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Biology subjects

Hefer, C. A.

Publications and source records attributed to Hefer, C. A..

2 recordsLinked to original sources

Geographic and Climatic Origins Shape the Leaf Metabolome of Populus trichocarpa

O_LIBackground and Aims: Chemodiversity is a fitness-relevant trait shaped by genetics, environment, and their interaction. Populus trichocarpa naturally inhabits broad climatic gradients and shows extensive variation in specialised metabolism. We investigated whether provenance and climate of origin imprint leaf chemodiversity and class-level relationships under common-garden conditions, and how these patterns relate to gene expression. C_LIO_LIMethods: Leaves from 87 P. trichocarpa genotypes representing 22 provenances from the west coast of North America growing in a common garden were profiled by untargeted FT-ICR-MS (1030 features) and targeted LC-MS/MS. A subset of 41 genotypes was subject to RNA-seq analyses. We tested whether provenance influenced multivariate patterns and whether metabolomic differences were related to geographic and climatic distance, where chemodiversity was quantified as Functional Hill Diversity. C_LIO_LIKey Results: P. trichocarpa metabolomes differed among origins despite shared growth conditions and showed distance-decay with both geography and climate. North-south extremes were well separated, and within-drainage samples shared high similarity. Flavonoid and isoprenoid pools strongly co-varied across individuals, whereas isoprene synthase activity did not predict total isoprenoids. Transcriptomes showed within-pathway coherence but limited overall provenance separation. C_LIO_LIConclusions: Leaf chemistry in P. trichocarpa retains signatures of geographic origin even under common-garden conditions. Coordinated investment in flavonoids and isoprenoids, together with among-origin differences in functional chemodiversity, reveals provenance-linked chemical fingerprints that complement genomic and metabolic trait data for climate-informed deployment. C_LI

plant biology↗

Lysis cassette-mediated exoprotein release in Yersinia entomophaga is controlled by a PhoB-like regulator

Secretion of exoproteins is a key component of bacterial virulence and is tightly regulated in response to environmental stimuli and host-dependent signals. The entomopathogenic bacterium Yersinia entomophaga MH96 produces a wide range of exoproteins including its main virulence factor, the 2.46 MDa insecticidal Yen-Tc toxin complex. Previously, a high-throughput transposon-based screening assay identified the region of exoprotein release (YeRER) as essential to exoprotein release in MH96. The current study defines the role of the YeRER-associated ambiguous holin/endolysin-based lysis cluster (ALC) and the novel RoeA regulator in the regulation and release of exoproteins in MH96. A mutation in the ALC region abolished exoprotein release and caused cell elongation, a phenotype able to be restored through trans-complementation with an intact ALC region. Endogenous ALC did not impact cell growth of the wild type, while artificial expression of an optimised ALC caused cell lysis. Using HolA-sfGFP and Rz1-sfGFP reporter, Rz1 expression was observed in all cells while HolA expression was limited to a small proportion of cells, which increased over time. Transcriptomic assessments found expression of the genes encoding the prominent exoproteins, including the Yen-Tc, was reduced in the roeA mutant and identified a 220 ncRNA of the YeRER intergenic that, when trans complemented in the wildtype, abolished exoprotein release. A model for Y. entomophaga mediated exoprotein regulation and release is proposed. ImportanceWhile theoretical models exist, there is not yet any empirical data that links ALC phage-like lysis cassettes with the release of large macro-molecular toxin complexes such as Yen-Tc in Gram-negative bacteria. In this study, we demonstrate that the novel Y. entomophaga RoeA activates the production of exoproteins (including Yen-Tc) and the ALC at the transcriptional level. The translation of the ALC holin is confined to a subpopulation of cells that then lyse over time, indicative of a complex hierarchical regulatory network. The presence of orthologous RoeA orthologue and a HolA like holin 5 of an eCIS Afp element in Pseudomonas chlororaphis combined with the presented data suggests a shared mechanism is required for the release of some large macromolecular protein assemblies such as the Yen-Tc, and further supports classification of phage-like lysis clusters as type 10 secretion systems.

microbiology↗