The Potential Use of Ectoine Produced by a Moderately Halophilic Bacteria Chromohalobacter salexigens KT989776 for Enhancing Germination and Primary Seedling of Flax “Linum usitatissimum L.” under Salinity Conditions

Main Article Content

Tamer A. Elsakhawy
Nashwa, A. H. Fetyan
Azza A. Ghazi

Abstract

The similarity between plant and microbial cells encourage the use of microbial metabolites of halophilic bacteria for the alleviation of salt stress in plants. In the current research work, a compatible solute ectoine extracted from a moderately halophilic bacteria Chromohalobacter salexigens KT989776 was used to enhance flax germination and primary seedling under different levels of salinity. Two successive experiments including germination in Petri plates under six levels of salinity (0, 3, 5, 7, 9 and 11 dS.m-1) and a pot experiment under three irrigating water salinity levels (2, 3 and 4) with two types of ectoine application (spray and soil addition) were conducted. Germination parameters were recorded for the first experiment while a fresh and dry weight of plants and peroxidase activity in addition to sodium-potassium ratio were estimated in the pot experiment. Also, ectoine accumulation in plants was detected using HPLC. Results of LC-MS proved the production of ectoine by C. salexigens KT989776 and ectoine enhanced significantly all germination parameters of flax seeds, decreased sodium accumulation in the plant, increased potassium content, and lowered peroxidase and phenoloxidase activity. Also, HPLC analysis proved that ectoine was detected in all treated samples while not detected in non-treated control.

Keywords:
Halophilic, chromohalobacter, compatible solutes, ectoine, flax, germination.

Article Details

How to Cite
Elsakhawy, T. A., Fetyan, N. A. H., & Ghazi, A. A. (2019). The Potential Use of Ectoine Produced by a Moderately Halophilic Bacteria Chromohalobacter salexigens KT989776 for Enhancing Germination and Primary Seedling of Flax “Linum usitatissimum L.” under Salinity Conditions. Biotechnology Journal International, 23(3), 1-12. https://doi.org/10.9734/bji/2019/v23i330078
Section
Original Research Article

References

Hernández JA. Salinity tolerance in plants: Trends and perspectives. Int. J. Mol. Sci. 2019;20: 2408.
DOI: 10.3390/ijms20102408

Boyer JS. Plant productivity and environment. Science. 1982;218:443–448.

Wani SH, Kumar V, Shriram V, Sah SK. Phytohormones and their metabolic engineering for abiotic stress tolerance in crop plants. Crop J. 2016;4(3):162–176.
DOI: 10.1016/j. cj.2016.01.010

Brown AD. Microbial water stress. Bacteriol Rev. 1976;40:803–846.

Nakayama H, Yoshida K, Ono H, Murooka Y, Shinmyo A. Ectoine, the compatible solute of Halomonas elongata, confers hyperosmotic tolerance in cultured tobacco cells. Plant Physiol. 2000;122: 1239–1248.
Available:https://doi.org/10.1104/pp.122.4.1239

Kathura H, Giri J, Nataraja KN, Murata N, Udayakumar M, Tyagi AK. Glycinebetaine-induced water-stress tolerance in coda- expressing transgenic indica rice is associated with up-regulation of several stress responsive genes. Plant Biotechnol. J. 2009;7:512-26.

Kraegeloh A, Kunte HJ. Novel insights into the role of potassium for OSMO regulation in Halomonas elongate. Extremophiles. 2002;6:453–462.

Galinski EA, Truper HG. Microbial behaviour in salt-stressed ecosystems. FEMS Microbiol. Rev. 1994;15:95–108.

Kai MC, Sowers KR, Robertson DE, Roberts MF, Gunsalus RP. Distribution of compatible solutes in the halophilic methanogenic archaebacteria. Bacteriol. 1991;173:5352–5358.

Bremer E, Kramer R. Coping with osmotic challenges: Osmoregulation through accumulation and release of compatible solutes in bacteria. Storz, G. Hengge-Aronis, R. (Eds.), Bact. Stress responses, ASM Press Washington D C. 2000;79–97.

Singh BP, Rateb ME, Rodriguez-Couto S, Polizeli Md LTDM, Li WJ. Microbial secondary metabolites: Recent developments and technological challenges. Front. Microbiol. 2019;10:914.
DOI: 10.3389/fmicb.2019.00914

Grammann K, Volke A, Kunte HJ. New type of osmoregulated solute transporter identified in halophilic members of the bacteria domain: TRAP transporter TeaABC mediates uptake of ectoine and hydroxyectoine in Halomonas elongata DSM2581T. Bacteriol. 2002;184:3078–3085.

Oren A, Larimer F, Richardson P, Lapidus A, Csonka LN. How to be moderately halophilic with broad salt tolerance: Clues from the genome of Chromohalobacter salexigens. Extremophiles. 2005;9:275–279.

Berti M, Fischer R, Wilckens F, Hevia B J. Adaptation and genotype × environment interaction of flax seed (Linum usitatissimum L.) genotypes in South Central Chile. Chil J Agri Res. 2010; 70:345–356.

El-Nagdy GA, Nassar EA. et al. Response of flax plant (Linum usitatissimum L.) to treatments with mineral and bio-fertilizers from nitrogen and phosphorus. Ame. Sci. 2010;6:207–217.

Kurt O, Bozkurt D. Effect of temperature and photoperiod on seedling emergence of flax (Linum usitatissimum L.). Agro. 2006;5:541–545.

Isayenkov SV, Maathuis FJM. Plant salinity stress: Many unanswered questions remain. Front., Plant Sci. 2019;10:80.
DOI: 10.3389/fpls.2019.00080

Muhammad Z, Husain F. Effect of NaCl salinity on the germination and seedling growth of some medicinal plants. Pakistan J Bot. 2010;42:889–897.

Mondal P, Remme RN, Das D, Ali Y, Kabir E. Germination and seedling growth of indigenous Aman rice under NaCl salinity. Int J Multidiscip Res Dev. 2015;2:251–257.

Nasri NR, Kaddour H, Mahmoudi O, Baâtour NB, Lachaâl M. The effect of osmopriming on germination, seedling growth and phosphatase activities of lettuce under saline condition. Afr J Biotech. 2011;10:14366–14372.

Husseiny SM, Sheref F, Amer H, Elakhawy TA. Biological activity of chemically modified levan produced by moderately halophilic chromohalobacter salexigens KT989777. Middle East J Appl Sci. 2015;5:812–822.

Sehgal SN, Gibbons NE. Effect of metal ions on the growth of Halobacter iumcutirubrum. Can J Microbiol. 1960;5:165–169.

Zhang LH, Lang YJ, Nagata S. Efficient production of ectoine using ectoine-excreting strain. Extremophiles. 2009;13:717–724.
Available:https://doi.org/10.1007/s00792-009-0262-2

Kader MA. A comparison of seed germination calculation formulae and the associated interpretation of resulting data. R Soc New South Wales. 2005;138:65–75.

Black AC, Evans DD, White JL, Ensminyer EL, Clark EF. Methods of soil analysis Amer. Soc Agro Inc Madison Wisconsin, USA; 1965.

Moghaieb RE, Saneoka H, Yossef SS, El-sharkawy AM, Fujita K. Improvement of salt tolerance in tomato plant (Lycopersicon esculentum) by transformation with ectoine biosynthetic genes. Transgenic Plant J. 2007;1:228–232.

Kar M. Mishra D. Catalase, peroxidase, and polyphenoloxidase activities during rice leaf senescence. Plant Physiol. 1976; 57(2):315-9.

Galinski EA, Pfeiffer HP, Trüper HG. 1,4,5,6,-Tetrahydro-2-methyl-4- pyridinecarboxylic acid. A novel cyclic amino acid from halophilic phototrophic bacteria of the genus Ectothiorhodospira. Eur J Biochem. 1985;149:135–139.

Krasilnikov N. The role of soil bacteria in plant nutrition. J Gen Appl. Microbiol. 1961;7:128–144.
Available:https://doi.org/10.2323/jgam.7.128

Bradáčová K, Weber NF, Morad-Talab N, Asim M, Imran M, Weinmann M, Neumann G. Micronutrients (Zn/Mn), seaweed extracts, and plant growth-promoting bacteria as cold-stress protectants in maize. Chem Biol Technol Agric. 2016;3:1–10.
Available:https://doi.org/10.1186/s40538-016-0069-1

Mehta P, Walia A, Kulshrestha S, Chauhan A, Shirkot CK. Efficiency of plant growth-promoting P-solubilizing Bacillus circulans CB7 for enhancement of tomato growth under net house conditions. J Basic Microbiol. 2015;55:33–44.
Available:https://doi.org/10.1002/jobm.201300562

Singh R, Kumar M, Mittal A, Mehta PK. Microbial metabolites in nutrition, healthcare and agriculture. Biotech. 2017;7:1–14.
Available:https://doi.org/10.1007/s13205-016-0586-4

Chen THH, Murata N. Enhancement of tolerance of abiotic stress by metabolic engineering of betaines and other compatible solutes. Curr Opin Plant Biol. 2002;5:250–257.
Available:https://doi.org/10.1016/S1369-5266(02)00255-8

da Costa MS, Santos H, Galinski EA. An overview of the role and diversity of compatible solutes in bacteria and archaea. Adv Biochem Eng Biotechnol. 1998;61:117–153.

Lentzen G, Schwarz T. Extremolytes: Natural compounds from extremophiles for versatile applications. Appl Microbiol Bio- Technol. 2006;72:623–634.

Lippert K, Galinski EA. Enzyme stabilization by ectoine type compatible solutes: Protection against heating, freezing and drying. Appl Microbiol Biotechnol. 1992;37:61-65.

Pastor JM, Salvador M, Argandoña M, Bernal V, Reina-Bueno M, Csonka LN, Iborra JL, Vargas C, Nieto JJ, Cánovas M. Ectoines in cell stress protection: Uses and biotechnological production. Biotechnol Adv. 2010;28:782–801.
Available:https://doi.org/10.1016/j.biotechadv.2010.06.005

He YZ, Gong J, Yu HY, Tao Y, Zhang S, Dong ZY. High production of ectoine from aspartate and glycerol by use of whole-cell biocatalysis in recombinant Escherichia coli. Microb Cell Fact. 2015;14:1–10.
Available:https://doi.org/10.1186/s12934-015-0238-0

Moghaieb RE, Nakamura A, Saneoka H, Fujita K. Evaluation of salt tolerance in ectoine-transgenic tomato plants (Lycopersicon esculentum) in terms of photosynthesis, osmotic adjustment, and carbon partitioning. GM Crops. 2011;2:58–65.
Available:https://doi.org/10.4161/gmcr.2.1.15831