Abstract
A system of randomly amplified polymorphic DNA (RAPD) markers was developed to facilitate the transfer of S. bulbocastanum (blb) genes into the S. tuberosum (tbr) genome by hybridization and backcrossing. DNA from tbr, blb and the hexaploid hybrid was used as a template for polymerase chain reaction (PCR) amplification. Polymorphic RAPD products, originating from 10-mer primers, specific for blb were cloned and sequenced at their ends to allow the synthesis of 18-mer primers. The 18-mer primers allowed a more reproducible assay than the corresponding RAPDs. Of eight 18-mer primer pairs, four amplified the expected products specific for blb. However, the stringency of the primer annealing conditions needed to be carefully optimized to avoid amplification of the homeologous tbr product, suggesting that the original RAPD polymorphisms were due to single base-pair changes rather than deletions or insertions. Two primers used for amplification of backcross 2 progeny segregated in a 1∶1 (presence:absence) ratio; the other two were unexpectedly absent. The most likely explanation for the loss of these markers is irregular meiosis in the original hexaploid hybrid and subsequent elimination of chromosomes. Cytological analysis of the meiosis in the hybrid demonstrated widespread irregular pairing and the presence of lagging univalents. In addition, the first backcross individual used as the parent for the second backcross had 54 chromosomes instead of the predicted 60. In conclusion, our results demonstrate that PCR technology can be used for the efficient isolation of taxon-specific markers in Solanum. Furthermore, by the use of these markers we detected the loss of chromosomes that was subsequently shown by cytological analysis to be caused by irregular meiosis of the somatic hybrid.
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Akkaya MS, Bhagwat AA, Cregan PB (1992) Length polymorphisms of simple sequence repeat DNA in soybean. Genetics 132:1131–1139
Austin S, Pohlman JD, Brown CR, Mojtahedi H, Santo GS, Douches DS, Helgeson JP (1993) Interspecific somatic hybridization between Solanum tuberosum L. and S. bulbocastanum Dun. as a means of transferring nematode resistance. Am Pot J 70:485–496
Barua UMM, Chalmers KJ, Hackett CA, Thomas WTB, Powell W, Waugh R (1993) Identification of RAPD markers linked to a Rhynchosporium secalis resistance locus in barley using near-isogenic lines and bulked segregant analysis. Heredity 71: 177–184
Benavente E, Orellana J (1991) Chromosome differentiation and pairing behaviour of polyploids:an assessment on preferential metaphase associations in colchicine-induced autotetraploid hybrids within the genus Secale. Genetics 128:433–442
Bonierbale MW, Plaisted RL, Tanksley SD (1988) RFLP maps based on a common set of clones reveal modes of chromosomal evolution in potato and tomato. Genetics 120:1095–1103
Coleman LC (1938) Preparation of leucobasic fuchsin for use in the Feulgen's reaction. Stain Technol 13:123–124
Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version II. Plant Mol Biol Rep 1:19–21
Dvorak J (1983) Evidence for genetic suppression of heterogenetic chromosome pairing in polyploid species of Solanum, sect. petota, Can J Genet Cytol 25:530–539
Ehlenfeldt MK, Helgeson JP (1987) Fertility of somatic hybrids from protoplast fusions of Solanum brevidens and S. tuberosum. Theor Appl Genet 73:395–402
Gebhardt C, Ritter E, Schachtschabel U, Walkemeier B, Uhrig H, Salamini F (1988) RFLP analysis and linkage mapping in Solanum tuberosum. Theor Appl Genet 78:65–75
Hawkes JG (1990) The potato-evolution, biodiversity and genetic resources. Smithsonian Institute Press, Washington, D.C.
Hermsen JG, Ramanna MS (1969) Meiosis in different F1-hybrids of Solanum acaule Bitt. x S. bulbocastanum Dun. and its bearing on genome relationship, fertility and breeding behaviour. Euphytica 18:27–35
Hermsen JG, Ramanna MS (1976) Barriers to hybridization of Solanum bulbocastanum Dun. and S. verrucosum Schlechtd. and structural hybridity in their F1 plants. Euphytica 25:1–10
Klein-Lankhorst RM, Vermunt AR, Liharaska T, Zabel P (1991) Isolation of molecular markers of tomato (L. esculentum) using random polymorphic DNA (RAPD). Theor Appl Genet 83:108–114
Matsubayashi M (1991) Phylogenetic relationships in the potato and its related species. In: Tsuchiya T, Gupa PK (eds) Chromosome engineering in plants: genetics, breeding, evolution, part B. Elsevier Science Publ, Amsterdam, pp 93–118
McGrath JM, Wielgus SM, Uchytil TF, Kim-Lee H, Haberlach GT, Williams CE, Helgeson JP (1994) Recombination of Solanum brevidens chromosomes in the second backcross generation from a somatic hybrid with S. tuberosum. Theor Appl Genet 88:917–924
Michelmore RW, Paran I, Kesseli RV (1991) Identification of markers linked to disease-resistance genes by bulked segregant analysis:a rapid method to detect markers in specific genomic regions by using segregating populations. Proc Natl Acad Sci USA 88:9828–9832
O'Brien S (1993) Genetic maps: locus maps of complex genomes vol 6, 6th edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.
Olson M, Hood L, Cantor C, Botstein D (1989) A common language for physical mapping of the human genome. Science 245:1434–1435
Paran I, Michelmore W (1993) Development of reliable PCR-based markers linked to downy mildew resistance genes in lettuce. Theor Appl Genet 85:985–993
Penner GA, Chong J, Wight CP, Molnar SJ, Fedak G (1993) Identification of a RAPD marker for the crown rust resistance gene Pc68 in oats. Genome 36:818–820
Sears ER (1976) Genetic control of chromosome pairing in wheat. Annu Rev Genet 10:31–51
Spooner DM, van der Ber K (1992) An analysis of recent taxonomic concepts in wild potatoes (Solanum sect. Petota). Genet Resources Crop Evol 39:23–38
Watanabe K (1994) Molecular genetics. In: Bradshaw JE, Mackay GR (eds) Potato genetics. CAB International Wallingford, UK, pp 213–235
Williams CE, Wielgus SM, Haberlach GT, Guenther C, Kim-Lee H, Helgeson JP (1993) RFLP analysis of chromosomal segregation in progeny from an interspecific hexaploid somatic hybrid between Solanum brevidens and Solanum tuberosum. Genetics 135:1167–1173
Williams JGK, Kubelik AR, Livak KJ, Rafalsky A, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18:6531–6535
Wu KK, Burnquist W, Sorrells ME, Tew TL, Moore PH, Tanksley SD (1992) The detection and estimation of linkage in polyploids using single-dose restriction fragments. Theor Appl Genet 83:294–300
Xu YS, Clark MS, Pehu E (1993) Use of RAPD markers to screen somatic hybrids between Solanum tuberosum and S. brevidens. Plant Cell Rep 12:107–109
Yu KF, Pauls KP (1993) Segregation of random amplified polymorphic DNA markers and strategies for molecular mapping in tetraploid alfalfa. Genome 36:844–851.
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Masuelli, R.W., Tanimoto, E.Y., Brown, C.R. et al. Irregular meiosis in a somatic hybrid between S. bulbocastanum and S. tuberosum detected by species-specific PCR markers and cytological analysis. Theoret. Appl. Genetics 91, 401–408 (1995). https://doi.org/10.1007/BF00222966
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DOI: https://doi.org/10.1007/BF00222966