Optimization and genetic induction of hairy roots in kelus (Kelussia odoratissima Mozaff.)

Document Type : Research Paper

Authors

1 M.Sc. Graduated in Biotechnology, Faculty of Agriculture, Dept. Production Engineering and plant Genetic, Lorestan University, Khorramabad, Iran

2 Prof. Faculty of Agriculture, Dept. Production Engineering Engineering and plant Genetic, Lorestan University, Khorramabad, I.R. Iran.

3 Ph.D graduated in plant breeding, Faculty of Agriculture, Dept. Production Engineering and plant Genetic, Lorestan University, Khorramabad, I.R. Iran

10.22092/ijrfpbgr.2022.356510.1395

Abstract

Kelus (Kelussia odoratissima Mozaff.) is one of the medicinal, spicy, aromatic and endemic herb that grows in restricted regions of Iran,. Due to the importance of its pharmaceutical compounds in traditional medicine, the excessive harvesting of kelus has put it at the risk of extinction. To protect kelus from extinction, plant tissue culture can be used to produce useful compounds and secondary metabolites. The hairy root culture is an efficient system for the production of medicinal compounds and valuable secondary metabolites in plants. For this purpose, two pathogenic strains of Agrobacterium rhizogenes (A4 and ATCC15834), two types of explants (cotyledon and hypocotyl), three levels of acetosringon (a1= 0 µmol, a2=50 µmol, and a3= 100 µmol), and three levels of glucose (b1=0, b2=0.5%, b3=1%) were applied using a factorial experiment based on a completely randomized design with three replications. The effect of such different factors on the induction and production of hairy roots in kelus was investigated. The results showed that both Agrobacterium strains were able to induce hairy roots in kelus. Also, ATCC15834 strain in MS medium with 100 µmol of acetosringon in the presence of 0.5% glucose had the highest efficiency in the production of hairy roots in cotyledon explants. The lowest amount of hairy roots was observed for A4 strain in MS medium without both acetosringon and glucose in both explants. PCR analysis was performed with rolC specific primers to ensure hairy root induction has occurred by A. rizhogenes. The results of this research showed that for the first time in Iran, it was possible to induce hairy roots in kelus explants.

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  • Ahmadi, F., Kadivar, M. and Shahedi, M. 2007. Antioxidant activity of Kelussia odoratissima Mozaff. in model and food systems. Food chemistry, 105(1): 57-64.
  • Akramian, M., Tabatabaei, S.M.F. and Mirmasoumi, M. 2008. Virulence of different strains of Agrobacterium rhizogenes on genetic transformation of four Hyoscyamus species.American-Eurasian journal of Agricultural & Environmental Sciences, 3(5): 759-763.
  • Brijwal, L. and Tamta, S. 2015. Agrobacterium rhizogenes mediated hairy root induction in endangered Berberis aristata DC. SpringerPlus, 4(1): 1-10.
  • Dewick, P.M. 2002. Medicinal natural products: a biosynthetic approach, John Wiley & Sons,507 pages.
  • Dini torkamani, M.R., Abaspour, N., Jafari, M. And Samadi, A. 2014. Induction and optimization of hairy root growth condition for Valeriana officinalis, through inoculation by Agrobacterium rhizogenes, Journal of cell and tissue, 5(1):23-30.
  • Ebrahimi, M., Mokhtari, A. and Amirian, R 2018. A highly efficient method for somatic embryogenesis of Kelussia odorotissima an endangered medicinal plant. Plant Cell, Tissue and Organ Culture, 132(1): 99-110.
  • Gawel, N. and Jarret, R. 1991. A modified CTAB DNA extraction procedure for Musa and Ipomoea. Plant Molecular Biology Reporter, 9(3):262-266.
  • Godwin, I., Todd, G., Ford-Lloyd, B. and Newbury, H.J. 1991. The effects of acetosyringone and pH on Agrobacterium-mediated transformation vary according to plant species. Plant Cell Reports, 9(12): 671-675.
  • Gelvin, Stanton B. Agrobacterium and plant genes involved in T-DNA transfer and integration. Annual Review of Plant Physiology and Plant Molecular Biology, June 2000, vol. 51, p. 223-256.
  • Haj hashemi, V.A., Ghanadi, A. and Soltani, L. 2003. Analgesic and antiinfammatory effects of amirkabiria adrotissima. Journal of Reserch in Medical Sciences (JRMS), 7(2):121-125.
  • Hasanloo, T., Rezazadeh, S. and Rahnama, H. 2009. Hairy roots as a source for production of valuable pharmaceutical materials. Journal of Medicinal Plants, 8(29): 1-190.
  • Jafari Hajati, R., Payamnoor, V., Ghasemi Bezdi, K. and Ahmadian Chashmi, N 2016. Production of pharmaceutical active ingredients via hairy root induction of Birch (Betula pendula). Iranian Journal of Rangelands and Forests Plant Breeding and Genetic Research, 24(2): 165-176 ).In persian).
  • Jalilian, A. 2017.Induction of Transgenic Hairy Roots in Medicinal Plant Poppy (Papaver Somniferum) by Agrobacterium Rhizogenes-Mediated Transformation. Journal of Plant Productions (Agronomy, Breeding and Horticulture), 39(4): 1-14. (In persian).
  • Khademi, M., Nazarian-Firouzabadi, F. and Ismaili, A. 2019. The effect of phosphorus and nitrogen on hairy roots production in Nicotiana tobaccum as a model plant. Journal of Plant Productions (Agronomy, Breeding and Horticulture), 44(1):13-24.
  • Kim, Y., Wyslouzil, B.E. and Weathers, P.J. 2002. Secondary metabolism of hairy root cultures in bioreactors. In Vitro Cellular & Developmental Biology-Plant, 38(1): 1-10.
  • Kumar, V., Sharma, A., Narasimha Prasad, B.C., Bhaskar Gururaj, H. and Aswathanarayana Ravishankar, G. 2006. Agrobacterium rhizogenes mediated genetic transformation resulting in hairy root formation is enhanced by ultrasonication and acetosyringone treatment. Electronic Journal of Biotechnology, 9(4): 349-357.
  • Levee, V., Garin, E., Klimaszewska, K. and Seguin, A. 1999. Stable genetic transformation of white pine (Pinus strobus L.) after cocultivation of embryogenic tissues with Agrobacterium tumefaciens. Molecular Breeding, 5(5): 429-440.
  • Li, W., Asada, Y. and Yoshikawa, T. 2000. Flavonoid constituents from Glycyrrhiza glabra hairy root cultures. Phytochemistry, 55(5): 447-456
  • Maroufi, A. and Majdi, M. 2015. Assessment of hairy roots induction of the medicinal plant Alecost (Tanacetum balsamita) using Agrobacterium rhizogenes. Genetic Engineering and Biosafety Journal, 4(2): 103-111.
  • Montazeri, F., Omidi, M., Khialparast, F. and Sabokdast, M. 2015. Hairy root induction by Agrobacterium rhizogenes in galbanum (Ferula gummosa). Iranian Journal of Rangelands and Forests Plant Breeding and Genetic Research. 22(2):251-260. (In persian).
  • Nader, B.L., Taketa, A.T.C., Pereda-Miranda, R. and Villarreal, M.L. 2006. Production of triterpenoids in liquid-cultivated hairy roots of Galphimia glauca. Planta Medica. 72(9): 842-844.
  • Oksman-Caldentey, K.-M. and Hiltunen, R. 1996. Transgenic crops for improved pharmaceutical products. Field crops research, 45(1): 57-69.
  • Omidbaigi, R., Sefidkon, F. and Saeedi, K. 2008. Essential oil content and composition of Kelussia odoratissima as an Iranian endemic plant. Journal of Essential Oil Bearing Plants. 11(6): 594-597.
  • Petrova, M., Zayova, E., Dincheva, I., Badjakov, I. and Vlahova, M. (2015). Influence of carbon sources on growth and GC-MS based metabolite profiling of Arnica montana L. hairy roots. Turkish Journal of Biology, 39(3): 469-478.
  • Rao, S.R. and Ravishankar, G. 2002. Plant cell cultures: chemical factories of secondary metabolites. Biotechnology advances. 20(2): 101-153.
  • Rolland, F., Baena-Gonzalez, E. and Sheen, J. 2006. Sugar sensing and signaling in plants: conserved and novel mechanisms. Annual Review of Plant Biology, 57(1): 675-709.
  • Salimi, M., Ebrahimi, A.E., Shojaei Asadieh, Z. And Saei Dehkordi, S. 2010. Essential oil composition of kelussia adrotissima Iranian Journal of Medicinal and Aromatics Plants. 26(2):147-156. (Inpersian).
  • Shen, W.-H., Escudero, J., Schläppi, M., Ramos, C., Hohn, B. and Koukolíková-Nicola, Z. 1993. T-DNA transfer to maize cells: histochemical investigation of beta-glucuronidase activity in maize tissues. Proceedings of the National Academy of Sciences, 90(4): 1488-1492.

Sivakumar, G., Yu, K., Hahn, E. and Paek, K. 2005. Optimization of organic nutrients for ginseng hairy roots production in large-scale bioreactors. Current Science, 89(4): 641-649.