Low tissue-water content and increased osmotic concentration of cell sap are associated with frost resistance. Changes in total osmotic concentration of cell sap are due mainly to changes in concentration of sugars. Generally, sugar content increases with hardening and decreases with dehardening. This study examined the effect of elevated sucrose levels (3% to 15%) in the medium on the cold hardiness of `Festival' red raspberry (Rubus idaeus L.) shoots in vitro. To determine whether expected hardening is caused by elevated sucrose levels or by osmotic stress, different levels of mannitol in the media have been tested. After growing raspberry shoots on media with different levels of sucrose and mannitol for 2 weeks, shoot moisture content (percent) was determined. Cold hardiness of the shoots was determined by using differential thermal analysis or artificially freezing the shoots and assessing the survival by regrowth test and visual rating.
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