The present study established a protocol for generating embryogenic cell suspension (ECS) cultures from an early stage embryogenic callus of Musa acuminata cv. ‘Berangan’. The necessary baseline data for morphohistological recognition and the selected gene expression for differentiating embryogenic and non-embryogenic cells were defined to enable early selection, as well as for quality assessment, which could aid in scaling up the production of these cells. In this study, male bud-derived callus on Murashige and Skoog (MS) medium containing 23 μM 2,4-dichlorophenoxyacetic acid (2,4-D) was transferred to M1 medium containing different concentrations of 2,4-D and picloram for somatic embryo initiation. ECSs established in liquid MS medium (M2a) containing 4.1 μMbiotin, 10 mg/l ascorbic acid, 100 mg/l glutamine, 100 mg/l malt extract, 4.5 μM 2,4-D, 1.0 μM zeatin, and 20 g/l sucrose were used to study embryogenic development. The frequency of embryo development and maturation was influenced by the culture media. The highest frequency of embryo development (96.4 %) and maturation (66.4 %) was achieved on solid MS hormone-free medium. Almost 69 % of embryos were induced to form shoots on the MS medium containing 34 μM 6-benzylaminopurine (BAP). Histological study indicated that only embryos with distinct layer of protoderm having cells with prominent nucleus and dense cytoplasm were able to regenerate into plantlets. Endochitinase and phenylalanine ammonia-lyase (PAL) genes were expressed at all the developmental stages, and their expression level was tightly regulated during embryo development and reached its maximum in germinating embryos. Expression of these genes in embryogenic cells was higher than in nonembryogenic cells. The study findings suggested that these genes play an important role in the developmental stage and regeneration pathway of somatic embryos of banana.
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