Отримання та культивування бородатих коренів рослин Bidens pilosa L.

Aim. The aim of the work was the establishment of Bidens pilosa L. “hairy” root culture using Agrobacterium rhizogenes -mediated transformation. Methods . A. rhizogenes strain А4 carried pCB161 and pCB124 vectors with the selective nptII (nos promoter and terminator) and ifn-α2b (Mll or 35S promoter respectively, and ocs terminator) genes were used for genetic transformation. Explants were cocultivated with bacterial suspension for 30 min and transferred to 1/2MS solidified medium. PCR analysis with primers specific for rolB, nptII, and ifn-α2b genes was used to confirm the transgenic nature of obtained roots. Results . The growth of roots started in 7-10 days after the transformation. The number of roots generated on the leaf explants was significantly lower than of those formed on the stem explants (2-8 and 12-22, respectively). The frequency of root formation for leaf and stem explants was 56.7±6.5% and 84.4±5.8%, respectively. The roots demonstrated the typical phenotype with significant branching, negative geotropism and were able to grow without growth regulators in the nutrient medium. The presence of rolB, ifn-α2b, and nptII genes was confirmed by PCR analysis. Conclusions . An effective system for genetic transformation of B. pilosa plants using A. rhizogenes -mediated transformation method was developed and "hairy" root culture was established. The presence of transferred genes was confirmed by PCR analysis. The obtained "hairy" root lines differed in growth rate. Keywords: Bidens pilosa L., Agrobacterium rhizogenes , transformation, "hairy" root culture.