Abstract
Protoplasts were isolated at high yields from actively growing callus and cell suspensions of cotyledons and needles of mature trees. The best protoplast growth response was obtained from cell suspensions of cotyledon and needle callus. Lower protoplast yields were obtained directly from young needles of flushing buds on explants from mature shoots (30-year-old trees) growing in vitro. In all cases, the first divisions, promoted by dimethyl sulfoxide, were observed in 10–45% of the protoplasts by 7–10 days. After 25–30 days, colonies of 8–10 cells were established. Browning of protoplast-derived cell cultures was observed within 40–45 days (cotyledons) and 20–25 days (mature tree sources).
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Abbreviations
- BA:
-
N6-Benzyladenine
- DCR:
-
Douglas-fir cotyledon revised medium
- 2,4-D:
-
2,4-dichlorophenoxyacetic acid
- DMSO:
-
dimethyl sulfoxide
- FDA:
-
Fluorescein diacetate
- Mes:
-
2-(N-morpholino) ethanesulfonic acid
- NAA:
-
α-naphthaleneacetic acid
References
Ahuja MR (1984) Silvae Genetica 33:32–37.
Boller T, Kende T (1976) Plant Physiol. 63:1123–1132.
David A, David H (1979) z. Pflanzenphysiol 94:1652–1657
David H, Jarlet E, David A (1984) Physiol. Plant. 61:477–482.
Durzan DJ (1980) In 50th Anniversary, May 8–10, 1979, The Institute of Paper Chemistry, Appleton, WI, pp. 31–60.
Gupta PK, Durzan DJ (1986) Bio/Technology, 4:643–645.
Gupta PK, Durzan DJ (1985) Plant Cell Rept. 4:177–179.
Hahne G, Hoffman F (1984) Proc. Nat. Acad. Sci. USA 81:5449–5453.
Hakman, IC, von Arnold S (1983) Plant Cell Rept. 2:92–96.
Hakman I, von Arnold S (1985) J. Plant Physiol. 121:149–158.
Jensen WA (1962) In Botanical histochemistry, principles and practice. W.A. Freeman, San Francisco.
Kinloch BB Jr., Parks GK, Fowler CW (1970) Science 167:193–195.
Kirby EG, Cheng TY (1979) Plant Sci. Lett. 14:145–154.
Patel KR, Shekhawat RS, Berlyn AP, Thorpe TA (1984) Plant Cell Tiss. Organ Cult. 3:85–90.
Rao PS, Atkins PO (1985) Protoplasma 124:80–86.
Steward FC, Krikorian AD (1970) In WR Sharp, PO Larsen, EF Paddock, V Raghavan (eds.). Plant cell and tissue culture, principles and application. Ohio State Univ. Press, Columbus, pp. 221–262.
Vasil IK, Vasil V (1980) In IK Vasil (ed.), Perspectives in plant cell and tissue culture. Intl. Rev. Cytol., Academic Press, New York, pp. 1–19.
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Communicated by O. L. Gamborg
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Gupta, P.K., Don Durzan, J. Isolation and cell regeneration of protoplasts from sugar pine (Pinus lambertiana). Plant Cell Reports 5, 346–348 (1986). https://doi.org/10.1007/BF00268598
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DOI: https://doi.org/10.1007/BF00268598