Abstract
A method has been developed which allows the recovery of calli from a high proportion of individual, injected, mesophyll protoplasts of Nicotiana tabacum c.v. Xanthi. A small drop of low melting point agarose is used both to hold protoplasts during microinjection and for their subsequent culture in feeder dishes. The feeder dishes consist of "beads" of protoplasts at a high density set in agarose to "feed" the infected protoplasts across a liquid medium.
The method has been used successfully both with normal protoplasts and protoplasts from which the vacuole has been removed.
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Abbreviations
- NT medium:
-
Nagata-Takebe medium (Nagata and Takebe, 1971)
- MS medium:
-
Murashige-Skoog medium (Murashige and Skoog, 1962)
- NAA:
-
1-Naphthaleneacetic acid
- BAP:
-
6-Benzylaminopurine
- LMP agarose:
-
low melting point agarose
References
Griesbach RJ Sink KC (1983) Plant Sci Lett 30: 297–301
Murashige T Skoog F (1962) Physiol Plant 15: 473–497
Nagata T Takebe I (1971) Planta 99: 12–20
Shillito RD Paszkowski J Potrykus I (1983) Plant Cell Reps 2: 244–247
Steinbiss HH Stabel P (1983) Protoplasma 116: 223–227
Watts JW Dawson JRO King JM (1980) In: Ingram DS Helgeson JP (eds) Tissue Culture Methods for Plant Pathologists, Oxford, Blackwell, pp 97–101
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Communicated by M. H. Zenk
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Lawrence, W.A., Davies, D.R. A method for the microinjection and culture of protoplasts at very low densities. Plant Cell Reports 4, 33–35 (1985). https://doi.org/10.1007/BF00285500
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DOI: https://doi.org/10.1007/BF00285500