Recreating in vitro tripartite mycorrhizal associations through functional bacterial biofilms.

Arbuscular mycorrhizal fungi (AMF) and beneficial bacteria are found naturally associated with most terrestrial plant roots. While it is now well known that bacteria colonize AMF and can form aggregates and biofilms, little is known about how interactions between bacterial communities and AMF take place under both in situ and in vitro conditions. We investigated the impact of inoculation with AMF-associated bacteria (AABs) of AMF by in vitro recreation of the interaction on synthetic growth media in a two-compartment Petri plate system. The inoculated AABs were found to be associated with the mycorrhizal co-culture and were found to migrate along growing AMF hyphae and to be associated with the spore surface. AABs differentially influenced the growth of the AMF and their functional capability demonstrated by analysis of phosphate solubilization, nitrogen fixation, and biofilm formation. We have thus characterized these important interactions adding to a further understanding of the synergistic relationship between the two cross-kingdom microbial partners. KEY POINTS: • An in vitro assay was utilized to recreate functional biofilms with AMF-associated bacteria. • AMF-associated bacteria formed a biofilm and enhanced sporulation of Rhizophagus irregularis. • AMF-bacterial interactions through biofilm formation influence the functional capability of both partners.

Diversity and Functionalities of Unknown Mycorrhizal Fungal Microbiota.

Beneficial ecosystem services provided by arbuscular mycorrhizal fungi (AMF) are the outcome of their synergistic actions with diverse bacterial communities (AMF-associated bacteria; AAB) living in strict association with AMF hyphae and spores. Herein, bacterial diversity associated with 6 AMF species from 33 different co-cultures belonging to order Glomerales and Diversisporales were identified, using a combination of culture-dependent functional analyses and amplicon sequencing. Overall, 231 bacterial strains were isolated from the AMF spores and hyphae which covered 30 bacterial genera and 52 species. Hierarchical clustering based on plant growth promoting traits identified 9 clades comprising diverse bacterial genera with clades 7, 8 and 9 representing the most functionally rich AAB. High-throughput amplicon sequencing across a small subset of 8 AMF co-cultures spread across 3 AMF genera identified Operational Taxonomic Units belonging to 118 bacterial genera. The study revealed a greater diversity of AAB from spores of in vitro transformed AMF root co-cultures rather than in situ, pot AMF cultures. Functionally active, culturable AABs with multiple plant growth promoting traits such as phosphate solubilisation, nitrogen fixation, biofilm formation, enzyme and plant growth regulator production along with biocontrol activity were identified. These properties could be utilized individually and/or as consortia with AMF, as biological growth enhancers.

Review of patents for agricultural use of arbuscular mycorrhizal fungi.

Mycorrhizal biotechnology has emerged as a major component of sustainable agriculture and allied activities. Innovations related to its role in agriculture, land reclamation, forestry, and landscaping are well recognized. This review presents the evolution of innovations worldwide related to arbuscular mycorrhizal fungi (AMF) in the past two decades, from 2000 to April 2020, and maintains that such innovations must continue in the future. An analysis of 696 patents showed that AMF have been used consistently as a biofertilizer and bioremediator over that period, although an upsurge was noted in propagation technologies, next-generation production methods, and formulation technologies. This review will familiarize mycorrhizologists with novel and evolving trends and will convince them of the importance of applying for patents to safeguard their innovations and the use of those innovations by industry.