Biological nitrogen fixation and plant growth promotion of lodgepole pine by an endophytic diazotroph Paenibacillus polymyxa and its GFP-tagged derivative

Abstract

Paenibacillus polymyxa P2b-2R is an endophytic diazotroph originally isolated from lodgepole pine. It is reported to fix significant amounts of nitrogen (N) and promote plant growth. To evaluate the endophytic colonization sites, a green fluorescent protein (GFP) derivative of P2b-2R was generated (P2b–2Rgfp), but the effects of GFP modification on the functioning of P2b-2R have not been fully elucidated yet. In this study, we wanted to confirm and contrast the N-fixing and plant-growth-promoting abilities of P2b-2Rgfp with those of the wild-type P2b-2R in lodgepole pine. Pine seedlings were grown in an N-limited environment and harvested 2, 4, 8, and 12 months after inoculation to evaluate the endophytic and rhizospheric colonization by both strains, the amount of N-fixed, and seedling growth enhancement. Both P2b-2R and P2b-2Rgfp strains formed persistent rhizospheric and endophytic populations; fixed N; and enhanced seedling growth continuously after 4 months. P2b-2Rgfp-treated seedlings outperformed the P2b-2R-treated seedlings in terms of biomass only during the initial stages of plant development, but the differences decreased during the trial and were not significant towards the end. To the best of our knowledge, this is the first study that reports the effects of GFP-tagging of an endophyte when inoculated into a gymnosperm tree species.