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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
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
Author information
Authors and Affiliations
Additional information
Communicated by M. H. Zenk
Rights and permissions
About this article
Cite this article
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
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00285500