Effects of sterilization, plant growth regulator and media for upscaling in vitro propagation of Musa balbisiana Colla cv. Abu nipah using temporary immersion bioreactor systems

Abstract

The expansion of banana production, especially cv. Abu Nipah is limited due to shortage of quality planting materials available to the farmers. Hence, the study aims to establish a protocol for micropropagation of M. balbisiana cv. Abu Nipah and subsequent upscaling via temporary immersion bioreactor system (TIBs). The objectives of this study are; 1) to study the effect of different ethanol and Clorox ratio on surface sterilization, 2) to study the effect of cytokinins on multiplication and elongation, 3) to study the effect of single and half-cut explant on multiplication, 4) to upscale the plantlet production via shake flask and TIB system and 5) to determine suitable potting media for acclimatization of cv. Abu Nipah. The sword suckers were surface sterilized with different ratio of ethanol to Clorox. Multiplication and elongation of cv. Abu Nipah plantlets were carried out using plant growth regulators; 6-benzylaminopurine (BAP), kinetin, thidiazuron (TDZ) and 2-isopentenyl adenine (2iP) at 5 mg/L and different explant types; single and half-cut explant. The culture was up-scaled with different systems; shake flask, TIB and solid media, and different media formulations; Murashige and Skoog (MS) and Gamborg B5 (B5) media. In vitro rooting was conducted using different auxins; 1-naphthaleneacetic acid (NAA), indole-3-butyric acid (IBA) and indole-3-acetic acid (IAA) at 1 mg/L. Lastly, plantlets were acclimatized to ex vitro environment using different potting mediums. Surface sterilization using 100%:100% ethanol to Clorox produced the highest explants survival (90%). Shoot multiplication and elongation were optimum in MS media supplemented with 5 mg/L of BAP using single explant. Upscaling using RITA® system produced the highest number of shoot (5.93) and shoot length (3.80 cm) while the highest number of root (11.06) and root length (10.70 cm) obtained in 1 mg/L NAA. Lastly, potting medium of peat moss : perlite (1:1 ratio) produced 92% plant survival. It can be concluded that this protocol can be used to mass propagate Musa balbisiana cv. Abu Nipah

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