Optimization of induction and culture conditions of transgenic hairy roots in the medicinal plant Scrophularia deserti

Abstract

Scrophularia deserti is an important medicinal plant belonging to the Scrophulariaceae family. Research has demonstrated that extracts of the plant have antidiabetic and anticancer effects. This species is a valuable source of biologically active compounds, specially iridoid glycosides and flavonoids. Hairy root system can be used for the in vitro production of pharmaceutically valuable compounds. In this research, transgenic hairy root induction was established through the mediation of the A13 strain of Agrobacterium rhizogenes. The effects of various explants (seedling without radicle, cotyledon and leaf) prepared from in vitro grown plants at different ages (10, 18, 25, 50 and 72-day old) and three different inoculation times (5, 15 and 30 min) on the efficiency of hairy root induction were investigated. The maximum hairy roots (22.33 roots per explant) induced from 18-day-old seedlings explants inoculated with bacterial suspension for 5 min. The transgenic status of hairy roots was confirmed by PCR using rolA and rolB genes-specific primers. Four different culture media (MS, half strength MS, B5 and PGoB) were used to determine the best suitable media composition for the optimum cell growth of two hairy root lines (G and H). The results revealed that MS medium was superior for growth and high biomass production of hairy roots . Hairy root line H showed maximum biomass production (16.33 mg/30 ml culture medium, in terms of fresh weight) after 4 weeks of culture. Hairy root lines developed in this study can be used to investigate the production of pharmaceutically important metabolites of S. deserti.

Keywords

• scrophularia deserti
• transgenic hairy root
• explant
• culture medium