Pepper is one of the most cultivated vegetables worldwide and also consumed substantially as aflavouring ingredient in different culinary traditions. Therefore, many researchers have focused onits breeding to develop new varieties. One of plant breeding aims is to attain disease and pestresistance. The use of tissue culture methods in plant breeding has many advantages. The responseof two nematode resistant pepper genotypes to the anther culture and the effect of cold pre-treatmentto the floral buds have been investigated in this study. Alata 2095 and Alata 2096 both specified asresistant to the nematode by Alata Horticultural Research Institute (Alata, Mersin, Turkey) wereused as plant material. Two pre-treatments were used in this study: cold and no cold. In coldtreatment, flower buds were kept in fridge at 4°C for 24 hours prior to the anther culture. Murashigeand Skoog medium contained 0.25% activated charcoal, 6.5 g L-1agar, 0.5 mg L-1 6-benzyl-aminopurine (BAP), 4 mg L-1 naphthalene-acetic-acid (NAA), 15 mg L-1silver nitrate (AgNO3), and 30 gL-1sucrose was used. After assessment, the highest mean of plant number was 39.08 per 100 anthersfor Alata 2095 genotype. It was 46.61 and 31.56 in cold and no cold treatment, respectively. ForAlata 2096 genotype, the mean was 1.96 per 100 anthers (1.68 and 2.25 in cold and no cold treatment,respectively). Statistical analyses confirmed that there was significant difference betweentreatments, genotypes and also significant interaction between those factors. At the end of the study,we can say that Alata 2095 genotype has a good androgenic response and it can be beneficial infurther pepper breeding studies.