Alfalfa Photosynthesis Under Partial Root-Zone Drying: Diurnal Patterns and Its Non-Stomatal Limitations

The effects of stomatal factors of plant leaves under partial root-zone drying (PRD) have been widely studied. However, the non-stomatal factors and the relationship between photosynthesis with soil moisture have not been explored. In this study, four treatments over-irrigation, full irrigation, moderate water deficit, and severe water deficit were investigated, aiming to evaluate the effects on the diurnal variation of alfalfa leaf photosynthesis under PRD and its relationship with stomatal and non-stomatal limitations, as well as soil moisture. The results showed that any levels of water deficit led to a decrease in the photosynthetic rate (<i>P_n</i>) of alfalfa leaves. Leaves under moderate and severe water deficit displayed a pronounced midday "photosynthetic lunch break," while those under over- and full irrigation did not display this phenomenon. Before 11:30 a.m., the reduction in <i>P_n</i> was primarily due to stomatal limitations, as evidenced by reduced stomatal conductance (<i>G_s</i>) and decreased intercellular CO₂ concentration (<i>C_i</i>). After 11:30 a.m., non-stomatal limitations became the dominant factor, with both <i>G_s</i> and transpiration rate (<i>T_r</i>) continuing to decrease, while <i>C_i</i> increased, indicating a shift in the limiting factors. Under PRD with moderate water deficit, alfalfa experienced both stomatal and non-stomatal limitations within a single day, leading to a hay yield reduction of 18.6%. Additionally, over-irrigation helped to maintain higher <i>P_n</i> and <i>T_r</i>, increasing alfalfa yield and thus improving water productivity by 33.1%. The correlation coefficients between soil moisture content at 10 cm depths with alfalfa leaf <i>P_n</i>, <i>T_r</i>, and <i>G_s</i> on the photosynthetic measurement day were 0.9864, 0.8571, and 0.8462, respectively. At 20 cm, the correlation coefficients were 0.8820, 0.6943, and 0.6951, respectively. The study concluded that both stomatal and non-stomatal mechanisms contributed to reduced alfalfa <i>P_n</i> in water deficit of PRD. Furthermore, shallow soil moisture also played a crucial role in influencing photosynthetic performance.