The genus Ferulabelongsto the Apiaceaefamily and mostly growsin arid climates such as central Asia, the Middle East, and central Europe(Pimenov et al., 2004; Pimenov and Leonov, 1993). In Iranian flora, the Apiaceaefamily consists of 112 genera, 316 species and 75 endemic plant species.
The genus Ferula contains 30 species including 15 endemic plants and is typically called koma or kema and (Sattar and Iranshahi, 2017). Many researchershave studied secondary metabolites of plants in the genus Ferula(Murray et al., 1982). Many species such as Ferula asafetida, F. gumosa and F.
latisecta have been used in traditionalmedicine to treat stomachache, hysteria, infant colitis, and asthma(Eigner and Scholz, 1999; Zargari, 1997). Furthermore,research related to the cytotoxic activity of main groups of chemical compounds from the genus Ferula has been carried out including research on umbelliprenin from F. szowitsiana, farnesiferol C from F. assa-foetida, and umbelliprenin, farnesiferol A, gummosin and badrakemone from F. persica var.persica(Barthomeuf et al.
, 2008; Lee et al., 2010; Shahverdi et al., 2006). In other words, most species of this genus have been known as a beneficial source of biologically active compounds such as coumarins, sesquiterpenes, sesquiterpenecoumarins, sesquiterpene lactones, sulphur-containing derivatives, monosaccharides and daucane esters and aromatic resins which are used in traditional medicine(Asghari et al., 2016; El-Razek et al.
, 2001; Iranshahi et al., 2007; Kanani et al., 2011; Kapoor, 2000; Razavi and Janani, 2015). Amongst these compounds, sesquiterpenecoumarinhas frequent biological usage and is formedfrom the common coumarin group. More extensive and promising biological properties can be expected from this class of natural compound.
Sesquiterpenecoumarins isolated from the genus Ferula have different biological activities such as antibacterial, antiviral (anti HIV), anti-inflammatory, spasmolytic, anticoagulant, P-glycoprotein (P-gp) inhibitory and cytotoxic properties(Abd El-Razek, 2003; Dastan et al., 2014; Gudarzi et al., 2015; Nazari and Iranshahi, 2011; Zarei et al., 2013).F. pseudalliaceais endemic speciefound in the Kurdish (Sanandaj) mountains of western Iran used in traditional medicine for various purposes.
One aspectof the phytochemical study of native plants of Iran is finding new antiprotozoal natural products(Adams et al., 2009). Recently some new disesquiterpene and sesquiterpenecoumarins were investigated from F. pseudalliacea. Sesqui- and disesquiterpenecoumarins of F. pseudalliaceawhich were found to have anti-bacterial, anti-cancer, anti-plasmodial, and phytotoxic effects(Dastan et al.
, 2016; Dastan et al., 2014; Dastan et al., 2012).F. pseudalliacea is one of the most important and endangered medicinal plants in western Iran which is found in a narrow area of Kurdistan Province.
F. pseudalliacea is spread in very low frequency in a mountains area due to poor seed germination. Poor seed germination in Apiaceaehas beenreported previously(Golmohammadi, 2013; Moghadam et al., 2014). For the sustainable use of native medicinal plants with seed dormancy underlaboratory conditions,special treatments are neededto increase seed germination rate and decrease the time of germination. Seed dormancy is a common phase of the Ferulaspecies life cycle(Keshtkar et al., 2008; Nadjafi et al.
, 2006; Zare et al., 2011). Agrobacterium rhizogenes is a soil-borne gram-negative bacterium which causes hairy roots on monocot and eudicot plants. In other words,hairy root disease is a pathological syndrome of dicotyledonous plants following wounding and infection with A. rhizogenes. Its root-inducing (Ri)plasmid of A.
rhizogenes containing transfer of DNA encoding root locus (rol) gene loci (rolA, rolB, and rolC) is responsible for the permanent introduction of genetic material into host cells(Lima et al., 2009).Hairy roots have been used for various purposes inthe last 30 years ranging from recombinant protein production and metabolic engineering to analyses of rhizosphere physiology and biochemistry(Ono and Tian, 2011).
Recently, A. rhizogenesmediated hairy root production has been utilized as a biotechnology tool in a variety of plant species to produce plant important secondary metabolites. Mostapplications of cultured hairy roots have focusedon the large scale production of useful products or secondary metabolites (Xu et al., 2006).Todate, there has beenno available report on optimization of factors affecting seed germination, hairy root induction and production of farnesiferol B content in F. pseudalliacea.
Thus, optimizing an efficient germination condition and Agrobacterium rhizogenes-mediated hairy root culture system could be a promising approach to supply farnesiferol B from F. pseudalliacea. The aim of the present study was to optimize seed germination in in vitro conditions and to induce and optimize hairy roots for F. pseudalliacea and check the hairy roots for production of secondary metabolites, in particularfarnesiferol B