SCHOOL OF GRADUATE STUDIES COLLEGE OF AGRICULTURE AND NATURAL RESOURCE IN VITRO REGENERATION OF Brassica spp, A LOCAL LANDRACE SIMUARE

Abstract

Simuare (Brassica spp), a leafy vegetable resembling kale, is widely grown and consumed in Ethiopia’s Gurage Zone. Due to its inability to flower and set seed, Simuare is propagated solely through cuttings, a method that is inefficient for producing sufficient planting material. This study was conducted to develop an efficient and cost-effective in vitro micro-propagation protocol to address these limitations. Explants were surface sterilized using ethanol (70% for 30 seconds), mancozeb (2.5mg/l), and bleach (2% for 20 minutes), achieving a survival rate of 99.6%. Murashige and Skoog (MS) medium supplemented with various concentrations of auxins and cytokinins was used for culture initiation, shoot multiplication, and root induction. Optimal shoot initiation was observed with 1 mg/L BAP and 1 mg/L kinetin, while the highest shoot multiplication occurred on MS medium containing 3 mg/L BAP and 0.1 mg/L GA3, producing 6.2 ± 1.68 leaves, 4.33 ± 1.87 shoots per explant, and a shoot height of 4.44 ± 0.83 cm. Root induction was most effective with 0.3 mg/L IBA and 0.2 mg/L NAA, yielding 5.04 ± 0.07 roots per plantlet and a root length of 3.06 ± 0.47 cm. The developed protocol successfully overcame challenges of conventional propagation, achieving high explant survival, efficient shoot multiplication, and effective root induction. These findings provide a reliable method for large-scale Simuare propagation, and the protocol is recommended for producing high-quality planting materials to meet growing demand.