An endophytic diazotroph, Paenibacillus polymyxa P2b-2R, previously isolated from internal tissues of lodgepole pine tree was tested for its ability to colonize, fix nitrogen, and promote growth of an important oilseed crop, canola. For this, we inoculated canola seeds with P. polymyxa P2b-2R and grew them for up to 60 days. Canola seedlings were harvested thrice, 20, 40, and 60 days after inoculation, to determine the rhizospheric and endophytic P2b-2R population (colony-forming units (cfu)) and quantify the biological nitrogen fixation (15N dilution assay) and growth promotion (height and biomass). The entire experiment was repeated to confirm the treatment effects. P2b-2R colonized canola rhizosphere with a population abundance of 1.41 × 108 cfu/g dry root in the first growth trial and 7.49 × 107 cfu/g dry root in the second growth trial. P2b-2R developed a persistent endophytic population in canola root, comprising 3.48 × 105 cfu/g fresh weight in the first growth trial and 2.36 × 103 cfu/g fresh weight in the second growth trial. In the first growth trial, P2b-2R inoculation enhanced seedling height by 25 % and biomass by 30 % after 60 days of inoculation. Similarly, in the second growth trial, P2b-2R inoculation promoted seedling height by 21 % and biomass by 24 %. P2b-2R inoculated seedlings fixed 22 and 19 % of nitrogen from the atmosphere in the first and second growth trials, respectively. These results clearly indicate that P. polymyxa P2b-2R, a lodgepole pine endophyte, is capable of fixing nitrogen and promoting growth of an important oilseed crop canola, which is an indication of its broad range host capability.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Anand R, Chanway C (2013) N2-fixation and growth promotion in cedar colonized by an endophytic strain of Paenibacillus polymyxa. Biol Fertil Soils 49:235–239
Anand R, Grayston S, Chanway C (2013) N2-fixation and seedling growth promotion of lodgepole pine by endophytic Paenibacillus polymyxa. Microb Ecol 66:369–374
Bal A, Chanway CP (2012a) Evidence of nitrogen fixation in lodgepole pine inoculated with diazotrophic Paenibacillus polymyxa. Botany 90:891–896
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
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
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
Barret M, Morrissey JP, O’Gara F (2011) Functional genomics analysis of plant growth-promoting rhizobacterial traits involved in rhizosphere competence. Biol Fertil Soils 47:729–743
Bøckman OC (1997) Fertilizers and biological nitrogen fixation as sources of plant nutrients: perspectives for future agriculture. Plant Soil 194:11–14
Canola Council of Canada (n.d.) What is canola? http://www.canolacouncil.org/oil-and-meal/what-is-canola/. Accessed 3 August 2015
Chanway CP, Nelson LM, Holl FB (1988) Cultivar-specific growth promotion of spring wheat (Triticum aestivum L.) by coexistent Bacillus species. Can J Microbiol 34:925–929
Dobereiner J (1992) Recent changes in concepts of plant bacteria interactions: endophytic N2 fixing bacteria. Sci Cult 44:310–313
Glick BR, Patten CL, Holguin G, Penrose DM (1999) Biochemical and genetic mechanisms used by plant growth promoting bacteria. Imperial College Press, London, UK, p 267
Hallmann J, Quadt-Hallmann A, Mahaffee WF, Kloepper JW (1997) Bacterial endophytes in agricultural crops. Can J Microbiol 43:895–914
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
Herridge DF, Peoples MB, Boddey RM (2008) Global inputs of biological nitrogen fixation in agricultural systems. Plant Soil 311:1–18
Kandel SL, Herschberger N, Kim SH, Doty SL (2015) Diazotrophic endophytes of poplar and willow for growth promotion of rice plants in nitrogen-limited conditions. Crop Sci 55:1765–1772
Kim S, Lowman S, Hou G, Nowak J, Flinn B, Mei C (2012) Growth promotion and colonization of switchgrass (Panicum virgatum) cv. Alamo by bacterial endophyte Burkholderia phytofirmans strain PsJN. Biotechnol Biofuels 5:1–10
Ladha JK, de Bruijn FJ, Malik KA (1997) Introduction: assessing opportunities for nitrogen fixation in rice—a frontier project. Plant Soil 194:1–10
Lal S, Tabacchioni S (2009) Ecology and biotechnological potential of Paenibacillus polymyxa: a minireview. Indian J Microbiol 49:2–10
Long HH, Schmidt DD, Baldwin IT (2008) Native bacterial endophytes promote host growth in a species-specific manner; phytohormone manipulations do not result in common growth responses. PLoS ONE 3, e2702
Malik KA, Bilal R, Mehnaz S, Rasul G, Mirza MS, Ali S (1997) Association of nitrogen-fixing, plant-growth-promoting rhizobacteria (PGPR) with kallar grass and rice. Plant Soil 194:37–44
Nelson LM (2004) Plant growth promoting rhizobacteria (PGPR): prospects for new inoculants. Crop Manage 3. doi:10.1094/CM-2004-0301-05-RV
Palacios OA, Bashan Y, de Bashan LE (2014) Proven and potential involvement of vitamins in interactions of plants with plant growth-promoting bacteria—an overview. Biol Fertil Soils 50:415–432
Pii Y, Mimmo T, Tomasi N, Terzano R, Cesco S, Crecchio C (2015) Microbial interactions in the rhizosphere: beneficial influences of plant growth-promoting rhizobacteria on nutrient acquisition process. A review. Biol Fertil Soils 51:403–415
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
Quecine MC, Araujo WL, Rossetto PB et al (2012) Sugarcane growth promotion by the endophytic bacterium Pantoea agglomerans 33.1. Appl Environ Microbiol 78:7511–7518
Ravi AV, Musthafa KS, Jegathammbal G, Kathiresan K, Pandian SK (2007) Screening and evaluation of probiotics as a biocontrol agent against pathogenic Vibrios in marine aquaculture. Lett Appl Microbiol 45:219–223
Rennie RJ, Larson RI (1979) Dinitrogen fixation associated with disomic chromosome substitution lines of spring wheat. Can J Bot 57:2771–2775
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, Austria, pp 107–131
Rennie RJ, DeFreitas JR, Ruschel AP, Vose PV (1983) 15N isotope dilution to quantify dinitrogen (N2) fixation associated with Canadian and Brazilian wheat. Can J Bot 61:1667–1671
Rosenblueth M, Martínez-Romero E (2006) Bacterial endophytes and their interactions with hosts. Mol Plant-Microbe Interact 19:827–837
Timmusk S, Grantcharova N, Wagner EGH (2005) Paenibacillus polymyxa invades plant roots and forms biofilms. Appl Environ Microbiol 71:7292–7300
United States Department of Agriculture, Economic Research Service (USDA ERS) (n.d.) Soybeans and oil crops: canola. http://www.ers.usda.gov/topics/crops/soybeans-oil-crops/canola.aspx. Accessed 3 August 2015
Weyens N, Van der Lelie D, Taghavi S, Vangronsveld J (2009) Phytoremediation: plant-endophyte partnerships take the challenge. Curr Opin Biotechnol 20:248–254
Yang, H (2015) Effects of Paenibacillus polymyxa and soil nitrogen on lodgepole pine (Pinus contorta var. Latifolia (dougl.) Engelm.) seedling growth. Master’s thesis, University of British Columbia, Canada. http://hdl.handle.net/2429/53004. Accessed 3 August 2015
The authors are grateful to the late Mr. Darshan K. Puri for his immense support in completion of this study. This work was supported through funding from the Natural Sciences and Engineering Research Council of Canada (NSERC) Discovery Grant (RGPIN 41832-13) to Dr. Chris P Chanway. The authors would also like to acknowledge “SeCan Association” for generously providing canola seeds for this research work.
Electronic supplementary material
Below is the link to the electronic supplementary material.
(PDF 410 kb)
About this article
Cite this article
Puri, A., Padda, K.P. & Chanway, C.P. 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 (2016). https://doi.org/10.1007/s00374-015-1051-y
- Biological nitrogen fixation
- Plant growth promotion
- Paenibacillus polymyxa