Plant regeneration from embryogenic cells-derived protoplasts of Bupleurum falcatum |
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Authors: | Bang Jae W In Dong S Chung Sung H Liu Jang R |
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Institution: | (1) Department of Biology, College of Natural Science, Chungnam National University, 200 Koong-dong, Yusong-gu, Taejon, 305-764, Korea;(2) Plant Cell and Molecular Biology Research Unit, Korea Research Institute of Bioscience and Biotechnology (KRIBB), P.O. Box 115, Taejon, 305-606, Korea |
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Abstract: | 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 × 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.
This revised version was published online in June 2006 with corrections to the Cover Date. |
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Keywords: | protoplast somaclonal variation somatic embryo tissue culture Umbelliferae |
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