Plant regeneration from embryogenic cells-derived protoplasts of Bupleurum falcatum

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Plant regeneration from embryogenic cells-derived protoplasts of Bupleurum falcatum
Jae W Bang; Dong S In; Sung H Chung; Jang Ryol Liu
Bibliographic Citation
Plant Cell Tissue and Organ Culture, vol. 55, no. 2, pp. 151-154
Publication Year
Hypocotyl segments of Bupleurum falcatum L. formed embryogenic calluses when cultured on Murashige and Skoog's (MS) medium supplemented with 9.0 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Suspension cultures were initiated by placing calluses into medium with 0.45 μM 2,4-D. Protoplasts were enzymatically isolated from suspension cultures. They were plated at a density of 5 x 104 protoplasts per ml on MS medium supplemented with 9% mannitol, 9.0 μM 2,4-D, 4.4 μM BA, 4.6 μM kinetin, and 0.6% Seaplaque agarose. After four weeks of culture, microcalluses were formed and subsequently transferred to MS solid medium with 18.1 μM 2,4-D. Upon transfer to MS basal medium, microcalluses gave rise to somatic embryos at a frequency of approximately 10%. They subsequently developed into plantlets. The regenerants were successfully transplanted to potting soil and grown to maturity in a greenhouse. The regenerants had the normal chromosome number of 2n=2x=20 and did not show morphological aberrancy.
protoplastsomaclonal variationsomatic embryotissue cultureUmbelliferae
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