Summary
Protoplasts of young Pisum Sativum L. seedlings from 7 different genotypes were isolated and regenerated to the callus stage. Germinating embryos were cultivated with cotyledons removed, thus avoiding intracellular starch accumulation in donor tissue. The first lateral shoots provided a source of homogenous meristematic cells which gave rise to sustained protoplast division and resulted in callus formation within 4, weeks. Root formation occurred on hormone-free medium and shoots developed on medium containing kinetin, 2iP or zeatin in the third subculture, when subcultured in monthly intervals.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
von Arnold, S.; Eriksson, T. Factors influencing the growth and division of pea mesophyll protoplasts. Physiol. Plant. 36:193–196; 1976.
von Arnold, S.; Eriksson, T. A revised medium for pea mesophyll protoplasts. Physiol. Plant. 39:257–260; 1977.
Constabel, F.; Kirkpatrick, J. W.; Gamborg, O. L. Callus formation from mesophyll protoplasts of Pisum sativum. Can. J. Bot. 51:2105–2106; 1973.
Gaff, F.; O'Kong Ogola. The use of non-permeation pigments for testing the survival of cells. J. Exp. Bot. 22:756–758; 1971.
Gamborg, O. L.; Miller, R. A.; Ojima, K. Nutrient requirements of suspension cultures of soybean root cells. Exp. Cell Res. 50:151–158; 1968.
Gamborg O. L.; Shyluk, J.; Kartha, K. K. Factors affecting the isolation and callus formation in protoplasts from the shoot apices of Pisum sativum L. Plant Science Letters 4:285–292; 1975.
Gupta, P. K.; Durzan, D. J. Biotechnology of somatic polyembryogenesis and plantlet regeneration in loblolly pine. Bio/Technology 5:147–151; 1987.
Jia, S. Factors affecting the division frequency of pea mesophyll protoplasts. Can. J. Bot. 60:2192–2196; 1982.
Kao, K. N.; Michayluk, M. R. Nutritional requirements for growth of Vicia hajastana cells and protoplasts at a very low population density in liquid media. Planta 126:105–110; 1975.
Landgren, C. R. The influence of culture conditions on mitotic activity in protoplasts derived from Pisum root cortical explants. Protoplasma 87:49–69; 1976.
Landgren, C. R. Gibberellin enhancement of the enzymatic release of Pisum root cell protoplasts. Physiol. Plant. 52:349–352; 1981.
Murashige, T.; Skoog, F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15:473–497; 1962.
Puonti-Kaerlas, J.; Eriksson, T. Improved protoplast culture and regeneration of shoots in pea (Pisum sativum L.). Plant Cell Reports 7:242–245; 1988.
Tan, M. M. C.; Rietveld, E. M.; Marrewijk, G. A. M., et al. Regeneration of leaf mesophyll protoplasts of tomato cultivars (L. esculentum): factors important for efficient protoplast culture and plant regeneration. Plant Cell Reports 6:172–175; 1987.
Author information
Authors and Affiliations
Additional information
This work was made possible by a joint GFP/BMFT—project to H.-J. Jacobsen (grant A 24/87-ZF).
Rights and permissions
About this article
Cite this article
Lehminger-Mertens, R., Jacobsen, HJ. Protoplast regeneration and organogenesis from pea protoplasts. In Vitro Cell Dev Biol 25, 571–574 (1989). https://doi.org/10.1007/BF02623570
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02623570