Summary
We have used random amplified polymorphic DNA (RAPD) markers to study genetic variation in Alstroemeria. The first objective was to examine the discriminatory power of RAPD markers in different genotypes of Alstroemeria obtained by traditional breeding. All genotypes examined, including commercial Alstroemeria varieties, could be distinguished on the basis of their RAPD profiles. Progeny plants could be distinguished from their parents. A second objective of this study was to investigate whether RAPD markers can be used as a routine tool to detect mutant plants, as an alternative to glasshouse testing. To address this objective, we analysed Alstroemeria plants that carried phenotypically visible mutations that either were induced by irradiation using X-rays or were the result of somaclonal variation. In eight out of a total of 13 mutant Alstroemeria plants obtained after irradiation or tissue culture we detected no polymorphisms when compared to control plants that were considered to be non-mutated. Only in five of the mutant plants analysed we detected one to two polymorphisms. These results suggest that frequent genome rearrangements had not occurred in the mutant plants analysed. These results also demonstrate that the RAPD technique is an inappropriate tool for the rapid screening of Alstroemeria for induced variation. It that the RAPD technique is an inappropriate tool for the rapid screening of Alstroemeria for induced variation. It seems probable that this conclusion would be equally applicable in other plant genera in which induced variation has occurred. However, the RAPD technique is a simple and effective tool for genetic fingerprinting of Alstroemeria varieties, provided their differences are due to sexual propagation.
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References
Anderberg M.R., 1973. Cluster Analysis for Applications. Academic Press Inc. New York.
Bailey L.H. & E.Z. Bailey, 1976. Hortus Third: A Concise Dictionary of Plant Cultivars in the United States and Canada. Macmillan Publishing Co., New York.
Broerties C. & H. Verboom, 1974. Mutation breeding of Alstroemeria. Euphytica 23: 39–44.
Brown P.T.H., (1989) DNA methylation in plants and its role in tissue culture. Genome 31: 717–729.
Brown P.T.H., E. Göbel & H. Lörz, 1991. RFLP analysis of Zea mays callus cultures and their regenerated plants. Theor Appl Genet 81: 227–232.
Brown P.T.H., F.D. Lange, E. Kranz & H. Lörz, 1993. Analysis of single protoplasts and regenerated plants by PCR and RAPD technology. Mol Gen Genet 237: 311–317.
Caetano-Anollés G., B.J. Bassam & P.M. Gresshoff, 1991. DNA amplification fingerprinting using very short arbitrary oligonu-cleotide primers. Bio/Technology 9: 553–557.
Dallas J.F., 1988. Detection of DNA “fingerprints” of cultivated rice by hybridization with a human minisatellite probe. Proc Natl Acad Sci USA 85: 6831–6835.
Doyle J.I. & J.L. Doyle, 1990. Isolation of plant DNA from fresh tissue. Focus 12: 13–15.
Evans D.A., 1989. Somaclonal variation: genetic basis and breeding applications. Trends Genet 5: 46–50.
Falkenhagen E.R., 1985. Isozyme studies in provenance research of forest trees. Theor Appl Genet 69: 335–347.
Gonzalez-Benito M.E. & P.G. Alderson, 1992. Callus induction and plant regeneration in Alstroemeria. J Exp Bot 43: 205–211.
Heinze B. & J. Schmidt, 1994. Clonal fidelity of norway spruce somatic embryos analyzed by RAPD. In: J.A. Pardos, M.R. Ahuja & R.E. Rossello (Eds.), Biotechnology of Trees, Conf. Proc., 18–22 Oct. 1993, Valsain, Spain, pp. 143–148 INIA, Madrid.
Hu J. & C.F. Quiros, 1991. Identification of broccoli and cauliflower cultivars with RAPD markers. Plant Cell Rep 10: 505–511.
Isabel N., L. Tremblay, M. Michaud, F.M. Tremblay & J. Bousquet, 1993. RAPDs as an aid to evaluate the genetic integrity of somatic embryogenesis-derived populations of Picea mariana (Mill.) B.S.P. Theor Appl Genet 86: 81–87.
Keil M. & A.R. Griffin, 1994. Use of random amplified polymorphic DNA (RAPD) markers in the discrimination and verification of genotypes in Eucalyptus. Theor Appl Genet 89: 442–450.
Kuhns L.J. & T.A. Fretz, 1978. Distinguishing rose cultivars by polyacrylamide gel electrophoresis. I. Extraction and storage of protein and active enzymes from rose leaves. J Mol Biol 80: 575–599.
Larkin P.J. & W.R. Scowcroft, 1981. Somaclonal variation-a novel source of variability from cell cultures for plant improvement. Theor Appl Genet 60:, 197–214.
Levall M.W., K. Bengtsson, N.-O. Nilsson, A. Hjerdin & C. Halldén, 1994. Molecular characterization of UV-treated sugar beet somaclones using RFLP markers. Physiologia Plantarum 90: 216–220.
Lin W.C. & P.L. Monette, 1987. In vitro propagation of Alstroemeria ‘Alsaan’. Plant Cell, Tissue and Organ Culture 9: 29–35.
Loubaka L. & M. Keil, 1995. RAPD markers for use in discrimination and verification of genotypes in Eucalyptus-Training of a Congolese forester in RAPD technology. In: B.M. Potts, N.M.G. Borralho, J.B. Reid, R.N. Cromer, W.N. Tibbits & C.A. Raymond (Eds.), Eucalypt Plantations: Improving Fibre Yield and Quality, pp. 444–447, Proc. CRC-IUFRO Conf Hobart, 19–24 Feb., CRC fro Temperate Hardwood Forestry: Hobart, Australia.
Müller E., P.T.H. Brown, S. Hartke & H. Lörz, 1990. DNA variation in tissue-culture-derived rice plants. Theor Appl Genet 80: 673–679.
Nagamine T., G.A. Todd, K.P. McCann, H.J. Newbyry & B.V. Ford-Lloyd, 1989. Use of restriction fragment length polymorphism to fingerprint beets at the genotype and species levels. Theor Appl Genet 78: 847–851.
Nordenstam B., 1978. Pilgrim from Lima-the route of alstroemeria to Sweden. Fauna-Flora (Stockh) 73: 134–136.
Nybom H. (1991) Applications of DNA fingerprinting in plant breeding. In: T. Burke, G. Dolf, A.J. Jeffreys & R. Wolff (Eds.), DNA Fingerprinting Approaches and Applications, pp. 294–311. Birkhäuser Verlag, Basel, Switzerland.
Pierik R.L.M., A.van Voorst, G. Booy, C.A.M.van Acker, C.L.C. Lelivelt & J.C.de Wit, 1988. Vegetative propagation of Alstroemeria hybrids in vitro. Acta Horticult 226: 81–89.
Riedy M.F., W.J. HamiltonIII & C.F. Aquadro, 1992. Excess of non-parental bands in offspring from known primate pedigrees assayed using RAPD PCR. Nucl Acids Res 20: 918.
Rogstad S.H., J.C. PattonII & B.A. Schaal, 1988. M13 repeat probe detects DNA minisatellite-like sequences in gymnosperms and angiosperms. Proc Natl Acad Sci USA 85: 9176–9178.
Sabir A., H.J. Newbury, G. Todd, J. Catty & B.V. Ford-Lloyd, 1992. Determination of genetic stability using isozymes and RFLPs in beet plants regenerated in vitro. Theor Appl Genet 84: 113–117.
Sambrook, J., E.F., Fritsch & T., Maniatis, 1989. Molecular Cloning: A Laboratory Manual. 2nd ed. Cold-Spring Harbor, New York.
Santi F. & M. Lemoine, 1990. Genetic markers for Prunus avium L. 2. Clonal identifications and discrimination from P. cerasus and P. cerasus x P. avium. Ann Sci For 47: 219–227.
Shenoy V.B. & I.K. Vasil, 1992. Biochemical and molecular analysis of plants derived from embryogenic tissue cultures of napier grass (Pennisetum purpureum K. Schum). Theor Appl Genet 83: 947–955.
Staub J.E. & V. Meglic, 1993. Molecular genetic markers and their legal relevance for cultivar discrimination: a case study in cucumber. Hort Technology 3: 291–300.
Stephens J.L. & H.G. Hughes, 1993. Isozyme characterization in Alstroemeria species and cultivars. HortSci 28: 222.
Taylor P.W.J., J.R. Geijskes, H.-L. Ko, T.A. Fraser, R.J. Henry & R.G. Birch, 1995. Sensitivity of random amplified polymorphic DNA analysis to detect change in sugar cane during tissue culture. Theor Appl Genet 90: 1169–1173.
Tobolski J.J. & R.D. Kemery, 1992. Identification of red maple cultivars by isozyme analysis. HortSci 27: 169–171.
Torres A.M., T. Millán & J.I. Cubero, 1993. Identifying rose cultivars using random amplified polymorphic DNA markers. HortSci 28: 333–334.
Vallés M.P., Z.Y. Wang, P. Montavon, I. Potrykus & G. Spangenberg, 1993. Analysis of genetic stability of plants regenerated from suspension cultures and protoplasts of meadow fescue (Festuca pratensis Huds.). Plant Cell Rep 12: 101–106.
Vyskot B., J. Fajkus, P. Kuglik, B. Koukalova & V. Kuhrova, 1991. Genome modifications in protoplast-derived tobacco plants: phenotypic evaluation and RFLP analysis. Biologia Plantarum (Praha) 33: 455–460.
Weising K., F. Weigand, A.J. Driesel, G. Kahl, H. Zischler & J. Epplen, 1989. Polymorphic simple GATA/GACA repeats in plant gnomes. Nucl Acids Res 17: 10128.
Weising K., J. Ramser, D. Kaemmer, G. Kahl & J.T. Epplen, 1991. Oligonucleotide fingerprinting in plants and fungi. In: T. Burke, G. Dolf, A.J. Jeffreys & R. Wolff (Eds.), DNA Fingerprinting Approaches and Applications, pp. 312–329. Birkhäuser Verlag. Basel, Switzerland.
Williams J.G.K., A.R. Kubelik, K.J. Livak, J.A. Rafalski & S.V. Tingey, 1990. DNA polymophisms amplified by arbitrary primers are useful as genetic markers. Nucl Acids Res 18: 6531–6535.
Welsh J., C. Peterson & M. McClelland, 1991. Polymorphisms generated by arbitrarily primed PCR in the mouse: application to strain identification and genetic mapping. Nucl Acids Res. 19: 303–306.
Wolff K., E.D. Schoen & J. Peters-Van Rijn, 1993. Optimizing the generation of random amplified polymorphic DNAs in chrysanthemum. Theor Appl Genet 86: 1033–1037.
Zabeau, M. & P. Vos, 1993. Selective restriction fragment amplification: a general method for DNA fingerprinting. European Patent Application, No. 92402629.7.
Zietkiewicz E., A. Rafalski & D. Labuda, 1994. Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics 20: 176–183.
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Anastassopoulos, E., Keil, M. Assessment of natural and induced genetic variation in Alstroemeria using random amplified polymorphic DNA (RAPD) markers. Euphytica 90, 235–244 (1996). https://doi.org/10.1007/BF00023864
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DOI: https://doi.org/10.1007/BF00023864