Skip to main content

Assessment of genetic fidelity of micropropagated date palm (Phoenix dactylifera L.) plants by RAPD and ISSR markers assay

Abstract

RAPD (Random Amplified Polymorphic DNA) and ISSR (Inter-Simple Sequence Repeats) markers assay were employed to validate the genetic stability of date palm (Phoenix dactylifera L.) plants multiplied through somatic embryogenesis with upto forty two in vitro subcultures. Out of the 160 RAPD and 21 ISSR primers screened, 30 RAPD and 12 ISSR primers produced a total of 347 (246 RAPDs + 101 ISSRs) clear, distinct and reproducible amplicons, which were monomorphic across all micropropagated plants (27) studied. Thus, a total 8592 bands (number of plants analysed x number of amplicons with all the primers) were generated which exhibited homogeneous banding patterns with both RAPD and ISSR markers. These results indicate that the micropropagation protocol developed by us for rapid in vitro multiplication is appropriate and suitable for clonal propagation of date palm and corroborated the fact that somatic embryogenesis can also be used as one of the safest modes for production of true-to-type plants.

This is a preview of subscription content, access via your institution.

References

  1. Ahmed O, Chokri B, Noureddine D, Mohamed M and Mokhtar T (2009). Regeneration and molecular analysis of date palm (Phoenix dactylifera L.) plantlets using RAPD markers. African J.Biotech. 8: 813–820

    CAS  Google Scholar 

  2. Andrea B, Maria A and Giovanni V (2004). Genetic stability and uniformity of Foeniculum vulgare Mill. regenerated plants through organogenesis and somatic embryogenesis. Plant sci. 166: 221–227

    Article  Google Scholar 

  3. Bekheet SA, Saker MM, Taha HS and Moursy HA (2001). Plant regeneration via somatic embryogenesis in date palm (Phoenix dactylifera L.). Arab J. Biotechnol. 4: 111–118

    Google Scholar 

  4. Dass HC, Kaul RK, Joshi SP and Bhansali RR (1989). In vitro regeneration of date palm plantlets. Curr. Sci. 58: 22–24

    Google Scholar 

  5. Devarumath R, Nandy S, Rani V, Marimuthu S, Muraleedharan N and Raina S (2002). RAPD, ISSR and RFLP fingerprints as useful markers to evaluate genetic integrity of micropropagated plants of three diploid and triploid elite tea clones representing Camellia sinensis (China type) and C. assamica ssp. assamica (Assam-India type). Plant Cell Rep. 21: 166–173

    CAS  Article  Google Scholar 

  6. Doyle JJ and Doyle JL (1990). Isolation of plant DNA from fresh tissue. Focus. 12: 13–15

    Google Scholar 

  7. Eke CR, Akomeah P and Asemota O (2005). Somatic embryogenesis in date palm (Phoenix dactylifera L.) from apical meristem tissues from ‘zebia’and ‘loko’ landraces. Afr. J. Biotechnol. 4: 244–246

    Google Scholar 

  8. Goto S, Thakur RC and Ishii K (1998). Determination of genetic stability in long-term micropropagated shoots of Pinus thunbergii Parl. using RAPD markers. Plant Cell Rep. 18: 193–197

    CAS  Article  Google Scholar 

  9. Hassan S, Bakhsh K, Gill ZA., Maqbool A and Ahmad W (2006). Economics of growing date palm in Punjab, Pakistan. Int. J. Agri. Biol. 8: 788–792

    Google Scholar 

  10. Hirochika H (1993). Activation of tobacco retrotransposons during tissue culture. EMBO J. 12: 2521–2528

    CAS  PubMed  Google Scholar 

  11. Huang WJ., Ning GG, Liu GF and Bao MZ (2009). Determination of genetic stability of long-term micropropagated plantlets of Platanus acerifolia using ISSR markers. Biol. Plant. 53: 159–163

    Article  Google Scholar 

  12. Joshi P and Dhavan V (2007). Assessment of genetic fidelity of micropropagated Swertia chirayita plantlets by ISSR marker assay. Biol. Plant. 51: 22–26.

    CAS  Article  Google Scholar 

  13. Lakshman V, Venkataramareddy SR and Neelwarne B (2007). Molecular analysis of genetic stability in long-term micropropagated shoots of banana using RAPD and ISSR markers. Electronic J. of Biotechnol. 10: 1–8

    Google Scholar 

  14. Leroy XJ, Leon K, Charles G and Branchard M (2000). Cauliflower somatic embryogenesis and analysis of regenerant stability by ISSRs. Plant Cell Rep. 19: 1102–1107

    CAS  Article  Google Scholar 

  15. Lopes T, Pinto G, Loureiro G, Costa A and Santos C (2006). Determination of genetic stability in cork oak long term somatic embryogenic cultures and derived plantlets using microsatellite markers. Tree Physiol. 26: 1145–1152

    CAS  PubMed  Google Scholar 

  16. Martins M, Sarmento D and Oliveira MM (2004). Genetic stability of micropropagated almond plantlets as assessed by RAPD and ISSR markers. Plant Cell Rep. 23: 492–496

    CAS  Article  PubMed  Google Scholar 

  17. Matthes M, Singh R, Cheah SC and Karp A (2001). Variation in oil palm (Elaeis guineensis Jacq.) tissue culturederived regenerants revealed by AFLPs with methylationsensitive enzymes. Theor. Appl. Genet. 102: 971–979

    CAS  Article  Google Scholar 

  18. Murashige T and Skoog F (1962). A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15: 473–497

    CAS  Article  Google Scholar 

  19. Phillips RL, Kaeppler SM and Olhoft P (1994). Genetic instability of plant tissue cultures: Breakdown of normal controls. Proc. Natl. Acad. Sci. USA. 91: 5222–5226

    CAS  Article  PubMed  Google Scholar 

  20. Rahman MH and Rajora OP (2001). Microsatellite DNA somaclonal variation in micropropagated trembling aspen (Populus tremuloides). Plant Cell Rep. 20: 531–536

    CAS  Article  Google Scholar 

  21. Reddy MP, Sarla N and Siddiq EA (2002). Inter simple sequence repeat (ISSR) polymorphism and its application in plant breeding. Euphytica. 128: 9–17

    Article  Google Scholar 

  22. Santos MDM, Buso GCS and Torres AC (2008). Evaluation of genetic variability in micropropagated propagules of ornamental pineapple [Ananas comosus var. bracteatus (Lindley) Coppens and Leal] using RAPD markers. Genet. Mol. Res. 7: 1097–1105

    CAS  Article  PubMed  Google Scholar 

  23. Sharma DR, Dawra S and Chowdhury JB (1984). Somatic embryogenesis and plant regeneration in Date palm (Phoenix dactylifera L.) cv Khadravi through tissue culture. Ind. J. Exp. Biol. 22: 596–598

    Google Scholar 

  24. Sharma SD, Ghosh SA, Mondal BB and Srivastava PS (2004). Somatic embryogenesis in a range of genotypes and genetic stability of the plants derived from somatic embryos using morphological and RAPD markers in Sweet potato. J. Biotech. 6: 119–124

    Google Scholar 

  25. Taha HS, Bakheet SA and EL-Bahr (2003). Alternative approach for micropropagation of the date palm c.v. Zaghlool. Arab J. of Biotechnol. 6: 103–112

    Google Scholar 

  26. Thakur R, Goto S, Ishi K and Jain SM (1999). Monitoring Genetic Stability in Quercus serata Thunb. Somatic Embryogenesis Using RAPD Markers. J. For. Res. 4: 157–160

    CAS  Article  Google Scholar 

  27. Tisserat B (1979). Propagation of date palm (Phoenix dactylifera L) in vitro. J. Exp. Bot. 30: 1275–1283

    CAS  Article  Google Scholar 

  28. Vasil IK (1987). Developing cell and tissue culture systems for improvement of cereal and grass crops. J. Plant Physiol. 128: 193–218.

    Google Scholar 

  29. Vendrame WA, Kochertm G and Wetzstein HY (1999). AFLP analysis of variation in pecan somatic embryos. Plant Cell Rep. 18: 853–857

    CAS  Article  Google Scholar 

  30. Venkatachalam L, Sreedha RV, 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 Reg. 51: 193–205

    CAS  Article  Google Scholar 

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to Nitish Kumar.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Kumar, N., Modi, A.R., Singh, A.S. et al. Assessment of genetic fidelity of micropropagated date palm (Phoenix dactylifera L.) plants by RAPD and ISSR markers assay. Physiol Mol Biol Plants 16, 207–213 (2010). https://doi.org/10.1007/s12298-010-0023-9

Download citation

Keywords

  • Date palm
  • genetic stability
  • ISSR
  • molecular marker
  • RAPD