Isolation and characterization of intergeneric somatic hybrids in the Apocynaceae family

• Published in: Theoretical and applied genetics 1991-10, Vol.82 (6), p.713-716
• Main Authors: Kostenyuk, I. (Inst. of Cell Biology and Genetic Engineering, Kiev (Russian Federation)), Lubaretz, O, Borisyuk, N, Voronin, V, Stoeckigt, J, Gleba, Y
• Format: Article
• Language: English
• Subjects: ADN; ALCALOIDE; ALCALOIDES; ALKALOIDS; APOCYNACEAE; Biological and medical sciences; Classical genetics, quantitative genetics, hybrids; DNA; Fundamental and applied biological sciences. Psychology; FUSION DE PROTOPLASTES; FUSION DEL PROTOPLASTO; Genetics of eukaryotes. Biological and molecular evolution; HIBRIDACION SOMATICA; HYBRIDATION SOMATIQUE; INDOL; Indole alkaloid; INDOLES; ISOENZIMAS; ISOENZYME; ISOENZYMES; Isozyme spectrum; PROTOPLAST FUSION; Pteridophyta, spermatophyta; Ribosomal DNA; RIBOSOMAS; RIBOSOME; RIBOSOMES; Somatic hybrid; SOMATIC HYBRIDIZATION; Vegetals
• ISSN: 0040-5752; 1432-2242
• DOI: 10.1007/BF00227315

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.