The application of biochar is mostly used to improve soil fertility, water retention capacity and nutrient uptake. The present study was conducted in order to study the impact of biochar at water deficiency conditions on the physiological and biochemical processes of Medicago ciliaris seedlings. Seedlings were cultivated under greenhouse conditions in pots filled with a mixture of soil and sand mixed in the presence or absence of 2% biochar. Plants of uniform size were subjected after a pretreatment phase (72 days) either to low (36% water holding capacity, water potential low) or high soil water potential (60% water holding capacity, water potential high). Pots were weighed every day to control and maintain a stable water holding capacity. In Medicago ciliaris, drought led to a significant reduction in plant growth and an increase in the root/shoot ratio. The growth response was accompanied by a decreased stomatal conductance and a reduction of the net CO₂ assimilation rate and water use efficiency. The associated higher risk of ROS production was indicated by a high level of lipid peroxidation, high antioxidant activities and high proline accumulation. Soil amendment with biochar enhanced the growth significantly and supported the photosynthetic apparatus of Medicago ciliaris species by boosting chlorophyll content and A_net both under well and insufficient watered plants and water use efficiency in case of water shortage. This increase of water use efficiency was correlated with the biochar-mediated decrease of the MDA and proline contents in the leaves buffering the impact of drought on photosynthetic apparatus by increasing the activity of enzymatic antioxidants SOD, APX, GPOX and GR and non-enzymatic antioxidants, such as AsA and DHAsA, giving the overall picture of a moderate stress response. These results confirmed the hypothesis that biochar application significantly reduces both the degree of stress and the negative impact of oxidative stress on Medicago ciliaris plants. These results implied that this species could be suitable as a cash pasture plant in the development of agriculture on dry wasteland in a future world of water shortages.

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