Influence of Agrobacterium rhizogenes strains on hairy roots induction in Trigonella foenum-graecum L. and secondary metabolites production

Document Type: Research Paper

Authors

Department of Horticultural Sciences, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran

Abstract

Fenugreek(Trigonella foenum-graecum L.) is a rich source of important medicinal metabolites. This plant belongs to the Fabaceae family. Induced hairy roots by Agrobacterium rhizogenes are a suitable tissue for the production of secondary metabolites, due to the stability and high production of roots without phytohormone in a short time. Different strains of Agrobacterium rhizogenes (A4, ATCC11325 and ATCC15834) were evaluated for induction of transformed hairy roots in T. foenum-graecum L. using seedling explants. The application of hairy root culture may become an alternative method for increase secondary metabolites. Transgenic status of the roots was confirmed by PCR using rolB specific primers. All of the A. rhizogenes strains led to hairy roots induction. The maximum frequency of transformation (97.87%) was obtained using A4 strain in 7-days-old seedling. The 7-days-old explants were inoculated using A4 strain result in highest fresh (0.166 g) and dry (0.080 g) weight of roots. The explants were inoculated by ATCC11325 strain produced hairy roots with highest amount of total phenol (8.113 mg/g DW) and flavonoid content (3.215 µg/g DW).

Keywords

Main Subjects


[1] Aasim, M., Hussain, N., Umer, E.M., Zubair, M., Hussain, S.B., Saeed, S.H., Rafique, T.S. and Sancak, C. 2010. In vitro shoot regeneration of fenugreek (Trigonellafoenum-graecum L.) using different cytokinins. Afr. J. Biotechnol, 9:( 42) 7174-7179.

[2] Akbarian, R., Hasanloo, T. and Khosroshahi, M. 2011. Evaluation of trigonelline production in Trigonella foenum-graecum hairy root cultures of two Iranian masses. Plant Omics J, 4: 408-412.

[3] Al-Mahdawe, M.M., Al-Mallah, M.K. and Al-Attrakchii, A.O. 2013. Genetically transformed hairy roots producing agropine induced on Trigonella foenum-graecum L. plant by Agrobacterium rhizogenes 1601. J of Biotechnology Research Center, 7: 91-98.

[4] Banihashemi, O., Khavari, R.A., Yassa, N. and Najafi, F. 2015. Induction of hairy roots in Atropa komarovii using Agrobacterium rhizogenes. Indian Journal of Fundamental and Applied Life Sciences, 5(3): 2014-2020.

[5] Bertani, G. 1952. Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. J Bacteriol, 62: 293–300.

[6] Chang, C., Yang, M., Wen, H. and Chern, J. 2002. Estimation of total flavonoid content in propolis by two complementary colorimetric methods. JFDA, 10: 178-182.

[7] Christey, M.C. and Braun, R.H. 2005. Production of hairy root cultures and transgenic plants by Agrobacterium rhizogenes – mediated transformation. Transgenic Plants: Methods and Protocols, 286: 47–60.

 [8] Doyle, J.J. and Doyle, J.L. 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin, 19: 11-15.

[9] Gamborg, O.L., Miller, R.A. and Ojima, K. 1968. Nutrient requirements of suspension cultures of soybean root cells. Experimental Cell Research, 50: 151–158.

[10] Georgiev, M., Pavlov, A. and Bley, T. 2007. Hairy root type plant in vitro systems as sources of bioactive substances. Applied Microbiology and Biotechnology, 74: 1175–1185.

[11] Kabirnotaj, S., Zolalla, J., Nematzadeh, G. and SHokri, E. 2013. Optimization of hairy root culture establishment in Chicory plants (Cichorium intybus) through inoculation by Agrobacterium rhizogenes. J. Agri. Biotec, 4(2): 61-75.

[12] Kayser, O. and Quax, W.G. 2007. Medicinal plant biotechnology.Vol. 1, WILEY-VCH Verlag GmbH & Co., Weinheim, 604 pp.

[13] Kim, S.I., Veena, A. and Gelvin, S.B. 2007. Genome-wide analysis of Agrobacterium T-DNA integration sites in the Arabidopsis genome generated under non-selective conditions. The plant Journal, 51: 779-791.

[14] Manuhara, Y.S.W., Kristanti, A.N., Utami, E.S.W., Yachya, A. 2015. Effect of sucrose and potassium nitrate on biomass and saponin content of Talinum paniculatum Gaertn. Hairy root in balloon-type bubble bioreactor. J Trop Biomed, 5(12): 1027- 1032.

 [15] Meda, A., Lamien, C.E., Romito, M., Millogo, J. and Nacoulma, O.G. 2005. Determination of the total phenolic, flavonoid and pralin contents in Burkina Fasan honey, as well as their scavenging activity. Food Chemistry,91: 571-577.

[16] Merkli, A., Christen, P. and Kapetanidis, I. 1997 Production of diosgenin by hairy root cultures of Trigonella foenum.graecum L. Plant Cell Reports, 16: 632-636.

[17] Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15: 473- 497.

[18] Peraza-Luna, F., Rodrı´guez-Mendiola, M., Arias-Castro, C., Bessiere, J.M. and Calva-Calva, g. 2001. Sotolone production by hairy root cultures of Trigonella foenum-graecum in Airlift with Mesh Bioreactors. J. Agric. Food Chemistry, 49: 6012-6019.

[19] Qaderi, A., Akbari, Z., Kalateh-jari, S, Fatehi, F., Tolyat, M., Jalali Moghadam, M. and Naghdi Badi, H. 2014. Improving Trigonelline Production in Hairy Root Culture of Fenugreek (Trigonella foenum-graecum). Journal of Medicinal Plants,15: 73-80.

[20] Sevon, N. and Oksman-Caldentey, K.M. 2002. Agrobacterium rhizogenes-Mediated Transformation: Root Cultures as a Source of Alkaloids. Planta Med, 68(10): 859-868.

[21] Shahabzadeh, Z., Heidari, B. and Faramarzi Hafez, R. 2013. Induction of Transgenic Hairy Roots in Trigonella foenumgraceum Co-cultivated with Agrobacterium Rhizogenes Harboring a GFPGene. Journal of Crop Science and Biotechnology, 16 (4): 263- 268.

[22] Sharafi, A., Hashemi, Sohi H., Mirzaee, H. and Azadi, P. 2014. In vitro regeneration and Agrobacterium mediated genetic transformation of Artemisia aucheri Boiss. Physiol Mol Biol Plants, 20(4): 487–494.

[23] Sharafi, A., Hashemi, S. H., Mousavi, A., Azadi, P., Razavi, K. and Ntui, V.O. 2012. A reliable and efficient protocol for inducing hairy roots in Papaver bracteatum. PCTOC, 113: 1-9.

[24] Srivastava, V., Kaur, R., Chattopadhyay, A.K. and Banerjee, S. 2013. Production of industrially important cosmaceutical and pharmaceutical derivatives of betuligenol by Atropa belladonna hairy root mediated biotransformation. Industrial Crops and Products, 44: 171–175.

[25] Valimehr, S., Sanjarian, F., Hashemi sohi, H., Sharafi, A. and Sabouni, F. 2014. A reliable and efficient protocol for inducing genetically transformed roots in medicinal plant Nepeta pogonosperma. Physiol Mol Biol Plants, 20(3): 351–356.