A Natural Defender: Endophytic Bacillus amyloliquefaciens AsL-1 from Alstonia scholaris Latex Effectively Controls Colletotrichum gloeosporioides in Mango

Biological control using beneficial microbes offers a sustainable alternative to chemical fungicides for managing postharvest diseases. This study reports the isolation and characterization of Bacillus amyloliquefaciens AsL-1 from the latex of Alstonia scholaris (L.) R. Br., unconventional ecological niche. The cell-free supernatant (CFS) of AsL-1 showed strong antifungal activity, inhibiting the growth of Colletotrichum musae (48.89 ± 0.57%), Glomerella cingulata (52.22 ± 0.00%), Fusarium graminearum (47.78 ± 0.57%), and Colletotrichum gloeosporioides (47.78 ± 0.00%) in vitro. Microscopy revealed that the CFS disrupted fungal development by blocking conidial germination and appressorium formation, and in C. gloeosporioides caused melanization defects linked to reduced virulence. In vivo tests on mango fruit confirmed that AsL-1 significantly decreased anthracnose lesion size and disease incidence. Protein analyses (SDS-PAGE, gel overlay, and LC-MS/MS) identified two antifungal proteins (24 and 16 kDa), corresponding to β-1,3-1,4-glucanase and flagellin. The detected β-1,3-1,4-glucanase activity indicates its role in degrading fungal cell walls and interfering with melanin biosynthesis pathways essential for pathogenicity. Overall, these findings highlight B. amyloliquefaciens AsL-1 as a promising protein-based biocontrol agent and show that latex-associated microbes may serve as valuable sources of new antifungal strategies.