Skip to main content
SpringerLink
Log in

Enhanced in vitro Regeneration in Sugarcane (Saccharum officinarum L.) by Use of Alternate High-Low Picloram Doses and Thidiazuron Supplementation

  • Published:
Cytology and Genetics Aims and scope Submit manuscript

Abstract

Sugarcane is an important domestic and industrial crop and considerable efforts have been made to improve its yield through conventional as well as biotechnological approaches. Genetic manipulation of sugarcane is dependent on an efficient reproducible in vitro regeneration regime. In the current study, the role of explant position, induced desiccation, picloram (PIC) levels for callogenesis as well as thidiazuron (TDZ) addition during regeneration phase has been appraised. Using an optimum combination of mentioned factors, an enhanced in vitro regeneration system has been established for two elite sugarcane cultivars. Embryogenesis was stimulated in cv. HSF-240 by MS medium augmented with 12.42 μM PIC while cv. CPF-237 exhibited sometic embryo formation when PIC supplementation was combined with induced desiccation (using 12 g L–1 agar). A decrease in embryogenesis frequency was recorded from base towards tip. The explants cultured alternately on high (12.42 μM) and low (4.14 μM) PIC medium produced the highest number of nodular calli which later exhibited maximum regeneration potential. Optimal shoot initiation was observed with 9.08 μM TDZ followed by medium having 2.27 μM 2,4-dichlorophenoxyacetic acid plus 4.43 μM benzylaminopurine. However, the shoots produced with former medium composition showed frailty as compared to the one regenerated on later medium. Healthy roots were initiated with 16.11 μM naphthalene acetic acid in the presence of 0.5% activated charcoal. Malondialdehyde content, catalase and peroxidase activity of in vitro and field grown sugarcane plants were analogous, indicating that the in vitro regenerated plants were equally fit for subsequent growth in natural conditions. The reported protocol can be helpful in devising strategies for a robust sugarcane genetic engineering regime.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1.
Fig. 2.
Fig. 3.
Fig. 4.
Fig. 5.
Fig. 6.

Similar content being viewed by others

REFERENCES

  1. Aebi, H., Catalase in vitro, Methods Enzymol., 1984, vol. 105, pp. 121–126.

    Article  CAS  Google Scholar 

  2. Aftab, F. and Iqbal, J., Plant regeneration from protoplasts derived from cell suspension of adventive somatic embryos in sugarcane (Saccharum sp hybrid cv CoL-54 and cv CP-43/33), Plant Cell Tiss. Organ Cult., 1999, vol. 56, pp. 155–162.

    Article  CAS  Google Scholar 

  3. Aftab, F., Zafar, Y., Malik, K.A., and Iqbal, J., Plant regeneration from embryogenic cell suspensions and protoplasts in sugarcane (Saccharum spp. hybrid cv CoL-54), Plant Cell Tiss. Organ Cult., 1996, vol. 44, pp. 71–78.

    Article  CAS  Google Scholar 

  4. Ali, A., Naz, S., Siddiqui, F.A., and Iqbal, J., Rapid clonal multiplication of sugarcane (Saccharum officinarum) through callogenesis and organogenesis, Pak. J. Bot., 2008, vol. 40, pp. 123–138.

    CAS  Google Scholar 

  5. Altpeter, F., Baisakh, N., Beachy, R., Bock, R., Capell, T., Christou, P., Daniell, H., Datta, K., Datta, S., Dix, P.J., Fauquet, C,et al., Particle bombardment and the genetic enhancement of crops: myths and realities, Mol. Breed., 2005, vol. 15, pp. 305–327.

    Article  Google Scholar 

  6. Chance, B. and Maehly, A.C., Assay of catalases and peroxidases, Methods Enzymol., 1955, vol. 2, pp. 764–775.

    Article  Google Scholar 

  7. Charleson, R.P., Stephen, C.W., and Matthew, A.J., Adventitious shoot regeneration of scotch spearmint (Mentha x Gracilis Sole), In Vitro Cell. Dev. Biol.–Plant, 2006, vol. 42, pp. 354–358.

    Article  Google Scholar 

  8. Chengalrayan, K. and Gallo-Meagher, M., Effect of various growth regulators on shoot regeneration of sugarcane, In Vitro Cell. Dev. Biol.–Plant, 2001, vol. 37, pp. 434–439.

    Article  CAS  Google Scholar 

  9. Chengalrayan, K., Abouzid, A., and Gallo-Meagher, M., In vitro regeneration of plants from sugarcane seed derived callus, In Vitro Cell. Dev. Biol–Plant, 2005, vol. 41, pp. 477–482.

    Article  Google Scholar 

  10. Croft, B.J., Magarey, R.C., Allsopp, P.G., Cox, M.C., Willcox, T.G., Milford, B.J., Wal, E.S., et al., Sugarcane smut in Queensland: arrival and emergency response, Austral. Plant Pathol., 2001, vol. 37, pp. 26–34.

    Article  Google Scholar 

  11. Dhawan, A.K., Moudgil, R., Dendsay, J.P.S., and Mandhan, R.P., Low thidiazuron levels promote and sustain shootlet multiplication in sugarcane, Ind. J. Plant Physiol., 2004, vol. 9, pp. 354–359.

    CAS  Google Scholar 

  12. Economic Survey of Pakistan, Government of Pakistan, Ministry of Food and Agriculture Islamabad, 2017-18.

  13. Gallomeagher, M., English, R.G., and Abouzid, A., Thidiazuron stimulates shoot regeneration of sugarcane embryogenic callus, In Vitro Cell. Dev. Biol.–Plant, 2000, vol. 36, pp. 37–40.

    Article  CAS  Google Scholar 

  14. Garcia, R., Cidade, D., Castellar, A., Lip, A., Magioli, C., Callado, C., and Mansur, E., In vitro morphogenesis patterns from shoot apices of sugar cane are determined by light and type of growth regulator, Plant Cell Tiss. Organ Cult., 2007, vol. 90, pp. 181–190.

    Article  Google Scholar 

  15. Gill, R., Malhotra, P.K., and Gosal, S.S., Direct plant regeneration from cultured young leaf segments of sugarcane, Plant Cell Tiss. Organ Cult., 2006, vol. 84, pp. 100205–100209.

    Article  Google Scholar 

  16. Grossmann, K., Mode of action of auxin herbicides: a new ending to a long, drawn out story, Trends Plant Sci., 2000, vol. 5, pp. 506–509.

    Article  CAS  Google Scholar 

  17. Heath, R.L. and Packer, L., Photoperoxidation in isolated chloroplasts: kinetics and stoichiometry of fatty acid peroxidation, Arch. Biochem. Biophys., 1968, vol. 125, pp. 189–198.

    Article  CAS  Google Scholar 

  18. Hotta, C.T., Lembke, C.G., Domingues, D.S., et al., The biotechnology roadmap for sugarcane improvement, Trop. Plant Biol., 2010, vol. 3, pp. 75–87.

    Article  CAS  Google Scholar 

  19. Jain, M., Chengalrayan, K., Abouzid, A., and Gallo-Meagher, M., Prospecting the utility of a PMI/mannose selection system for the recovery of transgenic sugarcane (Saccharum sp. hybrid) plants, Plant Cell Rep., 2007, vol. 26, pp. 581–590.

    Article  CAS  Google Scholar 

  20. Kaur, A. and Gosal, S.S., Desiccation of callus enhances somatic embryogenesis and subsequent shoot regeneration in sugarcane, Ind. J. Biotechnol., 2009, vol. 8, pp. 332–334.

    CAS  Google Scholar 

  21. Kaur, R. and Kapoor, M., Plant regeneration through somatic embryogenesis in sugarcane, Sugar Tech., 2016, vol. 18, pp. 93–99.

    Article  CAS  Google Scholar 

  22. Kumari, K., Lal, M., and Saxena, S., Enhanced micropropagation and tiller formation in sugarcane through pretreatment of explants with thidiazuron (TDZ), Biotech., 2017, vol. 7, p. 282.

  23. Lakshmanan, P., Geijskes, R.J., Aitken, K.S., Grof, C.L.P., Bonnett, G.D., and Smith, G.R., Sugarcane biotechnology: the challenges and opportunities, In Vitro Cell. Dev. Biol.–Plant, 2005, vol. 41, pp. 345–363.

    Article  CAS  Google Scholar 

  24. Lakshmanan, P., Geijskes, R.J., Wang, L.F., Elliott, A., Grof, C.P.L., Berding, N., and Smith, G.R., Developmental and hormonal regulation of direct shoot organogenesis and somatic embryogenesis in sugarcane (Saccharum sp interspecific hybrids) leaf culture, Plant Cell Rep., 2006, vol. 5, pp. 1007–1015.

    Article  Google Scholar 

  25. Mamun, M.A., Sikdar, M.B.H., Paul, D.K., Rahman, M.M., and Islam, M.R., In vitro micropropagation of some important sugarcane varieties of Bangladesh, Asian J. Plant Sci., 2004, vol. 3, pp. 666–669.

    Article  Google Scholar 

  26. Munir, N. and Aftab, F., The role of polyethylene glycol (PEG) pretreatment in improving sugarcane’s salt (NaCl) tolerance, Turk. J. Bot., 2009, vol. 33, pp. 407–415.

    Google Scholar 

  27. Murashige, T. and Skoog, F., A revised medium for rapid growth and bioassays with tobacco tissue culture, Physiol. Plant., 1962, vol. 15, pp. 473–497.

    Article  CAS  Google Scholar 

  28. Ramanand Kureel, N., Subhanand, N., Lal, M., and Singh, S.B., Plantlet regeneration through leaf callus culture in sugarcane, Sugar Tech., 2006, vol. 8, pp. 85–87.

    Article  Google Scholar 

  29. Rikiishi, K., Matsuura, T., Maekawa, M., and Takeda, K., Light control of shoot regeneration in callus cultures derived from barley (Hordeum vulgare L.) immature embryos, Breed Sci., 2008, vol. 58, pp. 129–135.

    Article  CAS  Google Scholar 

  30. Snyman, S.J., Meyer, G.M., Richards, J.M., Haricharan, N., Ramgareeb, S., and Huckett, B.I., Refining the application of direct embryogenesis in sugarcane: effect of the developmental phase of leaf disc explants and the timing of DNA transfer on transformation efficiency, Plant Cell Rep., 2006, vol. 25, pp. 1016–1023.

    Article  CAS  Google Scholar 

  31. Taiz, L. and Zeiger, E., Plant Physiology, Sunderland Massachusetts, USA: Sinauer Associates Inc., 2010, 5th ed.

    Google Scholar 

  32. Thomas, T.D., The role of activated charcoal in plant tissue culture, Biotechnol. Adv., 2008, vol. 26, pp. 618–631.

    Article  CAS  Google Scholar 

  33. Tiel, K., Enríquez, G.A., Ceballo, Y., Soto, N., Fuentes, A.D., Ferreira, A., Coll, Y., and Pujol, M., Development of a system for rapid plant regeneration from in vitro sugarcane (Saccharum officinarum L.) meristematic tissue, Biotecnol. Appl., 2006, vol. 23, pp. 22–24.

    Google Scholar 

Download references

ACKNOWLEDGMENTS

We are greatly thankful to Dr. Shahid Afghan, Ayub Agricultural Research Institute Faisalabad, for provision of sugarcane germplasm used in the current study.

Author information

Authors and Affiliations

  1. School of Biological Sciences, University of the Punjab, 54590, Lahore, Pakistan

    Muhammad Sohail Akram & Javed Iqbal

  2. Department of Botany, Government College University, 38000, Faisalabad, Pakistan

    Muhammad Sohail Akram

  3. Department of Botany, Government College Women University, Faisalabad, Pakistan

    Ambreen Khadija Alvi

Authors
  1. Muhammad Sohail Akram
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ambreen Khadija Alvi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Javed Iqbal
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

JI perceived the idea/s and MSA executed the experiments. MS and AKA helped in statistical analysis of the data and preparation of the manuscript. All the authors read/approved the manuscript for submission.

Corresponding author

Correspondence to Muhammad Sohail Akram.

Ethics declarations

The authors declare that they have no conflict of interests. This article does not contain any studies involving animals or human participants performed by any of the authors.

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Akram, M.S., Alvi, A.K. & Iqbal, J. Enhanced in vitro Regeneration in Sugarcane (Saccharum officinarum L.) by Use of Alternate High-Low Picloram Doses and Thidiazuron Supplementation. Cytol. Genet. 55, 566–575 (2021). https://doi.org/10.3103/S0095452721060025

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.3103/S0095452721060025

Keywords:

Search

Navigation