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Sugar Tech

, Volume 20, Issue 1, pp 50–59 | Cite as

Ethyl Methanesulfonate Mutagenesis and In Vitro Polyethylene Glycol Selection for Drought Tolerance in Sugarcane (Saccharum spp.)

  • M. Masoabi
  • J. Lloyd
  • J. Kossmann
  • C. van der Vyver
Research article
  • 508 Downloads

Abstract

Drought is a serious agronomic problem, and urgent attention to overcome drought stress is vital to eradicate or minimize its effects on crop production. Random induction of genomic mutation is a technique that can enhance genetic diversity leading to useful traits such as enhanced drought tolerance. In this study, sugarcane callus was exposed to different concentrations of the chemical mutagen, ethyl methanesulfonate (EMS). Concentrations of 20 mM and lower were identified as useful to induce genomic mutations without compromising in vitro sugarcane plant regeneration abilities. Furthermore, sugarcane callus was exposed to varying concentrations of polyethylene glycol (PEG) for different time periods in order to identify a suitable in vitro osmotic selection regime to simulate drought stress in vitro. The optimal in vitro osmotic selection treatment was identified as callus exposed to 20% (w/v) PEG6000 for 8 weeks, followed by a 2 weeks osmotic recovery period without PEG and ending with a further 8 week PEG selection period during somatic embryo regenerations. Sugarcane callus from the NCo310 cultivar was subsequently mutagenized with 16 mM EMS and in vitro selected on 20% (w/v) PEG6000, which resulted in the survival of 18 plantlets. These in vitro selected lines were subjected to preliminary greenhouse pot trials to confirm drought tolerance. Pot trials identified seven lines that outlived NCo310 control plants. In addition, when re-watered after the drought stress period, plants from one mutant line recovered and were able to form new shoots. The results from this study indicate, therefore, that EMS mutagenesis and in vitro selection for osmotic pressure using PEG can be successfully applied to cultivate sugarcane plants with improved morphological and physiological responses to water stress.

Keywords

Sugarcane Mutagenesis Drought Polyethylene glycol Ethyl methanesulfonate 

Notes

Acknowledgement

Research was funded by the South African Sugar Association (SASA; grant S004120), Mount Edgecombe, Durban, South Africa.

Author contribution statement

Miss M. Masoabi is responsible for conducting all the relevant laboratory experimental work and written the first draft of the manuscript. Dr J. Lloyd is responsible for supervising student experimental work, and editing of manuscript. Prof J. Kossmann is grant holder of the relevant research funding. Dr C. van der Vyver is responsible for research conceptualization, main student supervision and manuscript writing.

Compliance with Ethical Standards

Conflict of interest

Prof J Kossmann is the grant holder of the SASA, S004120 research grant. All other authors declare that they have no conflict of interest.

Supplementary material

12355_2017_524_MOESM1_ESM.pdf (1.3 mb)
Supplementary material 1 (PDF 1375 kb)

References

  1. Ali, A., S. Naz, S.S. Alam, and J. Iqbal. 2007. In vitro induced mutation for screening of red rot (Colletotrichum falcatum) resistance in sugarcane (Saccharum officinaruum). Pakistan Journal of Botany 39: 1979–1994.Google Scholar
  2. Bhat, R., N. Upadhyaya, A. Chaudhury, C. Raghavan, and F. Qiu. 2007. Chemical- and Irradiation- Induced mutants and TILLING. In Rice Functional Genomics: Challenges, Progress and Prospects 8: 148–180, ed. N.M. Upadhyaya. New York: Springer.Google Scholar
  3. Bidabadi, S.S., S. Meon, Z. Wahab, S. Subramaniam, and M. Mahmood. 2012. In vitro selection and characterization of water stress tolerant lines among ethyl methanesulphonate (EMS) induced variants of banana (Musa spp., with AAA genome). Australian Journal of Crop Science 6: 567–575.Google Scholar
  4. Blum, A. 2004. Use of PEG to induce and control plant water deficit in experimental hydroponics culture. http://www.plantstress.com/methods/PEG.htm. Accessed March 2015.
  5. Blum, A. 2014. Genomics for drought resistance—Getting down to earth. Functional Plant Biology 41: 1191–1198.CrossRefGoogle Scholar
  6. Carmo-Silva, A.E., A. Francisco, S.J. Powers, J. Alfred, A.J. Keys, et al. 2009. Grasses of different C4 subtypes reveal leaf traits related to drought tolerance in their natural habitats: Changes in structure, water potential, and amino acid content. American Journal of Botany 96: 1222–1235.CrossRefPubMedGoogle Scholar
  7. Comai, L., and S. Henikoff. 2006. Tilling: Practical single- nucleotide mutation discovery. The Plant Journal 45: 84–694.CrossRefGoogle Scholar
  8. El-Haris, M.K., and M.N. Barakat. 1998. Evaluation of the in vitro selected drought tolerant wheat lines under drought stress conditions. Journal of Agricultural Research 43: 293–302.Google Scholar
  9. Errabii, T., C.B. Gandonou, H. Essalmani, J. Abrini, M. Idaomar, and N. Skali-Senhaji. 2007a. Growth, proline and ion accumulation in sugarcane callus cultures under drought-induced osmotic stress and its subsequent relief. African Journal of Biotechnology 5: 1488–1493.Google Scholar
  10. Errabii, T., C.B. Gandonou, H. Essalmani, J. Abrini, M. Idaomar, and N.S. Senhaji. 2007b. Effects of NaCl and mannitol induced stress on sugarcane (Saccharum sp.) callus cultures. Acta Physiologiae Plantarum 29: 95–102.CrossRefGoogle Scholar
  11. FAO/IAEA database of mutant variety and genetic stocks. http://mvd.iaea.org/. Accessed November 2015.
  12. Fuller, M.P., E.M.R. Metwali, M.H. Eed, and A.J. Jellings. 2006. Evaluation of abiotic stress resistance in mutated populations of cauliflower (Brassica oleracea var. botrytis). Plant Cell, Tissue and Organ Culture 86: 239–248.CrossRefGoogle Scholar
  13. Gentile, A., L.I. Dias, R.S. Mattos, T.H. Ferreira, and M. Menossi. 2015. MicroRNAs and drought responses in sugarcane. Frontiers in Plant Science 6: 1–13.CrossRefGoogle Scholar
  14. Glantz, S.A., and B.K. Slinker. 2001. Primer of applied regression and analyses of variance. New York: McGraw-Hill Inc.Google Scholar
  15. Hasson, N.S., L.D. Shaaban, A.H. El-Sayed, and E.E. Seleem. 2004. In vitro selection for water stress tolerant callus line of Heliathus annus L Cv Myak. International Journal of Agricultural Biology 6: 13–18.Google Scholar
  16. Kenganal, M., R.R. Hanchnal, and H.L. Nadaf. 2008. Ethyl methane sulfonate (EMS) induced mutation and selection for salt tolerance in sugarcane in vitro. Indian Journal of Plant Physiology 13: 405–410.Google Scholar
  17. Koch, A.C., S. Ramgareeb, R.S. Rutherford, S.J. Snyman, and M.P. Watt. 2012. An in vitro mutagenesis protocol for the production of sugarcane tolerant to the herbicide imazapyr. In Vitro Cellular and Developmental Biology-Plant 48: 417–427.CrossRefGoogle Scholar
  18. Kumar, A., M. Bakshi, M. Kaur, S. Kapruwan, and M. Singh. 2015. Effects of mutagen treatment on the seed germination and callus induction in Dendrocalamus hamiltonii. Journal of Environmental and Applied Bioresearch 3: 234–237.Google Scholar
  19. Lu, G., C. Gao, X. Zeng, and B. Han. 2009. Identification of OsbZIP72 as a positive regulator of ABA response and drought tolerance in rice. Planta 229: 605–615.CrossRefPubMedGoogle Scholar
  20. Lu, H.F., H.T. Dong, C.B. Sun, D.J. Qing, N. Li, et al. 2011. The panorama of physiological responses and gene expression of whole plant of maize inbred line YQ7-96 at the three-leaf stage under water deficit and re-watering. Theoretical and Applied Genetics 123: 943–958.CrossRefPubMedGoogle Scholar
  21. Murashige, T., and F. Skoog. 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum 15(3): 473–497CrossRefGoogle Scholar
  22. Nikam, A.A., R.M. Devarumath, M.G. Shitole, V.S. Ghole, P.N. Tawar, and P. Suprasanna. 2014. Gamma radiation, in vitro selection for salt (NaCl) tolerance, and characterization of mutant in sugarcane (Saccharum officinarum L.). In Vitro Cellular and Developmental Biology-Plant 50: 766–776.CrossRefGoogle Scholar
  23. Oloriz, M.I., V. Gil, L. Rojas, N. Veitia, M. Hofte, and E. Jimenez. 2011. Selection and characterization of sugarcane mutants with improved resistance to brown rust obtained by induced mutation. Crop and Pasture Science 62: 1037–1044.CrossRefGoogle Scholar
  24. Parry, M.A.J., P.J. Madgwick, C. Bayon, K. Tearall, A. Hernandez-Lopez, et al. 2009. Mutation discovery for crop improvement. Journal of Experimental Botany 60: 2817–2825.CrossRefPubMedGoogle Scholar
  25. Patade, V.Y., P. Suuprasanna, and V.A. Bapat. 2008. Gamma irradiation of embryogenic callus cultures and in vitro selection for salt tolerance in sugarcane (Saccharum officinarum L.). Agricultural Sciences in China 7: 1147–1152.CrossRefGoogle Scholar
  26. Rao, S., and F.T.Z. Jabeen. 2013. In vitro selection and characterization of polyethylene glycol (PEG) tolerant callus lines and regeneration of plantlets from the selected callus lines in sugarcane (Saccharum officinarum L.). Physiology and Molecular Biology of Plants 19: 261–268.CrossRefPubMedPubMedCentralGoogle Scholar
  27. Reis, R.R., B.A.D.B. da Cunha, P.K. Martins, M.T.B.M. Martins, J.C. Alekcevetch, A. Chalfun-Junior, et al. 2014. Induced over-expression of AtDREB2A CA improves drought tolerance in sugarcane. Plant Science 221–222: 59–68.CrossRefPubMedGoogle Scholar
  28. Royan, M., P. Govender, M. Binedell, S. McFarlane, G. Leslie, R. Rhodes, R. Stranack, G. Maher, and S. Berry. 2011. Coping with drought. In The Link, 20 (1). South Africa: South African Sugarcane Research Institute. http://www.sasa.org.za.
  29. Rutherford, R.S., S.J. Snyman, and M.P. Watt. 2014. In vitro studies on somaclonal variation and induced mutagenesis: progress and prospects in sugarcane (Saccharum spp.)—A review. Journal of Horticultural Science and Biotechnology 89: 1–16.CrossRefGoogle Scholar
  30. Serrat, X., R. Esteban, N. Guibourt, L. Moysset, S. Nogués, and E. Lalanne. 2014. EMS mutagenesis in mature seed-derived rice calli as a new method for rapidly obtaining TILLING mutant populations. Plant Methods 10: 1–13.CrossRefGoogle Scholar
  31. Silue, S., N. Diarrassouba, I.J. Fofana, Y. Muhovski, A. Toussaint, G. Mergeai, J.M. Jacquemin, and J.P. Baudoin. 2013. Description of Phaseolus vulgaris L. aborting embryos from ethyl methanesulfonate (EMS) mutagenized plants. Biotechnology Agronomy Society Environment 17: 563–571.Google Scholar
  32. Snyman, S.J., P. Mhlanga, and M.P. Watt. 2016. Rapid screening of sugarcane plantlets for in vitro mannitol-induced stress. Sugar Tech 18: 437–440.CrossRefGoogle Scholar
  33. Smit, M.A., and A. Singels. 2006. The response of sugarcane canopy development to water stress. Field Crops Research 3: 91–97.CrossRefGoogle Scholar
  34. Talebi, A.B., A.B. Talebi, and B. Shahrokhifar. 2012. Ethyl methane sulphonate (EMS) induced mutagenesis in Malaysian rice (cv. MR219) for lethal dose determination. American Journal of Plant Sciences 3: 1661–1665.CrossRefGoogle Scholar
  35. Tsago, Y., M. Andargie, and A. Takele. 2014. In vitro selection of sorghum (Sorghum bicolor (L) Moench) for polyethylene glycol (PEG) induced drought stress. Plant Science Today 1: 62–68.CrossRefGoogle Scholar
  36. Vadez, V., J. Palta, and J. Berger. 2014. Developing drought tolerant crops: hopes and challenges in an exciting journey. Functional Plant Biology 41: 5–6.CrossRefGoogle Scholar
  37. Van Harten, A.M. 1998. Mutation breeding: Theory and Practical Applications. London: Cambridge University Press.Google Scholar
  38. Verslues, P.E., and E.A. Bray. 2004. LWR1 and LWR2 are required for osmoregulation and osmotic adjustment in Arabidopsis. Plant Physiology 136: 2831–2842.CrossRefPubMedPubMedCentralGoogle Scholar
  39. Wang, Y., L. Lin, and H. Chen. 2015. Assessing the economic impacts of drought from the perspective of profit loss rate: A case study of the sugar industry in China. Natural Hazards and Earth System Sciences 15: 1603–1616.CrossRefGoogle Scholar
  40. Wani, S.H., P.A. Sofi, S.S. Gosal, and N.B. Singh. 2010. In vitro screening of rice (Oryza sativa L.) callus for drought tolerance. Communications in Biometry and Crop Science 5: 108–115.Google Scholar
  41. Wiedenfeld, R.P. 2000. Water stress during different sugarcane growth periods on yield and response to N fertilization. Agricultural Water Management 43: 173–182.CrossRefGoogle Scholar
  42. Zambrano, A.Y., J.R. Demey, M. Fuchs, V. Gonzalez, R. Rea, O. De Sousa, and Z. Gutierrez. 2003. Selection of sugarcane plants resistant to SCMV. Plant Science 165: 221–225.CrossRefGoogle Scholar

Copyright information

© Society for Sugar Research & Promotion 2017

Authors and Affiliations

  • M. Masoabi
    • 1
  • J. Lloyd
    • 1
  • J. Kossmann
    • 1
  • C. van der Vyver
    • 1
  1. 1.Department of Genetics, Institute for Plant BiotechnologyUniversity of StellenboschStellenboschSouth Africa

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