biologia plantarum

Biologia plantarum 2010, 54:566-570 | DOI: 10.1007/s10535-010-0101-4

Hairy roots formation in recalcitrant-to-transform plant Chenopodium rubrum

S. Dmitrović¹, N. Mitić^1,*, S. Zdravković-Korać¹, B. Vinterhalter¹, S. Ninković¹, L. J. Ćulafić¹

¹ Institute for Biological Research "Siniša Stanković", University of Belgrade, Belgrade, Serbia

Susceptibility of C. rubrum to Agrobacterium-mediated transformation was demonstrated by inoculating the petioles of in vitro grown plants with A. rhizogenes strain A4M70GUS. Hairy roots were produced in 8 % of explants. They were isolated and maintained on plant growth regulator-free solid or liquid half-strength Murashige and Skoog medium for two years. Hairy root fresh mass increased 30 - 90 folds when grown in liquid medium, which was superior to solid medium, where most of the hairy roots produced calli. When these calli were grown on medium supplemented with 0.5 mg dm-3 thidiazuron, embryo-like structures were obtained. Transgenic status of long-term callus and hairy root cultures was confirmed by histochemical GUS assay, by PCR specific to the uidA, rolA&B and ags genes and by Southern hybridization.

Keywords: Agrobacterium rhizogenes; embryo-like structures; GUS activity; thidiazuron

Subjects: Agrobacterium rhizogenes; Chenopodium rubrum; hairy roots; polymerase chain reaction (PCR); Southern blot analysis; transgenic plants

Received: November 25, 2008; Accepted: May 15, 2009; Published: September 1, 2010Show citation