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KMID : 0368419850280030233
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Journal of Plant Biology
1985 Volume.28 No. 3 p.233 ~ p.241
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Isolation and Culture of Protoplasts from Hypocoty-derived Callus of Soybean(Glycine max)
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Abstract
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The isolation and culture of protoplasts from hypocotyl-derived calluses of Glycine max (L.) Merr. cv. Jangyeop were carried out. The maximum protoplast yield of 5.3×10 exp (6) per gram fresh callus was obtained by digestion for 6 hrs in an enzyme solution containing 3.5% cellulase, 1.5% macerozyme, 10% sorbitol and 0.1% CaCl_2·2H_2O at pH 5.8. Newly formed cell wall of protoplasts cultured in MS agar medium containing 10 μM α-naphthaleneacetic acid (NAA) and 32 μM N^6-benzylaminopurine(BAP) could be observed after 24 hrs culture. The first cell division of the protoplasts was observed after 3 days of culture; cell clusters after 2 weeks of culture. When transferred to solid media, the protoplasts formed cell clusters gave rise to proliferating calluses.
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