Standardization of an In Vitro Seed Germination Protocol Compared to Acid Scarification and Cold Stratification Methods for Different Raspberry Genotypes

In raspberry (Rubus idaeus L.) breeding programs, seed double dormancy causes delayed and low germination rates. In this study, an in vitro germination protocol was developed to increase raspberry seed germination rate and decrease germination time. This protocol was optimized on the cultivars "Polka" and "Tulamagic", specifically, their open-pollinated seeds, by applying two different cutting types (transverse, or longitudinal), followed by two different treatments (direct in vitro culture at 24 degrees C, or 2 weeks at 4 degrees C followed by in vitro culture at 24 degrees C). The best results for both cultivars were obtained when seeds were cut transversally and directly cultured in vitro at 24 degrees C (germination rate 73-78%, starting germination time 4 days after treatment). The optimized in vitro protocol was applied to 14 different combination crosses, observing a germination frequency around 87%, and speed in line with "Polka" and "Tulamagic". These results were compared to those from the 14 cross combinations having undergone acid scarification (95% H2SO4 for 30 min) and cold stratification (3 months), which, in comparison, only yielded a 25% average germination rate. The optimized in vitro protocol favors high germination efficiency for all raspberry genotypes tested, and presents advantages for the standardization of germination time and reduction in the parental genotypes' impact on germination rate.