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Fraser, T.

Publications and source records attributed to Fraser, T..

5 recordsLinked to original sources

Plantago lanceolata and Lolium perenne metabolite profiles, their impact on soil microbial community structures and soil biological nitrification inhibition

Background and aimsExcess nitrate (NO3-), from fertilizer overuse and intensive agriculture, can pollute water and contribute to greenhouse gas production (nitrous oxide - N2O). Plant metabolites from pastural herbs such as Plantago lanceolata (plantain) can inhibit microbial nitrification of ammonium to NO3- (biological nitrification inhibition - BNI) and change soil nitrogen cycle dynamics (lower potential nitrification rate - PNR). The main aim was to investigate differential plant metabolite expression associated with BNI and lowered PNR in different soil types. MethodsSix plantain cultivars were tested for BNI potential and screened for metabolites that correlated with inhibition of the ammonia oxidising bacterium (AOB) Nitrosospira multiformis. PNR and microbiome change was then investigated in four different New Zealand soils under the plantain cultivar Agritonic and ryegrass cultivar One50. ResultsPNR under plantain was 11 to 41% lower than fallow soil while PNR under ryegrass was 0 to 39% lower. In addition to verbascoside and aucubin, plantain metabolites associated with lower PNR included plantamajoside, riboflavin 3- and 5-sulfate, plantagoguanidinic acid. Chlorogenic acid was associated with lowered PNR under ryegrass. PNR reductions, microbiome structure and the ratio of ammonia oxidising archaea (AOA) relative to AOB was modulated by soil type. ConclusionPlantain and ryegrass lowered the PNR in four different soils and was correlated with metabolites beyond just aucubin and verbascoside. Based on candidate BNI-associated metabolites identified, it was hypothesised that lowered PNR is likely indirect through mechanisms such as chelation and appears to be dependent on both plant physiology and soil physicochemistry.

systems biology↗

Linking plantain derived metabolites in sheep urine with nitrification inhibition in soil

Metabolites from Plantago lanceolata (plantain) biomass have been linked with biological nitrification inhibition (BNI) in soil. After grazing, leaf metabolite chemistry is altered via digestion, and a suite of secondary metabolites are then delivered onto soil via dung and urine. The purpose of this study was to establish if urine from sheep grazed on plantain had BNI activity when added to pasture soil, and to identify the metabolite profile(s) that most likely contribute to the BNI effects observed. Groups of sheep (n=5 sheep per group) were grazed on one of nine different plantain cultivars in autumn and spring with analysis of leaf material, urine, soil incubation and BNI bioassay data used to identify potential metabolite candidates implicated with BNI. The urinary nitrogen and metabolite composition of sheep fed plantain varied significantly between cultivars and season. After 28 days of incubation, all soil microcosms treated with plantain-derived urine had up to 35% less nitrate than comparative ryegrass urine controls in both seasons, except one in autumn. The key phytochemistry associated with lower soil nitrate concentrations was phenylethanoid and iridoid glycosides resulting in a higher output of glucuronidated, methylated and sulfated secondary metabolites in the urine. Among 19 secondary metabolites identified in the urine, hydroxytyrosol-related metabolites as well as catechol glucuronide, 2-methoxyphenyl sulfate and guaiacol-{beta}-D-glucuronide appear to be the most likely target compounds with respect to the BNI effects observed. Variation in metabolites from different plantain cultivars affected the ratio of metabolite derivatives in urine, which ultimately affected soil nitrification rates. Cultivar phytochemistry is therefore an important consideration with respect to BNI under urine patches. HighlightsO_LISheep grazing different plantain cultivars had different urine compositions C_LIO_LIUrines elicited biological nitrification inhibition (BNI) in soil and in vitro C_LIO_LIDifferent BNI response was related to differential expression of urine metabolites C_LIO_LIKey urine metabolites associated with BNI are derived from glycosidic compounds C_LI

systems biology↗

Machine learning-based image analysis of Parkinson's disease iPS-derived neurons predicts genotype and reveals mitochondria-lysosome abnormalities

Mitochondrial and lysosomal dysfunction are central features of Parkinsons disease (PD) across major genetic forms including PRKN, SNCA, and LRRK2. We applied cell morphomics, a machine-learning-based framework combining high-content imaging with quantitative feature extraction, to analyse mitochondrial and lysosomal morphology at single-cell resolution in iPS cell-derived cortical neurons from PD patients and healthy controls (13 lines total). Supervised machine-learning models distinguished PD neurons from controls with high accuracy (AUC = 0.87) and reliably separated individual genotypes. Feature importance and attribution analysis revealed genotype-specific organelle biases, with mitochondrial features dominating classification in PRKN neurons, balanced mitochondrial and lysosomal contributions in SNCA neurons, and a greater lysosomal contribution in LRRK2 neurons. Multi-class models retained strong performance, and findings were reproduced across two independent laboratories using different dyes and imaging conditions. These results demonstrate that morphomics provides a robust and scalable framework to quantify genotype-specific organelle abnormalities in PD neurons and supports its application for cellular stratification and biomarker discovery.

neuroscience↗

Dopamine and cortical iPSC-derived neurons with different Parkinsonian mutations show variation in lysosomal and mitochondrial dysfunction: implications for protein deposition versus selective cell loss

BackgroundMutations causing Parkinsons disease (PD) give diverse pathological phenotypes whose cellular correlates remain to be determined. For example, those with PRKN loss of function mutations have significantly earlier selective vulnerability of dopamine neurons, those with SNCA mutations have increased alpha-synuclein deposition, while those with LRRK2 mutations have additional deposition of tau. Yet all three mutation types are implicated in mitochondrial and/or lysosomal dysfunction. Direct comparison of cell models with these mutations would clarify the relative cellular dysfunctions associated with these different pathological phenotypes. MethodsAn unbiased high-content imaging platform using orthogonal probes to assess both lysosomal and mitochondrial dysfunction, along with alpha-synuclein and tau protein deposition was established using induced pluripotent stem cell (iPSC) derived cortical and ventral midbrain neurons. Three mutation types, SNCA A53T, LRRK2 R1441G and PRKN loss of function (lof), were selected as exemplars of divergent PD pathological phenotypes and compared to each other, and to control iPSC from subjects without PD. ResultsDifferent PD mutations caused cell type specific dysfunctions, likely to impact on both selective neuronal vulnerability and the pathologies observed in PD. Comparison of dopamine neurons identified that both lysosomal and mitochondrial dysfunction were predominant with PRKN lof mutations, whereas immunofluorescent staining revealed that SNCA A53T and LRRK2 R1441G mutations had increased tau deposition. In contrast, cortical neurons with SNCA and LRRK2 mutations both had mitochondrial and autophagy impairments without protein deposition, with LRRK2 cells additionally showing decreased glucocerebrosidase activity and increased alpha-synuclein phosphorylation. ConclusionsLysosomal and mitochondrial dysfunction are predominant in dopamine neurons with PRKN lof mutations, and may drive the early selective loss of dopamine neurons in PRKN mutation carriers. More subtle cellular abnormalities in the SNCA A53T cell lines are likely to predispose to alpha-synuclein aggregation and tau protein deposition over time. The LRRK2 R1441G may also predispose to tau deposition, but despite substantial lysosomal dysfunction with increased alpha-synuclein phosphorylation, pathological alpha-synuclein accumulations were not observed. Understanding the mechanistic differences in how lysosomal and mitochondrial dysfunction impact on PD pathogenesis in different disease subtypes may be important for therapeutic development.

neuroscience↗

Meta-exoproteomics identifies active plant-microbe interactions operating in the rhizosphere

The advance of DNA sequencing technologies has drastically changed our perception of the complexity and structure of the plant microbiome and its role in augmenting plant health. By comparison, our ability to accurately identify the metabolically active fraction of soil microbiota and their specific functional role is relatively limited. Here, we combined our recently developed protein extraction method and an iterative bioinformatics pipeline to enable the capture and identification of extracellular proteins (meta-exoproteomics) expressed in the rhizosphere of Brassica spp. First, we validated our method in the laboratory by successfully identifying proteins related to the host plant (Brassica rapa) and a bacterial inoculant, Pseudomonas putida BIRD-1, revealing the latter expressed numerous rhizosphere specific proteins linked to the acquisition of plant-derived nutrients. Next, we analysed natural field-soil microbial communities associated with Brassica napus L (Oil Seed rape). By combining deep-sequencing metagenomics with meta-exoproteomics, a total of 1882 proteins were identified in bulk and rhizosphere samples. Importantly, meta-exoproteomics identified a clear shift (p<0.001) in the metabolically active fraction of the soil microbiota responding to the presence of B. napus roots that was not apparent in the composition of the total microbial community (metagenome). This metabolic shift was associated with the stimulation of rhizosphere-specialised bacteria, such as Gammaproteobacteria, Betaproteobacteria and Flavobacteriia and the upregulation of plant beneficial functions related to phosphorus and nitrogen mineralisation. By providing the first meta-proteomic level assessment of the active plant microbiome at the field-scale, this study demonstrates the importance of moving past a genomic assessment of the plant microbiome in order to determine ecologically important plant: microbe interactions driving plant growth.

microbiology↗