The Study on the Cultivable Microbiome of the Aquatic Fern Azolla Filiculoides L. as New Source of Beneficial Microorganisms
<p>(<b>a</b>) Culture of <span class="html-italic">A. filiculoides</span> under laboratory conditions on IRRI medium; (<b>b</b>) filaments of <span class="html-italic">A. azolla</span>e in a leaf cavity; (<b>c</b>) close up of <span class="html-italic">A. azollae</span>; both pictures taken from the light microscope at magnifications of 10x and 100x, respectively (Nikon Eclipse 80i, Nikon Instruments Europe B.V., Amsterdam, The Netherlands). Photo: A. Banach.</p> "> Figure 2
<p>UV microphotograph of the cyanobiont culture obtained during passage (Nikon Eclipse 80i microscope, magnification 4x, UV2A filter, Nikon Instruments Europe B.V., Amsterdam, The Netherlands). Photo: A. Banach.</p> "> Figure 3
<p>Examples of the most common isolates: (<b>a</b>) cream epiphyte, (<b>b</b>) yellow endophyte, and (<b>c</b>) white-cream filamentous form of epiphyte no. 37. Photo: A. Banach.</p> "> Figure A1
<p>Growth curves for the cultured microorganisms selected for phenotyping. <span class="html-italic">X</span>-axis presents time of incubation (hours) and <span class="html-italic">Y</span>-axis values of OD<sub>600</sub>. Logarithmic curves are fitted to the data.</p> ">
Abstract
:1. Introduction
2. Results
2.1. Azolla Cyanobiont
2.2. The Cultured Microbiome of A. filiculoides
2.3. Identification of Isolates
2.4. Synthesis of Plant Growth Promoters
3. Discussion
4. Materials and Methods
4.1. Plant Material
4.2. Azolla Cyanobiont
4.3. Isolation of Microorganisms
4.4. Cultivation and Description of Isolated Microorganisms
4.5. Identification of the Cyanobiont
4.6. Molecular Techniques
4.7. Phenotypic Characterization
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
Abbreviations
IAA | indole-3-acetic acid |
P | phosphorus |
PGPB | Plant Growth-Promoting Bacteria |
BUT | Butyrous |
MUC | Mucoid |
BRIT | Brittle |
OPQ | opaque |
TRANS | translucent |
IRID | iridescent |
F | flat |
R | raised |
C | convex |
U | umbonate |
sdH2O | deionized and sterilized water |
OD | optical density |
SD | standard deviation |
CMC | carboxylmethylcellulose |
PMB | P-mineralizing bacteria |
PSM | P-solubilizing bacteria |
PMPSB | phosphate mineralizing, phosphate solubilizing bacteria |
Appendix A
Cultivation of A. filiuloides
Isolation of Bacterial DNA
Phenotypic Analysis
Appendix B
Code 1 | Similar to | Similarity | Accession | c DNA (µg/mL) |
---|---|---|---|---|
EP1 | Achromobacter sp. IR27 | 97% | GU726513.1 | 55 |
EP2 | Bacillus cereus strain F2-2-21 | 99% | KX350029.1 | 1465 |
EP3 | Bacillus simplex strain Se2 | 99% | HQ432812.1 | 380 |
EP4 | Bacillus subtilis strain SUT2 | 99% | GU971415.1 | 940 |
EP5 | Bacillus subtilis strain RW134 | 99% | MH010185.1 | 50 |
EP6 | Microbacterium oxydans strain CanS-105 | 99% | KT580637.1 | 3640 |
EP7 | Delftia acidovorans isolate RI41 | 99% | DQ530127.1 | 20 |
EP8 | Bacillus thuringiensis strain WY9 | 100% | JQ936681.1 | 2288 |
EP9 | Bacillus subtilis strain BGR261 | 99% | KT074466.1896 | 25 |
EP10 | Achromobacter sp. strain SYP-B562 | 99% | KY636382.1 | 1560 |
EP11 | Bacillus subtilis strain RW134 | 99% | MH010185.1 | 170 |
EP12 | Achromobacter sp. strain SYP-B562 | 98% | KY636382.1 | 35 |
EP13 | Agrobacterium tumefaciens strain BF-R21 | 100% | KY292437.1 | 3280 |
EP14 | Achromobacter spanius strain 2S9 | 96% | KM374759.1 | 980 |
EP15 | n/a 2 | n/a | n/a | 655 |
EP16 | Bacillus sp. R-45540 | 100% | FR774944.1 | 260 |
EP17 | Bacillus subtilis strain RW134 | 99% | MH010185.1 | 120 |
EP18 | Bacillus pumilus strain IHBB 11092 | 99% | KR085935.1 | 3325 |
EP19 | Bacillus thuringiensis strain F9 | 99% | HQ432809.1 | 285 |
EP20 | Agrobacterium tumefaciens strain BF-R21 | 100% | KY292437.1 | 820 |
EP21 | Bacillus cereus strain AM11 | 99% | JQ435688.1 | 5965 |
EP22 | Bacillus cereus strain F1-1-1 | 99% | KX349989.1 | 340 |
EP23 | Alcaligenes sp. DH1f | 99% | KF557586.1 | 510 |
EP24 | Achromobacter sp. strain SYP-B562 | 100% | KY636382.1 | 560 |
EP25 | Microbacterium oxydans strain AE038-20 | 100% | KX369591.1 | 160 |
EP26 | Achromobacter sp. ATY31 | 98% | HQ219950.1 | 1698 |
EP27 | n/a | n/a | n/a | 80 |
EP28 | Bacillus cibi strain AIMST Ngme2 | 98% | JF939005.1 | 30 |
EP29 | Agrobacterium tumefaciens strain BF-R21 | 100% | KY292437.1 | 1590 |
EP30 | Achromobacter sp. strain SYP-B562 | 98% | KY636382.1 | 100 |
EP31 | Agrobacterium tumefaciens strain BF-R21 | 100% | KY292437.1 | 358 |
EP32 | Achromobacter sp. strain SYP-B562 | 99% | KY636382.1 | 2510 |
EP33 | n/a | n/a | n/a | 70 |
EP34 | Achromobacter marplatensis strain EY-T10 | 99% | KR476417.1 | 240 |
EP35 | Bacillus subtilis strain BGR261 | 99% | KT074466.1 | 120 |
EP36 | Bacillus sp. strain APNK5 | 94% | MG193758.1 | 340 |
EP37 | Bacillus weihenstephanensis strain P2 | 99% | HQ432810.1 | 3320 |
EP38 | Bacillus sp. strain Bac7 | 96% | KX500240.1 | 650 |
EP39 | n/a | n/a | n/a | 430 |
EP40 | n/a | n/a | n/a | 30 |
EP41 | Bacillus foraminis strain skuast2 | 97% | KY548645.1 | 755 |
EP42 | Bacillus sp. M16-1 | 99% | EF690408.1 | 2580 |
EP43 | Bacillus thuringiensis strain AHL1 | 99% | KT456534.1 | 40 |
EN1 | Bacillus subtilis strainBGR261 | 98% | KT074466.1 | 190 |
EN2 | Staphylococcus epidermidis strain HNL22 | 99% | EU373364.1 | 878 |
EN3 | Staphylococcus sp. iMSN20 | 99% | DQ401244.1 | 1147 |
EN4 | Staphylococcus epidermidis strain iCTE621 | 99% | DQ122332.1 | 813 |
EN5 | Staphylococcus epidermidis strain JPR-05 | 99% | HE716945.1 | 427 |
EN6 | Micrococcus aloeverae strain PP-06 | 98% | KX082870.1 | 640 |
EN7 | Micrococcus luteus strain LHR-04 | 97% | HE716930.1 | 348 |
EN8 | Bacillus subtilis strain NB-01 | 99% | HM214542.1 | 50 |
EN9 | Bacillus pumilus strain U38 | 99% | KC551966.1 | 350 |
EN10 | Micrococcus luteus strain IARI-THW-25 | 98% | KF054946.1 | 160 |
EN11 | Bacillus sp. IHB B 4034 | 99% | HM233998.1 | 6180 |
EN12 | Bacillus subtilis subsp. subtilis strain RG | 99% | JQ045774.1 | 660 |
EN13 | Bacillus pumilus strain U38 | 99% | KC551966.1 | 280 |
EN14 | Acinetobacter lwoffii strain Cl-01 | 98% | KC178575.1 | 5665 |
EN15 | Bacillus aryabhattai strain IARI-PC4-6 | 91% | KT149746.1 | 871 |
Name | Sequence from 5′ to 3′ | Reference |
---|---|---|
27F | AGAGTTTGATCATGGCTCAG | [56] |
518R | GTATTACCGCGGCTGCTGG | [56] |
23S30R | CTTCGCCTCTGTGTGCCTAGGT | [57,58] |
CYA359F | GGGGAATYTTCCGCAATGGG | [58,59] |
nif-Df | GATTTTCADGADAADGATATT | [60] |
nif-Dr | CCAIGGIATICCDTATTTTC |
References
- Gdanetz, K. The wheat microbiome under four management strategies, and potential for endophytes in disease control. Phytobiomes 2017, 1, 158–168. [Google Scholar] [CrossRef]
- Rout, M.E. The plant microbiome. In Genomes of Herbaceous Land Plants, 1st ed.; Paterson, A., Ed.; Academic Press: London, UK, 2014; Volume 69, pp. 279–309. [Google Scholar]
- Hardoim, P.R.; van Overbeek, L.S.; Berg, G.; Pirttilä, A.M.; Compant, S.; Campisano, A.; Döring, M.; Sessitsch, A. The Hidden World within Plants: Ecological and Evolutionary Considerations for Defining Functioning of Microbial Endophytes. Microbiol. Mol. Biol. Rev. 2015, 79, 293–320. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Berg, G.; Rybakowa, D.; Grube, M.; Köberl, M. The plant microbiome explored: Implications for experimental botany. J. Exp. Bot. 2016, 67, 995–1002. [Google Scholar] [CrossRef] [PubMed]
- Croes, S.; Weyens, N.; Colpaert, J.; Vangronsveld, J. Characterization of the cultivable bacterial populations associated with field grown Brassica napus L.: An evaluation of sampling and isolation protocols. Environ. Microbiol. 2015, 17, 2379–2392. [Google Scholar] [CrossRef] [PubMed]
- Santoyo, G.; Moreno-Hagelsieb, G.; Orozco-Mosqueda, M.C.; Glick, B.R. Plant growth-promoting bacterial endophytes. Microbiol. Res. 2016, 183, 92–99. [Google Scholar] [CrossRef] [PubMed]
- Egamberdieva, D.; Abd-Allah, E.F.; da Silva, J.T.A. Microbially assisted phytoremediation of heavy metal–contaminated soils. In Plant Metal Interaction. Emerging Remediation Techniques; Parvaiz, A., Ed.; Elsevier Inc.: Amsterdam, The Netherlands, 2016; pp. 483–498. [Google Scholar]
- Złoch, M.; Thiem, D.; Gadzała-Kopciuch, R.; Hrynkiewicz, K. Synthesis of siderophores by plant-associated metallotolerant bacteria under exposure to Cd2+. Chemosphere 2016, 156, 312–325. [Google Scholar] [CrossRef]
- Glick, B.R. Plant Growth-Promoting Bacteria: Mechanisms and Applications. Scientifica (Cairo) 2012, 2012, 963401. [Google Scholar] [CrossRef] [PubMed]
- Carrapiço, F. Azolla as a Superorganism. Its Implication in Symbiotic Studies. In Symbioses and Stress. Cellular Origin, Life in Extreme Habitats and Astrobiology; Seckbach, J., Grube, M., Eds.; Springer: Dordrecht, The Netherlands, 2010; Volume 17, pp. 225–241. [Google Scholar]
- Banach, A.M.; Banach, K.; Stępniewska, Z. Phytoremediation as a promising technology for water and soil purification: Azolla caroliniana Willd. As a case study. Acta Agrophysica 2012, 19, 241–252. [Google Scholar]
- Sood, A.; Uniyal, P.L.; Prasanna, R.; Ahluwalia, A.S. Phytoremediation Potential of Aquatic Macrophyte, Azolla. Ambio 2012, 41, 122–137. [Google Scholar] [CrossRef] [PubMed]
- Plazinski, J.; Zheng, Q.; Taylor, R.; Croft, L.; Rolfe, B.G.; Gunning, B.E.S. DNA probes show genetic variation in cyanobacterial symbionts of the Azolla fern and a closer relationship to free-living Nostoc strains than to free-living Anabaena strains. Appl. Environ. Microbiol. 1990, 56, 1263–1270. [Google Scholar]
- Gebhardt, J.S.; Nierzwicki-Bauer, S.A. Identification of a common cyanobacterial symbiont associated with Azolla spp. through molecular and morphological characterization of free-living and symbiotic cyanobacteria. Appl. Environ. Microbiol. 1991, 57, 2141–2146. [Google Scholar] [PubMed]
- Baker, J.A.; Entsch, B.; McKay, D.B. The cyanobiont in an Azolla fern is neither Anabaena nor Nostoc. FEMS Microbiol. Lett. 2003, 229, 43–47. [Google Scholar] [CrossRef]
- Pereira, A.L.; Vasconcelos, V. Classification and phylogeny of the cyanobiont Anabaena azollae Strasburger: An answered question? Int. J. Syst. Evol. Microbiol. 2014, 64, 1830–1840. [Google Scholar] [CrossRef] [PubMed]
- Grilli, M. Infrastrutture di Anabaena azollae vivente nelle foglioline di Azolla caroliniana (in Italian). Ann. Microb. Enzim 1964, XIV, 69–90. [Google Scholar]
- Nierzwicki-Bauer, S.A.; Aulfinger, H. Occurrence and Ultrastructural Characterization of Bacteria in Association with and Isolated from Azolla caroliniana. Appl. Environ. Microb. 1991, 57, 3629–3636. [Google Scholar]
- Carrapiço, F. Are bacteria the third partner of the Azolla-Anabaena symbiosis? Plant Soil 1991, 137, 157–160. [Google Scholar] [CrossRef]
- Serrano, R.; Carrapiço, F.; Vidal, R. The Presence of Lectins in Bacteria Associated with the Azolla-Anabaena Symbiosis. Symbiosis 1999, 27, 169–178. [Google Scholar]
- Zagajewski, P. Influence of environmental factors on growth and toxin production by cyanobacterial species (in Polish). Ph.D. Thesis, Adam Mickiewicz University Poznań. Faculty of Biology, Department of Water Protection, Poznań, Poland, 2012; pp. 64–65. [Google Scholar]
- Kozlikova-Zapomelova, E.; Chatchawan, T.; Kastovsky, J.; Komárek, J. Phylogenetic and taxonomic position of the genus Wollea with the description of Wollea salina sp. nov. (Cyanobacteria, Nostocales). Fottea 2016, 16, 43–55. [Google Scholar] [CrossRef]
- Shishido, J.; Humisto, A.; Jokela, J.; Liu, L.; Wahlsten, M.; Tamrakar, A.; Fewer, D.P.; Permi, P.; Andreote, A.P.; Fiore, M.F.; et al. Antifungal compounds from cyanobacteria. Mar. Drugs 2015, 13, 2124–2140. [Google Scholar] [CrossRef] [PubMed]
- Gugger, M.; Lyra, C.; Henriksen, P.; Couté, A.; Humbert, J.-F.; Sivonen, K. Phylogenetic comparison of the cyanobacterial genera Anabaena and Aphanizomenon. Int. J. Syst. Evol. Microbiol. 2002, 52, 1867–1880. [Google Scholar] [CrossRef]
- Willame, R.; Boutte, C.; Grubisic, S.; Wilmotte, A.; Komárek, J.; Hoffmann, L. Morphological and molecular characterization of planktonic cyanobacteria from Belgium and Luxembourg. J. Phycol. 2006, 42, 1312–1332. [Google Scholar] [CrossRef]
- Dijkhuizen, L.W.; Brouwer, P.; Bolhuis, H.; Reichart, G.-J.; Koppers, N.; Huettel, B.; Bolger, A.M.; Li, F.W.; Cheng, S.; Liu, X.; et al. Is there foul play in the leaf pocket? The metagenome of floating fern Azolla reveals endophytes that do not fix N2 but may denitrify. New Phytol. 2018, 217, 453–466. [Google Scholar] [CrossRef] [PubMed]
- Komárek, J.; Anagnostidis, K. Modern approach to the classification system of cyanophytes. Nostocales. Arch. Hydrobiol. Suppl. Algol. Stud. 1989, 56, 247–345. [Google Scholar]
- Canter-Lund, H.; Lund, J.W.G. Freshwater Algae: Their Microscopic World Explored; Biopress Ltd.: Bristol, UK, 1995; p. 360. [Google Scholar]
- Guiry, M.D.; Guiry, G.M. AlgaeBase. World-wide electronic publication, National University of Ireland, Galway. Available online: http://www.algaebase.org/search/species/detail/?species_id=65456 (accessed on 15 April 2019).
- Forni, C.; Riov, J.; Caiola, M.G.; Tel-Or, E. Indole-3-acetic acid (IAA) production by Arthrobacter species isolated from Azolla. J. Gen. Microbiol. 1992, 138, 377–381. [Google Scholar] [CrossRef] [PubMed]
- Ghodsalavi, B.; Ahmadzadeh, M.; Soleimani, M.; Madloo, P.B.; Taghizad-Farid, R. Isolation and characterization of rhizobacteria and their effects on root extracts of Valeriana officinalis. Aust. J. Crop Sci. 2013, 7, 338–344. [Google Scholar]
- Dutta, J.; Handique, P.J.; Thakur, D. Assessment of Culturable Tea Rhizobacteria Isolated from Tea Estates of Assam, India for Growth Promotion in Commercial Tea Cultivars. Front. Microbiol. 2015, 6, 1252. [Google Scholar] [CrossRef] [PubMed]
- Morel, M.; Iriarte, A.; Jara, E.; Musto, H.; Castro-Sowinski, S. Revealing the biotechnological potential of Delftia sp. JD2 by a genomic approach. AIMS Bioeng 2016, 3, 156–175. [Google Scholar] [CrossRef]
- Cho, K.M.; Hong, S.Y.; Lee, S.M.; Kim, Y.H.; Kahng, G.G.; Lim, Y.P.; Kim, H.; Yun, H.D. Endophytic Bacterial Communities in Ginseng and their Antifungal Activity Against Pathogens. Microb. Ecol. 2007, 54, 341–351. [Google Scholar] [CrossRef]
- Jorquera, M.A.; Hernández, M.T.; Rengel, Z.; Marschner, P.; de la Luz Mora, M. Isolation of culturable phosphobacteria with both phytate-mineralization and phosphate-solubilization activity from the rhizosphere of plants grown in a volcanic soil. Biol. Fertil. Soils 2008, 44, 1025–1034. [Google Scholar] [CrossRef]
- Chen, Y.P.; Rekha, P.D.; Arun, A.B.; Shen, F.T.; Lai, W.-A.; Young, C.C. Phosphate solubilizing bacteria from subtropical soil and their tricalcium phosphate solubilizing abilities. Appl. Soil. Ecol. 2006, 34, 33–41. [Google Scholar] [CrossRef]
- Liu, Y.; Tie, B.; Li, Y.; Lei, M.; Wei, X.; Liu, X.; Du, H. Inoculation of soil with cadmium-resistant bacterium Delftia sp. B9 reduces cadmium accumulation in rice (Oryza sativa L.) grains. Ecotoxicol. Environ. Saf. 2018, 163, 223–229. [Google Scholar] [CrossRef] [PubMed]
- Xu, X.; Liu, W.; Wang, W.; Tian, S.; Jiang, P.; Qi, Q.; Li, F.; Li, H.; Wang, Q.; Li, H.; Yu, H. Potential biodegradation of phenanthrene by isolated halotolerant bacterial strains from petroleum oil polluted soil in Yellow River Delta. Sci Total Environ 2019, 664, 1030–1038. [Google Scholar] [CrossRef] [PubMed]
- Braña, V.; Cagide, C.; Morel, M.A. The sustainable use of Delftia in agriculture, bioremediation, and bioproducts synthesis. In Microbial Models: From Environmental to Industrial Sustainability, 1st ed.; Castro-Sowinski, S., Ed.; Springer: Singapore, 2016; pp. 227–247. [Google Scholar]
- Wu, W.; Huang, H.; Ling, Z.; Yu, Z.; Jaing, Y.; Liu, P.; Li, X. Genome sequencing reveals mechanisms for heavy metal resistance and polycyclic aromatic hydrocarbon degradation in Delftia lacustris strain LZ-C. Ecotoxicology 2016, 25, 234–247. [Google Scholar] [CrossRef]
- Hou, Q.; Wang, C.; Guo, H.; Xia, Z.; Ye, J.; Liu, K.; Yang, Y.; Hou, X.; Liu, H.; Wang, J.; Du, B.; Ding, Y. Draft genome sequence of Delftia tsuruhatensis MTQ3, a strain of plant growth-promoting rhizobacterium with antimicrobial activity. Genome Announc 2015, 3, e00822-15. [Google Scholar] [CrossRef]
- Johnston, C.W.; Wyatt, M.A.; Li, X.; Ibrahim, A.; Shuster, J.; Southham, G.; Magarvey, N.A. Gold biomineralization by a metallophore from a gold-associated microbe. Nat. Chem. Biol. 2013, 9, 241–243. [Google Scholar] [CrossRef]
- Li, G.-X.; Zhou, S.-Y.-D.; Ren, H.-Y.; Xue, X.-M.; Xu, Y.-Y.; Bao, P. Extracellular Biomineralization of Gold by Delftia tsuruhatensis GX-3 Isolated from a Heavy Metal Contaminated Paddy Soil. ACS Earth Space Chem 2018, 2, 1294–1300. [Google Scholar] [CrossRef]
- Jangir, Y.; French, S.; Momper, L.M.; Moser, D.P.; Amend, J.P.; El-Naggar, M.Y. Isolation and Characterization of Electrochemically Active Subsurface Delftia and Azonexus Species. Front Microbiol 2016, 7, 756. [Google Scholar] [CrossRef] [PubMed]
- Shetty, A.; Chen, S.; Tocheva, E.I.; Jensen, G.J.; Hickey, W.J. Nanopods: A New Bacterial Structure and Mechanism for Deployment of Outer Membrane Vesicles. PLoS ONE 2011, 6, e20725. [Google Scholar] [CrossRef] [PubMed]
- Ahemad, M. Phosphate-solubilizing bacteria-assisted phytoremediation of metalliferous soils: A review. 3 Biotech 2015, 5, 111–121. [Google Scholar] [CrossRef]
- Watanabe, I.; Roger, P.A.; Ladha, J.K.; Van Hove, C. Biofertilizer germplasm collections at IRRI; International Rice Research Institute: Manila, Philippines, 1992; p. 8. [Google Scholar]
- Komárek, J. Cyanoprokaryota: 3rd Part: Heterocystous Genera. In Süßwasserflora von Mitteleuropa; Büdel, B., Gärtner, G., Krienitz, L., Schagerl, M., Eds.; Springer Spektrum: Berlin/Heidelberg, Germany, 2013; Volume 19, pp. 1–1130. [Google Scholar]
- Hindák, F. Fotografický atlas mikroskopických sinic (in Slovak); VEDA: Bratislava, Solvakia, 2001; p. 257. [Google Scholar]
- Stępniewska, Z.; Goraj, W.; Kuźniar, A.; Łopacka, N.; Małysza, M. Enrichment culture and identification of endophytic methanotrophs isolated from peatland plants. Folia Microbiol. 2017, 62, 381–391. [Google Scholar] [CrossRef] [Green Version]
- Truyens, S.; Weyens, S.; Cuypers, A.; Vangronsveld, J. Changes in the population of seed bacteria of transgenerationally Cd-exposed Arabidopsis thaliana. Plant Biol. (Stuttg) 2013, 15, 971–981. [Google Scholar] [CrossRef]
- Kasana, R.C.; Salwan, R.; Dhar, H.; Dutt, S.; Gulati, A. A Rapid and Easy Method for the Detection of Microbial Cellulases on Agar Plates Using Gram’s Iodine. Curr. Microbiol. 2008, 57, 503–507. [Google Scholar] [CrossRef]
- Alexander, D.B.; Zuberer, D.A. Use of chrome azurol S reagents to evaluate siderophore production by rhizsphere bacteria. Biol Fertil Soils 1991, 12, 39–45. [Google Scholar] [CrossRef]
- Sambrook, J.; Russell, R.W. Molecular cloning: A laboratory manual, 3rd ed.; Cold Spring Harbor Laboratory Press: New York, NY, USA, 2001; p. 2100. [Google Scholar]
- Gordon, S.; Weber, R. Colorimetric estimation of indoleacetic acid. Plant Physiol. 1951, 26, 192–195. [Google Scholar] [CrossRef] [PubMed]
- Lee, T.K.; Doan, T.V.; Yoo, K.; Choi, S.; Kim, C.; Park, J. Discovery of commonly existing anode biofilm microbes in two different wastewater treatment MFCs using FLX Titanium pyrosequencing. Appl. Microbiol. Biotechnol. 2010, 87, 2335–2343. [Google Scholar] [CrossRef] [PubMed]
- Taton, A.; Grubisic, S.; Brambilla, E.; De Wit, R.; Wilmotte, A. Cyanobacterial Diversity in Natural and Artificial Microbial Mats of Lake Fryxell (McMurdo Dry Valleys, Antarctica): A Morphological and Molecular Approach. Appl. Microbiol. Biotechnol. 2003, 69, 5157–5169. [Google Scholar] [CrossRef]
- Koskenniemi, K.; Lyra, Ch.; Rajaniemi-Wacklin, P.; Jokela, J.; Sivonen, K. Quantitative Real-Time PCR Detection of Toxic Nodularia Cyanobacteria in the Baltic Sea. Appl. Environ. Microb. 2007, 73, 2173–2179. [Google Scholar] [CrossRef]
- Nübel, U.; Garcia-Pichel, F.; Muyzer, G. PCR Primers To Amplify 16S rRNA Genes from Cyanobacteria. Appl. Environ. Microb. 1997, 63, 3327–3332. [Google Scholar]
- Dedysh, S.; Ricke, P.; Liesack, W. NifH and NifD phylogenies: An evolutionary basis for understanding nitrogen fixation capabilities of methanotrophic bacteria. Microbiology 2004, 150, 1301–1313. [Google Scholar] [CrossRef]
Genus | Potential Microorganism | Similarity | Accession no. | Reference |
---|---|---|---|---|
Anabaena | A. sp. 6-HorLes10 | 94% | KT290350.1 | [22] |
A. sp. HAN21/1 | 93% | KP701032.1 | [23] | |
A. cf. cylindrica 133 | 93% | AJ293110.1 | [24] | |
A. oscillarioides 0RO34S1 | 90% | DQ264246.1 | [25] |
No. | Type | Size 1 | Form 2 | Surface 3 | Texture 4 | Opacity 5 | Pigmentation 6 | Elevation 7 | Margin 8 | Gram Staining |
---|---|---|---|---|---|---|---|---|---|---|
1 | Epiphyte | ++ | i | d/r | BUT | OPQ | cream | F | Ent | G- |
2 | ++ | i | g/r | BUT | OPQ | w-c | R | Und | G+ | |
3 | ++ | i | g/r | BUT | OPQ | cream | R | Und | G+ | |
4 | + | c | d/s | BRIT | OPQ | white | F | Ent | G+ | |
5 | ++ | c | d/s | BRIT | OPQ | white | F | Und | G+ | |
6 | ++ | i | d/s | BUT | OPQ | yel-c | F | Und | G+ | |
7 | . | c | g/s | BUT | TRANS | white | R | Ent | G- | |
8 | . | c | g/r | BUT | TRANS | cream | R | Ent | G+ | |
9 | + | c | g/r | BUT | TRANS | yel-c | R | Ent | G+ | |
10 | + | c | g/r | BUT | TRANS | cream | R | Ent | G- | |
11 | ++ | i | d/r | BRIT | OPQ | yel-c | F | Und | G+ | |
12 | + | i | g/s | BUT | IRID | cream | C | Und | G- | |
13 | + | c | g/s | BUT | OPQ | yel-c | U | Ent | G- | |
14 | . | o | g/r | MUC | OPQ | cream | R | Ent | G- | |
15 | +++ | c | g/s | MUC | TRANS | yel-org | C | Ent | G- | |
16 | ++ | c | d/s | BUT | OPQ | w-c | R | Ent | G+ | |
17 | ++ | o | d/s | BRIT | OPQ | w-c | F | Ent | G+ | |
18 | + | i | d/s | BUT | OPQ | cream | F | Und | G+ | |
19 | +++ | o | d/s | BRIT | OPQ | cream | R | Und | G+ | |
20 | . | c | g/r | BUT | OPQ | cream | U | Ent | G- | |
21 | +++ | o | d/s | BUT | OPQ | beige | F | Und | G+ | |
22 | +++ | c | d/r | BUT | OPQ | beige | R | Und | G+ | |
23 | + | c | g/s | BUT | TRANS | cream | R | Ent | G- | |
24 | . | c | g/r | BUT | TRANS | beige | R | Ent | G- | |
25 | . | o | g/s | MUC | TRANS | yellow | R | Ent | G+ | |
26 | . | o | g/s | MUC | TRANS | cream | R | Ent | G- | |
27 | + | c | g/s | BUT | TRANS | w-c | F | Und | G+ | |
28 | + | i | g/s | BUT | OPQ | yel-org | R | Und | G+ | |
29 | . | c | g/s | BUT | TRANS | cream | U | Ent | G- | |
30 | + | c | g/r | BUT | OPQ | beige | R | Ent | G- | |
31 | + | c | d/s | MUC | IRID | beige | U | Ent | G- | |
32 | + | c | g/r | BUT | TRANS | beige | U | Ent | G- | |
33 | . | c | g/r | BUT | TRANS | beige | U | Ent | G- | |
34 | . | o | g/s | BUT | TRANS | cream | R | Ent | G- | |
35 | ++ | o | d/r | BRIT | OPQ | cream | F | Ent | G+ | |
36 | . | c | g/s | BUT | TRANS | cream | U | Ent | G+ | |
37 | +++ | f | g/s | BUT | TRANS | w-c | F | Fili | G+ | |
38 | . | c | g/s | MUC | OPQ | yellow | R | Ent | G+ | |
39 | + | c | g/s | MUC | TRANS | yellow | C | Ent | G- | |
40 | + | i | g/r | BUT | OPQ | cream | U | Ent | G- | |
41 | + | o | d/r | BUT | TRANS | cream | U | Ent | G+ | |
42 | +++ | o | d/s | BRIT | TRANS | beige | F | Ent | G+ | |
43 | +++ | o | d/s | BRIT | OPQ | beige | F | Und | G+ | |
1 | endophyte | . | c | g/r | BUT | OPQ | beige | U | Ent | G+ |
2 | . | c | g/s | BUT | TRANS | cream | R | Ent | G+ | |
3 | . | c | g/s | BUT | OPQ | cream | R | Ent | G+ | |
4 | . | c | g/s | BUT | TRANS | w-c | R | Ent | G+ | |
5 | + | c | g/s | BUT | TRANS | w-c | R | Ent | G+ | |
6 | . | c | g/s | MUC | IRID | yellow | R | Ent | G+ | |
7 | . | c | g/s | BUT | IRID | yellow | R | Und | G+ | |
8 | . | c | g/s | MUC | OPQ | yellow | R | Ent | G+ | |
9 | + | c | g/s | BUT | OPQ | cream | R | Ent | G+ | |
10 | ++ | c | g/s | MUC | IRID | yellow | R | Ent | G+ | |
11 | ++ | c | g/s | MUC | OPQ | yellow | R | Ent | G+ | |
12 | +++ | c | g/s | BUT | TRANS | cream | R | Ent | G+ | |
13 | ++ | c | g/s | BUT | OPQ | white-cream | R | Ent | G+ | |
14 | ++ | i | g/s | BUT | OPQ | white-cream | F | Und | G- | |
15 | + | i | g/s | BUT | OPQ | white-cream | F | Und | G+ |
No. | Isolate name | Accession no. | No. | Isolate name | Accession no. |
---|---|---|---|---|---|
Epiphytes | Epiphytes–continuation | ||||
1 | Achromobacter sp. AzoEpi1 | MG881884 | 29 | Agrobacterium sp. AzoEpi25 | MG881908 |
2 | Bacillus sp. AzoEpi2 | MG881885 | 31 | Agrobacterium sp. AzoEpi34 | MH605442 |
3 | Bacillus sp. AzoEpi3 | MG881886 | 34 | Achromobacter sp. AzoEpi26 | MG881909 |
4 | Bacillus sp. AzoEpi4 | MG881887 | 35 | Bacillus sp. AzoEpi27 | MG881910 |
5 | Bacillus sp. AzoEpi5 | MG881888 | 36 | Bacillus sp. AzoEpi28 | MG881911 |
6 | Microbacterium sp. AzoEpi6 | MG881889 | 37 | Bacillus sp. AzoEpi29 | MG881912 |
7 | Delftia sp. AzoEpi7 | MG881890 | 38 | Bacillus sp. AzoEpi30 | MG881913 |
8 | Bacillus sp. AzoEpi33 | MH605441 | 41 | Bacillus sp. AzoEpi35 | MH605443 |
9 | Bacillus sp. AzoEpi8 | MG881891 | 42 | Bacillus sp. AzoEpi31 | MG881914 |
10 | Achromobacter sp. AzoEpi9 | MG881892 | 43 | Bacillus sp. AzoEpi32 | MG881915 |
11 | Bacillus sp. AzoEpi10 | MG881893 | Endophytes | ||
12 | Achromobacter sp. AzoEpi11 | MG881894 | 1 | Bacillus sp. AzoEndo1 | MG859252 |
13 | Agrobacterium sp. AzoEpi12 | MG881895 | 2 | Staphylococcus sp. AzoEndo10 | MH605510 |
14 | Achromobacter sp. AzoEpi13 | MG881896 | 3 | Staphylococcus sp. AzoEndo11 | MH605511 |
16 | Bacillus sp. AzoEpi14 | MG881897 | 4 | Staphylococcus sp. AzoEndo12 | MH605512 |
17 | Bacillus sp. AzoEpi15 | MG881898 | 5 | Staphylococcus sp. AzoEndo13 | MH605513 |
18 | Bacillus sp. AzoEpi16 | MG881899 | 6 | Micrococcus sp. AzoEndo9 | MG881919 |
19 | Bacillus sp. AzoEpi17 | MG881900 | 7 | Micrococcus sp. AzoEndo14 | MH605514 |
20 | Agrobacterium sp. AzoEpi18 | MG881901 | 8 | Bacillus sp. AzoEndo2 | MG859253 |
21 | Bacillus sp. AzoEpi19 | MG881902 | 9 | Bacillus sp. AzoEndo3 | MG859254 |
22 | Bacillus sp. AzoEpi20 | MG881903 | 10 | Micrococcus sp. AzoEndo7 | MG881917 |
23 | Alcaligenes sp. AzoEpi21 | MG881904 | 11 | Bacillus sp. AzoEndo4 | MG859255 |
24 | Achromobacter sp. AzoEpi22 | MG881905 | 12 | Bacillus sp. AzoEndo5 | MG859256 |
25 | Microbacterium sp. AzoEpi23 | MG881906 | 13 | Bacillus sp. AzoEndo6 | MG859257 |
28 | Bacillus sp. AzoEpi24 | MG881907 | 14 | Acinetobacter sp. AzoEndo8 | MG881918 |
Isolate (Genera) | IAA (µg mL−1) | Cellulase Activity | Protease Activity | ||
---|---|---|---|---|---|
h 1 (cm) | h:c 2 | h (cm) | h:c | ||
Staphylococcus sp. AzoEndo11 | n/a 3 | 0.98 (0.25) | 1.19 (0.33) | 1.25 (0.21) | 0.45 (0.23) |
Micrococcus sp. AzoEndo14 | 17.900 (0.201) | 1.50 (0.14) | 1.05 (0.25) | 1.00 (0.19) | 0.31 (0.08) |
Bacillus sp. AzoEndo3 | n/a | 1.06 (0.20) | 1.06 (0.05) | 1.76 (0.36) | 1.01 (0.89) |
Acinetobacter sp. AzoEndo8 | n/a | 1.20 (0.00) | 0.80 (0.00) | 1.32 (0.34) | 0.58 (0.19) |
Achromobacter sp. AzoEpi1 | n/a | 1.03 (0.13) | 0.67 (0.08) | 1.22 (0.19) | 0.40 (0.09) |
Bacillus sp. AzoEpi2 | n/a | 0.80 (0.08) | 0.47 (0.07) | 1.10 (0.19) | 0.28 (0.15) |
Delftia sp. AzoEpi7 | 3.575 (0.029) | 0.23 (0.13) | 0.27 (0.17) | 1.12 (0.26) | 0.41 (0.18) |
Alcaligenes sp. AzoEpi21 | n/a | 0.65 (0.06) | 1.20 (0.23) | 1.57 (0.31) | 0.45 (0.18) |
Microbacterium sp. AzoEpi23 | n/a | 0.50 (0.08) | 1.20 (0.29) | 1.33 (0.34) | 0.78 (0.35) |
Agrobacterium sp. AzoEpi25 | 6.390 (0.053) | 0.68 (0.15) | 1.33 (0.47) | 2.95 (0.28) | 1.10 (0.33) |
Isolate | P mineralization | P solubilization | Siderophores | |||
---|---|---|---|---|---|---|
h 1 (cm) | h:c 2 | h (cm) | h:c | h (cm) | h:c | |
Staphylococcus sp. AzoEndo11 | 0.26 (0.19) | 0.23 (0.17) | 1.95 (0.72) | 1.91 (0.93) | n/a 3 | n/a |
Micrococcus sp. AzoEndo14 | 0.39 (0.08) | 0.32 (0.12) | 0.29 (0.44) | 0.64 (0.62) | n/a | n/a |
Bacillus sp. AzoEndo3 | 0.62 (0.18) | 0.47 (0.18) | 1.15 (0.54) | 1.35 (0.43) | 3.56 (0.17) | 10.10 (1.79) |
Acinetobacter sp. AzoEndo8 | 0.31 (0.10) | 0.25 (0.09) | n/a | n/a | n/a | n/a |
Achromobacter sp. AzoEpi1 | 0.36 (0.09) | 0.33 (0.11) | n/a | n/a | 0.56 (0.29) | 0.07 (0.04) |
Bacillus sp. AzoEpi2 | 0.52 (0.18) | 0.43 (0.15) | n/a | n/a | n/a | n/a |
Delftia sp. AzoEpi7 | 0.49 (0.19) | 0.43 (0.16) | 0.79 (0.29) | 1.01 (0.39) | 0.98 (0.21) | 0.64 (0.10) |
Alcaligenes sp. AzoEpi21 | 0.35 (0.13) | 0.31 (0.13) | n/a | n/a | 0.10 (0.00) | 0.39 (0.15) |
Microbacterium sp. AzoEpi23 | 0.46 (0.13) | 0.43 (0.15) | n/a | n/a | n/a | n/a |
Agrobacterium sp. AzoEpi25 | 0.43 (0.13) | 0.39 (0.15) | n/a | n/a | 0.41 (0.14) | 0.17 (0.06) |
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Banach, A.; Kuźniar, A.; Mencfel, R.; Wolińska, A. The Study on the Cultivable Microbiome of the Aquatic Fern Azolla Filiculoides L. as New Source of Beneficial Microorganisms. Appl. Sci. 2019, 9, 2143. https://doi.org/10.3390/app9102143
Banach A, Kuźniar A, Mencfel R, Wolińska A. The Study on the Cultivable Microbiome of the Aquatic Fern Azolla Filiculoides L. as New Source of Beneficial Microorganisms. Applied Sciences. 2019; 9(10):2143. https://doi.org/10.3390/app9102143
Chicago/Turabian StyleBanach, Artur, Agnieszka Kuźniar, Radosław Mencfel, and Agnieszka Wolińska. 2019. "The Study on the Cultivable Microbiome of the Aquatic Fern Azolla Filiculoides L. as New Source of Beneficial Microorganisms" Applied Sciences 9, no. 10: 2143. https://doi.org/10.3390/app9102143