Abstract
Increased market demand for high-quality Nordmann fir (Abies nordmanniana) Christmas trees, achieving reduced production costs and improved pest resistance, necessitates the establishment of efficient mass micropropagation platforms for the species. Somatic embryogenesis is a powerful tool for clonal propagation of woody plants. Utilizing the advantages of liquid culture media over solid media, the scalability of Nordmann fir somatic embryo proliferation was investigated through developing a batch culture using the disposable WAVE bioreactor (DWB) system, without feedback controls for pH and dissolved oxygen. This study revealed that the inoculation of a 10-L cellbag with 1 g L−1 FW (fresh weight) of A. nordmanniana embryogenic callus can produce ~ 70,000–111,000 vigorous and synchronized somatic embryos. Such outcomes were achieved with the rock speeds of 25 and 35 rpm and rock angle of 6° for 25 days at 24 ± 1 °C, respectively. The gas mix was a combination of O2 (21%) and a stepwise reduction of CO2 ended by 2% (v/v), at a flow rate of 0.2 L min−1. The application of 500 mg L−1 of 2-(N-morpholino)ethanesulfonic acid (MES) effectively stabilized the pH between 5.3 and 5.4. Furthermore, DWB was found an efficient system for accelerating and scaling up the proliferation of A. nordmanniana somatic embryos, as it is a high-yielding, user-friendly, functional, and rapid alternative to conventional methods with solid media.
This is a preview of subscription content, log in via an institution to check access.
Source of the figures: Operator’s manual of the system 20/50EHT (series). Part # 450026, Version: 10-2007, Type: 20/50EHT CE. WAVE/GE Healthcare Bioscience BioProcess Corp, NJ, USA
Similar content being viewed by others
References
Abiri R, Maziah M, Shaharuddin NA et al (2017) Enhancing somatic embryogenesis of Malaysian rice cultivar MR219 using adjuvant materials in a high-efficiency protocol. Int J Environ Sci Technol 14(78):1–18. https://doi.org/10.1007/s13762-016-1221-y
Abrahamsson M, Valladares S, Larsson E et al (2012) Patterning during somatic embryogenesis in Scots pine in relation to polar auxin transport and programmed cell death. Plant Cell Tiss Organ Cult 109:391–400. https://doi.org/10.1007/s11240-011-0103-8
Aizawa P, Benemar C, Wingenblixt C (2010) Conventional stirred bioreactor control system for monitoring and controlling pH and DO in a wave bioreactor. In: Noll T (ed) Cells and culture. ESACT proceedings, vol 4. Springer, Dordrecht, pp 823–828. https://doi.org/10.1007/978-90-481-3419-9_144
Blombach B, Takors R (2015) CO2—intrinsic product, essential substrate, and regulatory trigger of microbial and mammalian production processes. Front Bioeng Biotechnol. https://doi.org/10.3389/fbioe.2015.00108
Boisson AM, Gout E, Bligny R et al (2012) A simple and efficient method for the long-term preservation of plant cell suspension cultures. Plant Methods 8:4. https://doi.org/10.1186/1746-4811-8-4
Bonga JM, Aderkas PV, Klimaszewska K (2009) Conifers: culture and genetic engineering. In: Flickinger MC (ed) Encyclopedia of industrial biotechnology: bioprocess, bioseparation, and cell technology. Wiley, Hoboken, pp 1–5. https://doi.org/10.1002/9780470054581.eib240
Businge E, Trifonova A, Schneider C et al (2017) Evaluation of a new temporary immersion bioreactor system for micropropagation of cultivars of eucalyptus, birch and fir. Forests 8:196. https://doi.org/10.3390/f8060196
Chen CC, Bates R, Carlson J (2015) Effect of environmental and cultural conditions in medium pH and plant growth performance of Douglas-fir (Pseudotsuga menziesii) shoot culture. F1000Res 3:298. https://doi.org/10.12688/f1000research.5919.2
Corredoira E, Ballester A, Vieitez AM (2003) Proliferation, maturation and germination of Castanea sativa Mill. somatic embryos originated from leaf explants. Ann Bot 92(1):129–136. https://doi.org/10.1093/aob/mcg107
Deb CR, Tandon P (2004) Establishment of an embryogenic suspension culture for Pinus kesiya (Khasi pine) from various explants. Indian J Biotechnol 3:445–448
De-la-Peña C, Nic-Can GI, Galaz-Ávalos RM (2015) The role of chromatin modifications in somatic embryogenesis in plants. Front Plant Sci 6:635. https://doi.org/10.3389/fpls.2015.00635
dos Santos ALW, Silveira V, Steiner N et al (2010) Biochemical and morphological changes during the growth kinetics of Araucaria angustifolia suspension cultures. Braz Arch Biol Technol 53:497–504. https://doi.org/10.1590/S1516-89132010000300001
Ducos JP, Lambot C, Petiard V (2007) Bioreactors for coffee mass propagation by somatic embryogenesis. Int J Plant Dev Biol 1(1):1–12
Ducos JP, Terrier B, Courtois D (2009) Disposable bioreactors for plant micropropagation and mass plant cell culture. Adv Biochem Eng Biotechnol 115:89–115. https://doi.org/10.1007/10_2008_28
Eibl R, Kaiser S, Lombriser R (2010) Disposable bioreactors: the current state-of-the-art and recommended applications in biotechnology. Appl Microbiol Biotechnol 86:41–49. https://doi.org/10.1007/s00253-009-2422-9
Eibl R, Meier P, Stutz I et al (2018) Plant cell culture technology in the cosmetics and food industries: current state and future trends. Appl Microbiol Biotechnol 102(20):8661–8675. https://doi.org/10.1007/s00253-018-9279-8
Etienne H, Dechamp E, Barry-Etienne D et al (2006) Bioreactors in coffee micropropagation. Braz J Plant Physiol 18(1):45–54. https://doi.org/10.1590/S1677-04202006000100005
Felle HH (2005) pH regulation in anoxic plants. Ann Bot 96:519–532. https://doi.org/10.1093/aob/mci207
Find J, Grace L, Krogstrup P (2002) Effect of anti-auxins on maturation of embryogenic tissue cultures of Nordmanns fir (Abies nordmanniana). Physiol Plant 116:231–237. https://doi.org/10.1034/j.1399-3054.2002.1160213.x
Gakpetor PM, Mohammed H, Moreti D et al (2017) Periclinal chimera technique: new plant breeding approach. Genet Mol Res 6(3):21. https://doi.org/10.4238/gmr16039790
Gardner M, Knees S (2011) Abies nordmanniana ssp. nordmanniana. The IUCN red list of threatened species. e.T196098A8995492. https://doi.org/10.2305/iucn.uk.2011-2.rlts.t196098a8995492.en
Ghasemi A, Bozorg A, Rahmatib F et al (2019) Comprehensive study on Wave bioreactor system to scale up the cultivation of and recombinant protein expression in baculovirus-infected insect cells. Biochem Eng J 143:121–130. https://doi.org/10.1016/j.bej.2018.12.011
Giusti S, Mazzei D, Cacopardo L et al (2017) Environmental control in flow bioreactors. Processes 5:16. https://doi.org/10.3390/pr5020016
Gorbatenko O, Hakman I (2001) Desiccation-tolerant somatic embryos of Norway spruce (Picea abies) can be produced in liquid cultures and regenerated into plantlets. Int J Plant Sci 162(6):1211–1218. https://doi.org/10.1086/323416
Guan Y, Li S-G, Fan X-F (2016) Application of somatic embryogenesis in woody plants. Front Plant Sci 7:938. https://doi.org/10.3389/fpls.2016.00938
Guillou C, Fillodeau A, Brulard E et al (2018) Indirect somatic embryogenesis of Theobroma cacao L. in liquid medium and improvement of embryo-to-plantlet conversion rate. Vitro Cell Dev Biol Plant 54:377–391. https://doi.org/10.1007/s11627-018-9909-y
Himanen K, Lilja A, Rytkönen A et al (2013) Soaking effects on seed germination and fungal infection in Picea abies. Scand J For Res 28(1):1–7. https://doi.org/10.1080/02827581.2012.683037
Hussain SS, Rao AQ, Husnain T et al (2009) Cotton somatic embryo morphology affects its conversion to plant. Biol Plant 53:307. https://doi.org/10.1007/s10535-009-0055-6
Hvoslef-Eide AK, Olsen OAS, Lyngved R (2005) Bioreactor design for propagation of somatic embryos. Plant Cell Tiss Org Cult 81:265–276. https://doi.org/10.1007/s11240-004-6647-0
Jalili M, Ghasemi MR, Pifloush AR (2018) Stiffness and strength of granular soils improved by biological treatment bacteria microbial cements. Emerg Sci J 2(4):219–227. https://doi.org/10.28991/esj-2018-01146
Kim YW, Newton R, Frampton J (2009) Embryogenic tissue initiation and somatic embryogenesis in Fraser fir (Abies fraseri [Pursh] Poir.). Vitro Cell Dev Biol Plant 45:400. https://doi.org/10.1007/s11627-008-9169-3
Kliphuis AMJ, Martens DE, Janssen M et al (2011) Effect of O2:CO2 ratio on the primary metabolism of Chlamydomonas reinhardtii. Biotechnol Bioeng 108(10):2390–2402. https://doi.org/10.1002/bit.23194
Konar S, Karmakar J, Ray A (2018) Regeneration of plantlets through somatic embryogenesis from root derived calli of Hibiscus sabdariffa L. (Roselle) and assessment of genetic stability by flow cytometry and ISSR analysis. PLoS ONE 13(8):e0202324. https://doi.org/10.1371/journal.pone.0202324
Kumar V, Van Staden J (2017) New insights into plant somatic embryogenesis: an epigenetic view. Acta Physiol Plant 39:194. https://doi.org/10.1007/s11738-017-2487-5
Lehmann N, Dittler I, Lämsä M et al (2014) Disposable bioreactors for cultivation of plant cell cultures. In: Paek KY, Murthy H, Zhong JJ (eds) Production of biomass and bioactive compounds using bioreactor technology. Springer, Dordrecht, pp 17–46. https://doi.org/10.1007/978-94-017-9223-3_2
Matsunaga N, Kano K, Maki Y (2009) Culture scale-up studies as seen from the viewpoint of oxygen supply and dissolved carbon dioxide stripping. J Biosci Bioeng 107(4):412–418. https://doi.org/10.1016/j.jbiosc.2008.12.016
Mikola M, Seto J, Amanullah A (2007) Evaluation of a novel Wave bioreactor cellbag for aerobic yeast cultivation. Bioprocess Biosyst Eng 30:231–241. https://doi.org/10.1007/s00449-007-0119-y
Naciri M, Kuystermans D, Al-Rubeai M (2008) Monitoring pH and dissolved oxygen in mammalian cell culture using optical sensors. Cytotechnology 57:245–250. https://doi.org/10.1007/s10616-008-9160-1
Nawrot-Chorabik K (2016) Plantlet regeneration through somatic embryogenesis in Nordmann’s fir (Abies nordmanniana). J For Res 27(6):1219–1228. https://doi.org/10.1007/s11676-016-0265-7
Niazian M, Sadat-Noori SA, Galuszk P (2017) Genetic stability of regenerated plants via indirect somatic embryogenesis and indirect shoot regeneration of Carum copticum L. Ind Crops Prod 97:330–337. https://doi.org/10.1016/j.indcrop.2016.12.044
Nic-Can GI, López-Torres A, Barredo-Pool F (2013) New insights into somatic embryogenesis: LEAFY COTYLEDON1, BABY BOOM1 and WUSCHEL-RELATED HOMEOBOX4 are epigenetically regulated in Coffea canephora. PLoS ONE 8(8):e72160. https://doi.org/10.1371/journal.pone.0072160
Nielsen UB, Chastagner GA (2005) Variation in postharvest quality among Nordmann fir provenances. Hort Sci 40(3):553–557. https://doi.org/10.21273/HORTSCI.40.3.553
Nielsen UB, Hansen OK (2010) Response to selfing in seed set, seedling establishment and nursery growth based on controlled crosses of Abies nordmanniana clones. Silvae Genetica 59:2–3. https://doi.org/10.1515/sg-2010-0011
Nielsen UB, Hansen OK (2012) Genetic worth and diversity across 18 years in a Nordmann fir clonal seed orchard. Ann For Sci 69:69–80. https://doi.org/10.1007/s13595-011-0159-y
Nielsen UB, Kirkeby-Thomsen A, Roulund H (2002) Genetic variation in resistance to Dreyfusia nordmannianae Eckst. infestations in Abies nordmanniana (Stev.) Spach. For Ecol Manag 165:271–283. https://doi.org/10.1016/S0378-1127(01)00659-4
Nielsen UB, Hansen JK, Kromann HK (2011) Impact of site and provenance on economic return in Nordmann fir Christmas tree production. Scand J For Res 26(2):74–89. https://doi.org/10.1080/02827581.2010.526955
Nodoushan EJ (2018) Monthly forecasting of water quality parameters within Bayesian networks: a case study of Honolulu, Pacific Ocean. Civil Eng J 4(1):188–199. https://doi.org/10.28991/cej-030978
Nørgaard JV (1997) Somatic embryo maturation and plant regeneration in Abies nordmanniana Lk. Plant Sci 124:211–221. https://doi.org/10.1016/S0168-9452(97)04614-1
Nørgaard JV, Krogstrup P (1991) Cytokinin induced somatic embryogenesis from immature embryos of Abies nordmanniana Lk. Plant Cell Rep 9:509–513. https://doi.org/10.1007/BF00232107
Paek KY, Chakrabarty D, Hahn EJ (2005) Application of bioreactor systems for large scale production of horticultural and medicinal plants. Plant Cell Tiss Org Cult 81:287–300. https://doi.org/10.1007/s11240-004-6648-z
Prahadeesh N, Sithambaresan M, Mathiventhan U (2018) A study on hydrogen peroxide scavenging activity and ferric reducing ability of simple coumarins. Emerg Sci J 2(6):417–427. https://doi.org/10.28991/esj-2018-01161
Pullman GS, Olson K, Fischer T (2016) Fraser fir somatic embryogenesis: high frequency initiation, maintenance, embryo development, germination and cryopreservation. New For 47:453–480. https://doi.org/10.1007/s11056-016-9525-9
Reeves C, Hargreaves C, Trontin JF et al (2018) Simple and efficient protocols for the initiation and proliferation of embryogenic tissue of Douglas-fir. Trees 32:175–190. https://doi.org/10.1007/s00468-017-1622-7
Salaj T, Blehová A, Salaj J (2007) Embryogenic suspension cultures of Pinus nigra Arn: growth parameters and maturation ability. Acta Physiol Plant 29:225–231. https://doi.org/10.1007/s11738-007-0028-3
Scholz J, Suppmann S (2016) A re-usable wave bioreactor for protein production in insect cells. MethodsX 3:497–501. https://doi.org/10.1016/j.mex.2016.08.001
Singh V (1999) Disposable bioreactor for cell culture using wave-induced agitation. Cytotechnology 30:149–158. https://doi.org/10.1023/A:1008025016272
Sluis CJ (2005) Integrating automation technologies with commercial micropropagation. In: DuttaGupta S, Ibaraki Y (eds) Plant tissue culture engineering. Springer, Dordrecht, pp 231–251. https://doi.org/10.1007/978-1-4020-3694-1_13
Sun L (2018) Effect of cyclic stress level and overconsolidation ratio on permanent deformation behaviour of clayey subsoil. Civil Eng J 4(8):1772–1782. https://doi.org/10.28991/cej-03091113
Terrier B, Courtois D, Hénault N et al (2007) Two new disposable bioreactors for plant cell culture: the wave and undertow bioreactor and the slug bubble bioreactor. Biotechnol Bioeng 96(5):914–923. https://doi.org/10.1002/bit.21187
Valdiani A, Hansen OK, Nielsen UB et al (2019) Bioreactor-based advances in plant tissue and cell culture: challenges and prospects. Critic Rev Biotech 39(1):20–34. https://doi.org/10.1080/07388551.2018.1489778
Vooková B, Kormut’ák A (2007) Abies biotechnology—research and development of tissue culture for vegetative propagation. Tree For Sci Biotechnol 1(1):39–46
Wamelink GWW, van Dobben HF, Goedhart PW et al (2018) The role of abiotic soil parameters as a factor in the success of invasive plant species. Emerg Sci J 2(6):308–365. https://doi.org/10.28991/esj-2018-01155
Wang H, Zeuschner J, Eremets M (2016) Stable solid and aqueous H2CO3 from CO2 and H2O at high pressure and high temperature. Sci Rep 6:19902. https://doi.org/10.1038/srep19902
Warren R, Johnson EW (1988) A guide to the firs (Abies spp.) of the Arnold Arboretum. Arnoldia 48:3–48
Weber W, Weber E, Geisse S (2002) Optimisation of protein expression and establishment of the Wave bioreactor for baculovirus/insect cell culture. Cytotechnology 38:77–85. https://doi.org/10.1023/A:1021102015070
Westbrook A, Scharer J, Moo-Young M et al (2014) Application of a two-dimensional disposable rocking bioreactor to bacterial cultivation for recombinant protein production. Biochem Eng J 88:154–161. https://doi.org/10.1016/j.bej.2014.04.011
Wetzstein HY, Baker CM (1993) The relationship between somatic embryo morphology and conversion in peanut (Arachis hypogaea L.). Plant Sci 92(1):81–89. https://doi.org/10.1016/0168-9452(93)90068-b
Yuk IH, Baskar D, Duffy PH (2011) Overcoming challenges in Wave bioreactors without feedback controls for pH and dissolved oxygen. Biotechnol Prog 27(5):1397–1406. https://doi.org/10.1002/btpr.659
Zhan C, Hagrot E, Brandt L et al (2019) Study of hydrodynamics in wave bioreactors by computational fluid dynamics reveals a resonance phenomenon. Chem Eng Sci 193:53–65. https://doi.org/10.1016/j.ces.2018.08.017
Acknowledgement
The authors would like to appreciate the Green Development and Demonstration Program [Grønt Udviklings- og Demonstrationsprogram (GUDP) - Grant number: 34009-16-1081] of the Danish Ministry of Food, Agriculture and Fisheries for creating an opportunity to conduct the present article. Furthermore, the deceased senior scientist Jens Find, formerly Associate Professor at the Section for Forest, Nature and Biomass – University of Copenhagen, is gratefully acknowledged for his effort in organizing the Cell and Tissue Culture Laboratory as well as for preparing the bioreactor platform. Our appreciation is extended to the lab technicians Lisbeth Hansen and Nanna Jacobsen as well as the laboratory coordinator of the section “Preben Frederiksen” for their assistance in the present project. Our special thanks go to Janne Simola from GE Healthcare for his sincere assistance in initial setup of the WAVE bioreactor.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Editorial responsibility: Parveen Fatemeh Rupani.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Valdiani, A., Hansen, O.K., Johannsen, V.K. et al. An efficient bioreactor platform for scaling up the proliferation of Nordmann fir’s (Abies nordmanniana) somatic embryos. Int. J. Environ. Sci. Technol. 17, 1425–1438 (2020). https://doi.org/10.1007/s13762-019-02556-4
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13762-019-02556-4