Low-temperature stress modulates pollen tube growth through temperature-dependent multi-level regulatory mechanisms...

Low temperature is a major environmental stress factor that limits male reproductive success in plants; however, the mechanistic basis of pollen responses to different degrees of low temperature stress remains poorly understood. In this study, we investigated the cytological, biochemical, and molecular responses of tea (Camellia sinensis) pollen grains to low-temperature stress by germinating pollen in vitro at 15, 10, and 5 °C. Although pollen germination rate and pollen tube length were reduced under all low-temperature treatments compared with the control, the underlying regulatory responses differed significantly with the severity of low-temperature stress. At 15 °C, decreases in non-enzymatic antioxidants and stress-related proteins indicated an overall metabolic weakening, while the concomitant accumulation of callose, cellulose, and methyl-esterified pectins reflected an early structural adjustment of the pollen tube cell wall. At 10 °C, the increasing stress load was partially counterbalanced by the induction of enzymatic antioxidant activities, and continued cell wall reinforcement identified this temperature as a transitional state between metabolic limitation and coordinated defense activation. In contrast, exposure to 5 °C resulted in pronounced metabolic suppression, together with a shift of stress-related proteins toward membrane fractions and enhanced deposition of callose, cellulose, and particularly de-esterified acidic pectins, leading to increased cell wall rigidity and mechanical restriction of pollen tube elongation. Overall, this study demonstrates that low-temperature stress constrains pollen tube growth not through a single limiting factor but via a temperature-dependent reorganization of interconnected cytological, biochemical, and molecular mechanisms.