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 This accession may then be withdrawn from the same clonal origin as the other three.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.
Similar content being viewed by others
References
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: 97–103
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: 846–853
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: 111–117
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, London
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: 33–40
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: 265–275
Goulão L & Oliveira CM (2001) Molecular characterisation of cultivars of apple (Malus domestica Borkh.) using microsatellite (SSR and ISSR) markers. Euphytica 122: 81–89
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: 259–270
Greshoff PM & Doy CH (1972) Development and differentiation of haploid Lycopersicon esculentum (tomato). Planta 107: 161–170
Gupta PK & Rao JK (2002) Molecular markers in crop improvement: Present status and future needs in India. Plant Cell Tiss. Org. Cult. 70: 229–234
Karp A (2000) Molecular tools for detecting genetic diversity. Acta Hort. 530: 17–29
Larkin PJ & Scowcroft WR (1981) Somaclonal variation - a novel source of variability from cell cultures for plant improvement. Theor. Appl. Genet. 60: 197–214
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: 517–522
Nei M & Li WH (1979) Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc. Natl. Acad. Sci. USA 76: 5269–5273
Oliveira CM, Mota M, Monte-Corvo L, Goulão L & Silva DM (1999) Molecular typing of Pyrus based on RAPD markers. Sci. Hortic. 79: 163–174
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: 617–622
Pierik LRM (1987) In vitro Culture of Higher Plants. Martinus Nijhof Publishers, Dordrecht (p. 344)
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: 159–163
Ramanatha Rao V & Hodgkin T (2002) Genetic diversity and conservation and utilization of plant genetic resources. Plant Cell Tiss. Org. Cult. 68: 1–19
Rieseberg LH (1996) Homology among RAPD fragments in interspecific comparisons. Mol. Ecol. 5: 99–105
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: 375–381
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: 295–301
Seabra RC, Cotrim H, Ribeiro G & Pais MS (1996) First approach for molecular characterisation of Castanea sativa Mill. by RAPDs. Silva Lusitana 4: 251–253
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: 28–32
Tingey SV & Tufo JP (1993) Genetic analysis with random amplified polymorfic DNA markers. Plant Physiol. 101: 439–452
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: 6531–6535
Vieitez AM, Ballester A, Vieitez ML & Vieitez E (1983) In vitro plantlet regeneration of mature chestnut. J. Hort. Sci. 58: 457–463
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Carvalho, L.C., Goulão, L., Oliveira, C. et al. RAPD Assessment for Identification of Clonal Identity and Genetic Stability of in vitro Propagated Chestnut Hybrids. Plant Cell, Tissue and Organ Culture 77, 23–27 (2004). https://doi.org/10.1023/B:TICU.0000016482.54896.54
Issue Date:
DOI: https://doi.org/10.1023/B:TICU.0000016482.54896.54