Effective sustainable algal cultivation techniques are essential for mass production of the marine microalga Tetraselmis for biofuel and array of co-products. The phycospheric communities affect the microalgal growth and metabolism through various allelochemical and nutrient interactions; hence, their potential to affect the quantity and quality of both biomass and bioproducts is significant. In the present study, we have screened the phycospheric communities of biofuel producing Tetraselmis striata (KCTC1432BP). A total of 26 bacterial strains were isolated and identified from the phycosphere of Tetraselmis striata mass culture. Then, each bacterial strain was tested in co-cultivation conditions with Tetraselmis striata for evaluating its growth promoting and inhibitory effects. Among these all strains, two promising strains (Pelagibaca bermudensis KCTC 13073BP and Stappia sp. KCTC 13072BP) were selected because of their maximum growth promoting effects and mutualistic interactions. The growth rate, biomass productivity, lipid contents and fatty acids were analyzed during their combined growth in O3 media and compared with axenic growth of Tetraselmis striata. Later, growth promoting mechanisms in the co-cultivation environment were investigated for these promising bacterial strains under replete and limited conditions of nutrients (nitrate, phosphate and vitamin B12). The growth promoting potential of P. bermudensis was illustrated by the two fold enhancement in biomass productivity. These bacteria are promising for microalgal cultivation without any negative effects on the native seawater bacterial communities, as revealed by NGS analysis. This study represents, to date, the first report highlighting the role of phycospheric growth promoting bacteria of promising biofuel feedstock Tetraselmis striata.

Amelioration of biofuel feedstock of microalgae using sustainable means through synthetic ecology is a promising strategy. The co-cultivation model (Tetraselmis striata and Pelagibaca bermudensis) was evaluated for the robust biofuel production under varying stressors as well as with the selected two-stage cultivation modes. In addition, the role of metabolic exudates including the quorum-sensing precursors was assessed. The co-cultivation model innovated in this study supported the biomass production of T. striata in a saline/marine medium at a broad range of pH, salinity, and temperature/light conditions, as well as nutrient limitation with a growth promotion of 1.2–3.6-fold. Hence, this developed model could contribute to abiotic stress mitigation of T. striata. The quorum-sensing precursor dynamics of the growth promoting bacteria P. bermudensis exhibited unique pattern under varying stressors as revealed through targeted metabolomics (using liquid chromatography–mass spectrometry, LC–MS). P. bermudensis and its metabolic exudates mutually promoted the growth of T. striata, which elevated the lipid productivity. Interestingly, hydroxy alkyl quinolones independently showed growth inhibition of T. striata on elevated concentration. Among two-stage cultivation modes (low pH, elevated salinity, and nitrate limitation), specifically, nitrate limitation induced a 1.5 times higher lipid content (30–31%) than control in both axenic and co-cultivated conditions. Pelagibaca bermudensis is established as a potential growth promoting native phycospheric bacteria for robust biomass generation of T. striata in varying environment, and two-stage cultivation using nitrate limitation strategically maximized the biofuel precursors for both axenic and co-cultivation conditions (T and T-PB, respectively). Optimum metabolic exudate of P. bermudensis which act as a growth substrate to T. striata surpasses the antagonistic effect of excessive hydroxy alkyl quinolones [HHQ, 4-hydroxy-2-alkylquinolines and PQS (pseudomonas quorum signal), 2-heptyl-3-hydroxy-4(1H)-quinolone].