Comparison of viability tests for assessing cross-adaptation to freezing, heat and salt stresses induced by abscisic acid in bromegrass (Bromus inermis Leyss) suspension cultured cells

摘要: Several viability assays were compared to determine the most sensitive and appropriate method for estimating freezing, heat salt tolerance of Bromus inermis Leyss cells cultured with or without 75 μM abscisic acid (ABA) 4–7 days at 25°C. The sensitivity reliability individual tests depended on type stress applied degree injury. Regrowth, amino (AA) leakage fluorescein diacetate (FDA) staining all gave comparable estimates freezing tolerance. Triphenyl-tetrazolium chloride (TTC) reduction slightly overestimated tolerance, but was convenient. Polypeptide from freeze-thawed cells, as determined by SDS-polyacrylamide electrophoresis (SDS-PAGE), revealed that protein only occurred in ABA-treated frozen −21°C colder contrast control which leaked proteins following −5°C. test assessing regrowth, followed FDA staining, when TTC. TTC other tests. overestimation greater than estimates. It partially improved washing prior assays. AA not due erroneously high values A280 readings obtained. For convenient close agreement regrowth measurements whereas tended overestimate it. In general, while reliable, All consistently showed ABA induced cross-adaptation stresses bromegrass a exposure any these stresses.