Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Assessment of genetic purity of micropropagated plants of sugarcane by isozyme and RAPD analysis

  • 137 Accesses

  • 8 Citations

Abstract

Shoot tip expiants of nine sugarcane varieties(Saccharum species hybrid.), CoLk 8102, CoLk 8001, CoS 767, CoLk 9606, CoLk 9617, CoS 95255, BO 91, CoJ 64 and Co 1148, were established on a solid MS medium containing 0.8% agar, 3% sucrose and 0.05mg/l of IAA, BAP and kinetin. Shoot regeneration has been achieved in liquid MS medium containing 0.5mg/l each of IAA, BAP and kinetin. Multiple shoots were obtained on liquid MS medium containing 0.01mg/l IAA, 0.5mg/l each of 6- benzyl aminopurine (BAP), kinetin and rooting occurred in 1/2 strength liquid MS medium with 5.0 mg/l NAA and 0.01 mg/l each of BAP and kinetin. To detect the genetic purity of in vitro raised plantlets, expression of iso-peroxidases in donor plants andin vitro raised plantlets was compared through native PAGE. Different genotypes showed marked variation regarding number, position and intensity of bands. Micropropagated plantlets showed identical pattern of peroxidases to their parental clones. The genetic stability of micro propagated plantlets was also tested by RAPD analysis. The banding pattern of PCR amplified products from micro propagated plantlets was monomorphic based on RAPD profile in all the varieties. RAPD analysis and peroxidase isozyme pattern confirmed the genetic purity of sugarcane plantlets derivedin vitro.

This is a preview of subscription content, log in to check access.

References

  1. Anita, P., Jain, R.K., Sehrawat, A.R. and Punia, A. (2000). Efficient and cost- effective micropropagation of two early maturing varieties of sugarcane (Saccharum spp.)Indian Sugar50: 611–618.

  2. Cloutier, S. and Landry, B.S. (1994). Molecular markers applied to plant tissue culture.In vitro Cell. Develop. Biol.30: 32–39.

  3. Ford-Lloyd, B.V., Sabir, A., Newbury, H.J., Todd, C. and Catty, J. (1992). Determination of genetic stability using isozymes and RFLPs in beet plants regeneratedin vitro.Theor. Appl. Genet.84:113–117.

  4. Irvine, J. E., Benda, G.T. A., Legendre, B. L. and Machado, Jr.,G. R. (1984). The frequency of marker changes in sugarcane plants regenerated from callus culture.Plant Cell Tissue Org. Cult.3: 201–209.

  5. Kazim, A. and Shahid, A. (2001). Rapid multiplication of sugarcane through micropropagation technique.Pak. Sug. J.16: 11–14.

  6. Laemmli, U.K. (1970) Cleavage and structural proteins during the assembly of the head of bacteriophage T4. Nature227: 680- 685.

  7. Larkin, P. and Scowcroft, W. R. (1981). Somaclonal variation — a novel source of variability from cell cultures for plant improvement.Theor. Appl. Genet.60:197–214.

  8. Lee, TSG. (1987). Micropropagation of sugarcane (Saccharum spp.).Pl. Cell Tiss. Org. Cult.10: 47–55

  9. Lowry, O. H., Rosenbrough, N. J., Farr, A. L., and Randall, R. J. (1951). Protein measurement with folin Phenol reagent.J. Bio. Chem.193: 265- 275.

  10. Miller, A. R., Kelley, T. J. and Mujer, C. V. (1990). Anodic peroxidase isoenzymes and polyphenol oxidase activity from cucumber fruit: Tissue and substrate specificity.Phytochemistry29: 705–709.

  11. Murashige, T. and Skoog, F. (1962). A revised medium for rapid growth and bioassay with tobacco tissue culture.Physiol. Plantarum15: 473–497.

  12. Rani, V., Parida, A. and Raina, S. N. (1995). Random amplified polymorphic DNA (RAPD) markers for genetic analysis in micropropagated plants ofPopulus deltoides Marsh.Plant Cell Rep.14: 459–462.

  13. Saini, N., Saini, M.L. and Jain, R.K. (2004). Large scale production, field performance and RAPD analysis of micropropagated sugarcane plants.IndianJ. Genet.64: 102–107.

  14. Singh, B., Yadav, G.C. and Lal, M. (2001). An efficient protocol for micropropagation of sugarcane using shoot tip expiants.Sugar Tech.3: 113–116.

  15. Zhing, D.S. (1987). Comparisons of 6 isozymes in 12 accessions of Sugarcane materials.J. Fujian Agricultural College16:101–110

Download references

Author information

Correspondence to Radha Jain or Sangeeta Srivastava or Jyotsna Singh or Prashant S. Gupta.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Jain, R., Srivastava, S., Singh, J. et al. Assessment of genetic purity of micropropagated plants of sugarcane by isozyme and RAPD analysis. Sugar Tech 7, 15–19 (2005). https://doi.org/10.1007/BF02942522

Download citation

Key words

  • Isozyme pattern
  • RAPD
  • micropropagation
  • sugarcane