Influence of culture vessel characteristics and agitation rate on gaseous exchange,hydrodynamic stress,and growth of embryogenic cork oak (Quercus suber L.) cultures

Somatic embryogenesis can be induced in the leaves of cork oak (Quercus suber L.) trees. The use of this propagation system in multivarietal forestry requires the mass production of cloned plants at low cost. Investigations were made into the influence of three types of Erlenmeyer flask and three orbiting speeds (60, 110, and 160 rpm) on oxygen transfer rate (K_L a), the shear force index (SFI), biomass production, and the proliferation of embryogenic clumps (EMCs) in cultures during the proliferation phase. K_L a varied between 0.11 and 1.47 h⁻¹ without biomass production being limited by oxygen availability. The EMCs grew even in hypoxic conditions, although the suppression of gaseous exchange strongly reduced biomass production. Cultures with different levels of hydrodynamic stress and SFI values (1.4·10⁻³–8.8·10⁻³ cm min⁻¹) were obtained. Proliferation rates of EMCs increased with agitation rate and the SFI. The largest number of EMCs was obtained in baffled flasks agitated at 160 rpm (K_L a of 1.47 h⁻¹, and SFI of 8.8·10⁻³ cm min⁻¹) with mild hydrodynamic stress enhancing growth. Biomass production increased with agitation and hydrodynamic stress, but only when the SFI value was below 5·10⁻³ cm min⁻¹. The greatest biomass production was obtained in smooth 100 ml flasks agitated at 160 rpm. The differentiation of embryos was favoured by the lowest K_L a (0.11 h⁻¹) and SFI (1.40·10³ cm min⁻¹) values, achieved using these flasks when agitated at 60 rpm.