An improved regeneration protocol for the establishment of in vitro cultures of Juniperus phoenicea L.

The study aims to present an improved plant regeneration protocol through the establishment of in vitro cultures of Juniperus phoenicea L. The initial cultures were established on Rugini Olive Medium (OM), and Driver and Kuniyuki Walnut (DKW) medium either hormone free (Hf) or supplemented with kinetin (KIN) or 6-(γ,γ-dimethylallylamino)purine (2iP), at 0.1 mg L^‑1. The effect of 2iP at 0 (control), 0.1, 0.2 and 1.0 mg L^‑1 was examined after the establishment of the initial cultures.
The multiplication stage involved the successive subcultures on OM containing a) 0.1 mg L^‑1 2iP, b) 0.1 mg L^‑1 2iP and 0.01 mg L^‑1 1-naphthaleneacetic acid (NAA), c) 0.1 mg L^‑1 2iP and NAA at 0.1 mg L^‑1, d) 0.01 mg L^‑1 NAA, and e) 0.1 mg L^‑1 NAA. During the initial culture, the percentage of explants forming shoots reached up to 80% on DKW with 0.1 mg L^‑1 2iP, which was not significantly different from the percentages recorded on DKW Hf (73%) or OM with 0.1 mg L^‑1 2iP (70%). The production of shoots was higher on OM media either Hf or containing 0.1 mg L^‑1 2iP (4.8 and 5.5, respectively), and DKW supplemented with 0.1 mg L^‑1 2iP (4.9). In the multiplication stage, the highest number of shoots per explant (6.2) was recorded into media containing 0.1/0.01 2iP/NAA (mg L^‑1). Moreover, the percentage of explants forming shoots reached 100% across all hormone- containing media.
The shoot elongation ranged between 0.8 and 1.1 cm.
The present study revealed that the combined use of 0.1 mg L^‑1 2iP and 0.01 mg L^‑1 NAA promoted shoot proliferation and shoot growth of J. phoenicea.

International Symposium on Biotechnological Tools in Horticulture

K. Bertsouklis, P. Aroniada, P. Stamatopoulou, A. Paraskevopoulou

acclimatization, DKW, NAA, micropropagation, tissue culture, 2iP

https://doi.org/10.17660/ActaHortic.2026.1454.52