KONIECZNY, R., OBERT, B., BLEHO, J., NOVÁK, Ondřej, HEYM, C., TULEJA, M., MUELLER, J., STRNAD, Miroslav, MENZEL, D., ŠAMAJ, J. Stable transformation of Mesembryanthemum crystallinum (L.) with Agrobacterium rhizogenes harboring the green fluorescent protein targeted to the endoplasmic reticulum. Journal of Plant Physiology. 2011, 168(7), 722-729. ISSN 0176-1617. E-ISSN 1618-1328

Stable transformation of Mesembryanthemum crystallinum L (common ice plant) with a green fluorescent protein (GFP) construct targeted to the endoplasmic reticulum was obtained. Seven and fourteen days after germination seedlings were infected with Agro bacterium rhizogenes strain ARqua1 either by direct coating of the cut radicles with bacteria growing on solid medium or by immersion of the cut surface in bacterial suspension at different optical densities. Both methods of infection resulted in production of GFP-positive roots with a frequency ranging from 6 to 20% according to the age of the explants and the application procedure. The green fluorescing roots displayed the typical hairy root phenotype and were easily maintained in liquid medium without growth regulators for over 2 years. Stable expression of the transgene in the roots was confirmed by polymerase chain reaction (PCR), immunoblotting and the capacity of roots to grow and produce callus on kanamycin-enriched medium.