In vitro Micropropagation of Duckweed (Lemna minor L) Plant with Temporary Immersion System Bioreactors

Abstract

Which is very rich in protein in plant Lemna minor L. abundant in Turkey ecologically plays a very important role in protecting the elimination of water pollution and aquaculture environment balance. In this study, in vitro propagation of this plant with Temporary Immersion System Bioreactor and determination of the effects of used plant growth regulators on the protein content of the plant were aimed. With this objective different variant of media with and without sucrose, varying pH and concentrations of BAP, kinetin, TDZ in medium were analyzed. Experiments for micropropagation were performed for 8 hours in the dark and 16 hours in white fluorescent light (150 µ mol photons m-2s-1) under photoperiod and at 24 ± 1 ° C. The highest plant growth was observed at pH 7.23 in sugar free liquid MS medium containing 0.2 mg/L BAP. 50.44 plants per explant were recorded in this medium. In addition, the maximum number of plants per explant in liquid MS medium containing 0.05 mg/L kinetin was calculated as 57,823 and the maximum number of plants per explant in liquid MS medium containing 0.6 mg/L TDZ was calculated as 50.74. As a result of nitrogen determination studies with Kjeldahl method, the protein value of the plant was determined as 25.5%. As a result of hormone application, it was seen that protein content in plant increased to 29.18% with 0.5 mg/L BAP. It was concluded that the aim of the study were fulfilled and positive effects of Temporary Immersion System Bioreactors on plant multiplication were found.

Keywords

• In vitro
• Temporary Immersion System Bioreactors
• Lemna minor L.
• Micropropagation

References

1. 1. Bornette G and Puijalon S (2011). Response of aquatic plants to abiotic factors: a review. Aquat. Sci., 73:1–14. doi: 10.1007/s00027-010-0162-7
2. 2. Khataee AR, Movafeghi A, Torbati S, Salehi Lisar SY, Zarei M (2012). Phytoremediation potential of duckweed (Lemna minör L.) in degradation of C.I. Acid Blue 92: Artificial neural network modeling. Ecotoxicol. Environ. Saf., 80:291–298
3. 3. Wang WC, Freemark K (1995). The use of plants for environmental monitoring and assessment. Ecotoxicol. Environ. Saf., 30: 289-301
4. 4. Steven MD, Egbert H van N, Rudi MMR (2005). Growth limitation of Lemna minor due to high plant density. Aquat. Bot., 81: 245–251
5. 5. Rusoff LL, Blakeney Jr. EW, Culley Jr. DD (1980). Duckweeds (Lemnaceae family): a potential source of protein and amino acids. J. Agric. Food Chem., 28(4): 848-850
6. 6. Nayyef MA and Amal AS(2012). Effıcıency of duckweed (Lemna mınor L.) in phytotreatment of wastewater pollutants from basrah oil refınery. J. of Appl. Phytotech. in Environ. Sanitation, 1 (4): 163-172
7. 7. Uysal Y (2013). Removal of chromium ions from wastewater by duckweed, Lemna minor L. by using a pilot system with continuous flow. J. Hazard. Mater., 263: 486–492
8. 8. Cox KM, Sterling DJ and Regan JT (2006).Glycan optimization of a human monoclonal antibody in the aquatic plant Lemna minor. Nat. Biotechnol., 12: 1591–1597

9. 9. Brown DCW and Thorpe TA (1995). Crop improvement through tissue culture. World Journal of Microbiology and Biotechnology, 11: 409–415
10. 10. Khvatkov P, Chernobrovkina M, Okuneva A, Dolgov S (2019). Creation of culture media for efficient duckweeds micropropagation (Wolffia arrhiza and Lemna minor) using artificial mathematical optimization models. Plant Cell Tiss Org Cult 136: 85–100
11. 11. Etienne H, Berthouly M (2002).Temporary immersion systems in plant micropropagation. Plant Cell Tiss. Org. Cult., 69: 215–231
12. 12. Aitken-Christie J and Davies HE (1988). Development of a semi-automated micropropagation system. Acta Hortic., 230, 81-88 doi: 10.17660/ActaHortic.1988.230.7
13. 13. Harris RE and Mason EBB (1983). Two machines for in vitro propagatıon of plants in liquid media. Can. J. Plant Sci., 63(1): 311-316, doi: 10.4141/cjps83-032
14. 14. Yenice Z (2010). Micropropagation of common duckweed (Lemna minor L.) plants using temporary immersion system bioreactors. Master Thesis, Ankara University Biotechnology Institute, Ankara, Turkey
15. 15. Bradstreet RB (1954). Kjeldahl method for organic nitrogen. Anal. Chem., 26(1): 185-187 doi: 10.1021/ac60085a028
16. 16. Snedecor GW and Cochran WG (1967) Statistical Methods. 6th edn. Ames Iowa, Iowa State University Press
17. 17. Scott AJ and Knott M (1974). A cluster analysis method for grouping means in the analysis of variance. Biometrics, 30: 507-512
18. 18. Duncan DB (1955). Multiple range and multiple F tests. Biometrics, 11:1-42
19. 19. Artan RO (2007). Use of duckweed (Lemna sp) for further treatment of heavy metal containing wastewater. Master's thesis. Çukurova University. Institute of Science. Adana, Turkey
20. 20 Memmon A (2008). Elimination of Oil, Hydrocarbon and Pollution by Plant, Algae and Microoganisms. Science and Technology, January number, p 7-8
21. 21. Cook DA, Decker DM and Gallagher JL (1989). Rejeneration of Kosteletzkya virginica (L.) Presl. (Seashore Mallow) from callus cultures. Plant Cell Tiss Org Cult., 17; 111-119
22. 22. Straub PF, Decker DM and Gallagher JL (1988). Tissue culture and long-term regeneration of Phragmites australis (Cav.) Trin. Ex Steud. Plant Cell Tiss Org Cult., 15; 73-78
23. 23. Agrawal A and Mohan Ram HY (1995). In vitro germination and micropropagation of water chestnut (Trapa sp.). Aquatic Botany, 51; 135-146 24. Simon D and Helliwell S (1998). Extraction and quantification of chlorophyll a from freshwater green algae, Wat. Res., 32:2220-2223
24. 25. Kim KW, Jang GW (2004). Micropropagation of Drosera peltata, a tuberous sundew, by shoot tip culture. Plant Cell Tiss Org Cult., 77: 211–214
25. 26. Perica MC and Berljak J (1996). In vitro growth and regeneration of Drosera spatulata Labill on various media. Hortscience 31: 1033–1034
26. 27. Simola LK (1978). The effect of several amino acids and some inorganic nitrogen sources on the growth of Drosera rotundifolia in long and short-day condition. Z. Pflanzenphysiol. 90: 61–68
27. 28. Houllou-Kido LM, Costa AF, Lira MA, Farias I, Santos DC, Silva KS, Rivas R, Dias ALF, Alves GD (2009).Viability of Nopalea cochenilifera (cv. Ipa Sertânia) photoautotrophic micropropagation. VI International Congress on Cactus Pear and Cochineal. Acta Hort., 811: 309-314, doi: 10.17660/ActaHortic.2009.811.42
28. 29. Steward FC, Caplin S & Millar FK (1952). Investigations on growth and metabolism of plant cells. I. New techniques for the investigation of metabolism, nutrition and growth in undifferentiated cells. Ann. Bot. 16: 57–77
29. 30. Escalant JV, Teisson C, Cote F (1994). Amplified somatic embriyogenesis from male flowers of triploid banana and plantain cultivars (Musa spp). In Vitro Cell. Dev. Biol. 30: 181-186