Abstract
Diazotrophic bacteria can reduce N2 into plant-available ammonium (NH4+), promoting plant growth and reducing nitrogen (N) fertilizer requirements. However, there are few systematic studies on the effects of diazotrophic bacteria on biological N2 fixation (BNF) contribution rate and host plant N uptake and metabolism. In this study, the interactions of the diazotrophic Paenibacillus beijingensis BJ-18 with wheat, maize, and cucumber were investigated when it was inoculated to these plant seedlings grown in both low N and high N soils, with un-inoculated plants as controls. This study showed that GFP-tagged P. beijingensis BJ-18 colonized inside and outside seedlings, forming rhizospheric and endophytic colonies in roots, stems, and leaves. The numbers of this bacterium in the inoculated plants depended on soil N levels. Under low N, inoculation significantly increased shoot dry weight (wheat 86.1%, maize 46.6%, and cucumber 103.6%) and root dry weight (wheat 46.0%, maize 47.5%, and cucumber 20.3%). The 15N-isotope-enrichment experiment indicated that plant seedlings derived 12.9–36.4% N from BNF. The transcript levels of nifH in the inoculated plants were 0.75–1.61 folds higher in low N soil than those in high N soil. Inoculation enhanced NH4+ and nitrate (NO3-) uptake from soil especially under low N. The total N in the inoculated plants were increased by 49.1–92.3% under low N and by 13–15.5% under high N. Inoculation enhanced activities of glutamine synthetase (GS) and nitrate reductase (NR) in plants, especially under low N. The expression levels of N uptake and N metabolism genes: AMT (ammonium transporter), NRT (nitrate transporter), NiR (nitrite reductase), NR, GS and GOGAT (glutamate synthase) in the inoculated plants grown under low N were up-regulated 1.5–91.9 folds, but they were not obviously changed under high N. Taken together, P. beijingensis BJ-18 was an effective, endophytic and diazotrophic bacterium. This bacterium contributed to plants with fixed N2, promoted plant growth and N uptake, and enhanced gene expression and enzyme activities involved in N uptake and assimilation in plants. However, these positive effects on plants were regulated by soil N status. This study might provide insight into the interactions of plants with beneficial associative and endophytic diazotrophic bacteria.
Highlights
Biological nitrogen (N) fixation (BNF) is the major natural process through which atmospheric N2 is reduced to bioavailable NH4+, providing a large amount of natural N into cultivated agricultural systems (Galloway et al, 2008)
We investigated the colonization pattern and contributions of N2 fixation by P. beijingensis BJ-18 to plants, and the plant responses (N uptake and metabolism processes) to the infection
This study revealed that wheat gained the highest N from BNF under both low N and high N levels, followed by cucumber and by maize, suggesting that BNF rate was related to host plant species
Summary
Biological nitrogen (N) fixation (BNF) is the major natural process through which atmospheric N2 is reduced to bioavailable NH4+, providing a large amount of natural N into cultivated agricultural systems (Galloway et al, 2008). In addition to symbiotic N2-fixing Rhizobia associated with legumes, the non-symbiotic diazotrophic bacteria are important contributors to the N nutrition of non-legumes (Gupta et al, 2006). The non-symbiotic diazotrophic bacteria are highly diverse and associated with plants in different ways. Some bacteria live in the rhizosphere and are designated rhizobacteria (Kloepper and Beauchamp, 1992). Paenibacillus polymyxa WLY78 live inside the plant without causing damage and are classified as endophytic diazotrophic bacteria (Hao and Chen, 2017). Endophytic diazotrophic bacteria may have an advantage over associative diazotrophic bacteria and rhizobacteria, since they live within plant tissues where better niches are established for N2 fixation and assimilation of fixed N2 by the plant (Reinhold-Hurek and Hurek, 1998, 2011)
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