Establishment of hairy root lines and analysis of iridoids and secoiridoids in the medicinal plant Gentiana scabra

Abstract

BackgroundGentiana scabra is commonly known as ‘Longdan’ is an important herb in traditional Chinese medicines, commonly used for the treatment of inflammation, anorexia, indigestion and gastric infections. Iridoids and secoiridoids are main bioactive compounds which attributed to the pharmacological properties of this plant. The use of hairy root cultures as an excellent alternative for the production of pharmaceutically important metabolites in less time period with ensured quality of raw materials.ResultsAn efficient hairy root culture system of Gentiana scabra and influence of different plant growth regulators (PGRs) on the production of gentiopicroside, swertiamarin and loganic acid constituents were described. Leaf explants were infected with Agrobacterium rhizogenes, which induced hairy roots up to 21%. The transformed hairy root lines were confirmed by PCR using rolB and rolC gene-specific primers. Among various solid and liquid media, B5 liquid medium resulted maximum root biomass (36- fold higher) in 4-weeks. Quantitative analysis showed loganic acid was 6.6- fold higher in the presence of zeatin (1 mg/l) and gentiopicroside accumulation was 1.8- fold higher in the presence of naphthaleneacetic acid (NAA, 1 mg/l), as compared to the roots of plants grown in greenhouse. On the other hand, 1.4- and 2.5- fold higher gentiopicroside and swertiamarin were observed in the presence of 1.0 mg/l NAA as compared to commercial Gentiana herb No. 2. The result also showed iridoid and secoiridoid contents affected greatly by age, physiology and growing environment of the plant.ConclusionsThe use of hairy root cultures is an excellent alternative to harvesting natural or in vitro grown plants to produce pharmaceutically important metabolites in less time with ensured quality.Electronic supplementary materialThe online version of this article (doi:10.1186/1999-3110-55-17) contains supplementary material, which is available to authorized users.

Methyl jasmonate elicits the solasodine production and anti-oxidant activity in hairy root cultures of Solanum trilobatum L.

“…Depending upon the target tissue, the transformation frequency varies among plant species, and hence, the identification of suitable explant is indispensible for successful transformation. Root explants superseded other explants in Taraxacum platycarpum (Lee et al, 2004) while explants such as petiole in W. somnifera (Saravanakumar et al, 2012), hypocotyls in Psoralea corylifolia L. (Abhyankar et al, 2005), leaf in Artemisia vulgaris (Sujatha et al, 2013) and stem in Gentiana scabra (Huang et al, 2014) were found to be highly susceptible to A. rhizogenes strains.…”

Influence of different Agrobacterium rhizogenes strains on hairy root induction and analysis of phenolic and flavonoid compounds in marshmallow (Althaea officinalis L.)

“…Compared to bacteria suspension, insulin syringe provided a lower level of wounded and faced with the less influx of bacteria. Similarly, a study was performed to induce hairy roots of Gentiana scabra, the results showed that stem explants directly injected by A. rhizogenes induced the formation of hairy roots from all inoculated explants and failed to form in control samples (Huang et al 2014).…”

Potential of the genetically transformed root cultures of Plumbago europaea for biomass and plumbagin production

“…In comparison, hypocotyl explants showed the lowest frequency of transformation. Stem explants have been widely used for A. rhizogenes-mediated transformation in other plant species 32,33 and they were often the preferred explant type. 34 Our result confirms preceding reports designated that bacterial strains and type of explants significantly influenced the generation of transgenic roots.…”