, Volume 101, Issue 1, pp 117–125 | Cite as

Inter-simple sequence repeats PCR for characterization of closely related grapevine germplasm

  • Santiago Moreno
  • Juan Pedro Martín
  • Jesús María Ortiz


ISSR-PCR was carried out on 12 accessions of the grapevine (Vitis vinifera L.) variety 'Garnacha' and one accession of a related variety. The analysis had two objectives: to determine if ISSR-PCR could detect polymorphisms within a variety unable to be revealed previously with RAPDs, and to quantify the intralaboratory reproducibility of the assay. Two electrophoretic systems and staining procedures were assessed. No intravarietal differences could be detected. However, the ISSR approach revealed highly reproducible polymorphisms among the two varieties tested. The reproducibility of the banding patterns ranged from 86 to 94 per cent, being maximum when polyacrylamide gel electrophoresis and silver staining procedures were followed and weak bands discarded.

clonal characterization reproducibility simple sequence repeats Vitis vinifera 


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  1. Bachmann, K., 1994. Molecular markers in plant ecology. New Phytol 126: 403-418.CrossRefGoogle Scholar
  2. Bassam, B. J., G. Caetano-Annol és & P. M. Gresshoff, 1991. Fast and sensitive silver staining of DNAin polyacrylamide gels. Anal Biochem 80: 81-84.Google Scholar
  3. Becker, J. & M. Heun, 1995. Mapping of digested and undigested random amplified microsatellite polymorphisms in barley. Genome 38: 991-998.PubMedGoogle Scholar
  4. Bourquin, J. C., A. Sonko, L. Otten & B. Walter, 1993. Restriction fragment length polymorphism and molecular taxonomy in Vitis viniferaL. Theor Appl Genet 87: 431-438.Google Scholar
  5. Bourquin, J. C., P. Tournier, L. Otten & B. Walter, 1992. Identification of sixteen grapevine rootstocks by RFLP and RFLP analysis of nuclear DNA extracted from the wood. Vitis 31: 157-162.Google Scholar
  6. Bowditch, B. M., D. G. Albright, J. G. K. Williams & M. J. Braun, 1993. Use of randomly amplified polymorphic DNA markers in comparative genome studies. Methods Enzymol 224: 294-309.PubMedGoogle Scholar
  7. Bowers, J. E., E. B. Bandman & C. P. Meredith, 1993. DNAfingerprint characterization of some wine grape cultivars. Am J Enol Viticult 44: 266-274.Google Scholar
  8. Bowers, J. E., G. S. Dangi, R. Vignani & C. P. Meredith, 1996. Isolation and characterization of new polymorphic simple sequence repeat loci in grape (Vitis viniferaL.). Genome 39: 628-633.PubMedGoogle Scholar
  9. Caetano-Anoll és, G., B. J. Bassam & P. M. Gresshoff, 1992. Primertemplate interactions during DNA amplifications fingerprinting with single arbitrary oligonucleotides. Mol Gen Genet 235: 157- 165.CrossRefGoogle Scholar
  10. Caló, A., A. Costacurta, S. Cancellier & R. Forti, 1990. Garnacha, Grenache, Cannonao, Tocai rosso, un unico vitigno. Vignevini 9: 45-48.Google Scholar
  11. Collins, G. G. & R. H. Symons, 1993. Polymorphisms in grapevine DNA detected by the RAPD PCR technique. Plant Mol Biol Rep 11: 105-112.Google Scholar
  12. Gogorcena, Y., S. Arulsekar, A. M. Dandekar & D. E. Parfitt, 1993. Molecular markers for grape characterization. Vitis 32: 183-185.Google Scholar
  13. Kantety, R. V., X. Zeng, J. L. Bennetzen & B. E. Zehr, 1995. Assessment of genetic diversity in dent and popcorn (Zea maysL.) inbred lines using intersimple sequence repeat (ISSR) amplification. Mol Breeding 1: 365-373.CrossRefGoogle Scholar
  14. Litt, M. & J. A. Luty, 1989. A hypervariable microsatellite revealed by in vitroamplification of a dinucleotide repeatwithin the cardiac muscle actin gene. Am J Human Genet 44: 397-401.Google Scholar
  15. Mulcahy, D. L., M. Cresti, S. Sansavini, G. C. Douglas, H. F. Linskens, G. Bergamini Mulcahy, R. Vignani & M. Pancaldi, 1993. The use of random amplified polymorphic DNAs to fingerprint apple genotypes. Sci Hort Amsterdam 54: 89-96.CrossRefGoogle Scholar
  16. Reynier, A., 1991. Manuel de Viticulture. Lavoisier, Paris.Google Scholar
  17. Royo, B., J. González, M. J. Laquidain & M. P. Larumbe, 1989. Caracterización mediante análisis isoenzimático de la vinífera' Garnacha' (Vitis viniferaL.). Investigación Agraria: Producción y Protección Vegetal 4: 343-354.Google Scholar
  18. Salimath, S. S., A. C. D. Oliveira, I. D. Godwin & J. L. Bennetzen, 1995. Assessment of genome origins and genetic diversity in the genus Eleusinewith DNA markers. Genome 38: 757-763.PubMedGoogle Scholar
  19. Sensi, E., R. Vignani, W. Rohde & S. Biricolti, 1996. Characterization of genetic biodiversity with Vitis viniferaL. Sangiovese and Colorino genotypes byAFLP and ISTRDNAmarker technology. Vitis 35: 183-188.Google Scholar
  20. Shah, F. H., O. Rashid, A. J. Simons & A. Dunsdon, 1994. The utility of RAPD markers for the determination of genetic variation in oil palm (Elaeis guineensis). Theor Appl Genet 89: 713-718.CrossRefGoogle Scholar
  21. Skroch, P. W. & J. Nienhuis, 1995. Qualitative and quantitative characterization of RAPD variation among snap bean (Phaseolus vulgaris) genotypes. Theor Appl Genet 91: 1078-1085.Google Scholar
  22. Stavrakakis, M. & M. Loukas, 1983. The between-and within-grape-cultivars genetic variation. Sci Hort Amsterdam 19: 321-334.CrossRefGoogle Scholar
  23. Thomas, M. R., S. Matsumoto, P. Cain & N. S. Scott, 1993. Repetitive DNA of grapevine: classes present and sequences suitable for cultivar identification. Theor Appl Genet 86: 173-180.Google Scholar
  24. Thomas, M. R. & N. S. Scott, 1993. Microsatellite repeats in grapevine reveal DNA polymorphisms when analysed as sequence-tagged sites (STSs). Theor Appl Genet 86: 985-990.Google Scholar
  25. Torres, A. M., N. F. Weeden & A. Martín, 1993. Linkage among isozyme, RFLP and RAPD markers in Vicia faba. Theor Appl Genet 85: 937-945.CrossRefGoogle Scholar
  26. Vos, P., R. Hogers, M. Bleeker, M. Reijans, T. van de Lee, M. Hornes, A. Frijters, J. Pot, J. Peleman, M. Kuiper & M. Zabeau, 1995.Google Scholar
  27. AFLP: a new technique for DNA fingerprinting. Nucl Acids Res 23: 4407-4414.Google Scholar
  28. Wang, Z., J. L. Weber, G. Zhong & S. D. Tanksley, 1994. Survey of plant short tandem DNA repeats. Theor Appl Genet 88: 1-6.Google Scholar
  29. Ward, E., 1995. Improved polymerase chain reaction (PCR) detection of Gaeumannomyces graminisincluding a safeguard against false negatives. Eur J Plant Pathol 101: 561-566.CrossRefGoogle Scholar
  30. Weber, J. L. & P. E. May, 1989. Abundant class of human DNA polymorphisms which can be typed using the polymerase chain reaction. Am J Human Genet 44: 388-396.Google Scholar
  31. Williams, J. G. K., A. R. Kubelik, K. J. Livak, J. A. Rafalski & S. V. Tingey, 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucl Acids Res 18: 6531- 6535.PubMedGoogle Scholar
  32. Williams, J. J. K., M. K. Hanafey, J. A. Rafalski & S. V. Tingey, 1993. Genetic analysis using random amplified polymorphic DNA markers. Methods Enzymol 218: 704-740.PubMedCrossRefGoogle Scholar
  33. Wolff, K.& J. PetersVanRijn, 1993. Rapid detection of genetic variability in chrysanthemum (Dendranthema grandifloraTzvelev) using random primers. Heredity 71: 335-341.PubMedGoogle Scholar
  34. Wolff, K., E. Zietkiewicz & H. Hofstra, 1995. Identification of Chrysanthemum cultivars and stability of DNA fingerprint patterns. Theor Appl Genet 91: 439-447.CrossRefGoogle Scholar
  35. Wu, K., R. Jones, L. Danneberger & P. A. Scolnik, 1994. Detection of microsatellite polymorphisms without cloning. Nucl Acids Res 22: 3257-3258.PubMedGoogle Scholar
  36. Xu, H., D. J. Wilson, S. Arulsekar & A. T. Bakalinsky, 1995. Sequence-specific polymerase chainreaction markers derived from randomly amplified polymorphic DNA markers for fingerprinting grape (Vitis) rootstocks. J Am Soc Hort Sci 120: 714-720.Google Scholar
  37. Zietkiewicz, E., A. Rafalski & D. Labuda, 1994. Genome fingerprinting by simple sequence repeat (SSR)anchored polymerase chain reaction amplification. Genomics 20: 176-183.PubMedCrossRefGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  • Santiago Moreno
    • 1
  • Juan Pedro Martín
    • 1
  • Jesús María Ortiz
    • 1
  1. 1.Dpto. Biología Vegetal, ETSI Agrónomos (U.P.M.)Ciudad UniversitariaMadridSpain

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