Skip to main content

Advertisement

SpringerLink
Log in

Inoculating plants with the endophytic bacterium Pseudomonas sp. Ph6-gfp to reduce phenanthrene contamination

Environmental Science and Pollution Research Aims and scope Submit manuscript

Abstract

Plant organic contamination poses a serious threat to the safety of agricultural products and human health worldwide, and the association of endophytic bacteria with host plants may decrease organic pollutants in planta. In this study, we firstly determined the growth response and biofilm formation of endophytic Pseudomonas sp. Ph6-gfp, and then systematically evaluated the performance of different plant colonization methods (seed soaking (SS), root soaking (RS), leaf painting (LP)) for circumventing the risk of plant phenanthrene (PHE) contamination. After inoculation for 48 h, strain Ph6-gfp grew efficiently with PHE, oxalic acid, or malic acid as the sole sources of carbon and energy. Moreover, strain Ph6-gfp could form robust biofilms in LB medium. In greenhouse hydroponic experiments, strain Ph6-gfp could actively colonize inoculated plants internally, and plants colonized with Ph6-gfp showed a higher capacity for PHE removal. Compared with the Ph6-gfp-free treatment, the accumulations of PHE in Ph6-gfp-colonized plants via SS, RS, and LP were 20.1, 33.1, and 7.1 %, respectively, lower. Our results indicate that inoculating plants with Ph6-gfp could lower the risk of plant PHE contamination. RS was most efficient for improving PHE removal in whole plant bodies by increasing the cell numbers of Ph6-gfp in plant roots. The findings in this study provide an optimized method to strain Ph6-gfp reduce plant PAH residues, which may be applied to agricultural production in PAH-contaminated soil.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price includes VAT (France)

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  • Afzal M, Yousaf S, Reichenauer TG, Kuffner M, Sessitsch A (2011) Soil type affects plant colonization, activity and catabolic gene expression of inoculated bacterial strains during phytoremediation of diesel. J Hazard Mater 186:1568–1575

    Article  CAS  Google Scholar 

  • Afzal M, Yousaf S, Reichenauer TG, Sessitsch A (2012) The inoculation method affects colonization and performance of bacterial inoculant strains in the phytoremediation of soil contaminated with diesel oil. Int J Phytoremediat 14:35–47

    Article  Google Scholar 

  • Afzal M, Khan S, Iqbal S, Mirza MS, Khan QM (2013) Inoculation method affects colonization and activity of Burkholderia phytofirmans PsJN during phytoremediation of diesel-contaminated soil. Int J Biodeterior Biodegrad 85:331–336

    Article  CAS  Google Scholar 

  • Afzal M, Khan QM, Sessitsch A (2014) Endophytic bacteria: prospects and applications for the phytoremediation of organic pollutants. Chemosphere 117:232–242

    Article  CAS  Google Scholar 

  • Andria V, Reichenauer TG, Sessitsch A (2009) Expression of alkane monooxygenase (alkB) genes by plant-associated bacteria in the rhizosphere and endosphere of Italian ryegrass (Lolium multiflorum L.) grown in diesel contaminated soil. Environ Pollut 157:3347–3350

    Article  CAS  Google Scholar 

  • Bacilio-Jimenez M, Aguilar-Flores S, Ventura-zapata E, Perez-Campos E, Bouquelet S, Zenteno E (2003) Chemical characterization of root exudates from rice (Oryza sativa) and their effects on the chemotactic response of endophytic bacteria. Plant Soil 249:271–277

    Article  CAS  Google Scholar 

  • Baract T, Weyens N, Oeyen L, Taghavi S, van der Lelie D, Dubin D, Spliet M, Vangronsveld J (2009) Field note: hydraulic containment of a BTEX plume using poplar trees. Int J Phytoremediat 11:416–424

    Article  Google Scholar 

  • Bhushan B, Samanta SK, Chauhan A, Chakraborti AK, Jain RK (2000) Chemotaxis and degradation of 3-methyl-4-nitrophenol by Ralstonia sp. SJ98. Biochem Biophys Res Commun 275:129–133

    Article  CAS  Google Scholar 

  • Collins C, Fryer M, Grosso A (2006) Plant uptake of non-ionic organic chemicals. Environ Sci Technol 40:45–52

    Article  CAS  Google Scholar 

  • Compant S, Reiter B, Sessitsch A, Nowak J, Clement C, Barka EA (2005) Endophytic colonization of Vitis vinifera L. by plant grown-promoting bacterium Burkholderia sp. strain PsJN. Appl Environ Microbiol 71:1685–1693

    Article  CAS  Google Scholar 

  • De Weert S, Vermeiren H, Mulders IH, Hendrickx N, Bloemberg GV, Vanderleyden J, Mot RD, Lugtenberg BJJ (2002) Flagella-driven chemotaxis towards exudate components is an important trait for tomato root colonization by Pseudomonas fluorescens. Mol Plant Microbe Interact 15:1173–1180

    Article  Google Scholar 

  • Dong YM, Iniguez AL, Ahmer BMM, Triplett EW (2003) Kinetics and strain specificity of rhizosphere and endophytic colonization by enteric bacteria on seedlings of Medicago sativa and Medicago truncatula. Appl Environ Microbiol 69:1783–1790

    Article  CAS  Google Scholar 

  • Errampalli D, Leung K, Cassidy MB, Kostrzynska M, Blears M, Lee H, Trevors JT (1999) Applications of the green fluorescent protein as a molecular marker in environmental microorganisms. J Microbiol Methods 35:187–199

    Article  CAS  Google Scholar 

  • Ferreira A, Quecine MC, Lacava PT, Oda S, Azevedo JL, Araujo WL (2008) Diversity of endophytic bacteria from Eucalyptus species seeds and colonization of seedlings by Pantoea agglomerans. FEMS Microbiol Lett 287:8–14

    Article  CAS  Google Scholar 

  • Gao YZ, Collins CD (2009) Uptake pathways of polycyclic aromatic hydrocarbons in white clover. Environ Sci Technol 43:6190–6195

    Article  CAS  Google Scholar 

  • Gao YZ, Zhu LZ (2004) Plant uptake, accumulation and translocation of phenanthrene and pyrene in soils. Chemosphere 55:1169–1178

    Article  CAS  Google Scholar 

  • Gao YZ, Xiong W, Ling WT, Wang H, Ren LL, Zhang ZY (2008) Partitioning of polycyclic aromatic hydrocarbons between plant roots and water. Plant Soil 311:201–209

    Article  CAS  Google Scholar 

  • Germaine K, Keogh E, Garcia-Cabellos G, Borremans B, van der Lelie D, Barac T, Oeyen L, Vangronsveld J, Moore FP, Moore ERB, Campbell CD, Ryan D, Dowling DN (2004) Colonisation of poplar trees by gfp expressing bacterial endophytes. FEMS Microbiol Ecol 48:109–118

    Article  CAS  Google Scholar 

  • Germaine KJ, Liu XM, Cabello GG, Hogan JP, Ryan D, Dowling DN (2006) Bacterial endophyte-enhanced phytoremediation of the organochlorine herbicide 2,4-dichlorophenoxyacetic acid. FEMS Microbiol Ecol 57:302–310

    Article  CAS  Google Scholar 

  • Germaine KJ, Keogh E, Ryan D, Dowling DN (2009) Bacterial endophyte-mediated naphthalene phytoprotection and phytoremediation. FEMS Microbiol Lett 296:226–234

    Article  CAS  Google Scholar 

  • Glick BR (2014) Bacteria with ACC deaminase can promote plant growth and help to feed the world. Microbiol Res 169:30–39

    Article  CAS  Google Scholar 

  • Harish S, Kavino M, Kumar N, Balasubramanian P, Samiyappan R (2009) Induction of defense-related proteins by mixtures of plant growth promoting endophytic bacteria against banana bunchy top virus. Biol Control 51:16–25

    Article  CAS  Google Scholar 

  • Iijima S, Washio K, Okahara R, Morikawa M (2009) Biofilm formation and proteolytic activities of Pseudoalteromonas bacteria that were isolated from fish farm sediments. Microb Biotechnol 2:361–369

    Article  CAS  Google Scholar 

  • Ishii S, Koki J, Unno H, Hori K (2004) Two morphological types of cell appendages on a strongly adhesive bacterium Acinetobacter sp. strain Tol 5. Appl Environ Microbiol 70:5026–5029

    Article  CAS  Google Scholar 

  • James EK, Reis VM, Baldani JI, Döbereiner J (1994) Infection of sugar cane by the nitrogen-fixing bacterium Acetobacter diazotrophicus. J Exp Bot 45:757–766

    Article  CAS  Google Scholar 

  • James EK, Gyaneshwar P, Mathan N, Barraquio WL, Reddy PM, Iannetta PPM, Olivares FL, Ladha JK (2002) Infection and colonization of rice seedlings by the plant growth-promoting bacterium Herbaspirillum seropedicae Z67. Mol Plant-Microbe Interact 15:894–906

    Article  CAS  Google Scholar 

  • Khan S, Afzal M, Iqbal S, Khan QM (2013) Plant-bacteria partnerships for the remediation of hydrocarbon contaminated soils. Chemosphere 90:1317–1332

    Article  CAS  Google Scholar 

  • Kvesitadze E, Sadunishvili T, Kvesitadze G (2009) Mechanisms of organic contaminants uptake and degradation in plants. World Acad Sci Eng Technol 55:458–468

    Google Scholar 

  • Liu J, Liu S, Sun K, Sheng YH, Gu YJ, Gao YZ (2014) Colonization on root surface by a phenanthrene-degrading endophytic bacterium and its application for reducing plant phenanthrene contamination. PLoS One 9, e108249

    Article  Google Scholar 

  • Lugtenberg BJ, Dekkers L, Bloemberg GV (2001) Molecular determinants of rhizosphere colonization by Pseudomonas. Annu Rev Phytopathol 39:461–490

    Article  CAS  Google Scholar 

  • Mckone TE, Maddalena RL (2007) Plant uptake of organic pollutants from soil: bioconcentration estimates based on models and experiments. Environ Toxicol Chem 26:2494–2504

    Article  CAS  Google Scholar 

  • Nelson EB (2004) Microbial dynamics and interactions in the spermosphere. Annu Rev Phytopathol 42:271–309

    Article  CAS  Google Scholar 

  • Newman LA, Reynolds CM (2005) Bacteria and phytoremediation: new uses for endophytic bacteria in plants. Trends Biotechnol 23:6–8

    Article  CAS  Google Scholar 

  • Porteous Moore F, Barac T, Borremans B, Oeyen L, Vangronsveld J, van der Lelie D, Campbell CD, Moore ERB (2006) Endophytic bacterial diversity in poplar trees growing on a BTEX-contaminated site: the characterisation of isolates with potential to enhance phytoremediation. Syst Appl Microbiol 29:539–556

    Article  Google Scholar 

  • Ramey BE, Koutsoudis M, von Bodman SB, Fuqua C (2004) Biofilm formation in plant-microbe associations. Curr Opin Microbiol 7:602–609

    Article  CAS  Google Scholar 

  • Ryan RP, Germaine K, Franks A, Ryan DJ, Dowling DN (2008) Bacterial endophytes: recent developments and applications. FEMS Microbiol Lett 278:1–9

    Article  CAS  Google Scholar 

  • Sarand I, Timonen S, Nurmiaho-Lassila EL, Koivula T, Haahtela K (1998) Microbial biofilms and catabolic plasmid harbouring degradative fluorescent pseudomonads in Scots pine mycorrhizospheres developed on petroleum contaminated soil. FEMS Microbiol Ecol 27:115–126

    Article  CAS  Google Scholar 

  • Siciliano SD, Fortin N, Mihoc A, Wisse G, Labelle S, Beaumier D, Ouellette D, Roy R, Whyle LG, Banks MK, Schwab P, Lee K, Greer CW (2001) Selection of specific endophytic bacterial genotypes by plants in response to soil contamination. Appl Environ Microbiol 67:2469–2475

    Article  CAS  Google Scholar 

  • Singh R, Paul D, Jain RK (2006) Biofilms: implication in bioremediation. Trends Microbiol 14:590–600

    Article  Google Scholar 

  • Strobel G, Daisy B, Castillo U, Harper J (2004) Natural products from endophytic microorganisms. J Nat Prod 67:257–268

    Article  CAS  Google Scholar 

  • Sun K, Liu J, Gao YZ, Jin L, Gu YJ, Wang WQ (2014a) Isolation, plant colonization potential, and phenanthrene degradation performance of the endophytic bacterium Pseudomonas sp. Ph6-gfp. Sci Rep 4:5462

    CAS  Google Scholar 

  • Sun K, Liu J, Jin L, Gao YZ (2014b) Utilizing pyrene-degrading endophytic bacteria to reduce the risk of plant pyrene contamination. Plant Soil 374:251–262

    Article  CAS  Google Scholar 

  • Taghavi S, Barac T, Greenberg B, Borremans B, Vangronsveld J, van der Lelie D (2005) Horizontal gene transfer to endogenous endophytic bacteria from poplar improves phytoremediation of toluene. Appl Environ Microbiol 71:8500–8505

    Article  CAS  Google Scholar 

  • Torres AR, Araújo WL, Coursino L, Rossetto PB, Mondin M, Hungria M, Azevedo JL (2013) Colonization of Madagascar periwinkle (Catharanthus roseus), by endophytes encoding gfp marker. Arch Microbiol 195:483–489

    Article  CAS  Google Scholar 

  • Wang YJ, Xiao M, Geng XL, Liu JY, Chen J (2007) Horizontal transfer of genetic determinants for degradation of phenol between the bacteria living in plant and its rhizosphere. Appl Microbiol Biotechnol 77:733–739

    Article  CAS  Google Scholar 

  • Weyens N, van der Lelie D, Artois T, Smeets K, Taghavi S, Newman L, Carleer R, Vangronsveld J (2009a) Bioaugmentation with engineered endophytic bacteria improves contaminant fate in phytoremediation. Environ Sci Technol 43:9413–9418

    Article  CAS  Google Scholar 

  • Weyens N, van der Lelie D, Taghavi S, Vangronsveld J (2009b) Phytoremediation: plant endophyte partnerships take the challenge. Curr Opin Biotechnol 20:248–254

    Article  CAS  Google Scholar 

  • Weyens N, Boulet J, Adriaensen D, Timmermans JP, Prinsen E, Oevelen SV, D’Hean J, Smeets K, van der Lelie D, Taghavi S, Vangronsveld J (2012) Contrasting colonization and plant growth promoting capacity between wild type and a gfp-derative of the endophyte Pseudomonas putida W619 in hybrid poplar. Plant Soil 356:217–230

    Article  CAS  Google Scholar 

  • Wilson D (1995) Endophyte-the evolution of a term, and clarification of its use and definition. Oikos 73:274–276

    Article  Google Scholar 

  • Yousaf S, Afzal M, Reichenauer TG, Brady CL, Sessitsch A (2011) Hydrocarbon degradation, plant colonization and gene expression of alkane degradation genes by endophytic Enterobacter ludwigii strains. Environ Pollut 159:2675–2683

    Article  CAS  Google Scholar 

  • Zakria M, Ohsako A, Saeki YC, Yamamoto A, Akao S (2007) Colonization and nitrogen-fixing ability of Herbaspirillum sp strain B501 gfp1 and assessment of its growth-promoting ability in cultivated rice. Microbes Environ 22:197–206

    Article  Google Scholar 

  • Zhu X, Ni X, Liu J, Gao YZ (2014) Application of endophytic bacteria to reduce persistent organic pollutants contamination in plants. CLEAN-Soil Air Water 42:306–310

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was financially supported by the National Science Foundation of China (41201501, 41171380, 51278252, 21477056), the Science Foundation of Jiangsu Province (BK2012370, BK20130030), and the Doctoral Fund of the Ministry of Education of China (20120097120012).

Author information

Authors and Affiliations

  1. Institute of Organic Contaminant Control and Soil Remediation, College of Resources and Environmental Sciences, Nanjing Agricultural University, Weigang Road 1, Nanjing, 210095, China

    Kai Sun, Juan Liu, Yanzheng Gao, Yuehui Sheng, Fuxing Kang & Michael Gatheru Waigi

Authors
  1. Kai Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Juan Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yanzheng Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yuehui Sheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Fuxing Kang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Michael Gatheru Waigi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yanzheng Gao.

Additional information

Responsible editor: Robert Duran

K. Sun and J. Liu contribute equally to this paper.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sun, K., Liu, J., Gao, Y. et al. Inoculating plants with the endophytic bacterium Pseudomonas sp. Ph6-gfp to reduce phenanthrene contamination. Environ Sci Pollut Res 22, 19529–19537 (2015). https://doi.org/10.1007/s11356-015-5128-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11356-015-5128-9

Keywords

Search

Navigation