Advertisement

Plant Cell, Tissue and Organ Culture

, Volume 77, Issue 1, pp 23–27 | Cite as

RAPD Assessment for Identification of Clonal Identity and Genetic Stability of in vitro Propagated Chestnut Hybrids

  • Luísa C. Carvalho
  • Luíis Goulão
  • Cristina Oliveira
  • José Carlos Gonçalves
  • Sara Amâncio
Article

Abstract

Randomly amplified polymorphic DNA (RAPD) was used as a tool to assess the clonal identity of four in vitro propagated chestnut rootstock hybrids (Castanea sativa × C. crenata) described as originally isolated from the same mother tree. To confirm genetic stability after in vitro multiplication for more than 4 years, RAPD patterns of in vitro and donor plants were compared. From 40 arbitrary 10-mer primers used to amplify DNA, 21 provided patterns and were chosen for comparisons. Although significant differences were found in growth parameters between in vitro material of the putative clones, RAPD profiling showed polymorphism in none but one. This accession may then be withdrawn from the same clonal origin as the other three. As expected, no polymorphism was detected between the material propagated in vitro and the donor plants they originated from.

Castanea sativa × C. crenata chestnut clones genetic stability in vitro propagation RAPD 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Bao PH, Granata S, Castiglione S, Wang G, Giordani C, Cuzzoni E, Damiani G, Bandi C, Datta SK, Datta K, Potrykus I, Callegarin A & Sala F (1996) Evidence for genomic changes in transgenic rice (Oryza sativa L.) recovered from protoplasts. Transgenic Res. 5: 97–103CrossRefPubMedGoogle Scholar
  2. Bogani P, Simoni A, Liò P, Scialpi A & Buiatti M (1996) Genome flux in tomato cell clones cultured in vitro in different physiological equilibria. II A RAPD analysis of variability. Genome 39: 846–853Google Scholar
  3. Bouman H & De Klerk G-J (2001) Measurement of the extent of somaclonal variation in begonia plants regenerated undervarious conditions. Comparison of three assays. Theor. Appl. Genet. 102: 111–117CrossRefGoogle Scholar
  4. Buiatti M & Bogani P (1998) Exploiting genome plasticity for the detection of hypervariable sequences. In: Karp A, Isaac P & Ingram D (eds) Molecular Tools for Screening Biodiversity (pp. 471–484). Chapman & Hall, LondonGoogle Scholar
  5. Carvalho L & Amâncio S (2002) Antioxidant defence system in plantlets transferred from in vitro to ex vitro: Effects of increasing light intensity and CO2 concentration. Plant Sci. 162: 33–40CrossRefGoogle Scholar
  6. Gonçalves JC, Diogo G & Amâncio S (1998) In vitro propagation of chestnut (Castanea sativa × C. crenata): Effects of rooting treatments on plant survival and anatomical changes during adventitious root formation. Sci. Hort. 72: 265–275Google Scholar
  7. Goulão L & Oliveira CM (2001) Molecular characterisation of cultivars of apple (Malus domestica Borkh.) using microsatellite (SSR and ISSR) markers. Euphytica 122: 81–89CrossRefGoogle Scholar
  8. Goulão L, Cabrita L, Oliveira CM & Leitão JM (2001) Comparing RAPD and AFLPTM analysis in discrimination and estimation of genetic similarities among apple (Malus domestica Borkh.) cultivars. Euphytica 119: 259–270CrossRefGoogle Scholar
  9. Greshoff PM & Doy CH (1972) Development and differentiation of haploid Lycopersicon esculentum (tomato). Planta 107: 161–170Google Scholar
  10. Gupta PK & Rao JK (2002) Molecular markers in crop improvement: Present status and future needs in India. Plant Cell Tiss. Org. Cult. 70: 229–234CrossRefGoogle Scholar
  11. Karp A (2000) Molecular tools for detecting genetic diversity. Acta Hort. 530: 17–29Google Scholar
  12. Larkin PJ & Scowcroft WR (1981) Somaclonal variation - a novel source of variability from cell cultures for plant improvement. Theor. Appl. Genet. 60: 197–214Google Scholar
  13. Monte-Corvo L, Goulão L & Oliveira CM (2001) ISSR analysis of cultivars of pear and suitability of molecular markers for clone discrimination. J. Am. Soc. Hort. Sci. 126: 517–522Google Scholar
  14. Nei M & Li WH (1979) Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc. Natl. Acad. Sci. USA 76: 5269–5273PubMedGoogle Scholar
  15. Oliveira CM, Mota M, Monte-Corvo L, Goulão L & Silva DM (1999) Molecular typing of Pyrus based on RAPD markers. Sci. Hortic. 79: 163–174CrossRefGoogle Scholar
  16. Olmos SE, Lavia G, Di Renzo M, Mroginski L & Echenique V (2002) Genetic analysis of variation in micropropagated plants of Melia Azedarach L. In vitro Cell. Dev. Biol. Plant 38: 617–622Google Scholar
  17. Pierik LRM (1987) In vitro Culture of Higher Plants. Martinus Nijhof Publishers, Dordrecht (p. 344)Google Scholar
  18. Piola F, Rohr R & Heizmann P (1999) Rapid detection of genetic variation within and among in vitro propagated cedar (Cedrus libani Loudon) clones. Plant Sci. 141: 159–163CrossRefGoogle Scholar
  19. Ramanatha Rao V & Hodgkin T (2002) Genetic diversity and conservation and utilization of plant genetic resources. Plant Cell Tiss. Org. Cult. 68: 1–19CrossRefGoogle Scholar
  20. Rieseberg LH (1996) Homology among RAPD fragments in interspecific comparisons. Mol. Ecol. 5: 99–105Google Scholar
  21. Rus-Kortekaas W, Smulders MJM, Arens P & Vosman B (1994) Direct comparison of levels of genetic variation in tomato detected by a GACA-containing microsatellite probe and by random amplified polymorphic DNA. Genome 37: 375–381Google Scholar
  22. Santana C, Oliveira CM & Valdiviesso T (1999) Molecular typing of rootstocks hybrids (Castanea sativa × Castanea crenata) and Portuguese cultivars based on RAPD markers. Acta Hortic. 494: 295–301Google Scholar
  23. Seabra RC, Cotrim H, Ribeiro G & Pais MS (1996) First approach for molecular characterisation of Castanea sativa Mill. by RAPDs. Silva Lusitana 4: 251–253Google Scholar
  24. Soneji JR, Rao PS & Mhatre M (2002) Somaclonal variation in micropropagated dormant axillary buds of pineapple (Ananas comosus L., Merr.) J. Hort. Sci. Biotech. 77: 28–32Google Scholar
  25. Tingey SV & Tufo JP (1993) Genetic analysis with random amplified polymorfic DNA markers. Plant Physiol. 101: 439–452CrossRefGoogle Scholar
  26. Williams JGK, Kubelik AR, Livark K, Rafalski A & Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 18: 6531–6535PubMedGoogle Scholar
  27. Vieitez AM, Ballester A, Vieitez ML & Vieitez E (1983) In vitro plantlet regeneration of mature chestnut. J. Hort. Sci. 58: 457–463Google Scholar

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Luísa C. Carvalho
    • 1
  • Luíis Goulão
    • 2
  • Cristina Oliveira
    • 2
  • José Carlos Gonçalves
    • 3
  • Sara Amâncio
    • 1
  1. 1.DBEB/CBAA, Instituto Superior de Agronomia, UTLLisboaPortugal
  2. 2.DPAA, Instituto Superior de Agronomia, UTLLisboaPortugal
  3. 3.Lab. Biologia Vegetal, Escola Superior Agrária, IPCBCastelo BrancoPortugal

Personalised recommendations