Diazotrophic Paenibacillus beijingensis BJ-18 Provides Nitrogen for Plant and Promotes Plant Growth, Nitrogen Uptake and Metabolism

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 <i>Paenibacillus beijingensis</i> 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 <i>P. beijingensis</i> 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 <i>nifH</i> 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: <i>AMT</i> (ammonium transporter), <i>NRT</i> (nitrate transporter), <i>NiR</i> (nitrite reductase), <i>NR, GS</i> and <i>GOGAT</i> (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, <i>P. beijingensis</i> 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.