Nutrient replenishment is one of the significant factors limiting the full development of extensive pasturelands. As a result, bio-inputs have been increasingly used under field conditions to improve plant growth in low-fertility soils. This study aimed to evaluate their effects on the productivity, physiological responses to and nitrogen (N) accumulation in marandu grass (Urochloa brizantha cv. marandu). The experiment was carried out using a randomized block design with four treatments and three replicates in a greenhouse under controlled conditions. In low-fertility soils, lower synthetic fertilizer inputs were simulated (40% N-P-K) (Control), and three bio-inputs were additionally applied (bio-inputs + 40% N-P-K): Azo (Azospirillum brasilense seed inoculation); HS+Herb (leaf application of humic substances combined with a cocktail consisting of Herbaspirillum species), and Coinoc (a combination of Azo and HS+Herb). We evaluated the plant biometrics (shoot and root dry matter, stem diameter and height) and physiological parameters (CO2 assimilation rate, internal CO2 concentration, transpiration, instantaneous carboxylation efficiency, and the maximum quantum yield of photosystem II), as well as the N accumulation 45 days after seedling emergence. Azo-treatment did not differ from the control in most of the variables studied. HS+Herb and Coinoc significantly (p ≤ 0.05) increased the dry matter of shoots (+237% and +255%, respectively) and roots (+106% and 83%, respectively) compared to the control. These treatments (HS+Herb and Coinoc) significantly (p ≤ 0.05) increased (almost double) the size and stem diameter, improved physiological parameters, and accumulated more N (up to 89%) compared to the control. Applying humic substance-based bio-inputs combined with the Herbaspirillum species cocktail was more efficient in stimulating the growth of marandu grass grown in soil of reduced fertility.