Elicitors and precursors, an effective strategy for increasing betulin an anticancer agent in Betula litwinowii callus cultures

Mehrirad, Nastaran; Payamnoor, Vahide; Nazari, Jamile

Elicitors and precursors, an effective strategy for increasing betulin an anticancer agent in Betula litwinowii callus cultures

Document Type : Original Research

Authors

Nastaran Mehrirad ¹

vahide Payamnoor ²

Jamile Nazari ¹

¹ Gorgan University of Agriculture Sciences and Natural Resources

² Gorgan University of Agriculture Sciences and Natural Resources,

Abstract

Betulin is a pentacyclic lupane-type triterpene, mainly obtained from Betula species plants with a variety of biological actions such as anti-HIV and anticancer properties. This study aimed to enhance the production of betulin in cultures of Betula litwinowii calli under the influence of concentrations and duration time of elicitors and precursors. Collected leaves from Sangdeh habitat in summer, were cultured in WPM medium containing 2,4-D and BAP hormones. Four month calli were sub-cultured in medium containing elicitors such as salicylic acid, chlorocholine chloride and cobalt chloride and precursors such as sucrose, vitamin and glucose. . Calli were harvested from new culture media after two, three and four weeks and their wet and dry weights were calculated. Data analysis was performed based on two-factor factorial experiment (the first factor, elicitors and precursors each in four levels and the second factor, time with three levels) in a completely randomized design with three replications with SPSS software. Duncan's multiple range test were performed to compare the mean (p≤0.01). The results showed that two week elicitation with chlorocholine chloride (1.5 mg/l for one week) was the best with a more than 3-fold increasing in betulin induction compared to the control treatment (0.068 mg / g). The use of vitamin precursor (ten-fold the normal amount in WPM culture medium) for three weeks, causes the highest increase in betulin induction compared to the control (0.1 mg / g, respectively). In general, in the present study, this treatment is introduced as the best.

Keywords

betulin

birch

chloro-choline chloride

vitamins

20.1001.1.23222115.1401.13.3.1.8

Subjects

Pharmaceutical Biotechnology

1. Krasutsky, P.A., 2006. Birch bark research and development. Natural Product Research, 23:919-942.
2. Zare, H., Akarinia, M., Hosseini, SM., Ejtehadi, H and Amini Eshkevari, T., 2010. A new Record of Betula Litwinowii (Betulaceae) and a review of the geographical distribution of genus Betula in Iran. Iranian journal of botany, 16:237-241.
3. Jalili, A., and Arzani, H., 1999. A preliminary survey of endemic, rare and endangered plant species in Iran (red data book of Iran). Research institute of forests and rangelands, Tehran, Iran, 764p.
4. Nazari, J., Payamnoor, V. and Kavousi, M.R., 2013. Seed born fungi associated with birch trees in Northern forests of Iran. Iranian Journal of Range and protection research. 10 (2):165-168.
5. Häggman H., Sutela S., Welander M., (2007) Micropropagation of Betula Pendula Roth including Genetically modified Material. In: Jain S.M., Häggman H. (eds) Protocols for Micropropagation of Woody Trees and Fruits. Springer, Dordrecht. Pp: 153-162.
6. Zhao, J., Davis, LC and Verpoorte, R., 2005. Elicitor signal transduction leading to production of plant secondary metabolites. Biotechnology Advances, 23(4): 283-333.
7. Andrea, V. and Ricardo, B., 2007. Molecular aspects of the early stages of elicitation of secondary metabolites in plants. Plant Science, 172:861-875.
8. Asghari, GH., Mostajeran, A., sadeghi aliabadi, H., and Nakhaei, A., 2011. Effect of salicylic Acid and Silver Nitrate on Taxol production of Taxus baccata. Journal of Medicinal Plants, 11(8): 74-82.
9. Vasconsuelo, A., and Boland, R., 2007. Molecular aspects of the early stages of elicitation of secondary metabolites in plants. Plant Science, 172: 861-875.
10. Esmaeil zadeh Bahabadi, S. and Sharifi, M., 2013. Increase secondary metabolite production plants using biological Elicitors. Journal of cells and tissues, 4 (2): 119-128.
11. Barrios, H., Zhang, Y.L., Sandoval, C., and Xiao, Z.A., 2009. Increase of Taxol Production in Taxus globosa Shoot Callus by Chlorocholine Chloride. Natural Products Journal, 2: 33-37.
12. Ali, M.B., Yu, K.W., Hahn, E.J. and Paek, K.Y. 2006. Methyl jasmonate and salicylic acid elicitation induces ginsenosidesaccumulation, enzymatic and non-enzymatic antioxidantin suspension culture Panax ginseng roots in bioreactors. Plant Cell Report, 25:613-620.
13. Rezaei, A., Ghanati, F., and Amini Dehaghi, M., 2013. Salicylic acid-Induced physiological effects and Taxol production in cell culture of Taxus baccata L. National Congress on Medicinal Plants, Kish Island, Iran.
14. Badrhadad, A., Piri, K.H., and Ghiasvand, T., 2013. Increasing of alpha-tocopherol in cell suspension cultures (Elaeagnus angustifolia L.). International Journal of Agriculture and Crop Sciences, 5(12): 1328-1331.
15. Rezaei, A., Ghanati, F., and Jamshidi, M., 2013. Cobalt induced Taxol production and release by cell culture of Hazel (Corylus avellana L.). National Congress on Medicinal Plants, Kish Island, Iran.
16. Jafari hajati, R., Payamnoor, V., Ghasemi bezdi, k. and Ahmadian Chashmi, N., 2016. Effects of methyl jasmonate and salicylic acid on production of Betulin and Betulinic acid in cell suspension of birch (Betula pendula Roth.). Iranian Journal of Medicinal and Aromatic Plants, 32)6( :1037-1047.
17. Jafari hajati, R., Payamnoor, V., Ahmadian Chashmi, N. and Ghasemi bezdi, k., 2018. Improved accumulation of betulin and betulinic acid in cell
suspension culture of Betula pendula roth by abiotic and biotic elicitors, Preparative Biochemistry and Biotechnology, DOI: 10.1080/10826068. 2018.1514514.
18. Jafari hajati, R., Payamnoor, V., Ahmadian Chashmi, N., 2019. Effect of biotic and abiotic elicitors on production of betulin and betulinic acid in the
hairy root culture of Betula pendula Roth, Preparative Biochemistry and Biotechnology, DOI: 10.1080/10826068.2019.1650372
19. Feria Romero, I., Lazo, E., Ponce Noyola, T., Cerda Garcia Rojas C.M. and Ramos Valdivia, A.C. 2005. Induced accumulation of oleanolic acid and ursolic acid in cell suspension cultures of Uncaria tomentosa. Biotechnolgy Letters, 27:839-43.
20. Hayashi, H., Hiraoka, N. and Ikeshiro, Y. 2005. Differential regulation of soyasaponin and betulinic acid production by yeast extract in cultured licorice cells. Plant Biotechnology, 22(3):241-244.
21. Habibi Khaniani B, Moieni A, Abdollahi M. 2005. Production of secondary metabolites and pharmaceutical constituents through tissue and cell culture. J. Med. Plants.; 4 (14) :1-6. URL: http://jmp.ir/article-1-703-fa.html
22. Ghorbanly, M., and Delavar, K., 2001. Effect of the Type and Concentration of sugars on Taxol Content in Callus Culture of Taxus Baccata L. Iranian Journal of Agriculture Science, 32(3): 575-583.
23. Hasan poor, H., Bernard, F., and Shaker, H., 2007. Optimizing callus culture in Zataria multiflora Boiss for rosmarinic acid production. Iranian Journal of Rangelands and Forests Plant Breeding and Genetic Research, 15: 1-9.
24. Cusido R M, Palazon J, Bonfill M, Naviaosorio A, Morales C, Pinol M T. 2002. Improved paclitaxel and baccatin Ш production in suspension cultures of Taxus media. Biotechnol. Prog, 18: 418-423.
25. Nazari, J. Payamnoor, V. Aliadeh, M. GHasemi edi, K. 2017. Micropropagation of birch (Betula litwinowii) from leaf callus. Forest and Wood Products, 70 (2): 199-207.
26. Mehrirad, N., Payamnoor, V. and Naari, J. 2015. Effect of trees age and light on Betula litwinowii callogenesis and botulin induced in vitro conditions. Iranian Journal of Rangelands and Forests Plant Breeding and Genetic Research, 23 (1): 93-102.
27. Van Ket, N., Thi Lan Anh, T., and Hoang Uyen Dung, N., 2012. Effecting of Sucrose Concentrations and Inoculum density on adventitious root growth in cell suspension culture of Panax vietnamensis and initially growth in a Bioreactor. Southeast-Asian Journal of Sciences: 1 (2): 215-222.
28. Prakrash, G. and Srivastava, AK., 2005. Statistical media optimization for cell growth and azadirachtin production in Azadirachta indica suspension cultures. Process Biotech, 40: 3795.
29. Iriti M, and Faoro F., 2009. Chemical diversity and defence metabolism: How Plants cope with pathogens and ozone pollution. International Journal of Molecular Sciences, 10: 3371-3399.
30. Khosroushahi, A., Valizadeh, M., Ghasempour, M., Naghdibadi, H., 2005. Improved Taxol production by combination of inducing factors in suspension cell culture of Taxus baccata. Cell Biology International, 30: 262-269.
31. Kovacik, J., Backor, M., Strnad, M., and Repcak, M., 2009. Salicylic acid-induced changes to growth and phenolic metabolism in Matricaria chamomilla plants. Plant Cell Report, 28:135–143.
32. Hashim, K., Al-oubaidi, M., and Aseel salih, M.A., 2014. Increasing secondary metabolites of Calendula officinalis L. using salicylic acid in vitro. World Journal of Pharmacy and Pharmaceutical Sciences, 3(5): 1146-1155.
33. Mehrabani, B., Nazeri, S., and Piri, K., 2012. Evaluation of total produced phenol in Chaei Koohi (Stachys lavandulifolia Vahi) callus culture and possibility of its enhancement using Elicitors, Journal Biotecnology Agriculture, 2(4): 77-88.
34. Alvarez, P.S., Spollansky, T.C. and Giulietti A.M. 2000. The influence of different biotic and abiotic elisitors on the production and profile of tropane alkaloids in hairy root cultures of Brugmensia candida. Enzyme and Microbial Technology, 26:254-258.
35. Cai, ZH., Kastel, A., Speiser, C., and Smetanska, I., 2013. Enhanced Resveratrol Production in Vitis vinifera Cell Suspension Cultures by Heavy Metals without Loss of Cell Viability. Applied Biochemistry and Biotechnology, 11p.