Conservação in vitro de morangueiro via crescimento lento e criopreservação

Abstract

In vitro conservation provides short and medium term storage options by reducing plant metabolism (called slow growth) or long-term inhibition of metabolic reactions (called cryopreservation), both of which maximize the interval between subcultures, avoiding the somaclonal variation, as occurs in the strawberry after successive subcultures. In this context, the effect of different osmotic regulators, temperatures, explant types and evaluation periods on the in vitro conservation of 'Benicia' strawberry by slow growth and the adaptation of cryopreservation protocols by different methods in different cultivars were evaluated. Explants from 'Benicia' strawberry plants were used in vitro and maintained in MS medium of multiplication. The medium used in the experiments was the basic MS supplemented with 20g L-1 or 40g L-1 of mannitol or sorbitol. In the control treatment there was no addition of osmotic regulator. Two types of explants were inoculated in the media: explant only with aerial part (PA) and whole plant (PI), maintained for 6 months at 24±2°C in growth room and 12 months in BOD with temperature of 4±2°C, both with photoperiod of 16 hours and luminous intensity of 45-55 μmol m-2 s-1. The experimental design was completely randomized in a 5x2x5 factorial scheme (culture medium, explant type and evaluation periods), with three replicates of 10 explants, totaling 30 explants per treatment. Growth parameters, biochemical and physiological parameters were evaluated. According to the results obtained, it can be concluded that the temperature of 4°C, the whole plant and the use of sorbitol provide a reduction in the growth of 'Benicia' strawberry explants during 12 months of in vitro conservation without affecting growth parameters, physiological and biochemical characteristics of the species. In all tested cryopreservation protocols the strawberry buds did not survive liquid nitrogen, but the temperature of 4°C during the preacclimation period for 2-8 weeks was significant. The sucrose solution at 0.75M resulted in a higher survival rate. The desiccation times were not efficient and the use of proline increased the tolerance of the egg yolks.