Regeneration and plantlet development from somatic tissues of Aristolochia fimbriata

Abstract

Aristolochia fimbriata is a small herbaceous perennial in the basal angiosperm family Aristolochiaceae. The family contains diverse floral forms ranging from radial to monosymmetric flowers with a wide variety of insect pollinators. Additionally, Aristolochia species contain secondary metabolites that are important natural toxins and traditional medicines, and are critical to the reproduction of swallowtail butterflies. These characteristics, in combination with the small genome size and short life cycle of A. fimbriata, have prompted further development of this species as a model system to study the evolution of basal angiosperms. As a prerequisite for developing a genetic transformation procedure for Aristolochia, we developed protocols for in vitro plant multiplication, shoot organogenesis, rooting, and acclimation of tissue culturederived plants. Two varieties of Aristolochia were multiplied in vitro and rooted with 100% efficiency. Shoot regeneration was achieved within 1 month of culture initiation from whole leaf, internodal stem, and petiole explants. The highest regeneration success (97%) was recorded for stem explants. Regenerated and rooted shoots were acclimated to greenhouse conditions and developed flowers within 4 weeks of transplanting.

Callus induction, high frequency shoot organogenesis and assessment of clonal fidelity in Torenia bicolor Dalzell.

“…During the present investigation, it has been clearly indicated that the highest root induction from shoot was obtained on IBA. These results are comparable with those described by other researchers for IBA induced rooting in other species (Bliss et al, 2009;Cheruvathur and Thomas, 2014b;Phulwaria et al, 2014).…”

Efficiency of shoot regeneration from leaf, stem, petiole and petal explants of six cultivars of Chrysanthemum morifolium

“…Therefore alternative cytokinins may need to be considered for our chrysanthemum cultivars that were difficult to regenerate. The effect of explant source on direct shoot organogenesis has been reported by others (Pereira et al 2000;Dhar and Joshi 2005;Sharma and Nautiyal 2009;Bliss Plant Cell Tiss Organ Cult (2011) 107:295-304 301 et al 2009). Such variations can be attributed to age (Palmer and Keller 2011) and physiological condition of the explant, and are likely to be determined by genetic factors (Baroncelli et al 1978;Nagarathna et al 1991).…”

Characterization of the basal angiosperm Aristolochia fimbriata: a potential experimental system for genetic studies

“…We have begun to develop inbred lines to facilitate genome mapping and large scale mutagenesis experiments. High-efficiency transformation with a GFP construct allows rapid, nondestructive identification of transformed tissues for subsequent processing, and our in vitro micropropagation and regeneration methods can yield greenhouse acclimated plants in three months [47]. Individual A. fimbriata plants survive indefinitely, providing ongoing access to mutant lines that can be cloned for distribution to the research community.…”

“…A protocol for genetic transformation of A. fimbriata was developed based on in vitro shoot regeneration [47] from leaf and stem explants coupled with Agrobacterium -mediated transformation. Leaf and internodal stem segments (2-3 cm long) from rooted tissue cultured A. fimbriata plants were excised and immediately immersed in induction media [46] to keep moist.…”

“…The explants were inoculated with Agrobacterium strain AGL containing plasmid pGH00.0131 as previously described for Theobroma cacao L. [46]. After the inoculation the explants were blotted on sterile paper towels and co-cultivated on callus initiation medium (CI) [47] (0.5 mg/L 6BA, 1 mg/L NAA and 1mg/L TDZ) for 64 h in the dark at 27°C. Following co-cultivation, the explants were transferred to CI supplemented with 50 mg/L Geneticin (G418) (Cellgro, Herndon, VA) and 200 mg/L Claforan (Aventis, New Jersey) and incubated at 27°C in the dark for the remainder of the 14 days.…”