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Biology subjects

Hardt, M.

Publications and source records attributed to Hardt, M..

5 recordsLinked to original sources

Shading does not lower thermal tolerance in the coral Montipora capitata: implications for conservation intervention

Marine heatwaves trigger severe coral bleaching events that result in dramatic losses of coral reefs worldwide. An increasingly common method used to mitigate coral bleaching is to shade portions of reefs. However, as shaded corals become less exposed to environmental stress, shading has also been hypothesized to lower their thermal tolerance, which would be detrimental for their survival. Here, we investigated how long-term shading modifies the response of Montipora capitata to both light and temperature stressors. After two years of growth, we used the Coral Bleaching Automated Stress System (CBASS) to compare responses of 73% shaded and unshaded corals during 1) temperature stress under ambient light, 2) temperature stress under low light, and 3) ambient temperature with light stress. Results show that shaded corals are less resistant than control ones to light stress and to heat stress under high light, but not to heat stress under low light, where no difference was detected. We further demonstrate that low light intensity during heat stress reduces the decline of photosynthetic efficiency when compared to heat stress at ambient light, regardless of the light history of the coral colonies. Our data support light-mediated stress, both independent of and synergistic with heat-induced stress, and suggest that light and heat stresses trigger different molecular and cellular pathways. We hereby confirm the benefits of coral shading under both short- and long-term temperature stress, and underline the importance of light acclimation in the conservation and restoration of coral reefs.

ecology↗

Highly resolved spatial transcriptomics for detection of rare events in cells

Single-cell spatial transcriptomics technologies leveraged the potential to transcriptionally landscape sophisticated reactions in cells. Current methods to delineate such complex interplay lack the flexibility in rapid target adaptation and are particularly restricted in detecting rare transcripts. We developed a multiplex single-cell RNA In-situ hybridization technique, called Molecular Cartography (MC) that can be easily tailored to specific applications and, by providing unprecedented sensitivity, specificity and resolution, is particularly suitable in tracing rare events at a subcellular level. Using a SARS-CoV-2 infection model, MC allows the discernment of single events in host-pathogen interactions, dissects primary from secondary responses, and illustrates differences in antiviral signaling pathways affected by SARS-CoV-2, simultaneously in various cell types.

molecular biology↗

Isolation and characterization of barley (Hordeum vulgare) extracellular vesicles and their role in RNAi-based crop protection

The demonstration that spray-induced gene silencing (SIGS) can confer strong disease resistance bypassing the laborious and time-consuming transgenic expression of double-stranded (ds)RNA to induce gene silencing of pathogenic targets was groundbreaking. However, future field applications will require fundamental mechanistic knowledge on dsRNA uptake, processing, and its transfer. There is increasing evidence that extracellular vesicles (EVs) mediate the transfer of transgene-derived small interfering (si)RNAs in host-induced gene silencing (HIGS) applications. Here, we examined the role of EVs regarding the translocation of sprayed dsRNA from barley (Hordeum vulgare) to the target fungus Fusarium graminearum. We found barley EVs with 156 nm in size containing predominantly 21 and 19 nucleotide (nt) siRNAs starting with a 5-terminal Adenine. Notably, barley EVs contain less siRNA compared to EVs isolated from transgenic HIGS Arabidopsis plants. Together our results further underpin mechanistic differences between HIGS and SIGS applications and a minor role of EVs in SIGS.

plant biology↗

Host-induced gene silencing involves transfer of dsRNA-derived siRNA via extracellular vesicles

Small (s)RNAs and their double-stranded (ds)RNA precursors have been adopted to control diseases in crop plants through expression in transgenic plants and targeted gene silencing (host-induced gene silencing, HIGS). While HIGS strategies proved to be effective, the mechanism of RNA transfer at the plant - pathogen interface is widely unknown. Here we show that extracellular vesicles (EVs) purified from Arabidopsis thaliana plants expressing CYP3RNA, a dsRNA originally designed to target the three CYP51 genes of the fungal pathogen Fusarium graminearum, contain CYP3RNA-derived small interfering (si)RNAs as shown by RNA sequencing (RNA-seq) analysis. These transgene specific siRNAs had a length of 21 and 22 nucleotides with a bias towards 5-uracil (U) and 5-adenine (A). Notably, stringent protease and RNase treated EV fractions contained >70% less CYP3RNA-derived siRNAs, suggesting the presence of co-purified extravesicular nucleoprotein complexes stabilizing siRNAs outside of EVs. In addition, mutants of the ESCRT-III complex showed a loss of HIGS-mediated disease resistance and EVs isolated from these mutants were free of CYP3RNA-derived siRNAs. Together, these findings support the view that endosomal vesicle trafficking is required for HIGS mediating the transfer of transgene-derived siRNAs between donor host cells and recipient fungal cells probably in an EV-independent manner.

plant biology↗

Identification of salivary and plasma biomarkers for obesity in children by non-targeted metabolomic analysis

Chemical compounds in the saliva most likely to be associated with obesity are identified in a metabolomic analysis of paired whole saliva and plasma samples from 68 children (10-year old) who have also been evaluated for their gingival redness.\n\nResultsThrough metabolomic analysis119 compounds were found only in saliva, 210 only in plasma and 126 in both. The most common plasma metabolites were lipids. The most common saliva metabolites were peptides. Amino acids and their metabolites were common in both samples.\n\nSurrogate indicators were identified by computing correlations between saliva and plasma. 29 of the 126 found in both saliva and plasma had significant positive correlation and only 4 of those (urate, creatinine, pipecolate and hydroxyproline) were associated with obesity as potential surrogate biomarkers. The uremic toxin N1-Methyl-2-pyridone-5-carboxamide (2PY) was also elevated in the saliva of obese children. 21 biochemicals were elevated in both obesity and gingivitis suggesting that some biochemical pathways for gingivitis and obesity are shared.\n\nOf the metabolites found only in saliva, 35 were associated with obesity (p<0.01). The most significant was phosphate. Saliva had 53 dipeptides, seven of which were associated with obesity. Two volatile amines (putrescine and cadaverine) and their amino acid precursors (ornithine and lysine) were also associated with obesity.\n\nOf the metabolites found only in plasma, 64 were associated with obesity (p<0.01). Significant increases in branched-chain and aromatic amino acids, significant reductions in serotonin, serine, and glycine and increased androgen metabolism are changes observed in 10-year old obese children that have also been reported with adult patients having type II diabetes.\n\nConclusionsSalivary urate may be a valuable measure of metabolic disease particularly in association with fructose consumption. Elevated salivary creatinine levels and presence of 2PY suggests a possible association of obesity with developing kidney disease in obese children. Though not a surrogate variable, salivary phosphate, (AUROC= 0.8) could also be an important indicator of obesity. Cadaverine, putrescine and dipeptides are also likely oral bacterial products that could help define the metabolic pathways responsible for obesity. The results of this study indicate that salivary metabolites can be important indicators of developing metabolic disease in children and provide a biochemical signature in the pathways that lead to obesity.

epidemiology↗