Detection of abnormal banana plantlets produced by high BAP concentration and number of subcultures using representational difference analysis

Abstract

The disadvantage of somaclonal variation is mainly related to its limitation to provide disease-free and uniform plantlet in a large quantity and within a short duration. However, this technique faced a limited number of subculture and the use of the plant growth regulator to induce the shoot formation during propagation time. Hence, the objective of this study is to investigate the variation occur during prolong subculture with the increase of 6-benzylaminopurine (BAP) concentration, at morphological and molecular level. The results showed that the prolong subculture at 15 mg L-1 BAP had inhibited the shoot formation. The higher number of shoots per clump was observed at 5 mg L-1 at the 20th subculture. Genetic variation between clones was assessed by Representation Difference Analysis (RDA) technique. It was found that there was no genetic variation that can be amplified even though between the control and the BAP-treated explant. This implied a very small genetic variation had occurred among clones. The findings of this study provided a basis for further investigation to a genomic region in banana, that was susceptible to stress raised in tissue culture stage, especially when used higher concentrations of BAP (15 mg L-1) and number of subcultures.