Agrobacterium rhizogenes-mediated transformation for the increase of polyfructans content in roots of Lactuca sativa L.

Abstract

Aims. The studing of Lactuca sativa L. “hairy” roots was the aim of the work. Methods. The roots were obtained via cocultivation of leaves with Agrobacterium rhizogenes with pCB161 vector (selective nptII gene, target ifn-α2b gene). Selection of transgenic roots was performed on Murashige and Skoog medium with twice reduced content of macrosalts, 600 mg/l cefatoxime and 25 mg/l kanamycine. Results. Transformed “hairy” root lines differed in biomass increase from 0,02±0,003 to 0,16±0,01 g fresh weight during 30 days and polyfructans content from 60,3±3,9 to 163,3±29,7 mg/g dry weight. The maximum total fructan content was found in the one line of Odessky Kucheriavets cv “hairy” roots and one line of Green Coral “hairy” roots cultivated during 30 days (0,137±0,002 mg and 0,15±0,01 mg respectively). Conclusions. Differences of biomass increase and polyfructans content was observed in both lines of roots. Probably, such differences connected by that each line is separate transgenic event.

Key words: transformation, Agrobacterium, Lactuca sativa, fructans.