Improvement of friable callus induction of Crocus sativus L. and establishment of a cell suspension culture system with high biomass

Amini, Somaye; Ziaratnia, Seyed Mahdi; Rajabzadeh, Ghadir

doi:10.22077/jhpr.2023.6850.1338

Document Type : Original Article

Authors

¹ Department of Horticultural Science, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran

² Department of Food Biotechnology, Research Institute of Food Science and Technology (RIFST), Mashhad, Iran

³ Department of Food Nanotechnology, Research Institute of Food Science and Technology (RIFST), Mashhad, Iran

https://doi.org/10.22077/jhpr.2023.6850.1338

Abstract

Purpose: This study aims to explore the potential of in vitro culture as a method for scaling up the production of saffron based medicinal compounds, the most expensive spice renowned. Emphasis is placed on the critical role of friable callus (FC) formation as a prerequisite for successful suspension culture. Research method: The research primarily investigates FC formation, focusing on the impact of varying strengths of Murashige and Skoog (MS) medium as well as combinations of NAA or 2,4-D and BA or Kin on compact callus. Subsequently, the study involves supplementing the MS medium with different concentrations of 2,4-D, kin, zeatin, glutamine, sucrose, and nitrogen to establish a cell suspension culture. Findings: The highest FC yield was achieved on a solid medium containing 2,4-D (1 mg l^-1)+Kin (0.2 mg l^-1), resulting in a fresh weight (FW) of 0.413 g. Furthermore, MS combined with 2,4-D (1 mg l^-1)+Kin (0.2 mg l^-1)+glutamine (10 mg l^-1), as well as MS+2,4-D (0.5 mg l^-1)+zeatin (0.3 mg l^-1)+glutamine (10 mg l^-1), demonstrated the highest FW under suspension conditions. The study also identified that 30 g l^-1 sucrose and 30 µM were optimal for inducing maximum FW. Research limitations: Cell biomass is influenced by several factors that should to be optimized. Originality/Value: This research concludes that a cell suspension system holds promise for rapidly generating sufficient cell biomass to produce valuable secondary metabolites within a limited timeframe and space. Notably, the system successfully increased biomass from 0.2 to 1.2 g, underscoring its potential for efficient saffron-based product development.

Keywords

Main Subjects

Biotechnology and Tissue culture

References

Amini, S., Ziaratnia, S., & Hemmati, Kh. (2022). Optimization of conditions for increasing of saffron cell biomass and crocin production in stirred bioreactor. Plant Cell, Tissue and Organ Culture, 149(10), 1-13. https://doi.org/10.1007/s11240-022-02233-4

Basu, P., & Chand, S. (1998). Tropane alkaloids from callus cultures, differentiated and shoots of Hyoscyamus muticus L. Journal of Plant Biochemistry and Biotechnology, 7, 39–42. https://doi.org/10.1007/BF03263031

Dar , S. A., Nawchoo, I. A., Tyub, S., & 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 Royle ex Lindl. Biotechnology Reports, 32, 1-5. https://doi.org/https://doi.org/10.1016/j.btre.2021.e00688

Dos Santos, M. R. A., De Souza, C. A., & Paz, E. S. (2017). Growth pattern of friable calluses from leaves of Capsicum annuum var. annuum cv. Iberaba Jalapeño1. Revista Ciencia Agronomica, 48, 523–530. https://doi.org/10.5935/1806-6690.20170061

Gueven, A., & Knorr, D. (2011). Isoflavonoid production by soy plant callus suspension culture. Journal of Food Engineering. 103, 237–243. https://doi.org/10.1016/j.jfoodeng.2010.10.019

Hamasaki, R. M., Purgatto, E., & Mercier, H. (2005). Glutamine enhances competence for organogenesis in pineapple leaves cultivated in vitro. Brazilian Journal of Plant Physiology,17, 383–389. https://doi.org/10.1590/s1677-04202005000400006

Jamil, S. Z. M. R., Rohani, E. R., Baharum, S. N., & Noor, N. M. (2018). Metabolite profiles of callus and cell suspension cultures of mangosteen. 3 Biotech, 8, 322. https://doi.org/10.1007/s13205-018-1336-6

Keng, C. L., Singaram, N., & Lim, B. P. (2010). Production of artemisinin from cell suspension culture of artemisia annua L. Asia Pacific Journal of Molecular Biology and Biotechnology, 18, 137–139.

Knobloch, K. H, & Berlin, J. (1980). Influence of medium composition on the formation of secondary compounds in cell suspension cultures of Catharanthus roseus (L.) G. Don. Society for Biotechnological Research, 35, 551–556. https://doi.org/10.1515/znc-1980-7-805

Leustek, T., & Kirby, E. G. (1988). The influence of glutamine on growth and viability of cell suspension cultures of Douglas-fir after exposure to polyethylene glycol. Tree Physiology,4, 371–380. https://doi.org/10.1093/treephys/4.4.371

Marques, H. P., Barbosa, S., Nogueira, D.A., Santos, M.H., Santos, B.R., & Santos-filho P.R. (2017). Proteic and phenolics compounds contents in Bacupari callus cultured with glutamine and nitrogen sources. Brazilian Journal of Biology, 78, 41–46. https://doi.org/10.1590/1519-6984.03416

Moshtaghi, N. (2020). Tissue and cell culture of saffron. In Saffron. (Koocheki A, Khajeh-Hosseini M eds). Woodhead Publishing, 229–246.

Murthy,H. N., Lee, E. J., & Paek, K. Y. (2014). Production of secondary metabolites from cell and organ cultures: strategies and approaches for biomass improvement and metabolite accumulation. Plant Cell, Tissue and Organ Culture, 118, 1–16. https://doi.org/10.1007/s11240-014-0467-7

Mustafa, N. R., De Winter, W., Van Iren, F., & Verpoorte, R. (2011). Initiation, growth and cryopreservation of plant cell suspension cultures. Nature Protocols, 6, 715–742. https://doi.org/10.1038/nprot.2010.144

Osman, N. I., Jaafar Sidik, N., & Awal, A. (2016). Effects of variations in culture media and hormonal treatments upon callus induction potential in endosperm explant of Barringtonia racemosa L. Asian Pacific Journal of Tropical Biomedicine, 6, 143–147. https://doi.org/10.1016/j.apjtb.2015.10.007

Pant, B. (2014). Application of plant cell and tissue culture for the production of phytochemicals in medicinal plants. Advances in Experimental Medicine and Biology, 808, 25–39. https://doi.org/10.1007/978-81-322-1774-9_3

Ramandi, A., Gholizadegan, A., & Seifi, A. (2022). Optimization of callogenesis and cell suspension culture in saffron. Journal of Saffron Research, 10(2), 276-284. https://doi.org/10.22077/jsr.2022.5718.1198

Rao, S.R., & Ravishankar, G.A. (2002). Plant cell cultures: chemical factories of secondary metabolites. Biotechnology Advances, 20, 101-153. https://doi.org/10.1016/s0734-9750(02)00007-1

Scragg, A.H., Ashton, S., & York, A. (1990). Growth of Catharanthus roseus suspensions for maximum biomass and alkaloid accumulation. Enzyme and Microbial Technology, 12, 292-298. https://doi.org/10.1016/0141-0229(90)90101-U

Shanjani, P. S. (2003). Nitrogen effect on callus induction and plant regeneration of Juniperus excelsa. International Journal of Agriculture and Biology, 5(4), 419–422.

Souza, J. M. M., Berkov, S., & Santos, A. S. (2014). Improvement of friable callus production of Boerhaavia paniculata rich and the investigation of its lipid profile by GC/MS. Brazilian Academy of Sciences, 86, 1015–1027. https://doi.org/10.1590/0001-3765201420130098

Taghizadeh, M., Nasibi, F., Kalantari, K. M., & Benakashani, F. (2020). Callogenesis optimization and cell suspension culture establishment of Dracocephalum polychaetum Bornm. and Dracocephalum kotschyi Boiss.: An in vitro approach for secondary metabolite production. The South African Journal of Botany, 132, 79–86. https://doi.org/10.1016/j.sajb.2020.04.015

Taherkhani, T., Asghari Zakaria, R., Omidi, M., & Zare, N. (2019). Effect of ultrasonic waves on crocin and safranal content and expression of their controlling genes in suspension culture of saffron (Crocus sativus L.). Natural Product Research, 33, 486–493. https://doi.org/10.1080/14786419.2017.1396598

Thacker, X., Thomas, K., Fuller, M., Smith, S. H.,& Dubois, J. (2018). Determination of optimal hormone and mineral salts levels in tissue culture media for callus induction and growth of industrial Hemp (Cannabis sativa L.). The Journal of Agricultural Science, 9, 1250–1268. https://doi.org/10.4236/as.2018.910088

Verma, S. K., Das, A. K., Cingoz, G. S., & Ekrem Gurel, E. U. (2016). Influence of nutrient media on callus induction, somatic embryogenesis and plant regeneration in selected Turkish crocus species. Biotechnology Reports, 10, 66–74. https://doi.org/10.1016/j.btre.2016.03.006

Ziaratnia, S. M. & Amini, S. (2021). The effect of developmental stages of corm, type of medium and plant growth regulators in callus induction of Crocus sativus L. Journal of Horticulture and Postharvest Research, 4(Special issue: recent advances in saffron), 43-56. https://doi.org/10.22077/jhpr.2021.3684.1166

Ziaratnia, S. M., Ohyama, K., Hussein, A. A. F., Muranaka, T., Lall, N., Kunert,K. J., & Marion Meyer, J. J. (2009). Isolation and identification of a novel chlorophenol from a cell suspension culture of Helichrysum aureonitens. Chemical and Pharmaceutical Bulletin, 57, 1282-1283. https://doi.org/10.1248/cpb.57.1282

Improvement of friable callus induction of Crocus sativus L. and establishment of a cell suspension culture system with high biomass

References

References

Volume 6, Issue 4 - Serial Number 23December 2023December 2023Pages 397-408

Volume 6, Issue 4 - Serial Number 23
December 2023
December 2023
Pages 397-408