Summary
Protoplasts were isolated from callus cultures of Rauwolfia serpentina Benth., Rhazya stricta Decaisne, and Catharanthus roseus (L.) G. Don, or from leaves of Vinca minor L. Protoplast isolation, culture, and fusion techniques as well as hybrid screening systems were developed for these species, and hybrids were obtained. Hybrid combinations were Rauwolfia + Vinca, Rauwolfia + Catharanthus, Rauwolfia + Rhazya, and Catharanthus + Vinca. For hybrid isolation, the physiological complementation method was utilized. Analyses of the material obtained included a cytogenetic study of the chromosomes, a study of multiple molecular forms of transferases and esterases, and the blot hybridization of restricted nuclear DNA using ribosomal DNA as a probe. Hybrids were identified in all species' combinations tried. A ten-fold increase in the accumulation of raucaffricine (relative to the parental Rauwolfia strain) was observed in one cell line of the Rauwolfia + Vinca hybrid. Our studies indicated the genetic stability of the great majority of the hybrid cell lines over a period of more than 20 months of in vitro growth. No shoot morphogenesis has so far been observed in this material.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Balsevich J (1988) Monoterpene indole alkaloids from Apocynaceae other than Catharanthus roseus. In: Constabel F, Vasil IK (eds) Cell culture and stomatic cell genetics. Vol 5. Phytochemicals in plant cell cultures. Academic Press, San Diego, pp 371–384
Brewer GJ (1970) An introduction to isozyme techniques. Academic Press, New York London
Caboche M (1980) Nutritional requirements of protoplastderived haploid tobacco cells grown at low cells densities in liquid medium. Planta 189:7–18
Chopra RN, Nazar SL, Chopra IC (1956) A glossary of Indian medicinal plants. C.S.I.R., New Delhi, pp 245–253
Cresswell RC, Fowler MW, Scragg AH (1988) Glyphosate-tolerance in Catharanthus roseus. Plant Sci 54:55–63
DeLuca V, Kurz WGW (1988) Monoterpene indole alkaloids (Catharanthus alkaloids). In: Constabel F, Vasil IK (eds) Cel lculture and somatic cell genetics. Vol 5. Phytochemicals in plant cell cultures. Academic Press, San Diego, pp 385–401
Fujita Y, Takahashi S, Yamada Y (1985) Selection of cell lines with high productivity of shikonin derivatives by protoplast culture of Lithospermum erythrorhizon cells. Agric Biol Chem 49: 1755–1759
Gleba Yu, Sytnik KM (1984) Protoplast fusion. Springer, Berlin Heidelberg New York
Kao KN, Michayluk MR, Constabel F, Gamborg OL (1974) Plant protoplast fusion and growth of intergeneric hybrid cells. Planta 120:215–227
Kobayashi S (1987) Uniformity of plants regenerated from orange (Citrus sinensis Osb.) protoplasts. Theor Appl Genet 74:10–14
Kolosha VO, Fodor II (1986) Structural heterogenity of Citrus lemon rDNA. Mol Biol (USSR) 20:656–659
Kutchan TM, Hampp N, Lottspeich F, Beyreuther K, Zenk MM (1988). The DNA clone for strictosidine synthase from Rauvolfia serpentina: DNA sequence determination and expression in Escherichia coli. FEBS Lett 237:40–44
Medgyesy P, Menczel L, Maliga P (1980) The use of cytoplasmic streptomycin resistance: chloroplast transfer from Nicotiana tabacum into Nicotiana sylvestris, and isolation of their somatic hybrids. Mol Gen Genet 179:693–698
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473
Nagy F, Lazar G, Menczel L, Maliga P (1983) A heteroplasmic state induced by protoplast fusion is a necessary condition for detecting rearrangements in Nicotiana mitochondrial DNA. Theor Appl Genet 66:203–207
Petiard V, Baubault C, Bariaud A, Hutin M, Courtois D (1985) Studies on variability of plant tissue cultures for alkaloid production in Catharanthus roseus and Papaver somniferum callus cultures. In: Neumann K-H, Barz W, Reinhard E (eds) Primary and secondary metabolism of plant cell cultures. Springer, Berlin Heidelberg New York, pp 133–142
Sakushima A, Nishibe S (1988) Mass spectrometry in the structural determination of the flavonol triglicosides from Vinca minor. Phytochemistry 27:915–920
Shure M, Wessler S, Fedoroff N (1983) Molecular identification and isolation of the Waxy locus in maize. Cell 35:225–233
Sidorov VA, Menczel L, Nagy F, Maliga P (1980) Chloroplast transfer in Nicotiana based on metabolic complementation between irradiated and iodoacetate treated protoplasts. Planta 152:341–345
Sidorov VA, Piven NM, Gleba YY, Sytnik KM (1985) Somatic hybridization of Solanaceae. Naukova Dumka, Kiev
Takeuchi Y, Komamine A (1981) Glucans in the cell walls regenerated from Vinca rosea protoplasts. Plant Cell Physiol 22:1585–1594
Zenk MH, Deus B (1982) Natural product synthesis by plant cell cultures. In: Fujiwara A (ed) Plant tissue culture. Maruzen, Tokyo, pp 391–394
Author information
Authors and Affiliations
Additional information
Communicated by G. Wenzel
Rights and permissions
About this article
Cite this article
Kostenyuk, I., Lubaretz, O., Borisyuk, N. et al. Isolation and characterization of intergeneric somatic hybrids in the Apocynaceae family. Theoret. Appl. Genetics 82, 713–716 (1991). https://doi.org/10.1007/BF00227315
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00227315