Features of Agrobacterium rhizogenes-mediated genetic transformation of Artemisia vulgaris L., Artemisia annua L. and Ruta graveolens L. medicinal plants

  • К. О. Дробот
  • Н. А. Матвєєва
  • А. М. Шаховський

Abstract

Aim. Аrtemisia vulgaris, Аrtemisia annua, Ruta graveolens "hairy" roots obtaining was the aim of the work. Methods. We used A4 A. rhizogenes wild strain for genetic transformation. Leaf, internode and root explants were cocultivated with bacterial suspension and transferred onto MS medium. Cefotaxime were added to the medium in 3–9 days after transformation. The presence of rolB gene were determined by PCR analysis. Results. Transformation frequency (TF) was found to be 15 % and 100 % (A. vulgaris); 10 % and 10 % (A. annua); 2 % and 3 % (R. graveolens) in case of internodes and leaves using respectively. This parameter depended in type of explants and time of it cultivation on the medium without cefotaxime. Prolongation of this period has led to TF increasing. PCR analysis proved the presence of rolB genes in obtained “hairy” roots. Conclusions. A. vulgaris, А. annua and R. graveolens "hairy" root cultures can be obtained using A4 A. rhizogenes-mediated transformation. Leaves were found to be the optimal explants for the effective “hairy” roots establishment.

Keywords: “hairy” roots, Artemisia vulgaris, Аrtemisia annua, Ruta graveolens.