Synergistic Effects of Non-Thermal Plasma Exposure Time and Drought on Alfalfa (

Applying non-thermal plasma (NTP) to seeds prior to sowing is recognized for its ability to enhance germination and promote plant growth. This study investigated the effects of NTP seed treatment on alfalfa seed surface characterization, germination, growth, and biochemical traits under varying water conditions. NTP modified seed surface properties by decreasing water contact angle, roughening the coat, and reducing O-H/N-H and C-H band intensities, while major functional groups remained intact. Short plasma exposures (<2 min) enhanced germination, whereas prolonged treatment (10 min) reduced viability, indicating embryo sensitivity. In pot experiments, both 1 and 5 min treatments improved fresh and dry weight, stem and root elongation, pigment accumulation, and protein content, particularly under normal and moderate water stress, while extended exposure (10 min) offered limited benefits and could be detrimental under severe drought. Root growth was most responsive, suggesting enhanced water and nutrient uptake. Plasma had modest effects on polyphenols and flavonoids but influenced early physiological responses and antioxidant activity. These findings highlight NTP as a promising seed priming tool to improve alfalfa performance, though further studies are needed to clarify the mechanisms and specific contributions of plasma components.