In earlier work on improvement of persistance in forage legumes, we selected genotypes from highly productive cultivars of alfalfa, Algonquin and Apica (Euphytica 45: 105-112, 1990) and cv. Florex red clover (Plant Cell Reports 8: 395-398, 1989) capable of in vitro regeneration from callus and cell culture. The alfalfa germplasm and its F1 progeny as well as an F2 red clover population were tested for cold stress tolerance. Plantlets were hardened in culture tubes at 2 or 5 ° C, 8 h photoperiod, for at least four weeks and then subjected to freezing temperatures, -16 or -10°C for alfalfa and red clover, respectively. Survival of regenerative genotypes was significantly higher than of the non-regenerative ones in both species. A strong positive correlation (r = 0.78) between the regenerative trait and plant survival was found in alfalfa. The experiments indicate that in vitro selection for regenerative trait may improve cold stress tolerance of alfalfa and red clover.
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