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Selection of monoploids for protoplast fusion and generation of intermonoploid somatic hybrids of potato

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Abstract

The union of potato monoploid genotypes (2n=1x =12) through protoplast fusion may result in vigorous somatic hybrids due to a reduction of the “genetic load” normally present in this highly heterozygous tetraploid (2n=4x=48) crop. More than 100 androgenic monoploids derived from diploid (2n=2x=24)Solanum phureja Juz. & Buk. andS. chacoense Bitt. xS. phureja clones were evaluated in field trials during 1996 and 1997 to identify the most promising genotypes for protoplast fusion experiments. Compared to the total population, the 1996 selected genotypes had higher means for tuber number (30.1 vs 11.2 tubers/plant), average tuber weight (3.0 vs 1.8 g/tuber) and total yield (66.1 vs 20.4 g/plant). Similarly, the 1997 selected genotypes had higher means for tuber number (42.8 vs 25.4 tubers/plant), average tuber weight (3.6 vs 2.5 g/tuber) and total yield (114.0 vs 63.4 g/plant) compared to the total population. The 31 selected monoploid genotypes from 1996-97 varied in their response to protoplast isolation and culture from no growth (9), cell enlargement (5), limited cellular divisions (8), callus formation (5) to plant regeneration from callus (4). Chemical fusion and electrofusion produced three groups of intermonoploid somatic hybrids. Polymorphic simple sequence repeat (SSR) loci enabled distinction of somatic hybrids from parental somaclones. Rapid DNA extraction with SSR analysis enabled screening of calluses to identify somatic hybrid tissue prior to plant regeneration. The somatic hybrids were highly polyploid, mostly hexaploid (2n=6x=72), possibly due to fusion of endopolyploid protoplasts and/or chromosomal doubling during plant regeneration.

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Literature Cited

  • Austin, S. and A.C. Cassells. 1983. Variation between plants regenerated from individual calli produced from separated potato stem callus cells. Plant Sci Letters 31:107–114.

    Article  Google Scholar 

  • Austin, S., M.K Ehlenfeldt, M.A Baer, and J.P. Helgeson. 1986. Somatic hybrids produced by protoplast fusion betweenSolanum tuberosum andSolanum brevidens: phenotypic variation under field conditions. Theor Appl Genet 71:682–690.

    Article  Google Scholar 

  • Baird, E., S. Cooper-Bland, R. Waugh, M. DeMaine, and W. Powell. 1992. Molecular characterisation of inter- and intra-specific somatic hybrids of potato using randomly amplified polymorphic DNA (RAPD) markers. Mol Gen Genet 233:469–475.

    Article  PubMed  CAS  Google Scholar 

  • Bhojwani, S.S. and M.K. Razdan. 1983. Cytogenetic studies.In: Plant Tissue Culture: Theory and Practice. Elsevier Science Publishers B.V., Amsterdam, pp. 159–180.

    Google Scholar 

  • Chaput, M.H., D. Sihachakr, G. Ducreux, D. Marie, and N. Barghi. 1990. Somatic hybrid plants produced by electrofusion between dihaploid potatoes: BF15 (H1), Aminca (H6) and Cardinal (H3). Plant Cell Rep 9:411–414.

    Article  Google Scholar 

  • Cheng, J. and R.E. Veilleux. 1991. Genetic analysis of protoplast culturability inSolanum, phureja. Plant Sci 75:257–265.

    Article  Google Scholar 

  • Cooper-Bland, S., MJ.De Maine, M.L.M.H. Fleming, M.S. Phillips, W. Powell, and A. Kumar. 1994. Synthesis of intraspecific somatic hybridsof Solanum tuberosum: assessments of morphological, biochemical and nematode(Globodera pallida) resistance characteristics. J Exp Bot 45:1319–1325.

    Article  CAS  Google Scholar 

  • Doyle, J.J. and J.L. Doyle. 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem. Bull. 19. Phytochemical Section of the Botanical Society of America, Irvine, CA pp. 11–15.

    Google Scholar 

  • Fish, N., A. Karp, and M.G.K Jones. 1988. Production of somatic hybrids by electrofusion inSolanum. Theor Appl Genet 76:260–266.

    Google Scholar 

  • Gilissen, L.J.W., M.J.van Staveren, H.A. Verhoeven, and K.S. Ramulu. 1992. Somatic hybridization between potato andNicotiana plumbaginifolia. 1. Spontaneous biparental chromosome elimination and production of asymmetric hybrids. Theor Appl Genet 84:73–80.

    Article  Google Scholar 

  • Haberlach, G.T., B.A. Cohen, N.A Reichert, MA Baer, L.E. Towill, and J.P. Helgeson. 1985. Isolation, culture and regeneration of protoplasts from potato and several relatedSolanum species. Plant Sci 39:67–74.

    Article  Google Scholar 

  • Matthey, W.M., R. Eijlander, J.R.Ade Koning, and KM. Louwes. 1992. Interspecific hybridization between the cultivated potatoSolanum tuberosum subspeciestuberosum L. and the wild speciesS. circaeifolium Bitter exhibiting resistance toPhytophthora infestans (Mont.) de Bary andGlobodera pallida (Stone) Behrens. 1. Somatic hybrids. Theor Appl Genet 83:459–466.

    Google Scholar 

  • Matthey, W.M. and K.J. Puite. 1992. Tetraploid potato hybrids through protoplast fusions and analysis of their performance in the field. Theor Appl Genet 83:807–812.

    Google Scholar 

  • M’Ribu, H.K and R.E. Veilleux. 1991. Phenotypic variation and correla tions between monoploids and doubled monoploids ofSolarium phureja. Euphytica 54:279–284.

    Google Scholar 

  • Murashige, T. and F. Skoog. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473- 497.

    Article  CAS  Google Scholar 

  • Möllers, C. and G. Wenzel. 1992. Somatic hybridization of dihaploid potato protoplasts as a tool for potato breeding.Bot Acta 105:133–139.

    Google Scholar 

  • Novy, R.G. and J.P. Helgeson. 1994. Somatic hybrids betweenSolarium etuberosum and diploid, tuber bearingSolarium clones. Theor Appl Genet 89:775–782.

    Google Scholar 

  • Ochatt, S.J., P.K Chand, E.L. Rech, M.R. Davey, and J.B. Power. 1988. Electroporation-mediated improvement of plant regeneration from colt cherryPrunus avium Pseudocerasus) protoplasts. Plant Sei 54:165–169.

    Article  Google Scholar 

  • Owen, H.R., R.E. Veilleux, D. Levy, and D.L. Ochs. 1988. Environmental, genotypic and ploidy effects on endopolyploidization within a genotype ofSolarium phureja and its derivatives. Genome 30:506–510.

    Google Scholar 

  • Paz, M.M. and R.E. Veilleux. 1997. Genetic diversity based on randomly amplified polymorphic DNA (RAPD) and its relationship with the performance of diploid potato hybrids. J Amer Soc Hort Sei 122:740–747.

    CAS  Google Scholar 

  • Pynacker, L.P., K Sree Ramulu, P. Dykhuis, and M.A Ferwerda. 1989. Flow cytometric and karyological analysis of polysomaty and polyploidization during callus formation from leaf segments of various potato genotypes. Theor Appl Genet 77:102–110.

    Article  Google Scholar 

  • Provan, J., W. Powell, and R. Waugh. 1996. Microsatellite analysis of rela tionships within cultivated potato(Solarium tuberosum). Theor Appl Genet 86:975–984.

    Google Scholar 

  • Rech, EX., S.J. Ochatt, P.K Chand, J.B. Power, and M.R. Davey. 1987. Electro-enhancement of division of plant protoplast-derived cells. Protoplasma 141:169–176.

    Article  Google Scholar 

  • Sambrook, J., E.F. Fritsch, and T. Maniatis. 1989. Molecular Cloning: a Laboratory Manual. 2nd ed. Cold Spring Laboratory Press, Cold Spring Harbor, NY, pp. 6.3–6.34.

    Google Scholar 

  • SAS Institute. 1985. SAS User’s guide: statistics. Version 5 éd. SAS Insti tute Inc, Cary, N.C.

    Google Scholar 

  • Sherraf, I., S. Tizroutine, M.H. Chaput, M. Allot, I. Mussio, D. Sihachakr, L Rossignol, and G. Ducreux. 1994. Production and characteriza tion of intergeneric somatic hybrids through protoplast electro fusion between potato(Solarium tuberosum) andLycopersicon penneUii.Plant Cell Tiss Org Cult 37:137–144.

    Article  CAS  Google Scholar 

  • Tan, M.M.C., E.M. Rietveld, AM. van Marrewyk Gysbert, and Ad J. Kool. 1987. Regeneration of leaf mesophyll protoplasts of tomato cul tivars(L. esculentum): factors important for efficient protoplast culture and plant regeneration. Plant Cell Rep 6:172–175.

    Article  Google Scholar 

  • Uytewaal, B.A, E. Posthumus, L.C.J.M. Suurs, E. Jacobsen, and J.G.Th. Hermsen. 1987a Relative performance of monohaploid potato clones and their diploid parents at plant level and after protoplast isolation and subsequent fusion. Theor Appl Genet 74:451458.

    Google Scholar 

  • Uytewaal, B.A, E. Jacobsen, and J.G.Th. Hermsen. 1987b.Morphology and vigour of monohaploid potato clones, their corresponding homozygous diploids and tetraploids and their heterozygous diploid parent. Euphytica 36:745–753.

    Article  Google Scholar 

  • Varrieur, J.M., R.J. Cutright and R.E. Veilleux. 1999. Selection pressures inherent in the monoploid-derivation mechanisms, androgenesis and gynogenesis. Am J Potato Res 76:386 (Abstr).

    Google Scholar 

  • Veilleux, R.E. and M.E. Compton. 1996. Isolation, culture and fusion of tobacco and potato protoplasts.In: Trigiano, R.N. and D.J. Gray (eds), Plant Tissue Culture Concepts and Laboratory Exercises. CRC Press, Boca Raton, FL. pp. 215–225.

    Google Scholar 

  • Veilleux, R.E., L.Y. Shen, and M.M. Paz. 1995. Analysis of the genetic composition of anther-derived potato by randomly amplified polymorphic DNA and simple sequence repeats. Genome 38:1153–1162.

    PubMed  CAS  Google Scholar 

  • Wang, H., M. Qi, and A.J. Cutler. 1993. A simple method of preparing plant samples for PCR. Nucleic Acids Res 21:4153–4154.

    Article  PubMed  CAS  Google Scholar 

  • Ward, A.C., J.St-J. Phelpstead, A.E. Gleadle, N.W. Blackhall, S. Cooper-Bland, A. Kumar, W. Powell, J.B. Power, and M.R. Davey. 1994. Interspecific somatic hybrids between dihaploidSolanum tuberosum L. and the wild species,S. pinnatisectum Dun. J Exp Bot 45:1433–1440.

    Article  CAS  Google Scholar 

  • Yu, Y.G., M.A. Saghai Maroof, G.R. Buss, P.J. Maughan, and S.A. Tolin. 1994. RFLP and microsatellite mapping of a gene for soybean mosaic virus resistance. Phytopathology 4:60–64.

    Article  Google Scholar 

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Correspondence to Richard E. Veilleux.

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Johnson, A.A.T., Piovano, S.M., Ravichandran, V. et al. Selection of monoploids for protoplast fusion and generation of intermonoploid somatic hybrids of potato. Am. J. Pot Res 78, 19–29 (2001). https://doi.org/10.1007/BF02874821

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