Direct Plant Regeneration From Immature Male Inflorescence of Banana (<em>Musa spp</em>.)

Banana (<i>Musa spp</i>.) is a globally important horticultural crop that faces significant challenges from pests and diseases, which threaten yield and long-term sustainability. The efficient production of clean, disease-free planting material is essential for both commercial plantations and small-holder systems. This paper presents a rapid and reproducible protocol for direct plant regeneration from immature male inflorescences of banana. The method involves surface sterilization of immature male flowers, longitudinal dissection, and culture on Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (BAP), enabling direct shoot regeneration from floral meristems without an intermediate regenerable callus phase. This approach offers several advantages over traditional embryogenic cell suspension (ECS) methods, including simplified sterilization, high regeneration efficiency, and scalability. The protocol was successfully applied to multiple banana cultivars, including Cavendish (AAA) and Lady Finger (AAB), achieving 100% shoot regeneration efficiency with plantlet production within 6-8 months. This protocol provides a reliable and efficient alternative for rapid mass propagation of banana plants, supporting sustainable production and research applications. Key features • The protocol can be performed in a standard tissue culture lab without expensive instruments or complex setup, making it accessible for labs in resource-limited settings. • Minimal contamination risk since immature male inflorescences enclosed within bracts are naturally protected, and the simplified sterilization procedure leads to consistently low contamination rates. • Potential for high multiplication where each immature male flower produces 50-100 shoots under optimized conditions, reducing the number of subcultures needed for large-scale propagation. • The method performed equally well in genetically distinct banana cultivars (AAA and AAB groups), suggesting broader applicability across diverse <i>Musa</i> genotypes.