Abstract
Metal whole-cell biosensors (WCBs) have been reported as very useful tools to detect and quantify the presence of bioavailable fractions of certain metals in water and soil samples. In the current work, two bacterial WCBs able to report Cr(VI) presence and plants growing on Cr(VI)-enriched soil/medium were used to assess the potential transfer of this metal to organisms of higher trophic levels, and the risk of transfer to the food chain. To do it, the functionality of the WCBs within tissues of inoculated plants in contact with Cr(VI)-contaminated soil and water was studied in vitro and in a controlled greenhouse environment. One WCB was the previously described Ochrobactrum tritici pCHRGFP2 and the second, Nitrospirillum amazonense pCHRGFP2, is a newly engineered naturally-occurring endophytic microorganism. Three rice varieties (IAC 4440, BRS 6 CHUÍ, IRGA 425) and one maize variety (1060) were tested as hosts and subjected to Cr(VI) treatments (25 μM), with different results obtained. Inoculation of each WCB into plants exposed to Cr(VI) showed GFP expression within plant tissues. WCBs penetrated the root tissues and later colonized the shoots and leaves. In general, a higher fluorescence signal was detected in roots, together with a higher Cr content and denser WCB colonization. Best fluorescence intensities per plant biomass of shoots were obtained for plant host IRGA 425. Therefore, by analyzing colonized tissues, both WCBs allowed the detection of Cr(VI) contamination in soils and its transfer to plants commonly used in crops for human diet.
Graphic abstract
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Amaro F, Turkewitz AP, Martín-González A, Gutiérrez J-C (2011) Whole-cell biosensors for detection of heavy metal ions in environmental samples based on metallothionein promoters from Tetrahymena thermophila. Microb Biotechnol 4:513–522. https://doi.org/10.1111/j.1751-7915.2011.00252.x
American Public Health Association (1998) Metals, part 3000. In: Clesceri LS, Greenberg AE, Eaton AD (eds) Standard methods for the determination of water and wastewater. American Public Health Association, Washington, pp 65–68
Branco R, Chung AP, Johnston T, Gurel V, Morais PV, Zhitkovich A (2008) The chromate-inducible chrBACF operon from the transposable element TnOtChr confers resistance to chromium (VI) and superoxide. J Bacteriol 190:6996–7003. https://doi.org/10.1128/JB.00289-08
Branco R, Cristóvão A, Morais PV (2013) Highly sensitive, highly specific whole-cell bioreporters for the detection of chromate in environmental samples. PLoS ONE 8:e54005. https://doi.org/10.1371/journal.pone.0054005
Branco R, Morais PV (2013) Identification and characterization of the transcriptional regulator ChrB in the chromate resistance determinant of Ochrobactrum tritici 5bvl1. PLoS ONE 8:e77987. https://doi.org/10.1371/journal.pone.0077987
Branco R, Morais PV (2016) Two superoxide dismutases from TnOtchr are involved in detoxification of reactive oxygen species induced by chromate. BMC Microbiol 16:27. https://doi.org/10.1186/s12866-016-0648-0
Chakraborty U, Chakraborty BN, Basnet M, Chakraborty AP (2009) Evaluation of Ochrobactrum anthropi TRS-2 and its talc based formulation for enhancement of growth of tea plants and management of brown root rot disease. J Appl Microbiol 107:625–634. https://doi.org/10.1111/j.1365-2672.2009.04242.x
Coelho C, Branco R, Natal-da-Luz T, Sousa JP, Morais PV (2015) Evaluation of bacterial biosensors to determine chromate bioavailability and to assess ecotoxicity of soils. Chemosphere 128:62–69. https://doi.org/10.1016/j.chemosphere.2014.12.026
Dary M, Chamber-Pérez MA, Palomares AJ, Pajuelo E (2010) “In situ” phytostabilisation of heavy metal polluted soils using Lupinus luteus inoculated with metal resistant plant-growth promoting rhizobacteria. J Hazard Mater 177:323–330. https://doi.org/10.1016/j.jhazmat.2009.12.035
de Guimarães OPE, Parentoni SN, Pacheco CAP, Meirelles WF, Guimarães LJM, da Silva AR et al (2009) Comun Téc 169: BRS 1060 – híbrido simples de milho. Embrapa Milho e Sorgo, Sete Lagoas, MG, Brazil
Dogo S, Razic S, Manojlovic D, Slavkovic L (2011) Analysis of the bioavailability of Cr(III) and Cr(VI) based on the determination of chromium in Mentha piperita by graphite furnace atomic absorption spectrometry. J Serb Chem Soc 76:143–153. https://doi.org/10.2298/JSC100401130D
Egener T, Hurek T, Reinhold-Hurek B (1998) Use of green fluorescent protein to detect expression of nif genes of Azoarcus sp. BH72, a grass-associated diazotroph, on rice roots. Mol Plant-Microbe Interact 11:71–75. https://doi.org/10.1094/MPMI.1998.11.1.71
EMBRAPA (1991) EMBRAPA 6 Chuí: nova cultivar de arroz irrigado. Embrapa Clima Temperado, Pelotas
European Commission (EC) (2016) Proposal for a regulation of the European Parliament and of the Council laying down rules on the making available on the market of CE marked fertilising products and amending Regulations (EC) No 1069/2009 and (EC) No 1107/2009. COM/2016/0157 final - 2016/084 (COD).
Földi C, Dohrmann R, Matern K, Mansfeldt T (2013) Characterization of chromium-containing wastes and soils affected by the production of chromium tanning agents. J Soils Sedim 13:1170–1179. https://doi.org/10.1007/s11368-013-0714-2
Francisco R, Alpoim MC, Morais PV (2002) Diversity of chromium-resistant and-reducing bacteria in a chromium-contaminated activated sludge. J Appl Microbiol 92:837–843. https://doi.org/10.1046/j.1365-2672.2002.01591.x
Francisco R, de Abreu P, Plantz BA, Schlegel VL, Carvalho RA, Morais PV (2011) Metal-induced phosphate extracellular nanoparticulate formation in Ochrobactrum tritici 5bvl1. J Hazard Mater 198:31–39. https://doi.org/10.1016/j.jhazmat.2011.10.005
Francisco R, Branco R, Schwab S, Baldani JI, Morais PV (2017) Impact of plant-associated bacteria biosensors on plant growth in the presence of hexavalent chromium. World J Microbiol Biotechnol 34:12. https://doi.org/10.1007/s11274-017-2389-0
Gui Q, Lawson T, Shan S, Yan L, Liu Y (2017) The application of whole cell-based biosensors for use in environmental analysis and in medical diagnostics. Sensors. https://doi.org/10.3390/s17071623
Gutiérrez JC, Amaro F, Martín-González A (2015) Heavy metal whole-cell biosensors using eukaryotic microorganisms: an updated critical review. Front Microbiol 6:48. https://doi.org/10.3389/fmicb.2015.00048
Hardoim PR, van Overbeek LS, van Elsas JD (2008) Properties of bacterial endophytes and their proposed role in plant growth. Trends Microbiol 16:463–471. https://doi.org/10.1016/j.tim.2008.07.008
Haydon MJ (2014) Getting a sense for zinc in plants. New Phytol 202:10–12. https://doi.org/10.1111/nph.12736
Hessels AM, Chabosseau P, Bakker MH, Engelen W, Rutter GA, Taylor KM, Merkx M (2015) eZinCh-2: aversatile, genetically encoded FRET sensor for cytosolic and intraorganelle Zn2+ imaging. ACS Chem Biol 10:2126–2134. https://doi.org/10.1021/acschembio.5b00211
Hoagland DR, Arnon DI (1950) The water-culture method for growing plants without soil. University of California, College of Agriculture, Agricultural Experiment Station, Berkeley, Calif.
Huang P, de Bashan L, Crocker T, Kloepper JW, Bashan Y (2017) Evidence that fresh weight measurement is imprecise for reporting the effect of plant growth-promoting (rhizo)bacteria on growth promotion of crop plants. Biol Fertil Soils 53:199–208
Hurek T, Reinhold-Hurek B, Van Montagu M, Kellenberger E (1994) Root colonization and systemic spreading of Azoarcus sp. strain BH72 in grasses. J Bacteriol 176:1913–1923. https://doi.org/10.1128/jb.176.7.1913-1923.1994
Ivask A, Virta M, Kahru A (2002) Construction and use of specific luminescent recombinant bacterial sensors for the assessment of bioavailable fraction of cadmium, zinc, mercury and chromium in the soil. Soil Biol Biochem 34:1439–1447. https://doi.org/10.1016/S0038-0717(02)00088-3
Joutey NT, Sayel H, Bahafid W, Ghachtouli NE (2015) Mechanisms of hexavalent chromium resistance and removal by microorganisms. In: Whitacre D (ed) Reviews of environmental contamination and toxicology. Springer: Cham, vol 233, pp 45–69. https://doi.org/10.1007/978-3-319-10479-9_2
Lanquar V, Grossman G, Vinkenborg JL, Merkx M, Thomine S, Frommer WB (2014) Dynamic imaging of cytosolic zinc in Arabidopsis roots combining FRET sensors and RootChip technology. New Phytol 202:198–208. https://doi.org/10.1111/nph.12652
Lebuhn M, Achouak W, Schloter M, Berge O, Meier H, Barakat M et al (2000) Taxonomic characterization of Ochrobactrum sp. isolates from soil samples and wheat roots, and description of Ochrobactrum tritici sp. nov. and Ochrobactrum grignonense sp. nov. Int J Syst Evol Microbiol 50:2207–2223. https://doi.org/10.1099/00207713-50-6-2207
Lin S-Y, Hameed A, Shen F-T, Liu Y-C, Hsu Y-H, Shahina M et al (2014) Description of Niveispirillum fermenti gen. nov., sp. nov., isolated from a fermentor in Taiwan, transfer of Azospirillum irakense (1989) as Niveispirillum irakense comb. nov., and reclassification of Azospirillum amazonense (1983) as Nitrospirillum amazonense gen. nov. Antonie Van Leeuwenhoek 105:1149–1162. https://doi.org/10.1007/s10482-014-0176-6
Lopes SIG, de Rosso AF, Kempf D, Lopes MCB, Carmona PS, Funck GRD, et al (2009) IRGA 425: Nova cultivar para o sistema de cultivo pré-germinado no Rio Grande do Sul. In: VI congresso Brasileiro de arroz irrigado, fitomelhoramento, biotecnologia, bioclimatologia e ecofisiologia, Porto Alegre, Brazil. SOSBAI (Sociedade Sul Brasileira de Arroz Irrigado), pp. 135 – 139. Available from: https://www.sosbai.com.br/docs/VI_CBAI_Fitomelhoramento.pdf
Ma Y, Rajkumar M, Zhang C, Freitas H (2016) Inoculation of Brassica oxyrrhina with plant growth promoting bacteria for the improvement of heavy metal phytoremediation under drought conditions. J Hazard Mater 320:36–44. https://doi.org/10.1016/j.jhazmat.2016.08.009
Magalhães F, Baldani J, Souto S, Kuykendall J, Döbereiner J (1983) A new acid-tolerant Azospirillum species. An Acad Bras Cienc 55:417–430
Miller WG, Leveau JHJ, Lindow SE (2000) Improved gfp and inaZ broad-host-range promoter-probe vectors. Mol Plant-Microbe Interact 13:1243–1250. https://doi.org/10.1094/MPMI.2000.13.11.1243
Mitchell B (2002) Resource and environmental management. Pearson Education, Harlow
Morais PV, Branco R, Francisco R (2011) Chromium resistance strategies and toxicity: what makes Ochrobactrum tritici 5bvl1 a strain highly resistant. Biometals 24:401–410. https://doi.org/10.1007/s10534-011-9446-1
Ngom A, Nakagawa Y, Sawada H, Tsukahara J, Wakabayashi S, Uchiumi T et al (2004) A novel symbiotic nitrogen-fixing member of the Ochrobactrum clade isolated from root nodules of Acacia mangiu. J Gen Appl Microbiol 50:17–27. https://doi.org/10.2323/jgam.50.17
Qin Y, Dittmer PJ, Park JG, Jansen KB, Palmer AE (2011) Measuring steady-state and dynamic endoplasmic reticulum and Golgi Zn2+ with genetically encoded sensors. Proc Natl Acad Sci USA 108:7351–7356. https://doi.org/10.1073/pnas.1015686108
Ramos HJO, Roncato-Maccari LDB, Souza EM, Soares-Ramos JRL, Hungria M, Pedrosa FO (2002) Monitoring Azospirillum-wheat interactions using the gfp and gusA genes constitutively expressed from a new broad-host range vector. J Biotechnol 97:243–252. https://doi.org/10.1016/S0168-1656(02)00108-6
Reinhold-Hurek B, Hurek T (2011) Living inside plants: bacterial endophytes. Curr Opin Plant Biol 14:435–443. https://doi.org/10.1016/j.pbi.2011.04.004
Rodrigues EP, Rodrigues LS, Oliveira ALM, Baldani VLD, dos Teixeira KRS, Urquiaga S et al (2008) Azospirillum amazonense inoculation: effects on growth, yield and N2 fixation of rice (Oryza sativa L.). Plant Soil 302:249–261. https://doi.org/10.1007/s11104-007-9476-1
Roncato-Maccari LD, Ramos HJ, Pedrosa FO, Alquini Y, Chubatsu LS, Yates MG et al (2003) Endophytic Herbaspirillum seropedicae expresses nif genes in gramineous plants. FEMS Microbiol Ecol 45:39–47. https://doi.org/10.1016/S0168-6496(03)00108-9
Sant’Anna FAD, Débora T, Shana W, Irene S (2011) Tools for genetic manipulation of the plant growth-promoting bacterium Azospirillum amazonense. BMC Microbiol 11:107. https://doi.org/10.1186/1471-2180-11-107
Singh HP, Mahajan P, Kaur S, Batish DR, Kohli RK (2013) Chromium toxicity and tolerance in plants. Environ Chem Lett 11:229–254. https://doi.org/10.1007/s10311-013-0407-5
Sousa T, Branco R, Piedade AP, Morais PV (2015) Hyper accumulation of arsenic in mutants of Ochrobactrum tritici silenced for arsenite efflux pumps. PLoS ONE 10:e0131317. https://doi.org/10.1371/journal.pone.0131317
Thévenot DR, Toth K, Durst RA, Wilson GS (2001) Electrochemical biosensors: recommended definitions and classification. Biosens Bioelectron 16:121–131. https://doi.org/10.1081/AL-100103209
Tibazarwa C, Corbisier P, Mench M, Bossus A, Solda PV, Mergeay M et al (2001) A microbial biosensor to predict bioavailable nickel in soil and its transfer to plants. Environ Pollut 113:19–26. https://doi.org/10.1016/S0269-7491(00)00177-9
Trujillo ME, Willems A, Abril A, Planchuelo A-M, Rivas R, Ludena D et al (2005) Nodulation of Lupinus albus by strains of Ochrobactrum lupini sp. nov. Appl Environ Microbiol 71:1318–1327. https://doi.org/10.1128/AEM.71.3.1318-1327.2005
United States Environmental Protection Agency—USEPA (1998) SW-846: test methods for evaluating solid waste, physical and chemical methods. USEPA, Washington
Usberti Filho JA, Azzini LE, Camargo OAB, Soave J, Schmidt NC, Villela OV et al (1986) IAC-4440: novo cultivar de arroz irrigado para o Estado de São Paulo. Instituto Agronômico, Campinas, Brazil
Verma SC, Singh A, Chowdhury SP, Tripathi AK (2004) Endophytic colonization ability of two deep-water rice endophytes, Pantoea sp. and Ochrobactrum sp. using green fluorescent protein reporter. Biotechnol Lett 26:425–429. https://doi.org/10.1023/B:BILE.0000018263.94440.ab
World Health Organization (WHO) (2011) Chemical fact sheet. Chromium. In: World Health Organization (ed), Guidelines for drinking-water quality, 4th edn. WHO, Geneva, p. 340
Zurdo-Pineiro JL, Rivas R, Trujillo ME, Vizcaino N, Carrasco JA, Chamber M et al (2007) Ochrobactrum cytisi sp. nov., isolated from nodules of Cytisus scoparius in Spain. Int J Syst Evol Microbiol 57:784–788. https://doi.org/10.1099/ijs.0.64613-0
Acknowledgements
This work was financed by the “Science without Borders” program of Brazilian government through Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), process #400756/2012–9, and also R.F. “pós-doutorado júnior (PDJ)” fellowship #160619/2012–2, and R.B. “pesquisador visitante especial (PVE)” fellowship #304009/2012–1 and project ERA-MIN 02 2015 BioCriticalMetals. We thank Juliana Meneses, Isabel Alves, and Ednelson Gomes (Embrapa Agrobiologia) for help on sample digestion, and Nelson Moura Brasil and Jair Guedes (Universidade Federal Rural do Rio de Janeiro) for helping on atomic absorption spectrometry analyses.
Author information
Authors and Affiliations
Corresponding author
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Francisco, R., Branco, R., Schwab, S. et al. Two plant-hosted whole-cell bacterial biosensors for detection of bioavailable Cr(VI). World J Microbiol Biotechnol 35, 129 (2019). https://doi.org/10.1007/s11274-019-2703-0
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s11274-019-2703-0