The Effect of Temperature and Growth Regulators on Callus Induction in Watermelon Anthers

Callus induction is one of the pathways required for haploid plant regeneration through anther culture. Pollen viability, as well as the effect of growth regulators and cold pretreatment on anthers of two watermelon lines (Smile and Sugar Baby) to induce callus formation were herein evaluated. Pollen viability was estimated through the staining technique using 2% acetic carmine. Male flower buds were collected and disinfested to allow removal anthers. These anthers were placed on Murashige and Skoog medium, which was supplemented with 2,4-dichlorophenoxyacetic (2,4-D) at 0.0, 0.5, 1.0, 2.0 or 5.0 mu M or with 6-benzylaminopurine at 0.0, 0.5, 1.0, 1.5, or 2.0 mu M, in combination with 2.0 mu M of 2,4-dichlorophenoxyacetic. Anthers were pretreated at 4 degrees C, for two days and then placed in vitro. Both watermelon lines provided high pollen viability rates (from 93 to 98%). The 2.0 and 5.0 mu M concentrations of 2,4-D stimulated higher friable callus formation. The optimal concentration of 2,4-D was estimated at 3.78 mu M and 4.17 mu M, which had callus induction rates of 64% and 52%, respectively. The combination of 2.0 mu M of 2,4-D and 6-benzylaminopurine did not lead to increased anther response to callus induction. The pre-treatment applied to flower buds at 4 degrees C enabled callus induction and the anther response to callus induction was genotype-dependent.