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RAPD Fingerprint to Appraise the Genetic Fidelity of In Vitro Propagated Araucaria excelsa R. Br. var. glauca Plantlets

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Abstract

Randomly amplified polymorphic DNA (RAPD) was used as a tool to assess the genetic fidelity of in vitro propagated Araucaria excelsa R. Br. var. glauca with explants taken from orthotropic stem along with their related mother plants after treatment with kinetin, 2iP, BA (0.02–0.26 mg/l) and TDZ (0.001–1 mg/l) to produce axillary shoots. TDZ and kinetin induced more shoot and higher length per explant. Results showed a total of 1,676 fragments were generated with 12 RAPD primers in micropropagated plants and their donor mother plants. The number of loci ranged from 6 in OPB 12–18 in OPY 07 with a size ranging from 250 bp in OPH 19–3500 bp in OPH 11. Cluster analysis of RAPD data using UPGMA (unweighted pair group method with arithmetic average) revealed more than 92% genetic similarities between tissue cultured plants and their corresponding mother plant measured by the Jaccard’s similarity coefficient. Similarity matrix and PCoA (two dimensional principal coordinate analysis) resulted in the same affinity. Primers had shown 36% polymorphism. However, careful monitoring of tissue culture derived plants might be needed to determine that rooted shoots are adventitious in origin.

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References

  1. Burrows, G. E., Offord, C. A., Meagher, P. F., & Ashton, K. (2003). Axillary meristems and the development of epicormic buds in wollemi pine (Wollemia nobilis). Annals of Botany, 92, 835–844.

    Article  CAS  Google Scholar 

  2. Giri, C. C., Shyamkumar, B., & Anjaneyulu, C. (2004). Progress in tissue culture, genetic transformation and application of biotechnology to trees: An overview. Trees, 18, 115–132.

    Google Scholar 

  3. Burrows, G. E., Doley, D. D., Haines, R. J., & Nikles, D. G. (1988). In vitro propagation of Araucaria cunninghamii and other species of the araucariaceae via axillary meristems. Australian Journal of Botany, 36, 665–676.

    Article  Google Scholar 

  4. Traore, A., Xing, Z., Bonser, A., & Carlson, J. (2005). Optimizing a protocol for sterilization and in vitro establishment of vegetative bud from mature Douglas fir trees. HortScience, 40, 1464–1468.

    Google Scholar 

  5. Haines, R. J., & de Fossard, R. A. (1977). Propagation of Hoop pine (Araucaria cunninghamii AIT.). Acta Horticulturae, 78, 297–302.

    Google Scholar 

  6. Sehgal, L., Sehgal, O. P., & Khosla, P. K. (1989). Micropropagation of Araucaria columnaris Hook. Annals of Forest Science, 46, 158–160.

    Article  Google Scholar 

  7. Williams, J. G. K., Kubelik, A. R., Livak, K. J., Rafalski, J. A., & Tingey, S. V. (1990). DNA polymorphism amplified by arbitrary primers are useful as genetic marker. Nucleic Acids Research, 18, 6531–6565.

    Article  CAS  Google Scholar 

  8. Martin, G. B., Williams, J. G. K., & Tanskley, S. D. (1991). Rapid identification of markers linked to a Pseudomonas resistance gene in tomato by using random primers and near isogenic lines. Proceedings of the National Academy of Sciences of the United States of America, 88, 2230–2340.

    Google Scholar 

  9. Welsh, J., & McClelland, M. (1990). Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Research, 18, 7213–7218.

    Article  CAS  Google Scholar 

  10. Tulseiram, L. K., Glaubits, J. C., Kiss, G., & Carlson, J. E. (1992). Single tree genetic linkage mapping in conifers using haploid DNA from megagametophytes. Nature Biotechnology, 10, 686–690.

    Article  Google Scholar 

  11. Rieseberg, L. H. (1996). Homology among RAPD fragments in interspecific comparisons. Molecular Ecology, 5, 99–105.

    Article  CAS  Google Scholar 

  12. Olmos, S. E., Lavia, G., Di Renzo, M., Mroginski, L., & Echenique, V. (2002). Gentic analysis of variation in micropropagation plants of Melia Azedarach L. In Vitro Cellular and Developmental Biology Plant, 38, 617–622.

  13. Venkatachalam, L., Sreedhar, R. V., & Bhagyalakshmi, N. (2007). Micropropagation in banana using high levels of cytokinins does not involve any genetic changes as revealed by RAPD and ISSR markers. Plant Growth Regulation, 51, 193–205.

    Article  CAS  Google Scholar 

  14. Sen, S., Skaria, R., & Abdul Muneer, P. M. (2010). Genetic diversity analysis in Piper species (Piperaceae) using RAPD markers. Molecular Biotechnology, 46, 72–79.

    Article  CAS  Google Scholar 

  15. Piola, F., Rohr, R., & Heizmann, P. (1999). Rapid detection of genetic variation within and among in vitro propagated cedar (Cedrus libani Loudon) clones. Plant Science, 141, 159–163.

    Article  CAS  Google Scholar 

  16. Jayanthi, M., & Mandal, P. K. (2001). Plant regeneration through somatic embryogenesis and RAPD analysis of regenerated plants in Tylophora indica (Burm. F. Merrill.). In Vitro Cellular and Developmental Biology Plant, 37, 576–580.

  17. Mondal, T. K., & Chand, P. K. (2002). Detection of genetic variation among micropropagated tea Camellia sinensis (L.) by RAPD analysis. In Vitro Cellular and Developmental Biology Plant, 38, 296–299.

    Article  CAS  Google Scholar 

  18. Carvalho, L. C., Goulao, L., Oliveira, C., Goncalves, J. C., & Amancio, S. (2004). RAPD assessment for identification of clonal identity and genetic stability of in vitro propagated chesnut hybrids. Plant Cell, Tissue and Organ Culture, 77, 23–27.

    Article  CAS  Google Scholar 

  19. Abdi, Gh., Salehi, H., & Khosh-Khui, M. (2008). Nano silver: A novel nanomaterial for removal of bacterial contaminants in valerian (Valeriana officinalis L.) tissue culture. Acta Physiology Plantarum, 30, 709–714.

    Article  CAS  Google Scholar 

  20. Sarmast, M. K., Salehi, H., & Khosh-Khui, M. (2011). Nano silver treatment is effective in reducing bacterial contamination of Araucaria excelsa R. Br. var. glauca explants. Acta Biologica Hungarica, 62 (in press).

  21. Murashige, T., & Skoog, F. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15, 473–497.

    Article  CAS  Google Scholar 

  22. Stange, C., Prehn, D., & Arce-Johnson, P. (1998). Isolation of Pinus radiate genomic DNA suitable for RAPD analysis. Plant Molecular Biology Reporter, 16, 1–8.

    Article  Google Scholar 

  23. Rohlf, F. J. (1998). NTSYS-pc, numerical taxonomy and multivariate analysis, version 2.02. New York: Exeter software.

    Google Scholar 

  24. Jaccard, P. (1908). Nouvelles recherches surla distribution florale. Bulletin de la Societe Vaudoise des Sciences Naturelles, 44, 223–270.

    Google Scholar 

  25. Oliviera, E. J., Padua, J. G., Zucchi, M. I., & Venkovsky, R. (2006). Origin, evolution and genome distribution of microsatellites. Biology, 29, 294–307.

    Google Scholar 

  26. Pollefeys, P., & Bousquet, J. (2003). Molecular genetic diversity of the French-American grapevine hybrids cultivated in North America. Genome, 46, 1037–1048.

    Article  CAS  Google Scholar 

  27. Larkin, P. J., & Scowcroft, W. R. (1981). Somaclonal variation–a noval source of variability from cell culture for plant improvement. Theoretical and Applied Genetics, 60, 197–214.

    Article  Google Scholar 

  28. Smith, M. K. (1988). A review of factor influencing the genetic stability of micropropagated banana. Fruits, 43, 219–233.

    Google Scholar 

  29. Haisel, D., Hofman, P., Vágner, M., Lipavská, H., Tichá, I., Schäfer, C., & Čapková, V. (2001). Ex vitro phenotype stability if affected by in vitro cultivation. Biologia Plantarum, 44, 321–324.

    Article  CAS  Google Scholar 

  30. Schestibratov, K. A., Mikhailov, R. V., & Dolgov, S. V. (2003). Plantlet regeneration from subculturable nodular of Pinus radiate. Plant Cell, Tissue and Organ Culture, 72, 139–146.

    Article  CAS  Google Scholar 

  31. Bonga, J. M., Klimaszewska, K. K., & von Aderkas, P. (2010). Recalcitrance in clonal propagation, in particular of conifers. Plant Cell, Tissue and Organ Culture, 100, 241–254.

    Article  Google Scholar 

  32. Thorpe, T. A. (2007). History of plant tissue culture. Molecular Biotechnology, 37, 169–180.

    Article  CAS  Google Scholar 

  33. Hartmann, H. T., Kester, D. E., Davies, F. T., & Geneve, R. L. (2002). Plant propagation, principles and practices (p. 880). Upper Saddle River, NJ: Pearson Education, Inc.

    Google Scholar 

  34. Sarmast, M. K., Salehi, H., & Khosh-Khui, M. (2009). Using plagiotropic shoot explants in tissue culture of Araucaria excelsa R Br. var. glauca. Advances in Environmental Biology, 3, 191–194.

    CAS  Google Scholar 

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Acknowledgments

The authors thank two anonymous reviewers for their insightful comments and suggestions.

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Authors and Affiliations

  1. Department of Horticultural Science, College of Agriculture, Shiraz University, Shiraz, Iran

    Mostafa Khoshhal Sarmast, Hassan Salehi & Morteza Khosh-Khui

  2. Institute of Biotechnology, Shiraz University, Shiraz, Iran

    Amin Ramezani, Ali Asghar Abolimoghadam & Ali Niazi

Authors
  1. Mostafa Khoshhal Sarmast
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  2. Hassan Salehi
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  3. Amin Ramezani
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  4. Ali Asghar Abolimoghadam
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  5. Ali Niazi
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  6. Morteza Khosh-Khui
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Correspondence to Hassan Salehi.

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Sarmast, M.K., Salehi, H., Ramezani, A. et al. RAPD Fingerprint to Appraise the Genetic Fidelity of In Vitro Propagated Araucaria excelsa R. Br. var. glauca Plantlets. Mol Biotechnol 50, 181–188 (2012). https://doi.org/10.1007/s12033-011-9421-7

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