Sugar Tech

pp 1–7 | Cite as

Establishment of an Open, Sugar-Free Tissue Culture System for Sugarcane Micropropagation

  • Jia-ju Lu
  • Ahmad Ali
  • Er-qi He
  • Guo-qiong Yan
  • Tira-umphon ArakEmail author
  • San-Ji GaoEmail author
Research Article


Tissue culture has been used for many years to produce a large number of plant species by micropropagation. Unfortunately, microbial contamination and production costs are major problems associated with this technology. We have developed a low-cost, open tissue culture system for rapid sugarcane micropropagation that uses a combination of broad-spectrum antimicrobial compounds, a sugar-free medium, and CO2 gas fertilizer as carbon source. A novel antimicrobial compound called “QX1” that contains nicotine, garlic extract, carbendazim, and “Yipeilong” was developed in this study. Non-autoclaved induction medium containing 0.5% (v/v) QX1 had a significantly lower microbial contamination and a significantly higher explant survival rate in comparision to that of autoclaved induction medium. Sugar-free medium containing 0.5% (v/v) QX1 and 1.0 g/L CO2 gas fertilizer was used for multiplication and rooting under the open tissue culture environment. Propagation efficiencies were similar in the newly developed and conventional tissue culture systems, but contamination was significantly less in the new system during both multiplication and rooting. Also, plantlet survival rate, fresh weight, and net photosynthetic capacity were increased significantly, and production costs decreased dramatically using the new tissue culture system compared with the conventional method. Thus, the low-cost, open, sugar-free tissue culture system can be adapted to produce sugarcane plantlets on a commercial scale.


Sugarcane Tissue culture Micropropagation Antimicrobial CO2 gas 



This work was supported by grants from the Guizhou International Cooperation Base of Modern Agricultural Science and Technology (No. [2019]5804), the Guizhou Innovation Talent Base of Subtropical Crop Science and Technology (No. [2016]22), the National Sparking Plan Project of China (No. 2014GA820009), and the China Agriculture Research System (CARS-170302).

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

12355_2019_758_MOESM1_ESM.pptx (577 kb)
Supplementary material 1 (PPTX 577 kb)
12355_2019_758_MOESM2_ESM.pptx (300 kb)
Supplementary material 2 (PPTX 299 kb)


  1. Ainsworth, E.A., and S.P. Long. 2005. What have we learned from 15 years of free-air CO2 enrichment (FACE)? A meta-analytic review of the responses of photosynthesis, canopy properties and plant production to rising CO2. New Phytologist 165: 351–372.CrossRefPubMedGoogle Scholar
  2. Bakri, I.M., and C.W. Douglas. 2005. Inhibitory effect of garlic extract on oral bacteria. Archives of Oral Biology 50: 645–651.CrossRefPubMedGoogle Scholar
  3. Barnabas, L., A. Ramadass, R.S. Amalraj, M. Palaniyandi, and V. Rasappa. 2015. Sugarcane proteomics: an update on current status, challenges, and future prospects. Proteomics 15: 1658–1670.CrossRefPubMedGoogle Scholar
  4. Cheong, E.J., R. Mock, and R. Li. 2012. Elimination of five viruses from sugarcane using in vitro culture of axillary buds and apical meristems. Plant Cell, Tissue and Organ Culture 109: 439–445.CrossRefGoogle Scholar
  5. Cui, G., W.X. Dan, X. Qi, and Z.X. Sun. 2004. The preliminary study on plant open tissue culture. Journal of Shandong Agricultural University (Natural Science) 35: 529–533.Google Scholar
  6. Dan, M., S. Li, and M.M. Lu. 2017. Application of sugar free tissue culture on sugarcane rapid propagation. Journal of Anhui agriculture Science 45: 29–30.Google Scholar
  7. Davis, M.J., A.G. Gillaspie Jr., A.K. Vidaver, and R.W. Harris. 1984. Clavibacter: a new genus containing some phytopathogenic coryneform bacteria, including Clavibacter xyli subsp. xyli sp. nov., subsp. nov. and Clavibacter xyli subsp. cynodontis subsp. nov., pathogens that cause ratoon stunting disease of sugarcane and Ber. International Journal of Systematic Bacteriology 34: 107–117.CrossRefGoogle Scholar
  8. Davis, M.J., P. Rott, P. Baudin, and J.L. Dean. 1994. Evaluation of selective media and immunoassays for detection of Xanthomonas albilineans, causal agent of sugarcane leaf scald disease. Plant Disease 78: 78–82.CrossRefGoogle Scholar
  9. De Souza, A.P., M. Gaspar, E.A. Da Silva, E.C. Ulian, A.J. Waclawovsky, R.V. Dos Santos, M.M. Teixeira, G.M. Souza, and M.S. Buckeridge. 2008. Elevated CO2 increases photosynthesis, biomass and productivity, and modifies gene expression in sugarcane. Plant, Cell and Environment 31 (8): 1116–1127.CrossRefPubMedGoogle Scholar
  10. George, P., and J. Manuel. 2013. Low cost tissue culture technology for the regeneration of some economically important plants for developing countries. International Journal of Agriculture, Environment and Biotechnology 6 (Special Issue): 703–711.Google Scholar
  11. Heo, J., and T. Kozai. 1999. Forced ventilation micropropagation system for enhancing photosynthesis, growth and development of sweetpotato plantlets. Environmental Control in Biology 37: 83–92.CrossRefGoogle Scholar
  12. Hu, S. 2009. Research of the nicotine to disinfect pathogenic bacteria affects. Biotechnology 19: 73–75.Google Scholar
  13. Kozai, T. 2005. Photoautotrophic (sugar-free medium) micropropagation systems for large-scale commercialization. Propagation of Ornamental Plants 5: 23–34.Google Scholar
  14. Kozai, T., and Y. Iwanami. 1988. Effects of CO2 enrichment and sucrose concentration high photon fluxes on plantlet growth carnation (Dianthus caryophyllus L.) in tissue culture during the preparation stage. Journal of the Japanese Society for Horticultural Science 57: 279–288.CrossRefGoogle Scholar
  15. Kozai, T., and C. Kubota. 2005. Concepts, definitions, ventilation methods, advantages and disadvantages. In Photoautotrophic (sugar-free medium) micropropagation as a new propagation and transplant production system, ed. T. Kozai, F. Afreen, and S.M.A. Zobayed, 19–30. Dordrecht: Springer.CrossRefGoogle Scholar
  16. Lal, M., A.K. Tiwari, G.N. Gupta, and Kavita Kumari. 2015. Commercial scale micropropagation of sugarcane: constraints and remedies. Sugar Tech 17: 339–347.CrossRefGoogle Scholar
  17. Li, Z. 2009. Application of CO2 gas fertilizer in greenhouse for vegetables. Agricultural Technology and Equipment 22: 27–29.Google Scholar
  18. Li, J., and J. Zhou. 2004. Applied technology of CO2 gas fertilizer in greenhouse for vegetables. Northwest Horticulture 9: 7–9.Google Scholar
  19. Lu, J., A. Tira-Umphon, Z. Zhang, S. Lei, and L. Yu. 2014. Effect of bacteriostat (Qianxing No. 1) on open tissue culture of sugarcane. Agricultural Technology and Equipment 15: 1478–1481.Google Scholar
  20. Lu, J., Z. Zhang, S. Peng, X. Li, Z. Li, and X. Huang. 2012. Application of compound bacteriostat in the open tissue culture of sugarcane. Chinese Journal of Tropical Agriculturey 32: 68–71.Google Scholar
  21. Luo, Q., K. Ahmad, H.Y. Fu, J.D. Wang, R.K. Chen, and S.J. Gao. 2016. Genetic diversity and population structure of Sorghum mosaic virus infecting Saccharum spp. hybrids. Annals of Applied Biology 169: 398–407.CrossRefGoogle Scholar
  22. Millan-Almaraz, J.R., R.G. Guevara-Gonzalez, R. Romero-Troncoso, R.A. Osornio-Rios, and I. Torres-Pacheco. 2009. Advantages and disadvantages on photosynthesis measurement techniques: A review. African Journal of Biotechnology 8: 7340–7349.Google Scholar
  23. Murashige, T., and F. Skoog. 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum 15: 473–497.CrossRefGoogle Scholar
  24. Nowak, R.S., D.S. Ellsworth, and S.D. Smith. 2004. Functional responses of plants to elevated atmospheric CO2 do photosynthetic and productivity data from FACE experiments support early predictions? New Phytologist 162: 253–280.CrossRefGoogle Scholar
  25. Perez, A., L. Napoles, C. Carvajal, M. Hernandez, and J. Lorenzo. 2004. Effect of sucrose, inorganic salts, inositol, and thiamine on protease excretion during pineapple culture in temporary immersion bioreactors. Vitro Cellular and Developmental Biology-Plant 40: 311–316.CrossRefGoogle Scholar
  26. Rott, P.C., C. Kaye, M. Naranjo, J.M. Shine Jr., S. Sood, J.C. Comstock, and R.N. Raid. 2016. Controlling sugarcane diseases in Florida: A challenge in constant evolution. Proceedings of the International Society of Sugarcane Technologists 29: 595–600.Google Scholar
  27. Sawant, R.A., and P.N. Tawar. 2011. Use of sodium hypochlorite as media sterilant in sugarcane micropropagation at commercial scale. Sugar Tech 13: 27–35.CrossRefGoogle Scholar
  28. Vu, J.C., and L.H. Allen Jr. 2009. Stem juice production of the C4 sugarcane (Saccharum officinarum) is enhanced by growth at double-ambient CO2 and high temperature. Journal of Plant Physiology 166: 1141–1151.CrossRefPubMedGoogle Scholar
  29. Vu, J.C., L.H. Allen Jr., and R.W. Gesch. 2006. Up-regulation of photosynthesis and sucrose metabolism enzymes in young expanding leaves of sugarcane under elevated growth CO2. Plant Science 171: 123–131.CrossRefGoogle Scholar
  30. Xiao, Y., and T. Kozai. 2006. Photoautotrophic growth and net photosynthetic rate of sweet potato plantlets in vitro as affected by the number of air exchanges of the vessel and type of supporting material. Tsinghua Science and Technology 11: 481–489.CrossRefGoogle Scholar
  31. Xiao, Y., Y.H. Lok, and T. Kozai. 2003. Photoautotrophic growth of sugarcane plantlets in vitro as affected by photosynthetic photon flux and vessel air exchanges. Vitro Cellular and Developmental. Biology Plant 39 (2): 186–192.CrossRefGoogle Scholar
  32. Xiao, Y., G. Niu, and T. Kozai. 2011. Development and application of photoautotrophic micropropagation plant system. Plant Cell, Tissue and Organ Culture 105 (2): 149–158.CrossRefGoogle Scholar

Copyright information

© Society for Sugar Research & Promotion 2019

Authors and Affiliations

  1. 1.School of Crop Production Technology, Institute of Agricultural TechnologySuranaree University of TechnologyNakhon RatchasimaThailand
  2. 2.Guizhou Institute of Subtropical CropsXingyiChina
  3. 3.National Engineering Research Center for SugarcaneFujian Agriculture and Forestry UniversityFuzhouChina
  4. 4.Xingyi No. 2 High SchoolXingyiChina

Personalised recommendations