Summary
Somatic hybrids of diploid amylose-free (amf) Solanum tuberosum and diploid S. brevidens were made by Poly-Ethylene-Glycol (PEG) or electrofusion methods. For the isolation of interspecific hybrids the use of selection markers (kanamycin and hygromycin resistance) was useful but not essential. In this 2x+2x interspecific combination 4x and 6x somatic hybrids were obtained. Seed set was the best in 4x×4x (S. tuberosum) backcrosses, but seed germination was the best in 6x×4x combinations, using in vitro germination of unripe seeds harvested 25 days after pollination. A high degree of pollen stainability (30–40%) was observed in 7 tetraploid hybrids and very low in all hexaploids. After iodine staining, the recessive amf marker was expressed by a red colour instead of blue, visible in starch-containing cells like columella cells of root tips, (micro)tubers or microspores. As expected, complementation was observed in starch-containing cells of the fusion hybrids. Segregation of the amf marker was clearly observed in microspores of 4x and 6x hybrids. Segregation ratios in the 4x hybrids showed variable recombination frequencies. In the backcross progeny of hexaploid F12-5 with a tetraploid amf mutant one amylose-free recombinant among 67 plants was found, indicating the occurrence of meiotic recombination in the megaspore mother cells.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Austin, S., M.A.Baer & J.P.Helgeson, 1985. Transfer of resistance to potato leaf roll virus from Solanum brevidens into Solanum tuberosum by somatic fusion. Plant Sci. 39: 75–82.
Barsby, T.L., J.F.Shepard, R.J.Kemble & R.Wong, 1984. Somatic hybridisation in the genus Solanum: S. tuberosum and S. brevidens. Plant Cell Report 3: 165–167.
Bokelmann, G.S. & S.Roest, 1983. Plant regeneration from protoplasts of potato (Solanum tuberosum cv. ‘Bintje’). Z. Pflanzenphysiol. 109: 259–265.
Ehlenfeldt, M.K. & J.P.Helgeson, 1987. Fertility of somatic hybrids from protoplast fusions of Solanum brevidens and S. tuberosum. Theor. Appl. Genet. 73: 395–402.
Fish, N., A.Karp & M.G.K.Jones, 1988. Production of somatic hybrids by electrofusion in Solanum. Theor. Appl. Genet. 76: 260–266.
Flipse, E., N. Huisman, J.E.M. Bergervoet, B.J. de Vries, E. Jacobsen & R.G.F. Visser. Expression and inheritance of a wildtype GBSS-gene inserted into an amylose-free potato mutant by Agrobacterium tumefaciens. in press, TAG.
Gebhardt, C., E.Ritter, A.Barone, T.Debener, B.Walkemeier, U.Schachtschabel, H.Kaufmann, R.D.Thompson, M.W.Boierbale, M.W.Ganal, S.D.Tanksley & F.Salamini, 1991. RFLP maps of potato and their alignment with the homeologous tomato genome. Theor. Appl. Genet. 83: 49–57.
Gibson, R.W., M.G.K.Jones & N.Fish, 1988. Resistance to potato leaf roll virus and potato virus Y in somatic hybrids between dihaploid Solanum tuberosum and Solanum brevidens. Theor. Appl. Genet. 76: 113–117.
Hermsen, J.G.Th. & L.M.Taylor, 1979. Successful hybridisation of non-tuberous Solanum etuberosum Lind. and tuber bearing S. pinnatisectum Dun. Euphytica 28: 1–7.
Hovenkamp-Hermelink, J.H.M., E.Jacobsen, A.S.Ponstein, R.G.F.Visser, G.H.Vos-Scheperkeuter, E.W.Bijmolt, J.N.deVries, B.Witholt & W.J.Feenstra, 1987. Isolation of an amylose-free starch mutant of the potato (Solanum tuberosum L.). Theor. Appl. Genet. 75: 217–221.
Hovenkamp-Hermelink, J.H.M., J.N.deVries, P.Adamse, E.Jacobsen, B.Witholt & W.J.Feenstra, 1988. Rapid estimation of the amylose/amylopectin ratio in small amounts of tuber and leaf tissue of the potato. Potato Res. 31: 241–246.
Jacobsen, E., J.H.M.Hovenkamp-Hermelink, H.T.Krijgsheld, H.Nijdam, L.P.Pijnacker, B.Witholt & W.J.Feenstra, 1989. Phenotypic and genotypic characterisation of an amylose-free starch mutant of the potato. Euphytica 44: 43–48.
Jacobsen, E., P.Reinhout, J.E.M.Bergervoet, J.deLooff, P.E.Abidin & M.S.Ramanna, 1992. Isolation and characterisation of potato-tomato somatic hybrids using an amylose-free potato mutant as a parental genotype. Theor. Appl. Genet. 85: 159–164.
Jefferson, R.A., T.A. Kavanagh & M.W. Bevan, 1987. GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO. J. 3901–3907.
Johnston, S.A. & R.E.HannemanJr, 1982. Manipulations of Endoserm Balance Number overcome crossing barriers between diploid Solanum species. Science 217: 446–448.
Jones, M.G.K., R.Dunckley, S.Steele, A.Karp, R.Gibson, N.Fish, J.Valkonen, T.Poutala & E.Pehu, 1990. Transfer of resistance to PLRV and PVY from S. brevidens to potato by somatic hybridisation: characterisation and field evaluation. In: H.J.J.Nijkmap et al. (Eds), Progress in Plant Cellular and Molecular Biology, pp. 286–292. Kluwer Academic Publishers Dordrecht, Boston, London.
Neal, C.E. & L.D.Topoleski, 1983. Effects of the basal medium on growth of immature embryos in vitro. J. Amer. Hort. Sci. 108: 434–438.
Murashige, T. & F.Skoog, 1962. A revised medium for rapid growth and bioessays with tobacco tissue cultures. Physiol. Plant 15: 473–497.
Neal, C.E. & L.D.Topoleski, 1985. Hormonal regulation of growth and development of tomato embryos in vitro. J. Amer. Hort. Sci. 110: 869–873.
Pehu, E., A.Karp, K.Moore, S.Steele, R.Dunkley & M.G.K.Jones, 1989. Molecular, cytogenetic and morphologogical characterisation of somatic hybrids of dihaploid Solanum tuberosum and diploid S. brevidens. Theor. Appl. Genet. 78: 696–704.
Pereira, A., M.Aarts, S.vanAgtmaal, W.J.Stiekema & E.Jacobsen, 1991. Waxy variegation in transgenic potato. Maydica 36: 323–327.
Preiszner, J., A.Feher, O.Veisz, J.Sutka & D.Dudits, 1991. Characterisation of morphological variation and cold resistance in interspecific somatic hybrids between potato (Solanum tuberosum L.) and S. brevidens Phil. Euphytica 57: 37–49.
Ramanna, M.S. & R.Prakken, 1967. Structure of and homology between pachytene and somatic metaphase chromosomes of the tomato. Genetics 38: 115–133.
Suurs, L.C.J.M., E.Jongedijk & M.M.C.Tan, 1989. Polyacrylamide gradient gel electrophoresis: a routine method for high resolution isozyme electrophoresis of Solanum and Lycopersicon species. Euphytica 40: 181–186.
Tempelaar, M.J. & M.G.K.Jones, 1985. Directed electrofusion between protoplasts with different responses in a mass fusion system. Plant Cell Reports 4: 92–95.
Uijtewaal, B.A., L.C.J.M.Suurs & E.Jacobsen, 1987. Protoplast fusion of monohaploid (2n=x=12) potato clones; identification of somatic hybrids using malate dehydrogenase as a biochemical marker. Plant Sci. 51: 277–284.
Williams, C.E., G.J.Hunt & J.P.Helgeson, 1990. Fertile somatic hybrids of Solanum species: RFLP analysis of a hybrid potato and its sexual progeny from crosses with potato. Theor. Appl. Genet. 80: 545–551.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Jacobsen, E., Malvar, R., Huigen, D.J. et al. Isolation and characterisation of somatic hybrids of diploid Solanum tuberosum and Solanum brevidens and the use of amylose-free starch mutation for detection of introgression. Euphytica 69, 191–201 (1993). https://doi.org/10.1007/BF00022365
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00022365