Efficient Adventitious Shoot Organogenesis for the Conservation of Pseudolysimachion nakaianum (Ohwi) T. Yamaz., an Endemic Species in South Korea
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Pseudolysimachion nakaianum (Ohwi) T. Yamaz. is an endemic and important species in the forest ecosystems of South Korea. This study aimed to develop an in vitro propagation system for P. nakaianum to enhance the availability of this rare plant for commercial horticulture and restoration purposes. We utilized different explants like cotyledon (excised after 2 weeks), petiole and leaf (collected after 8 weeks) from in vitro seedlings to assess the impact of five different cytokinins on adventitious shoot organogenesis. In both cotyledon and petiole explants, half-strength Murashige and Skoog (MS) medium, supplemented with 2 mg L-1 thidiazuron (TDZ) and 0.05 mg L-1 1-naphthaleneacetic acid (NAA) evoked the highest rate of shoot induction (61.1%±9.6% and 26.7%±8.8%, respectively) and the highest number of shoots (3±0.6 and 4.4±0.5, respectively) per explant. In contrast, leaf explants showed no in vitro regeneration. Multiple shoots originating from the explants were separated individually and transferred to half-strength MS medium supplemented with 30 g L-1 sucrose and 3 g L-1 GelriteTM. P. nakaianum plantlets were transplanted into plastic pots filled with a mixture of coco peat, decomposed granite, peat, and perlite (5:1:1:3) and approximately 70% the plants were survived for 4 weeks. This protocol outlines an effective method for the direct in vitro regeneration of P. nakaianum from cotyledon and petiole explants providing valuable information for the conservation of this species.
Abstract
Pseudolysimachion nakaianum (Ohwi) T. Yamaz. is an endemic and important species in the forest ecosystems of South Korea. This study aimed to develop an in vitro propagation system for P. nakaianum to enhance the availability of this rare plant for commercial horticulture and restoration purposes. We utilized different explants like cotyledon (excised after 2 weeks), petiole and leaf (collected after 8 weeks) from in vitro seedlings to assess the impact of five different cytokinins on adventitious shoot organogenesis. In both cotyledon and petiole explants, half-strength Murashige and Skoog (MS) medium, supplemented with 2 mg L-1 thidiazuron (TDZ) and 0.05 mg L-1 1-naphthaleneacetic acid (NAA) evoked the highest rate of shoot induction (61.1%±9.6% and 26.7%±8.8%, respectively) and the highest number of shoots (3±0.6 and 4.4±0.5, respectively) per explant. In contrast, leaf explants showed no in vitro regeneration. Multiple shoots originating from the explants were separated individually and transferred to half-strength MS medium supplemented with 30 g L-1 sucrose and 3 g L-1 GelriteTM. P. nakaianum plantlets were transplanted into plastic pots filled with a mixture of coco peat, decomposed granite, peat, and perlite (5:1:1:3) and approximately 70% the plants were survived for 4 weeks. This protocol outlines an effective method for the direct in vitro regeneration of P. nakaianum from cotyledon and petiole explants providing valuable information for the conservation of this species.