Quantity and distribution of arbuscular mycorrhizal fungal storage organs within dead roots.

The formation of storage organs, such as spores and vesicles, is a central part of the life cycle of an arbuscular mycorrhizal fungus (AMF), but the conditions under which this occurs in AMF are not well understood. Here, quantity and distribution of storage organs formed by the arbuscular mycorrhizal fungus (AMF) Funneliformis mosseae within dead (excised) roots were characterised. 'Trap roots' (TR), separated from the growth substrate by a 30-µm mesh, supported hyphal growth and formation of storage organs of the AMF. Hyphae developed both inside and on the outside of the TR and also within air gaps of surrounding nylon mesh compartments, but formation of vesicles and spores was confined to the interior and to the surface of the TR. Up to 20 % of the TR length harboured newly formed storage organs, resulting in a number of about 60 per mg TR dry weight. The portion of TR length containing storage organs was greater in coarse (diameter >300 µm) than in thin (<150 µm) TR, irrespective of whether the TR were sourced from an AMF host or non-host plant. We conclude that the AMF's extraradical mycelium produces its storage organs within dead roots in preference to air space in the substrate. Dead roots may indirectly supply nutrients to AMF (once they have been mineralised) or represent a protected space for the fungal structures to develop. The experimental technique described here allows for the preparation of AMF spores and vesicles of F. mosseae free of any mineral substrate.

Stable, fluorescent markers for tracking synthetic communities and assembly dynamics.

BACKGROUND: After two decades of extensive microbiome research, the current forefront of scientific exploration involves moving beyond description and classification to uncovering the intricate mechanisms underlying the coalescence of microbial communities. Deciphering microbiome assembly has been technically challenging due to their vast microbial diversity but establishing a synthetic community (SynCom) serves as a key strategy in unravelling this process. Achieving absolute quantification is crucial for establishing causality in assembly dynamics. However, existing approaches are primarily designed to differentiate a specific group of microorganisms within a particular SynCom. RESULTS: To address this issue, we have developed the differential fluorescent marking (DFM) strategy, employing three distinguishable fluorescent proteins in single and double combinations. Building on the mini-Tn7 transposon, DFM capitalises on enhanced stability and broad applicability across diverse Proteobacteria species. The various DFM constructions are built using the pTn7-SCOUT plasmid family, enabling modular assembly, and facilitating the interchangeability of expression and antibiotic cassettes in a single reaction. DFM has no detrimental effects on fitness or community assembly dynamics, and through the application of flow cytometry, we successfully differentiated, quantified, and tracked a diverse six-member SynCom under various complex conditions like root rhizosphere showing a different colonisation assembly dynamic between pea and barley roots. CONCLUSIONS: DFM represents a powerful resource that eliminates dependence on sequencing and/or culturing, thereby opening new avenues for studying microbiome assembly. Video Abstract.

Ulva prolifera polysaccharide enhances the root colonisation by Bacillus amyloliquefaciens strain Cas02.

The application of biocontrol agent is important for the sustainable development of agriculture. Unsuccessful or limited colonisation by plant growth-promoting rhizobacteria (PGPR) has become an important constraint factor for their commercial application. Here, we report that Ulva prolifera polysaccharide (UPP) promotes root colonisation by Bacillus amyloliquefaciens strain Cas02. UPP serves as an environmental signal for bacterial biofilm formation and its glucose residue is used as a carbon source for the synthesis of the exopolysaccharides and poly-gamma-glutamate present in biofilm matrix. Greenhouse experiments demonstrated that UPP could effectively enhance the root colonisation by Cas02 in both the bacterial population and survival time under natural semiarid soil conditions. Furthermore, the microbiome analysis also indicated the promoted colonisation by Cas02, as well as the improved bacterial rhizosphere community structure, after combined treatment of UPP and Cas02. This study provides a practical approach to improve the biocontrol agent with seaweed polysaccharides.

Microbial Inoculation Improves Growth of Oil Palm Plants (Elaeis guineensis Jacq.).

Introduction of diazotrophic rhizobacteria to oil palm tissues during the in vitro micropropagation process establishes an early associative interaction between the plant cells and bacteria. In the association, the diazotrophs provide the host plants with phytohormones and fixed nitrogen. This study was conducted to observe growth of bacterised tissue cultured oil palm plants under ex vitro conditions after 280 days of growth. Root dry weight, shoot dry weight, root volume, bacterial colonisation, leaf protein and chlorophyll content of the host plants were observed. The results revealed that the inocula successfully colonised roots of the host plants. Plants inoculated with Acetobacter diazotrophicus (R12) had more root dry weight and volume than plants inoculated with Azospirillum brasilense (Sp7). Leaf protein and chlorophyll content were higher in the bacterised plants compared to Control 2 plants (inoculated with killed Sp7). These results suggest that the diazotrophs successfully improved the growth of the host plant (oil palm) and minimised the amount of N fertiliser necessary for growth.