In Vitro Shoot Multiplication of Elite Sugarcane (Saccharum officinarum L.) Genotypes Using Liquid Shake Culture System

Abstract

Study was carried out with an objective to determine the appropriate concentrations and combinations of 6-benzylaminopurine (BAP) and kinetin for in vitro shoot multiplication of elite sugarcane genotypes i.e., N52 and N53. Shoot tip was used as explant source. Shoot initiation from explant of the two genotypes was achieved at a combination of BAP, kinetin and NAA (0.5 mg/l each). For shoot multiplication, the regenerated and twice subcultured shoots on semi-solid medium were transferred on liquid Murashige and Skoog medium containing 3% sucrose, fortified with various concentrations and combinations of BAP (0, 0.5, 1, 1.5, 2 mg/l) and Kinetin (0, 0.5, 1, 1.5 mg/l).The cultures were agitated continuously on an orbital shaker moving at 80 rpm. Analysis of variance (ANOVA) showed that the interaction effects of 6-benzylaminopurine, kinetin and the sugarcane genotypes on number of shoots per explant, average shoot length and number of leaves per shoot was very highly significant (P < 0.0001). Genotype N52 showed a maximum of 6.95 ± 0.19 shoots per explant with 4.75 ± 0.06 cm shoot length and 5.65 leaves per shoot on liquid MS medium fortified with 2 mg/l BAP + 0.5mg/l kinetin while genotype N53 produced a maximum of 6.30 ± 0.26 shoots per explant with 3.94 ± 0.03 average shoot length and 5.83 leaves per shoots on liquid MS medium supplemented with 1.5 mg/l BAP + 0.5 mg/l kinetin. Finally, from study results we can deduce that the application of this protocol helps for rapid in vitro shoot multiplication of elite sugarcane genotypes.

Keywords: Murashige and Skoog, shoot tip, explant, 6-benzylaminopurine