Improvement of seed germination and in vitro propagation of Bupleurum latissimum Nakai through embryogenesis

Authors: KEE HWA BAE

Abstract: Abstract: Bupleurum latissimum Nakai is a critically important endangered plant belonging to the family Apiaceae. Seed germination was promoted by soaking in the dormancy breaker gibberellic acid (GA₃) (50% germination as compared with 4% of the control). An optimal temperature for germination of seed previously soaked in GA₃ solution was determined by incubation at various temperatures. Seed germination of 31.8% was observed at 25 °C. These results indicate that the seeds of B. latissimum are difficult to germinate, even when treated with GA₃. The greatest callus induction (94.8%) was observed in root explants of seedlings grown on MS medium containing a specific concentration of 2,4-dichlorophenoxyacetic acid (2,4-D) (1.0 mg L^-1) and N⁶-benzyladenine (BA) (3.0 mg L^-1). Induction of somatic embryo was observed in 78.5% of the root segment cultured on MS medium containing 3.0 mg L^-1 2,4-D alone. The highest shoot induction rate was obtained in MS medium containing 30 g L^-1 sucrose (number of shoots, 6.8; average length of shoot, 8.8 cm). After acclimation in artificial soil (1:1:1 mixture of peat moss, perlite, and sand) and transfer to the greenhouse, 98% of plantlets survived over 2 months. This in vitro propagation protocol will be very useful for the conservation of this critically endangered plant.

Keywords: Bupleurum latissimum Nakai, callus, endangered, endemic, GA3, regeneration

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