Background:The production of pure lines through androgenesis induction is a promising and alternative approach to classical breeding programs. However, this technology in tomato requires the development and optimization of methods.
Aim:The present study aimed to evaluate the feasibility of callus (haploid) production through pollen culture (indirect androgenesis) and the subsequent development of doubled haploid plants in two tomato cultivars under in vitro conditions.
Methodology:In the first stage, mother plants of tomato were grown from the seeds of two cultivars, 4129 and 8320 hybrids varieties, and a suitable culture medium was identified. Flower buds at different developmental stages were collected and surface sterilized by washing under running water and then treated with 25% and 50% sodium hypochlorite solutions for 5, 10, and 15 minutes. A three-factor factorial experiment in a completely randomized unbalanced design was conducted. The experimental factors included cultivar (4129 and 8320 hybrids varieties), sterilization treatment (25% and 50% sodium hypochlorite for 5 and 10 minutes), and days after planting (6 and 12 days).In the second stage, the sterilized anther explants were cultured on N&N medium. This experiment also followed a three-factor factorial experiment. Factors included cultivar (4129, 8320 hybrids varieties), plant growth regulators (2,4-D, BA, IAA), and the days after culture (5, 10, 15, 20, 25,and 30 days). In the third stage, doubled haploid plants were induced using media supplemented with different concentrations of colchicine and varying levels of IAA and BA. A four-factor factorial experiment in a completely randomized unbalanced design was used. Factors included cultivar (4129, 8320 hybrids varieties), colchicine concentrations (0 and 25 mg/L), plant growth regulators (IAA and BA), and days after culture (5, 10, 15, 20, 25, and 30 days).
Findings:In the first experiment, comparison of the means of percentage of disinfectant concen