[Ex vitro production of transgenic composite plants in Glycyrrhiza glabra via Agrobacterium rhizogenes-mediated transformation as a new method]

Background: Licorice, Glycyrrhiza glabra [belong to Leguminosae family] is one of the most popular medicinal plants in the world and it is widely used in many fields such as medical, pharmaceutical, confectionery and health industries. Different parts of licorice [shoots, leaves and roots] were had various components such as Glycyrrhzin that was used for some proposes

Objective: The current study was done with the aim of gene transfer via Agrobacterium rhizogenes by ex vitro method for hairy root production in licorice

Methods: The experiment was laid out as a completely randomized design [CRD] with five treatments in three replications. At first, root of young plantlets was eliminated and excited plantlets were putted in the glass wool contain suspension of bacteria. After 10 to 14 days of inoculation with Agrobacterium rhizogenes, the roots were appeared. The percentage of root induction by four strains of Agrobacterium [ATCC 15834, GMI 9534, A4 and A13] with check [without bacteria] was investigated

Results: The results of PCR analysis with specific primers for roots of composite plants [putative transgenic] was shown that three strains of bacteria [A4, A13 and GMI 9534] and strain ATCC 15834, were produced 100% and 66.66% transgenic roots respectively

Conclusion: Thus, production of composite licorice plants was remarked due to it has low cost, fast and simple

Genetic transformation studies and scale up of hairy root culture of Glycyrrhiza glabra in bioreactor

The study was undertaken to induce hairy roots in Glycyrrhiza glabra in leaf explants and to optimize the nutritional requirement for its growth kinetics at shake flask and bioreactor level. Pathogenecity of Agrobacterium depends upon transformation ability of strain and age, type, and physiological state of explants. Agrobacterium rhizogenes strain K599 was used to infect leaf explants of G. glabra. Explants of different age groups were obtained from 2 to 5 weeks old in vitro grown cultures. Bacterial strain K599 could induce hairy roots in 3 and 4 weeks old leaf explants cultured on B5, MS, NB and WP basal semi-solid medium. Leaf explants of 2 and 5 weeks old culture were not responsive to bacterial infection in terms of hairy root induction. Maximum transformation frequency (TF) of tested bacterial strain was 47 percent obtained in 3 weeks old explants after 25 days of incubation on MS basal semi solid medium. NB and B5 both media composition showed 20 percent of transformation frequency after 28 and 38 days respectively. WP medium did not support induction of roots in cultured leaf explants infected with A. rhizogenes strain K599even after 50 days of incubation. Further, when all the four media combinations were tested for root growth it was found that though WP was not responsive for hairy root induction, yet all four basal media supported hairy root growth and a gradual increase in fresh weight biomass was observed with an increase in culture duration. However amongst all, the NB medium composition supported best growth of hairy roots followed by MS, B5 and WP media. About 20 times increase in root biomass on fresh weight basis was recorded after 45days of culture in NB medium. Initial inoculum of roots (0.18 g. F.wt./ flask) containing 50 ml of liquid culture medium produced 3.59 g (F. wt.) biomass. A fast growing hairy root clone G6 was grown in a 5 l capacity mechanically agitated bioreactor provided with a nylon mesh septum. After 30 days of sterile...