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Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
Published on: October 15, 2019
Revealing and characterizing bacterial communities of in vitro Musa species through 16S rDNA metabarcoding and
Christopher A Sambolín-Pérez1, Sharelys M Montes-Jiménez1, Hirelys M Montes-Jiménez1
1Institute of Sustainable Biotechnology, Inter American University of Puerto Rico, Barranquitas, PR, 00794, USA.
Abstract:
Bananas and plantains, part of the Musa genus, are key global food crops that are threatened by various factors, including hurricanes and microbial infections. The production of phytopathogen-free plants in Temporary Immersion Bioreactors (TIB) has gained attention due to improved yield and health. However, the impact of TIB on Musa spp. microbiomes remains poorly understood. Thus, elucidating the role of in vitro Musa spp. microbiome is crucial for developing healthier plantlets with beneficial microbes, such as plant growth-promoting bacteria (PGPB), which are essential for plant development and might help Musa spp. thrive in abiotic and biotic stresses after in vitro development. To reveal the potential association of PGPB, we aimed to identify and characterize the bacterial communities from in vitro (TIB) Musa spp. varieties (Maiden, Dwarf, and Maricongo) pseudostems using both culture-independent (16S rDNA-metabarcoding) and culture-dependent methods to elucidate their diversity and roles in plant health. Our results identified four bacterial phyla, with Bacillota being the most dominant, followed by Pseudomonadota, Actinobacteriota, and Bacteroidota. Brevibacillus sp. and Xylella sp. were the dominant genera. The isolates included Lysobacteraceae and Terribacillus spp., and the microbiomes metabolic pathways featured cofactors and amino acid biosynthesis. These findings enhance the understanding of bacterial communities in Musa spp. under in vitro conditions, highlighting the potential effects of artificial environments on host microbiomes, and encouraging innovative research into bacterial-plant interactions. This may aid in identifying specific bacteria with potential PGPB traits in Musa spp., offering new ways to enhance production and protection.
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