Establishment of Bidens pilosa L. ‘hairy’ root culture
Journal Title: Вісник Українського товариства генетиків і селекціонерів - Year 2015, Vol 13, Issue 1
Abstract
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.
Authors and Affiliations
N. A. Matvieieva, A. M. Shakhovsky
Keywords
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