Euphorbia lathyris subsp. root, L.

Ricigliano, Vincent A., Sica, Vincent P., Knowles, Sonja L., Diette, Nicole, Howarth, Dianella G. & Oberlies, Nicholas H., 2020, Bioactive diterpenoid metabolism and cytotoxic activities of genetically transformed Euphorbia lathyris roots, Phytochemistry (112504) 179, pp. 1-9 : 2

2.1. Genetic transformation and establishment of E. lathyris root cultures

Transformed roots of E. lathyris were established via A. rhizogenes - mediated infection of greenhouse grown plants. Stem explants of E. lathyris were used as the target tissue for infection and root induction ( Fig. 1a ). Root primordia were observed at the infection sites, emerging from callus as early as 15 days after inoculation ( Fig. 1b ). Fully developed adventitious roots showed an abundance of root hairs along the axis that are characteristic of the “hairy root” phenotype ( Fig. 1c ). An isogenic root line displaying vigorous growth was propagated on hormone-free solid media ( Fig. 1d ), and then grown in quadruplicate liquid culture flasks on an orbital shaker to generate the experimental biomass used in subsequent experiments ( Fig. 1e ).

Figures

Fig. 1. Establishment and culture maintenance of transformed E. lathyris roots. (a) Stem explants of 3-week-old greenhouse grown plants were used for co-culture with A. rhizogenes. (b) Roots emerged from callus at the site of infection after 2–3 weeks.(c) Adventitious roots displaying the characteristic of the “hairy root” phenotype. (d) Growth characteristics of the isogenic root line used in this study on agar media and in (e) liquid media.