Identification of Bacteria Obtained from Dactylorhiza urvilleana Rhizoid Region, Metal Tolerances, Bioremidant Characteristics and Effects on Maize Germination in Copper Presence

Abstract

Rapidly increasing industrialization and technological developments cause hazardous wastes to spread to the environment at a high rate. In our study, three bacterial (5O1, 5O8, 112O1) strains isolated from the rhizoid region of the orchid plant (Dactylorhiza urvilleana) were characterized by conventional and molecular methods (nuclear 16S rDNA). In order to characterize the isolates, primarily macroscopic, microscopic, some biochemical and physical properties of the strains were investigated. The usability of the strains screened for their general properties as bioremediation strains, in the prevention of high copper accumulation in agricultural soils was investigated. With traditional and molecular studies, two of the strains were defined as species level (Bacillus mycoides, B. popilliae) and one at genus level. They were determined that strains were tolerant to the tested all metal salts (Fe, Zn, Cu, Pb and Ag salts in the 1-10 mM range) except for the 5O1 strain to Ag salt, and 112O1 strain to Zn salt. The highest copper tolerance was observed in 5O1, 112O1 and 5O8 strains, respectively. The strains were determined that the copper minimum inhibition concentration values were 12.5-25 mM and the minimum bactericidal concentration value was 50 mM. The examined in terms of properties such as Indole-3-acetic acid (IAA), 1-aminocyclopropane-1-carboxylic acid (ACC) Deaminase activities, and phosphate solubility, it was determined that they promoted plant germination and growth. When the germination success of maize seeds in the presence of copper was examined, it was concluded that positive results were obtained, there was no significant difference between strains and therefore strains could be used in copper bioremediation.

Keywords

• Bacillus
• Metal tolerance,
• Bioremediation,
• Corn germination

References

1. Afsharmanesh H, Ahmadzadeh M, Majdabadi A, Motamedi F, Behboudi K & Javan-Nikkhah M (2013). Enhancement of biosurfactants and biofilm production after gamma irradiation induced mutagenesis of Bacillus subtilis UTB1, a biocontrol agent of Aspergillus flavus. Archives of Phytopathology and Plant Protection 46: 1874-1884.
2. Aydoğan M N, Algur Ö M & Özdemir M (2013). Isolation and characterisation of some bacteria and microfungus solving tricalcium phosphate. Atatürk Üniversitesi Biyoloji Bölümü 1: 11-20.
3. Baker G C, Smith J J & Cowan D A (2003). Review and re-analysis of domain-specific 16S primers. Journal Microbiol Methods 55: 541-555.
4. Cetinkaya Dönmez G, Aksu Z, Ozturk A & Kutsal T (1999). A comparative study on heavy metal biosorption characteristics of some Algae. Process Biochemistry 34: 885-892.
5. Chen X H, Koumoutsi A & Scholz R (2007). Comparative analysis of the complete genome sequence of the plant growth-promoting bacterium Bacillus amyloliquefaciens FZB42. Nat Biotechnol 25(9), 1007-1014.
6. Cheung K H & Gu A J D (2005). Chromate reduction by Bacillus megaterium TKW3 isolated from marine sediments. World J Microbiol Biotechnol 21: 213-219.
7. Das S N, Dutta S & Kondreddy A (2010). Plant growth-promoting chitinolytic Paenibacillus elgii responds positively to Tobacco root exudates. J Plant Growth Regul 29(4): 409-418.
8. Das, S N & Chandran P (2011). Biodegradation of petroleum sludge and microbial degradation of petroleum hydrocarbon contaminants: an overview, review article. SAGE-Hindawi Access to Research Biotechnology Research International, 13 Article ID 941810.

9. Dworken M & Foster J (1958). Experiments with some microorganisms which utilize ethane and hydrogen. J Bacteriol 75: 592-601.
10. Esertaş Ü Z Ü, Uzunalioğlu E, Güzel Ş, Bozdeveci A, Karaoğlu Ş A (2020). Determination of bioremediation properties of soil borne Bacillus sp. 5O5Y11 and its effect on the development of Zea mays in the presence of copper. Archives of Microbiology 202: 1817-1829.
11. Francis I, Holsters M & Vereecke D (2010). The Gram positive side of plant–microbe interactions. Environ Microbiol 12(1): 1-12.
12. Fürnkranz M, Müller H & Berg G (2009). Characterization of plant growth promoting bacteria from crops in Bolivia. J Plant Dis Protect 116(4): 149-155. Gardner G (2009). Plant hormone signaling. Hort Science 44(1): 222-223.
13. Ghanashyam C & Jain M (2009). Role of auxin-responsive genes in biotic stress responses. Plant Signal Behav 4 (9): 846-848.
14. Ghojavand H, Vahabzadeh F, Roayaei E & Shahraki A K (2008). Production and properties of a biosurfactant obtained from a member of the Bacillus subtilis group (PTCC 1696). J Colloid Interface Sci 324: 172-176.
15. Gillespie I M M & Philp J C (2013). Bioremediation, an environmental remediation technology for the bioeconomy. Trends in Biotechnology 31: 329-332.
16. Ho Y S, Porter J F & McKay G (2002). Equilibrium isotherm studies for the sorption of divalent metal ıons onto peat: copper, nickel and lead single component systems. Water, Air Soil Pollut 141: 1-33.
17. Holt J G, Krieg N R, Sneath P H A, Stanley J T & Williams S T (1994). Bergey's Manual of Determinative Bacteriology (9th ed.), Baltimor: Williams & Wilkins, Co. ISBN-13: 978-0683006032.
18. Huang J W & Cunninghams S D (1996). Lead phytoextraction: species variation in lead uptake and tanslocation. New Phytol 134(1): 75-84.
19. Idris E E, Iglesias D J & Talon M (2007). Tryptophan-dependent production of indole-3-acetic acid (IAA) affects level of plant growth promotion by Bacillus amyloliquefaciens FZB42. Mol Plant-Microbe Interact 20(6): 619–626.
20. Jayaraj J, Yi H & Liang G (2004). Blattapplikation von Bacillus subtilis AUBS1 reduziert die blattscheidendurre und € induziert abwehrmechanismen in reis [foliar application of Bacillus subtilis AUBS1 reduces sheath blight and triggers defense mechanisms in rice]. J Plant Dis Prot 111(2): 115-125.
21. Jjemba P K (2004). Environmental Microbiology: Principles and Applications. Enfield (NH) USA: Science Publishers, Inc., 257- 265.
22. Kamnev A A, Lelie D (2000). Chemical and biological parameters as tools to evaluate and improve heavy metal phytoremediation. Biosci Rep 20:239-258.
23. Kandler O & Weiss N (1986). Genus Lactobacillus beijerinck 1901, 212AL. In Bergey’s Manual of Systematic Bacteriology, Edited by Sneath P H A, Mair N S, Sharpe M E & Holt J G. Baltimore: Williams & Wilkins. 2: 1209-1234.
24. Karunakaran G, Suriyaprabha R, Manivasakan P, Yuvakkumar R, Rajendran V, Prabu P & Kannan N (2013). Effect of nanosilica and silicon sources on plant growth promoting rhizobacteria, soil nutrients and maize seed germination. IET Nanobiotechnol 7(3): 70-77.
25. Leelasuphakul W, Sivanunsakul P & Phongpaichit S (2006). Purification, characterization and synergistic activity of B-1,3- glucanase and antibiotic extract from an antagonistic Bacillus subtilis NSRS 89-24 against rice blast and sheath blight. Enzyme Microb Technol 38(7): 990-997.
26. Naja G, Diniz V & Volesky B (2010). Predicting Metal Biosorption Performance. In Proceedings of the 16th International Biotechnology Symposium. ed. by Harrison S T L, Rawlings D E and Petersen J. IBS – Compress Co., Cape Town, South Africa, 553-562.
27. National Committee for Clinical Laboratory Standard (1999). Methods for Determining Bactericidal Activity of Antimicrobial Agents; Approved Guideline. NCCLS Willanova PA, M26-A, 19 (18).
28. Nies D H (1999). Microbial heavy-metal resistance. Appl Microbiol Biotechnol 51(6): 730-50.
29. Özer A, Özer D & Özer A (2004). The adsorption of copper (ıı); ions on to dehydrated wheat bran (DWB): determination of the equilibrium and thermodynamic parameters. Process Biochem 39: 2183-2191.
30. Rivas R, Velázquez E, Zurdo-Piñeiro J L, Mateos P F & Martínez Molina E (2004). Identification of microorganisms by PCR amplification and sequencing of universal amplified ribosomal region present in both prokaryotes and eukaryotes. Microbiol Methods 56(3): 413-26.
31. Saha P, Datta S & Sanyal S K (2010). Application of natural clayey soil as adsorbent for the removal of copper from wastewater. Journal of Environmental Engineering 1409-1417.
32. Sahan T, Ceylan H, Sahiner N & Aktas N (2010). Optimization of removal conditions of copper ions from aqueous solutions by trametes versicolor. Bioresource Technology 101: 4520-4526.
33. Sambrook J, Fritschi E F & Maniatis T (1989). Molecular Cloning: A Laboratory. ISBN-978-1936113-42-2.
34. Shafi J, Tian H & Ji M (2017). Bacillus species as versatile weapons for plant pathogens: a review. Biotechnology & Biotechnological Equipment 31(3): 446-459. DOI: 10.1080/13102818.2017.1286950.
35. Sinha S, Mukherjee SK (2009) Pseudomonas aeruginosa KUCD1, a possible candidate for cadmium bioremediation. Braz J Microbiol 40: 655-662.
36. Tanzadeh J & Shareghi M (2017). Isolatıon and identification of halotolerant microorganisms resistant to heavy metals in the city of Qom, Iran. Lebanese Science Journal 18(2): 180-185.
37. Velásquez L & Dussan J (2009). Biosorption and bioaccumulation of heavy metals on dead and living biomass of Bacillus sphaericus. Journal of Hazardous Materials 167: 713-716.
38. Vessey JK (2003). Plant growth promoting rhizobacteria as biofertilizers. Plant and Soil 255(2): 571-586.
39. Vidhyasekaran P, Kamala N & Ramanathan A (2001). Induction of systemic resistance by Pseudomonas fluorescens PF1 against Xanthomonas oryzae pv. oryzae in Rice Leaves. Phytoparasitica 29(2): 155-166.
40. Villegas L C, Llamado A L, Catsao K V, Raymundo A K (2018). Removal of heavy metals from aqueous solution by biofilm-forming bacteria isolated from mined-out soil in Mogpog, Marinduque, Philippines. Philippine Science Letter 11: 18-27.