In Vitro Cellular and Developmental Biology - Plant, 2024 (SCI-Expanded)
In this study, a rapid and efficient biotechnological method was developed for the critically endangered (CR) endemic species Tripleurospermum ziganaense (Asteraceae) in Turkey to reintroduce it to its natural habitat. Murashige and Skoog basal medium (MS) supplemented with 1.0 mg L−1 gibberellic acid (GA3) was highly effective with 80% germination percentage. MS medium containing 1.0 mg L−1 6-BA and 0.1 mg L−1 IBA was superior for the highest average shoot number of 11.73-fold per explant. The same basal medium strengthened with 1.0 mg.L−1 2iP plus 0.1 mg L−1 IBA was highly favorable for the highest average shoot length of 47.77 mm. In vitro micropropagated plants had similar DNA ploidy levels (x) and chromosome number (2n = 18) compared to mother plants. Rooting success was achieved between 93.33 and 100% in all rooting media. MS medium strengthened with 1.0 mg L−1 IBA was remarkably effective in terms of the highest average root number with 11.71-fold per explant. On the other hand, MS medium fortified with 0.25 mg L−1 NAA was quite conspicuous with an average root length of 124.03 mm per explant. All rooted plantlets were initially acclimatized in climate room conditions and then transferred from optimal greenhouse conditions into a botanical garden with 100% and 90% survival rates, respectively. Afterward, plantlets were reintroduced into experimental plot near their wild population of T. ziganaense, with 90% survival. Flow cytometry and cytology-assisted cytogenetic fidelity assessments of the micropropagated plantlets exhibited ploidy as well as chromosomal stability within the plantlets and with coherence to their mother plant. The preliminary results obtained from the combination of micropropagation with reintroduction efforts offered an excellent opportunity for in situ and ex situ conservation efforts to bolster populations of threatened endemic T. ziganaense. This approach consisting of the components of conservation, propagation, and reintroduction (CPR) may potentially serve as a model for saving and enriching other species at risk.