The ophiostomatoid fungi are an assemblage of ascomycetes which are arguably best-known for their associations with bark and ambrosia beetles (Curculonidae) and blue stain (sap stain) of many economically important tree species. These fungi are considered a significant threat to coniferous forests, which has resulted in numerous studies characterising the diversity of bark beetles and their ophiostomatoid associates globally. The diversity of ophiostomatoid fungi present in Australian pine plantations, however, remains largely undetermined. The aims of this study were therefore to reconsider the diversity of ophiostomatoid fungi associated with Pinus in Australia, and to establish the baseline of expected taxa found within these plantation ecosystems. To achieve this, we reviewed Australian plant pathogen reference collections, and analysed samples collected during forest health surveillance programs from the major pine growing regions in south-eastern Australia. In total, 135 ophiostomatoid isolates (15 from reference collections and 120 collected during the current study) were assessed using morphological identification and ITS screening which putatively distinguished 15 taxonomic groups. Whole genome sequencing (WGS) of representative isolates from each taxon was performed to obtain high-quality sequence data for multi-locus phylogenetic analysis. Our results revealed a greater than expected diversity, expanding the status of ophiostomatoid fungi associated with Pinus in Australia to include 14 species from six genera in the Ophiostomatales and a single species residing in the Microascales. While most of these were already known to science, our study includes seven first records for Australia and the description of one new species, Graphilbum ipis-grandicollis sp. nov.. This study also provides an early example of whole genome sequencing (WGS) approaches replacing traditional PCR-based methods for taxonomic surveys. This not only allowed for robust multi-locus sequence extraction during taxonomic assessment, but also permitted the rapid establishment of a curated genomic database for ophiostomatoid fungi which will continue to aid in the development of improved diagnostic resources and capabilities for Australian biosecurity.

Plant growth-promoting bacteria can improve host plant traits including nutrient uptake and metabolism and tolerance to biotic and abiotic stresses. Understanding the molecular basis of plant–bacteria interactions using dual RNA-seq analyses provides key knowledge of both host and bacteria simultaneously, leading to future enhancements of beneficial interactions. In this study, dual RNA-seq analyses were performed to provide insights into the early-stage interactions between barley seedlings and three novel bacterial strains (two Paenibacillus sp. strains and one Erwinia gerundensis strain) isolated from the perennial ryegrass seed microbiome. Differentially expressed bacterial and barley genes/transcripts involved in plant–bacteria interactions were identified, with varying species- and strain-specific responses. Overall, transcriptome profiles suggested that all three strains improved stress response, signal transduction, and nutrient uptake and metabolism of barley seedlings. Results also suggested potential improvements in seedling root growth via repressing ethylene biosynthesis in roots. Bacterial secondary metabolite gene clusters producing compounds that are potentially associated with interactions with the barley endophytic microbiome and associated with stress tolerance of plants under nutrient limiting conditions were also identified. The results of this study provided the molecular basis of plant growth-promoting activities of three novel bacterial strains in barley, laid a solid foundation for the future development of these three bacterial strains as biofertilisers, and identified key differences between bacterial strains of the same species in their responses to plants.

Seed-associated microbiomes represent an underexplored frontier in synthetic community (SynCom) design, particularly in forage legumes such as lucerne (Medicago sativa L.), where early microbial assembly can shape plant development. Crop wild relatives (CWRs) harbour more diverse seed microbiomes and may contain microbes with greater functional potential than domesticated lucerne. To test this, SynComs were constructed from seed-borne bacteria isolated from M. laciniata (drought-resilient) and M. littoralis (salt-tolerant). Two three-strain SynComs were assembled from taxa consistently shared across lucerne and its CWRs, and a third six-strain ‘Mix’ SynCom combined both sets. The aim of this study was to assess whether these SynComs exert phenotypic effects on lucerne growth when used as seed inocula alongside the native microbiome during early development and later vegetative stages under well-watered and drought conditions. Inoculation enhanced germination and early growth, with the Mix SynCom producing the strongest gains. Microbiome profiling at 24 days revealed treatment-specific restructuring, with enrichment of beneficial taxa and microbial coalescence. While early-stage benefits diminished at later stages, and drought ultimately reduced biomass across all treatments, the findings demonstrate that CWR-derived SynComs can enhance lucerne establishment and early growth while restructuring host microbiomes, providing a framework for seed-applied microbial solutions in sustainable agriculture.

Research into understanding the structure, composition and vertical transmission of crop seed microbiomes has intensified, although there is much less research into the seed microbiomes of crop wild relatives. Our previous study showed that the standard seed storage procedures (e.g., seed drying and storage temperature) can influence the seed microbiome of domesticated Glycine max. In this study, we characterized the seed microbiota of Glycine clandestina, a perennial wild relative of soybean (G. max (L.) Merr.) to expand our understanding about the effect of other storage procedures such as the periodic regeneration of seed stocks to bulk up seed numbers and secure viability on the seed microbiome of said seed. The G. clandestina microbiota was analysed from Generation 1 (G1) and Generation 2 (G2) seed and from mature plant organs grown in two different soil treatments T (treatment [native soil + potting mix]) and C (control [potting mix only]). Our dataset showed that soil microbiota had a strong influence on next generation seed microbiota, with an increased contribution of root microbiota by 90% and seed transmissibility by 36.3% in G2 (T) seed. Interestingly, the G2 seed microbiota primarily consisted of an initially low abundance of taxa present in G1 seed. Overall, our results indicate that seed regeneration can affect the seed microbiome composition and using native soil from the location of the source plant can enhance the conservation of the native seed microbiota.

The bird cherry-oat aphid, Rhopalosiphum padi (L.), is an economically significant pest of pasture grasses, the latter being capable of hosting several fungal endophyte–perennial ryegrass symbiota rich in alkaloids and toxic to vertebrates and invertebrates. Measuring aphid feeding behaviour can provide insights into the effectiveness and mode of action of different fungal endophytes. This study investigated the effects of different Epichloë–perennial ryegrass symbiota on the feeding behaviour of R. padi using the electrical penetration graph technique while also assessing the aphid life history. In most cases, endophytes had significant feeding deterrence and paired fecundity and mortality effects. But, in some instances, endophytes with the highest aphid mortality did not significantly deter feeding, suggesting a more complicated scenario of interactions between the relative concentration of metabolites, e.g., host plant defence response metabolites and alkaloids, and/or physical changes to leaf morphology. Overall, this study sheds light on the mode of action of Epichloë endophytes against aphids and highlights the importance of Epichloë–perennial ryegrass symbiota in the management of insect pests such as aphids in pasture-based grazing systems.

Seed-associated microbiomes represent a critical yet underutilised frontier in synthetic community (SynCom) design, particularly in forage legumes such as lucerne (Medicago sativa L.), where early microbial assembly can shape plant development. We developed ecologically-informed SynComs using seed-derived bacterial isolates from Medicago crop wild relatives (CWRs), prioritising genera that co-occur with the cultivated lucerne seed microbiota. Two three-strain SynComs and one six-strain ‘Mix’ SynCom were applied via seed inoculation. Plants were glasshouse-grown; a subset was transferred to a separate phenomics facility, exposed to contrasting water regimes, and later returned to the glasshouse for post-stress recovery. A multi-tiered approach combining phenotyping, high-throughput imaging, and 16S rRNA profiling captured plant–microbiome dynamics. SynCom inoculation consistently enhanced germination and early growth, with the ‘Mix’ SynCom producing the strongest early-stage phenotypic gains. Microbiome profiling revealed treatment-specific restructuring, showing selective enrichment and microbial coalescence. While SynCom effects were most pronounced early, their influence under drought and later stages was variable and context dependent. This study positions seed-based, CWR-derived SynComs as effective modulators of early plant–microbiome interactions. By integrating ecological selection with temporal phenotyping, we present a scalable framework for microbiome-informed crop improvement and a translational path toward seed-based microbial solutions in sustainable agriculture.