 0.3, and 0.5 mg/L) based on a completely randomized design (CRD) with three replicates. Furthermore, the potential interaction effects of three different tissues (hypocotyl, stem, and leaf) of W. coagulans and two different Agrobacterium rhizogenes strains (R1000 and GM) were scrutinized for the hairy root induction using a factorial experiment based on CRD. For callus induction, among nine different hormonal treatments resulted from multiplying three concentrations of BAP and three concentrations of 2,4-D, one of which (i.e., “0.1 mg/L 2,4-D + 0.5 mg/L BAP”) had the maximum amounts of fresh weight (3.024 g), dry weight (0.082 g), and callus volume (16.91 cm3), as it was significantly different from the remaining eight hormonal treatments for the three traits (Duncan's test, p< 0.05). To make clear discrimination among nine different hormonal treatments and determine the best one(s) for callus induction in W. coagulans, a hierarchical cluster analysis (HCA) was also applied. All the treatments were placed in three main groups of I (six members), II (one member), and III (two members), of which the second cluster containing only one member (i.e., the hormonal treatment of “0.1 mg/L 2,4-D + 0.5 mg/L BAP”) had the highest quantities of the three aforesaid traits, could be accordingly proposed towards acquiring higher biomass. Regarding hairy root induction, no significant differences were observed either for interaction or for single effects (p < 0.05). However, among six different combinations (two strains and three tissue), the hairy root induction ratios ranged from 38.33% (GM/Hypocotyl) to 59.23% (for R1000/Leaf). Therefore, applying R1000 strain and leaf explant seems to be more effective for hairy root induction in W. coagulans compared to the other five combinations.

Keywords

Main Subjects

References

  1. Rahman, M.H., Roy, B., Chowdhury, G.M., Hasan, A. and Saimun, M.S.R. Medicinal plant sources and traditional healthcare practices of forest-dependent communities in and around Chunati Wildlife Sanctuary in southeastern Bangladesh. Environmental Sustainability; 2022;5(2):207-241.
  2. Robinson M.M., Zhang X. The world medicines situation 2011, traditional medicines: Global situation, issues and challenges. World Health Organization, Geneva. 2011;31:1-2.
  3. Mirshekar M., Ebrahimi M., Ajorlo M. Ethnobotanical and Phytochemical Study of Withania coagulans (Stocks) Dunal in Khash City, Iran. J. Med Plant Res. 2022;11(2):277-285.
  4. Srivastava Y., Sangwan N.S. Improving medicinal crops through phytochemical perspective: Withania somnifera (Ashwagandha). In: Advancement in crop improvement techniques. Elsevier; 2020;275-295.
  5. Negi M.S., Singh A., Lakshmikumaran M. Genetic variation and relationship among and within Withania species as revealed by AFLP markers. Genome. 2000;43(6):975-80.
  6. Panwar J., Tarafdar J.C. Distribution of three endangered medicinal plant species and their colonization with arbuscular mycorrhizal fungi. J. Arid Environ. 2006;65(3):337-50.
  7. Hemalatha S., Wahi A.K., Singh P.N., Chansouria J.P. Hypoglycemic activity of Withania coagulans Dunal in streptozotocin induced diabetic rats. J. Ethnopharmacol. 2004;93(2-3):261-4.
  8. Bharti S.K., Kumar A., Sharma N.K., Krishnan S., Gupta A.K., Padamdeo S.R. Antidiabetic effect of aqueous extract of Withania coagulans flower in Poloxamer-407 induced type 2 diabetic rats. J. Med Plant Res. 2012; 25;6(45):5706-13.
  9. Naz A., Choudhary M.I. Withanolides from Withania coagulans. Phytochem. 2003;63(4):387-90..
  10. Ho T.T., Lee J.D., Ahn M.S., Kim S.W., Park S.Y. Enhanced production of phenolic compounds in hairy root cultures of Polygonum multiflorum and its metabolite discrimination using HPLC and FT-IR methods. Appl. Microbiol. Biotechnol. 2018;102:9563-75.
  11. Sivanandhan G., Kapil Dev G., Jeyaraj M., Rajesh M., Arjunan A., Muthuselvam M., Manickavasagam M., Selvaraj N., Ganapathi A. Increased production of withanolide A, withanone, and withaferin A in hairy root cultures of Withania somnifera (L.) Dunal elicited with methyl jasmonate and salicylic acid. Plant Cell, Tissue and Organ Culture (PCTOC). 2013;114:121-9.
  12. Saxena P., Ahlawat S., Ali A., Khan S., Abdin M.Z. Gene expression analysis of the withanolide biosynthetic pathway in hairy root cultures of Withania somnifera elicited with methyl jasmonate and the fungus Piriformospora indica. Symbiosis 2017;71(2):143-154.
  13. Pandey V., Srivastava R., Akhtar N., Mishra J., Mishra P., Verma, P.C. Expression of Withania somnifera steroidal glucosyltransferase gene enhances withanolide content in hairy roots. Plant Mol Biol Rep. 2016;34(3):681-689.
  14. Sivanandhan G., Arunachalam C., Selvaraj N., Sulaiman A.A., Lim Y.P., Ganapathi A. Expression of important pathway genes involved in withanolides biosynthesis in hairy root culture of Withania somnifera upon treatment with Gracilaria edulis and Sargassum wightii. Plant Physiol. Biochem. 2015;91:61-64.
  15. Thilip C., Soundar Raju C., Varutharaju K., Aslam A., Shajahan A. Improved Agrobacterium rhizogenes-mediated hairy root culture system of Withania somnifera (L.) Dunal using sonication and heat treatment. 3 Biotech. 2015;5:949-56.
  16. Dehdashti S.M., Acharjee S., Kianamiri, S. and Deka, M. An efficient Agrobacterium rhizogenes-mediated transformation protocol of Withania somnifera. Plant Cell, Tissue and Organ Culture (PCTOC). 2017;128:55-65.
  17. Namdeo A.G., Ingawale D.K. Ashwagandha: Advances in plant biotechnological approaches for propagation and production of bioactive compounds. J. Ethnopharmacol. 2021;271:113709.
  18. Hosseini S.M., Bahramnejad B., Douleti Baneh H., Emamifar A., Goodwin P.H. Hairy root culture optimization and resveratrol production from Vitis vinifera subsp. sylvesteris. World J. Microbiol Biotechnol. 2017;33:1-0.
  19. Carlín A.P., Tafoya F., Alpuche Solís A.G. and Pérez-Molphe-Balch E. Effects of different culture media and conditions on biomass production of hairy root cultures in six Mexican cactus species. In Vitro Cell Dev Biol-Plant. 2015;51:332-9.
  20. Ma, H., Meng, X., Xu, K., Li, M., Gmitter Jr, F.G., Liu, N., Gai, Y., Huang, S., Wang, M., Wang, N., Xu, H. Highly efficient hairy root genetic transformation and applications in citrus. Front. Plant Sci. 2022;27;13:1039094.
  21. Khezri M., Asghari Zakaria R., Zare N., Johari-Ahar M. Improving galegine production in transformed hairy roots of Galega officinalis L. via elicitation. AMB Express. 2022;12(1):65.
  22. Foti C., Pavli O.I. High-efficiency Agrobacterium rhizogenes-mediated transgenic hairy root induction of Lens culinaris. Agronomy. 2020;10(8):1170.
  23. Massah M., Rabiei M. Effect of Acetosyringone, Sucrose and Nutrients on Transgenic Hairy Root Induction in Chenopodium quinoa using different Rhizobium rhizogenes strains.
  24. Park C.H., Zhao S., Yeo H.J., Park Y.E., Baska T.B., Arasu M.V., Al-Dhabi N.A., Park S.U. Comparison of different strains of Agrobacterium rhizogenes for hairy root induction and betulin and betulinic acid production in Morus alba. Nat. Prod Commun. 2017;12(4):1934578X1701200403.
  25. Sathasivam R., Choi M., Radhakrishnan R., Kwon H., Yoon J., Yang S.H., Kim J.K., Chung Y.S., Park S.U. Effects of various Agrobacterium rhizogenes strains on hairy root induction and analyses of primary and secondary metabolites in Ocimum basilicum. Front. Plant Sci. 2022;13:983776.
  26. Rani G., Virk G.S., Nagpal A. Callus induction and plantlet regeneration in Withania somnifera (L.) Dunal. In Vitro Cellular & Developmental Biology-Plant. 2003;39:468-74.
  27. Chakraborty N., Banerjee D., Ghosh M., Pradhan P., Gupta N.S., Acharya K., Banerjee M. Influence of plant growth regulators on callus mediated regeneration and secondary metabolites synthesis in Withania somnifera (L.) Dunal. Physiol. Mol Biol. 2013;19:117-25.
  28. Adil M., Abbasi B.H., ul Haq I. Red light controlled callus morphogenetic patterns and secondary metabolites production in Withania somnifera L. Biotechnol. Rep. 2019;24:e00380.
  29. Mirjalili M.H., Esmaeili H. Callus induction and withanolides production through cell suspension culture of Withania coagulans (Stocks) Dunal. J. Medicinal Plants. 2022;21(81):79-91.
  30. Classic Murashige T., Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant. 1962;15:473-97.
  31. Metsalu T., Vilo J. ClustVis: a web tool for visualizing clustering of multivariate data using Principal Component Analysis and heatmap. Nucleic Acids Res. 2015;43(W1):W566-W570.
  32. Mishra B.N., Ranjan R. Growth of hairy‐root cultures in various bioreactors for the production of secondary metabolites. Biotechnol. Appl. Biochem. 2008;49(1):1-0.
  33. Halder M., Sarkar S., Jha S. Elicitation: A biotechnological tool for enhanced production of secondary metabolites in hairy root cultures. Eng. Life Sci. 2019;19(12):880-95.
  34. Thwe A., Valan Arasu M., Li X., Park C.H., Kim S.J., Al-Dhabi N.A., Park S.U. Effect of different Agrobacterium rhizogenes strains on hairy root induction and phenylpropanoid biosynthesis in tartary buckwheat (Fagopyrum tataricum Gaertn). Front. Microbiol. 2016;7:318.
  35. Pandey V., Misra P., Chaturvedi P., Mishra M.K., Trivedi P.K., Tuli R. Agrobacterium tumefaciens-mediated transformation of Withania somnifera (L.) Dunal: an important medicinal plant. Plant Cell Rep. 2010;29:133-41.
  36. Mishra M.K., Chaturvedi P., Singh R., Singh G., Sharma L.K., Pandey V., Kumari N., Misra P. Overexpression of WsSGTL1 gene of Withania somnifera enhances salt tolerance, heat tolerance and cold acclimation ability in transgenic Arabidopsis plants. PLoS One. 2013;8(4):e63064.
  37. Mirjalili H.M., Fakhr‐Tabatabaei S.M., Bonfill M., Alizadeh H., Cusido R.M., Ghassempour A., Palazon J. Morphology and withanolide production of Withania coagulans hairy root cultures. Eng. Life Sci. 2009;9(3):197-204.
  38. Attaran Dowom S., Abrishamchi P., Radjabian T., Salami S.A. Elicitor-induced phenolic acids accumulation in Salvia virgata Jacq. hairy root cultures. Plant Cell, Tissue and Organ Culture (PCTOC). 2022;148(1):107-117.
  39. Verma P., Mathur A.K., Shanker K. Growth, alkaloid production, rol genes integration, bioreactor up-scaling and plant regeneration studies in hairy root lines of Catharanthus roseus. Plant Biosyst. 2012;146(sup1):27-40.
  40. Nasiri J., Naghavi M.R., Alizadeh H., Fattahi Moghadam M.R., Mashouf A., Nabizadeh M. Modified AHP-based decision-making model toward accurate selection of eligible maintenance media for production of taxanes in Taxus baccata callus culture. Acta Physiol Plant. 2015;37:1-5.
Journal of Medicinal plants and By-Products
Volume 13, Issue 3
September 2024
Pages 669-679

Files

Share

How to cite

Statistics