Encapsulation of Centaurium Erythraea Rafn – An Efficient Method for Regeneration of Transgenic Plants

Abstract

An effective procedure for producing transformed Centaurium erythraea plants from synthetic seeds is described. Explants were encapsulated in 3% sodium alginate with 3% sucrose. Encapsulated hairy roots were cultured on half-strength Murashige and Skoog (1/2 MS) or Woody Plant (WPM) agar-solidified regeneration media supplemented with 6-benzylaminopurine (BAP) or without the cytokinin. The use of WPM with 0.5 mg/L of BAP gave the best shoot formation frequency (86%) and mean number of shoots (15) per root segment. Shoots rooted with 97% frequency on 1/2 MS without growth regulators. Encapsulated shoot buds were cultured on onethird-strength MS agar medium (1/3 MS) supplemented with indole-3-butyric acid (IBA) (0.05 mg/L). The plantlet conversion frequency was 32%. The encapsulated hairy roots and shoot buds were stored for 4, 6 or 14 weeks at 4°C. Synthetic seeds encapsulated with 3% sodium alginate with 3% sucrose stored at 4°C remained viable for 6 weeks but their developmental parameters significantly decreased. Adding nutrient medium and growth regulator to the alginate matrix increased plantlet recovery from both non-stored and stored synthetic seeds: synthetic seeds retained their viability and ability to form plantlets even after 14 weeks of storage. Regenerated transformed plantlets of C. erythraea were acclimatized in the greenhouse.K Ke ey y w wo or rd ds s: : Centaurium erythraea, hairy roots, transgenic plants, synthetic seeds.

Cryopreservation and effect of lighting conditions and cytokinins on in vitro multiplication of Miconia ligustroides (DC.) Naudin

“…The sodium alginate serves as artificial endosperm and must provide nutrients for the growth of the encapsulated explants (Kumar et al, 2005;Piatczak and Wysokinska, 2013). The addition of growth regulators can improve the development of plant material because the capsules provide a direct contact area with the explant with the small contact surface in test tubes or Petri dishes (Etienne and Berthouly, 2002;Polzin et al, 2014), thus increasing the desired effects.…”