The utilization of biomass ash in sustainable agriculture and increasing its fertilizing efficiency by biological agents, potentially sequestering CO2, have become important issues for the global economy. The aim of this paper was to investigate the effects of ash from sorghum (Sorghum bicolor L. Moench) and Jerusalem artichoke (Helianthus tuberosus L.) biomass, a biogas plant digestate, and a Spirodela polyrhiza extract, acting alone or synergistically, on soil fertility and the development, health and physiological properties of sorghum plants. The results show novel information concerning differences in the composition and impact of ash, depending on its origin, soil properties and sorghum plant development. Sorghum ash was more effective than that from Jerusalem artichoke. Ash used alone and preferably acting synergistically with the digestate and Spirodela polyrhiza extract greatly increased soil fertility and the growth, biomass yield and health of sorghum plants. These improvements were associated with an increased chlorophyll content in leaves, better gas exchange (photosynthesis, transpiration, stomatal conductance), greater enzyme activity (acid and alkaline phosphatase, RNase, and total dehydrogenase), and a higher biomass energy value. The developed treatments improved environmental conditions by replacing synthetic fertilizers, increasing the sequestration of CO2, solving the ash storage problem, reducing the need for pesticides, and enabling a closed circulation of nutrients between plant and soil, maintaining high soil fertility.