Abstract
Freeze tolerance is a complex agronomic trait that is difficult to evaluate in the field because of year‐to‐year variation in weather. Discovering plant characteristics closely related to freeze tolerance would enable more effective selection for this important trait. To explore possible physiological mechanisms and search for useful characteristics related to freeze tolerance in winter barley, we conducted field and growth chamber experiments with seven freeze‐tolerant and seven freeze‐susceptible genotypes that exhibited contrasting winter survival in preliminary field screenings. In a 2‐year field experiment, malondialdehyde, proline and water‐soluble carbohydrate concentrations were measured during cold acclimation and deacclimation to investigate differences in osmoregulation and membrane stability between freeze‐tolerant and freeze‐sensitive genotypes. All parameters varied by sampling year, and significant differences between freeze tolerance groups were found mainly during cold deacclimation in the spring. In growth chamber experiments, the size of xylem vessels and stomates was measured with and without cold acclimation. Freeze‐tolerant genotypes had smaller xylem vessels and stomates than freeze‐sensitive genotypes with and without cold acclimation, and small stomatal length was associated with a small xylem vessel area. Thus, it may be possible to improve freeze tolerance in winter barley by selecting smaller xylem and stomate cells. This study also validated germplasms of winter barley with differential freeze tolerance for future projects on breeding for improving winter hardiness and on plant physiology and genetics in response to freezing stress.