The turfgrass industry has attempted to adopt market available N fixing bacterial strains to supply exogenous N requirements. However, turfgrass inoculations with market available N fixing strains have led to inconclusive results. Research on naturally occurring N fixers has mainly focused on abundance and diversity on roots and below ground soil. These ecological niches are known to be impacted by regular management practices of turfgrass which also interfere with the colonization rates of microorganisms. Therefore, the use of endophytic N fixing bacteria which are naturally present in turfgrasses such as bermudagrass, creeping bentgrass, and tall fescue is an attractive alternative regarding a higher microbial stability and potential of colonization. In this study we evaluated the culturing potential of N fixing bacteria from three turfgrass and three common weed species, assayed their potential to grow under different ammonium chloride (NH4Cl ) concentrations, and compared the isolates with the dominant taxa from a previous study in the community composition of N fixing bacteria in ‘CitrablueTM’ St. Augustinegrass [Stenotaphrum secundatum (Waltz)]. The plant biomass was collected from ‘CitrablueTM’, ‘Celebration’ bermudagrass [Cynodon. dactylon (L.)], ‘Empire’ zoysiagrass [Zoysia japonica (Steud)], goosegrass [Eleusine indica (L.) Gaertn], crabgrass [Digitaria sanguinalis (L.) Scoop], and bull paspalum [Paspalum setaceum (Michaux)]. The biomass was washed with water and surface sterilized using consecutive immersions in ethanol 75%, bleach 50%, and three rinses with ultrapure water. After sterilization, N fixers were isolated using a N free enrichment technique with Carbon Combined Media (CCM) and streaking on CCM solid plates. Taxonomy of the isolates was assigned using 16S and nifH Illumina miseq sequencing, consequently, the isolates were grown in liquid CCM with NH4Cl concentrations ranging from 100 uM to 0.001 uM. The isolates, Sphingomonas trueperi, Kosakonia radicitans, Herbaspirillum rubrisulbicans, Agrobacterium salinotolerans, Stenotrophomonas maltophilia, Kosakonia oryzae, Enterobacter sp. were not able to grow below 1 uM of NH4Cl in liquid CCM, however, these strains were the best performing strains because were able to grow at lower NH4Cl concentrations. Lastly, the isolates were not genotype specific to either turfgrasses or weeds and were not correlated with the dominant taxa of N fixers on the ‘CitrablueTM’ study.
