In spite of its diverse limitations, the conventional propagation method is exclusively used for multiplication of sugarcane planting materials in the Ethiopian Sugar Estates since the establishment of the sugar industry in 1954. The present study was carried out to optimize in vitro shoot multiplication protocol for two selected sugarcane varieties (B41-227 and N14) widely grown in Ethiopian sugar estates to complement with the conventional propagation method. In the study, initiated aseptic shoot tip cultures of the two sugarcane varieties were treated with four concentrations (1.5, 2, 2.5 and 3 mgL-1) of 6- benzylaminopurine (BAP) and Indole-3-acetic acid (IAA) (0.25, 0.5, 0.75 and 1 mgL- 1) while Plant growth regulator free medium was used as free check (control). The experiment was set up in a completely randomized design (CRD) with three factor factorial treatment combinations arrangements. Data were collected on number of shoots per explant, average shoot length and number of leaves per shoot after 30 days. Data were subjected to three way analysis of variance. The study verified that medium fortified with 1.5 mgL-1 6-benzylaminopurine (BAP) and 0.5 mgL-1 indole-3-acetic acid (IAA) for B41-227 and 2 mgL-1 6-benzylaminopurine (BAP) and 0.25 mgL-1 indole-3-acetic acid (IAA) for N14 resulted in optimum multiplication responses. On these media, B41-227 produced 15.5 ± 2.90 shoots per explant with 5.93 ± 0.57 cm average shoot length and 6.4 ± 1.49 leaves per shoot while N14 gave 11 ± 00 shoots per explant with 6.32 ± 0.23 cm average shoot length and 5.8 ± 0.06 leaves. Thus, the optimized protocol can be used for rapid in vitro mass multiplication of the sugarcane varieties and hence minimize the limitations sugarcane planting materials.
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