The role of grain size and inoculum amount on biocrust formation by Leptolyngbya ohadii

Cyanobacteria are widespread prokaryotic organisms that represent feasible biotechnological tools to set up valid approaches to counteract desertification. Their peculiar physiological traits, and their resilience to abiotic stresses, allow their application on abiotically constrained soils to trigger their stabilization. A successful cyanobacteria inoculation results in the formation of cyanobacterial biocrusts, complex microbial communities characterized by tangled filament meshes imbued in a matrix of self-secreted extracellular polysaccharides (EPS) that keep loose sediments and aggregates firmly in place. However, the capability to form stable cyanobacterial biocrusts is not common to all the species, and a mix of factors can hamper the process, notably inoculum amount, and substrate characteristics. The aim of this work was to assess the influence of inoculum quantity and substrate granulometry on the physical stability of cyanobacterial biocrusts induced by inoculating the strain Leptolyngbya ohadii in a microcosm experiment, under laboratory conditions. After applying three different initial inoculum amounts on two different sand granulometries (medium and coarse sand), we assayed aggregate stability, physical stability and surface hydrophobicity on the resulting biocrusts during a 30-day incubation. Also, the features and the role of the EPS synthesized by L. ohadii were studied following their isolation, characterization, and direct application on the sand. The two EPS fractions produced by the strain, one more soluble and easily released in the surrounding medium (released polysaccharides, RPS) and one solidly attached to the filaments (glycocalyx EPS, G-EPS), were separately tested. Cyanobacterial biocrusts visibly formed in all the microcosms after 15 days. However, we observed a strong effect of sand granulometry in affecting aggregate stability and tensile strength, both of which appeared weaker on coarse sand. A higher amount of initial inoculum was necessary to produce stable biocrusts on coarse sand compared to medium sand. Also, we observed how the inoculation of EPS alone did not sort most of the significant effects that we detected by inoculating the whole culture, pointing at the importance of the action of the cyanobacterial filaments in soil conglomeration. However, a significant increase in physical stability was achieved by inoculating G-EPS on medium sand, suggesting the involvement of this fraction in biocrusts structuration. This work analyzes for the first time the effects of the variable grain size and inoculum amount in the achievement of physically stable biocrusts by cyanobacteria inoculation. The results that we obtained are useful in improving and optimizing the process of biomass preparation and dispersion for future indoor and outdoor studies.