Response of Alfalfa (Medicago sativa L.) to Abrupt Chilling as Reflected by Changes in Freezing Tolerance and Soluble Sugars

Abstract

Abrupt-chilling events threaten the survival of alfalfa plants, the ability to cope with such condition should be considered during cultivar selection in the production. To assess biochemical and molecular responses of alfalfa to abrupt chilling, the cultivars “WL440HQ” (WL) and “ZhaoDong” (ZD) were subjected to a five-phase experimental regime that included two abrupt-chilling events. The freezing tolerance of the crown was determined as the semi-lethal temperature (LT50) calculated from electrolyte leakage. Soluble sugar concentrations were quantified by ion chromatography. The mRNA transcript levels of four genes encoding enzymes (β-amylase, sucrose phosphate synthase, galactinol synthase, and stachyose synthase) involved in sugar metabolism and two cold-regulated genes (Cas15A and K3-dehydrin) were quantified using quantitative real-time PCR analysis. During the abrupt-chilling events, the LT50 decreased significantly in ZD but not in WL. The rapid response of ZD to abrupt chilling may have been due to the large increases in raffinose and stachyose concentrations, which were consistent with increased transcript levels of the galactinol synthase and stachyose synthase genes. Transcript levels of the cold-regulated genes Cas15A and K3-dehydrin were correlated with increased freezing tolerance under abrupt chilling. The results provide a reference for selection of appropriate cultivars to reduce the risk of crop damage in production areas where early autumn or late spring frosts are likely.