Metatranscriptomics and nitrogen fixation from the rhizoplane of maize plantlets inoculated with a group of PGPRs

Abstract

Plant roots are inhabited by a large diversity of microbes, some of which are beneficial for the growth of plants and known as plant growth promoting rhizobacteria (PGPR). In this work, we designed a multispecies inoculum of PGPRs containing Rhizobium phaseoli, Sinorhizobium americanum and Azospirillum brasilense nitrogen-fixing strains and other plant-growth promoting bacteria such as Bacillus amyloliquefaciens and Methylobacterium extorquens. We evaluated the effect of this group of bacteria on the growth of one-month-old maize plants. The multispecies inoculum exerted a beneficial effect on maize plants that was greater than that obtained with single-bacteria. Using the same multispecies inoculant, acetylene reduction was recorded in 5-day-old roots indicating active nitrogen fixation by bacteria in maize. Azospirillum nitrogen fixation was lower than that obtained with the multispecies inoculum. We focused on the analysis of R. phaseoli gene expression in presence of other PGPRs. Many R. phaseoli up- regulated genes in roots in the presence of other bacteria are hypothetical, showing our poor knowledge of bacteria-bacteria interactions. Other genes indicated bacterial nutrient competition and R. phaseoli stress. Differentially expressed transcriptional regulators were identified that may be key in bacteria-bacteria interaction regulation. Additionally, gene expression was analyzed from Azospirillum but not from sinorhizobia and methylobacteria due to the low number of transcripts obtained from maize roots. The metatranscriptomic analysis from maize roots showed expression of Azospirillum nif genes in the presence of PGPR bacteria. Our hypothesis is that other bacteria stimulate Azospirillum capacity to fix nitrogen and this should be further explored.

Keywords: Azospirillum; Bacillus; Biological nitrogen fixation; Rhizobium.