Advertisement

Symbiosis

, Volume 69, Issue 2, pp 123–129 | Cite as

Seedling growth promotion and nitrogen fixation by a bacterial endophyte Paenibacillus polymyxa P2b-2R and its GFP derivative in corn in a long-term trial

  • Akshit Puri
  • Kiran Preet Padda
  • Chris P. Chanway
Short Communication

Abstract

A plant growth promoting endophyte, Paenibacillus polymyxa P2b-2R, originally isolated form a lodgepole pine seedling and its green fluorescent protein (GFP) derivative, P2b-2Rgfp, were evaluated for their ability to survive, fix atmospheric nitrogen (N) and promote plant growth when inoculated into corn (Zea Mays L.) in a long-term trial. We were also interested to see the effects of GFP-tagging of P2b-2R on its ability to promote growth of corn seedlings in a long-term study. Corn seedlings were inoculated with either strain P2b-2R or P2b-2Rgfp and non-inoculated seedlings were treated as controls. Seedlings were harvested after 3 months and evaluated for plant growth promotion (length and biomass) and N fixation (15N foliar dilution assay). Colonization and survival of P2b-2R and P2b-2Rgfp outside (rhizosphere) and inside (internal tissues) the inoculated seedlings were also determined. Both strains survived inside and outside corn seedlings forming rhizospheric and endophytic colonies in stem and root tissues. Inoculation by P2b-2R strain promoted corn plant growth via enhancing seedling length and biomass by 52 % and 53 %, respectively. Similarly, P2b-2Rgfp inoculation enhanced seedling length by 68 % and biomass by 67 %. Corn seedlings inoculated with strain P2b-2R derived 30 % of foliar N from the atmosphere and seedlings inoculated with P2b-2Rgfp derived 32 % of foliar N from the atmosphere. But there was no statistically significant difference between P2b-2R and P2b-2Rgfp treated seedlings in terms of overall seedling length, biomass and amount of N fixed in this long-term trial. These results combined with the results from an earlier study suggest that P. polymyxa P2b-2R and its GFP-tagged derivative is capable of enhancing overall plant growth throughout the life cycle of corn plant.

Keywords

Corn Paenibacillus polymyxa Bacterial endophytes Plant growth promoting bacteria Nitrogen fixation Plant growth promotion 

Notes

Acknowledgments

The authors would like to dedicate this work to Late Mr. Darshan K. Puri, whose lovely memories were the motivation behind this project. This study was supported through funding from Natural Sciences and Engineering Research Council of Canada (NSERC) Discovery Grant (RGPIN 41832–13) to Dr. Chris P. Chanway.

References

  1. Anand R, Chanway CP (2013a) Detection of GFP-labeled Paenibacillus polymyxa in auto-fluorescing pine seedling tissues. Biol Fertil Soils 49:111–118. doi: 10.1007/s00374-012-0727-9 CrossRefGoogle Scholar
  2. Anand R, Chanway C (2013b) N2-fixation and growth promotion in cedar colonized by an endophytic strain of Paenibacillus polymyxa. Biol Fertil Soils 49:235–239. doi: 10.1007/s00374-012-0735-9 CrossRefGoogle Scholar
  3. Anand R, Chanway CP (2013c) nif gene sequence and arrangement in the endophytic diazotroph Paenibacillus polymyxa strain P2b-2R. Biol Fertil Soils 49:965–970. doi: 10.1007/s00374-013-0793-7 CrossRefGoogle Scholar
  4. Anand R, Grayston S, Chanway CP (2013) N2-fixation and seedling growth promotion of lodgepole pine by endophytic Paenibacillus polymyxa. Microb Ecol 66:369–374. doi: 10.1007/s00248-013-0196-1 CrossRefPubMedGoogle Scholar
  5. Bal A, Chanway CP (2012a) Evidence of nitrogen fixation in lodgepole pine inoculated with diazotrophic Paenibacillus polymyxa. Botany 90:891–896. doi: 10.1139/b2012-044 CrossRefGoogle Scholar
  6. Bal A, Chanway CP (2012b) 15N foliar dilution of western red cedar in response to seed inoculation with diazotrophic Paenibacillus polymyxa. Biol Fertil Soils 48:967–971. doi: 10.1007/s00374-012-0699-9 CrossRefGoogle Scholar
  7. Bal A, Anand R, Berge O, Chanway CP (2012) Isolation and identification of diazotrophic bacteria from internal tissues of Pinus contorta and Thuja plicata. Can J For Res 42:807–813. doi: 10.1139/x2012-023 CrossRefGoogle Scholar
  8. Baldani JI, Olivares FL, Hemerly AS et al (1998) Nitrogen-fixing endophytes: recent advances in the association with graminaceous plants grown in the tropics. In: Elmerich EC (ed) Biological nitrogen fixation for the 21st century. Springer, The Netherlands, pp 203–206. doi: 10.1007/978-94-011-5159-7_90 CrossRefGoogle Scholar
  9. Bressan W, Borges MT (2004) Delivery methods for introducing endophytic bacteria into maize. Biocontrol 49:315–322. doi: 10.1023/B:BICO.0000025372.51658.93 CrossRefGoogle Scholar
  10. Çakmakçi R, Dönmez F, Aydın A, Şahin F (2006) Growth promotion of plants by plant growth-promoting rhizobacteria under greenhouse and two different field soil conditions. Soil Biol Biochem 38:1482–1487. doi: 10.1016/j.soilbio.2005.09.019 CrossRefGoogle Scholar
  11. Chalfie M, Tu Y, Euskirchen G, Ward W, Prasher D (1994) Green fluorescent protein as a marker for gene expression. Science 263:802–805. doi: 10.1126/science.8303295 CrossRefPubMedGoogle Scholar
  12. Chanway CP, Holl FB, Nelson LM (1988) Cultivar-specific growth promotion of spring wheat (Triticum aestivum L.) by coexistent Bacillus species. Can J Microbiol 34:925–929. doi: 10.1139/m88-164 CrossRefGoogle Scholar
  13. Chanway C, Anand R, Yang H (2014) Nitrogen fixation outside and inside plant tissues. In: Ohyama T (ed) Advances in biology and ecology of nitrogen fixation. InTech, Croatia, pp 3–23. doi: 10.5772/57532 Google Scholar
  14. Compant S, Reiter B, Sessitsch A, Nowak J, Clément C, Barka EA (2005) Endophytic colonization of Vitis vinifera L. by plant growth-promoting bacterium Burkholderia sp. strain PsJN. Appl Environ Microbiol 71:1685–1693. doi: 10.1128/AEM.71.4.1685-1693.2005 CrossRefPubMedPubMedCentralGoogle Scholar
  15. Döbereiner J (1992) Recent changes in concepts of plant bacteria interactions: endophytic N2 fixing bacteria. Ciênc Cult 44:310–313Google Scholar
  16. Figueiredo MVB, Burity HA, Martínez CR, Chanway CP (2008) Alleviation of drought stress in the common bean (Phaseolus vulgaris L.) by co-inoculation with Paenibacillus polymyxa and Rhizobium tropici. Appl Soil Ecol 40:182–188. doi: 10.1016/j.apsoil.2008.04.005 CrossRefGoogle Scholar
  17. Freitas A, Vieira CL, Santos C, Stamford N, Lyra M (2007) Caracterização de rizóbios isolados de Jacatupé cultivado em solo salino do estado de Pernambuco, Brasil. Bragantia 66:497–504. doi: 10.1590/S0006-87052007000300017 CrossRefGoogle Scholar
  18. Glick BR (2012) Plant growth-promoting bacteria: mechanisms and applications. Scientifica 2012:963401. doi: 10.6064/2012/963401 CrossRefPubMedPubMedCentralGoogle Scholar
  19. Gouzou L, Burtin G, Philippy R, Bartoli F, Heulin T (1993) Effect of inoculation with Bacillus polymyxa on soil aggregation in the wheat rhizosphere: preliminary examination. Geoderma 56:479–491. doi: 10.1016/0016-7061(93)90128-8 CrossRefGoogle Scholar
  20. Haggag WM, Timmusk S (2008) Colonization of peanut roots by biofilm-forming Paenibacillus polymyxa initiates biocontrol against crown rot disease. J Appl Microbiol 104:961–969. doi: 10.1111/j.1365-2672.2007.03611.x CrossRefPubMedGoogle Scholar
  21. Lal S, Tabacchioni S (2009) Ecology and biotechnological potential of Paenibacillus polymyxa: a minireview. Indian J Microbiol 49:2–10. doi: 10.1007/s12088-009-0008-y CrossRefPubMedPubMedCentralGoogle Scholar
  22. Lebuhn M, Heulin T, Hartmann A (1997) Production of auxin and other indolic and phenolic compounds by Paenibacillus polymyxa strains isolated from different proximity to plant roots. FEMS Microbiol Ecol 22:325–334. doi: 10.1016/S0168-6496(97)00007-X CrossRefGoogle Scholar
  23. Meng X, Yan D, Long X, Wang C, Liu Z, Rengel Z (2014) Colonization by endophytic Ochrobactrum anthropi Mn1 promotes growth of Jerusalem artichoke. Microb Biotechnol 7:601–610. doi: 10.1111/1751-7915.12145 CrossRefPubMedPubMedCentralGoogle Scholar
  24. Nielsen P, Sørensen J (1997) Multi-target and medium-independent fungal antagonism by hydrolytic enzymes in Paenibacillus polymyxa and Bacillus pumilus strains from barley rhizosphere. FEMS Microbiol Ecol 22:183–192. doi: 10.1016/S0168-6496(96)00089-X CrossRefGoogle Scholar
  25. Padda KP (2015) Impact of GFP-modification of Paenibacillus polymyxa on its ability to enhance growth of corn, canola and tomato seedlings. Master’s Thesis, University of British Columbia, Canada. http://hdl.handle.net/2429/55019. Accessed 11 February 2016
  26. Padda KP, Puri A, Chanway CP (2015) Effect of GFP tagging of Paenibacillus polymyxa P2b-2R on its ability to promote growth of canola and tomato seedlings. Biol Fertil Soils. doi: 10.1007/s00374-015-1083-3 Google Scholar
  27. Puri A, Padda KP, Chanway CP (2015) Can a diazotrophic endophyte originally isolated from lodgepole pine colonize an agricultural crop (corn) and promote its growth? Soil Biol Biochem 89:210–216. doi: 10.1016/j.soilbio.2015.07.012 CrossRefGoogle Scholar
  28. Puri A, Padda KP, Chanway CP (2016) Evidence of nitrogen fixation and growth promotion in canola (Brassica napus L.) by an endophytic diazotroph Paenibacillus polymyxa P2b-2R. Biol Fertil Soils 52:119–125. doi: 10.1007/s00374-015-1051-y CrossRefGoogle Scholar
  29. Rennie RJ (1981) A single medium for the isolation of acetylene-reducing (dinitrogen-fixing) bacteria from soils. Can J Microbiol 27:8–14. doi: 10.1139/m81-002 CrossRefPubMedGoogle Scholar
  30. Rennie RJ, Fried M, Rennie DA (1978) Concepts of 15N usage in dinitrogen fixation studies. In: Isotopes in biological dinitrogen fixation. International Atomic Energy Agency, Vienna, pp 107–131Google Scholar
  31. Rodriguez H, Mendoza A, Antonia Cruz M, Holguin G, Glick BR, Bashan Y (2006) Pleiotropic physiological effects in the plant growth-promoting bacterium Azospirillum brasilense following chromosomal labeling in the clpX gene. FEMS Microbiol Ecol 57:217–225. doi: 10.1111/j.1574-6941.2006.00111.x CrossRefPubMedGoogle Scholar
  32. Timmusk S, Nicander B, Granhall U, Tillberg E (1999) Cytokinin production by Paenibacillus polymyxa. Soil Biol Biochem 31:1847–1852. doi: 10.1016/S0038-0717(99)00113-3 CrossRefGoogle Scholar
  33. Timmusk S, Grantcharova N, Wagner EH (2005) Paenibacillus polymyxa invades plant roots and forms biofilms. Appl Environ Microbiol 71:7292–7300. doi: 10.1128/AEM.71.11.7292-7300.2005 CrossRefPubMedPubMedCentralGoogle Scholar
  34. Urquiaga S, Cruz KHS, Boddey RM (1992) Contribution of nitrogen fixation to sugar cane: nitrogen-15 and nitrogen-balance estimates. Soil Sci Soc Am J 56:105–114. doi: 10.2136/sssaj1992.03615995005600010017x CrossRefGoogle Scholar
  35. Weyens N, Boulet J, Adriaensen D et al (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. doi: 10.1007/s11104-011-0831-x CrossRefGoogle Scholar
  36. Zimmer M (2002) Green fluorescent protein (GFP): applications, structure, and related photophysical behavior. Chem Rev 102:759–781. doi: 10.1021/cr010142r CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  1. 1.Department of Forest and Conservation Sciences, Faculty of ForestryUniversity of British ColumbiaVancouverCanada

Personalised recommendations