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Optimizing a novel micropropagation system for Poacea in a single, multifactor experiment

  • Jeffrey William AdelbergEmail author
  • Jacqueline Antionette Naylor-Adelberg
  • Rabia Fawzi El-hawaz
Original Article
  • 80 Downloads

Abstract

A disposable, flexible film vessel was demonstrated for liquid culture micrporopagation of Poacea × ‘Cape Breeze’ and a micropropagation process was optimized for this system. Process factors involving media volume and plant density; mineral factors of ammonium and phosphate concentrations; and the important organic chemicals sucrose and benzyladenine; were simultaneously varied in the presence and absence of an inert paper substrate (40 treatment combinations for the six continuous factors, duplicated for the nominal factor, paper). Plant multiplication was enhanced by having fewest plants per vessel (10) with the largest volume of medium (90 mL), with the greatest concentrations of benyzladenine (10 µM) and phosphate (6.25 mM), in the absence of the paper. Optimal ex vitro growth, measured by dry mass increase on the mist bed required plants that had been grown under very different in vitro conditions (the smallest volume of medium per vessel (30 mL), fewest plants per vessel (10), low benzyladenine (1.6 µM), lowest phosphate (1.25 mM), in the presence of paper). Media that produced the largest in vitro plants produced plants that rapidly increased in dry mass in the greenhouse. Increasing the number of plants per vessel (30) increases the economy of the system with greater numbers of plants, but they are smaller and grow more slowly than the low-density cultures. In a small experiment using 80 vessels, separate optimal conditions in both process and media formulation were determined for both stage II and stage III cultures of Poacea. The analysis identified most important factors that drive the responses, from other significant responses with less impact.

Key message

A liquid culture micropropagation system for Poacea in flexible-film vessels was optimized for process and nutritional factors in both stages II and III in a small, response surface method experiment.

Keywords

Flexible film vessel Liquid medium Response surface method Switchgrass 

Notes

Author contributions

JWA designed the experiment, collected and analyzed data, and drafted the manuscript. JNA conducted the research and collected the data. RFE collected and analyzed data and helped draft manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare they have no conflicts of interest.

Supplementary material

11240_2019_1604_MOESM1_ESM.xlsx (15 kb)
Supplementary material 1 (XLSX 15 kb)

References

  1. Adelberg J (2005) Efficiency in a thin-film system for micropropagation of Hosta. Plant Cell Tiss Organ Cult 81:359–368CrossRefGoogle Scholar
  2. Adelberg JW (2010) Sucrose water and nutrient use during stage II multiplication of two turmeric clones (Curcuma longa L.) in liquid medium. Sci Hortic 12:262–267CrossRefGoogle Scholar
  3. Adelberg J, Fari M (2010) Applied physiology and practical bioreactor designs for micropropagation of ornamental plants. Propagation of Ornamental Plants 10:205–219Google Scholar
  4. Adelberg J, Toler J (2004) Comparison of agar and an agitated, thin-film liquid system for micropropagation of ornamental elephant ears. HortScience 39:1088–1092CrossRefGoogle Scholar
  5. Adelberg JW, Delgado MP, Tomkins JP (2010) Spent medium analysis for liquid culture micropropagation of Hemerocallis on Murashige and Skoog medium. In Vitro Cell Dev Biol Plant 46:95–107CrossRefGoogle Scholar
  6. Adelberg J, Driesse T, Halloran S, Bridges WC (2013) Relationships between nutrients and plant density in liquid media during micropropagation and acclimatization of turmeric. In Vitro Cell Dev Biol Plant 49:724–736CrossRefGoogle Scholar
  7. Caesar L, Adelberg J (2015) Using a multifactor approach for improving stage II responses of Helleborus hybrids in micropropagation. Propag Ornam Plants 15:125–135Google Scholar
  8. Chu I (1995) Economic analysis of automated micropropagation. In: Aitken-Christie J, Kozai T, Smith MAL (eds) Automation and environmental control in plant tissue culture. Kluwer Academic, Dordrecht, pp 19–27CrossRefGoogle Scholar
  9. El-Hawaz RF, Bridges WC, Adelberg JW (2015) In vitro growth of Curcuma longa L. in response to five mineral elements and plant density in fed-batch culture systems. PLoS ONE 10(4):e0118912.  https://doi.org/10.1371/journal.pone.0118912 CrossRefGoogle Scholar
  10. El-Hawaz RF, Park D, Bridges WC, Adelberg JW (2016) Optimizing in vitro mineral nutrition and plant density increases greenhouse growth rate of Curcuma longa L. during greenhouse growth. Plant Cell Tiss Organ Cult 4:5.  https://doi.org/10.1007/s11240-016-0974-9 Google Scholar
  11. George EF, De Klerk GJ (2008) The components of tissue culture media II: macro- and micro-nutrients. In: George EF, Hall MA, De Klerk G-J (eds) Plant propagation by tissue culture, 3rd edn. Springer, Dordrecht, pp 65–114Google Scholar
  12. George EF, Debergh PC (2008) Micropropagation: uses and methods. In: George EF, Hall MA, De Klerk G-J (eds) Plant propagation by tissue culture, 3rd edn. Springer, Dordrecht, pp 33–34Google Scholar
  13. Jones JB, Murashige T (1974) Tissue culture propagation Achemea fasciata Baker and other bromeliads. Comb Proc Int Plan Propag 24:117–126Google Scholar
  14. Kane ME (2011) Propagation by shoot culture. In: Trigiano RN, Gray DJ (eds) Plant tissue culture, development and biotechnology. CRC Press, Taylor Francis Group, Boca Raton, pp 181–192Google Scholar
  15. Larkens D (2013) Calculating and reporting effect sizes to facilitate cumulative science: a practical primer for t-tests and ANOVAs. Front Psychol 4:863Google Scholar
  16. Leifert C, Lumsden PJ, Pryce S, Murphy KP (1991) Effects of mineral nutrition on the growth of tissue cultured plants. In: Goulding KH (ed.) Horticultural exploitation of recent biological developments. Proceedings of the Institute of Horticulture, pp 43–57Google Scholar
  17. Leifert C, Murphy KP, Lumsden PJ (1995) Mineral and carbohydrate nutrition of plant cell and tissue cultures. CRC Crit Rev Plant Sci 14:83–109CrossRefGoogle Scholar
  18. Miller LR, Murashige T (1976) Tissue culture propagation of foliage plants. In Vitro Cell Dev Biol 12:797–813CrossRefGoogle Scholar
  19. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  20. Murashige T, Shabde MN, Hasegawa PM, Takatori FH, Jones JB (1972) Propagation of asparagus through shoot apex culture. I. Nutrient media for formation of plantlets. J Am Soc Hortic Sci 97:158–161Google Scholar
  21. Nguyen TQ, Nguyen NH, Hoang NN, Pham MD, Nguyen TM, Huynh HD (2011) Photoautotrophic micropropagation for sustainable production of plant species. J Sci Technol 49:25–32Google Scholar
  22. Niedz RP, Evens TJ (2008) The effects of nitrogen and potassium nutrition on the growth of nonembryogenic and embryogenic tissue of sweet orange (Citrus sinensis (L.) Osbeck). BMC Plant Biol 8:126.  https://doi.org/10.1186/1471-2229-8-126 CrossRefGoogle Scholar
  23. Niedz RP, Evens TJ (2017) Design of experiments (DOE)—history, concepts, and relevance to in vitro culture. In Vitro Cell Dev Biol Plant. 52:547–562CrossRefGoogle Scholar
  24. Niedz RP, Evens TJ, Hyndman SE, Adkins S, Chellemi DO (2011) In vitro shoot growth of Brugmansia × candida Pers. Physiol Mol Biol Plants 18:69–78CrossRefGoogle Scholar
  25. Niedz RP, Hyndman SE, Evens TJ, Weathersbee AA (2014) Mineral nutrition and in vitro growth of Gerbera hybrida (Asteraceae). In Vitro Cell Dev Biol Plant 50:458–470CrossRefGoogle Scholar
  26. Petersen RG (1985) Response surfaces. Design and analysis of experiments. Marcel Decker Inc., New York, pp 252–301Google Scholar
  27. Reed BM, Wada S, DeNoma J, Niedz RP (2013) Improving in vitro mineral nutrition for diverse pear germplasm. In Vitro Cell Dev Biol Plant 49:343–355CrossRefGoogle Scholar
  28. Smith MAL, Spomer LA (1995) Vessels, gels, liquid media, and support systems. In: Aitken-Christie J, Kozai T, Smith MAL (eds) Automation and environmental control in plant tissue culture. Kluwer Academic, Dordrecht, pp 371–404CrossRefGoogle Scholar
  29. Tascan A, Adelberg J, Tascan M, Joshee N, Yadav A (2010) Polyester fiber controlled hyperhydricity of Scutellaria species in in vitro Liquid Culture Systems. HortScience 45:1723–1728CrossRefGoogle Scholar
  30. Thorpe TA, Murashige T (1968) Some histochemical changes underlying shoot initiation in tobacco callus. Am J Bot 55:710Google Scholar
  31. Thorpe TA, Murashige T (1970) Some histochemical changes underlying shoot initiation in tobacco callus. Can J Bot 48:277–285CrossRefGoogle Scholar
  32. Thorpe T, Stasolla C, Yeung EC, De Klerk G-J, Roberts A, George EF (2008) The components of tissue culture media II: organic sdditions, osmotic and pH effects and support systems. In: George EF, Hall MA, De Klerk G-J (eds) Plant propagation by tissue culture, 3rd edn. Springer, Dordrecht, pp 115–174Google Scholar
  33. Van Staaten J, Zazimilova E, George EF (2008) Plant growth regulators: cytokinins and their analogues and antagonists. In: George EF, Hall MA, De Klerk G-J (eds) Plant propagation by tissue culture, 3rd edn. Springer, Dordrecht, pp 205–226Google Scholar
  34. Walker PN, Harris JP, Gautz LD (1991) Optimal environment for sugarcane micropropagation. Trans ASAE 34:2609–2614CrossRefGoogle Scholar
  35. Ziv M (1995) In vitro acclimatization. In: Aitken-Christie J, Kozai T, Smith MAL (eds) Automation and environmental control in plant tissue culture. Kluwer Academic, Dordrecht, pp 493–516CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Jeffrey William Adelberg
    • 1
    • 2
    Email author
  • Jacqueline Antionette Naylor-Adelberg
    • 1
  • Rabia Fawzi El-hawaz
    • 1
  1. 1.Department of Plant and Environmental SciencesClemson UniversityClemsonUSA
  2. 2.E143 Poole Agricultural CenterClemson UniversityClemsonUSA

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