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.
This is a preview of subscription content, log in via an institution to check access.
References
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
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
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
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
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
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
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
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
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
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
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
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
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
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
Döbereiner J (1992) Recent changes in concepts of plant bacteria interactions: endophytic N2 fixing bacteria. Ciênc Cult 44:310–313
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
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
Glick BR (2012) Plant growth-promoting bacteria: mechanisms and applications. Scientifica 2012:963401. doi:10.6064/2012/963401
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
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
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
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
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
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
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
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
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
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
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
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 107–131
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
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
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
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
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
Zimmer M (2002) Green fluorescent protein (GFP): applications, structure, and related photophysical behavior. Chem Rev 102:759–781. doi:10.1021/cr010142r
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.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Puri, A., Padda, K.P. & Chanway, C.P. 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. Symbiosis 69, 123–129 (2016). https://doi.org/10.1007/s13199-016-0385-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13199-016-0385-z