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To ensure high yields of pollen calluses in float cultures of Hordeum vulgare anthers,inoculation densities of at least 60 pretreated stage 2 anthers are needed per ml of medium.This is shown for two spring barleys, Sabarlis and Delta, and one winter barley, Maris Otter.The optimal density varies from batch to batch, and may be as high as 120 per ml in someinstances. Exogenous hormones (kinetin and 2, 4-D) are not essential for callus productionbut without them higher anther densities are required. High densities can be achieved with relatively few anthers by culture in drops of medium(0.2ml). In this event, the volume of medium needs to be increased gradually in order to main-tain vigorous growth of the calluses. Alternatively, inoculation densities can be reduced topractical levels (10--20 per ml) by pre-conditioning of the medium with anthers. At low densi-ties with medium pre-conditioned by stages 5--6 anthers, that is, anthers older than those usedfor culture, better callus yields and better growth of the calluses are obtained than at peakdensities with unconditioned medium. It is suggested that the ’drop procedure’ combined with ’conditioning’ would be aprofitable approach to the testing of other species which, like barley, have small and suppos-edly poor responding anthers.
To ensure high yields of pollen calluses in float cultures of Hordeum vulgare anthers, inoculation densities of at least 60 pretreated stages 2 anthers are needed per ml of medium. This is shown for two spring barleys, Sabarlis and Delta, and one winter barley, Maris Otter.The optimal density varies from batch to batch, and may be as high as 120 per ml in someinstances. Exogenous hormones (kinetin and 2, 4-D) are not essential for callus production but without them higher anther densities are required. can this achieved with relatively few anthers by culture in drops of medium (0.2 ml). In this event, the volume of medium needs to be increased vig in growth to the calluses. At low densi-ties with medium pre-conditioned by stages 5--6 anthers, that is, anthers older than those used for culture, better callus yields and better growth of the calluses are obtained than at peakdensities with unconditioned medium. It is suggested that the ’drop procedure’ combined with ’conditioning’ would be aprofitable approach to the testing of other species which, like barley, have small and suppos-edly poor responding anthers.