Growth of a common planktonic diatom quantified using solid medium culturing

The ability to grow on solid culture medium is a pre-requisite for a successful microbial genetic model organism. Skeletonema marinoi, a bloom-forming, planktonic marine microalga, is widely used in ecological, evolutionary and population genetics studies. We have tested and confirmed the ability of this common organism to grow on solid culture medium (agar) under experimentally manipulated conditions. We established a protocol for quantifying growth characteristics - length of lag phase, growth rate, maximum biomass yield - on agar medium. The procedure was tested under experimental treatments and the resulting growth changes correlated with those observed in standard liquid culture. The ability to grow on solid medium broadens the use of S. marinoi as a molecular model, where agar is routinely used for various purposes (growth, selection, storage); and the possibility to quantify colony growth opens the way for high throughput, automated, or semi-automated phenotyping solutions

Infection by a foliar endophyte elicits novel arabidopside-based plant defence reactions in its host, Cirsium arvense

Endophytic fungi live asymptomatically within plants. They are usually regarded as non-pathogenic or even mutualistic, but whether plants respond antagonistically to their presence remains unclear, particularly in the little-studied associations between endophytes and nong-raminoid herbaceous plants. We investigated the effects of the endophyte Chaetomium cochlioides on leaf chemistry in Cirsium arvense. Plants were sprayed with spores; leaf material from both subsequent new growth and the sprayed leaves was analysed 2 wk later. Infection frequency was 91% and63% for sprayed and new growth, respectively, indicating that C. cochlioides rapidly infects new foliage. Metabolomic analyses revealed marked changes in leaf chemistry with infection, especially in new growth. Changes in several novel oxylipin metabolites were detected, including arabi-dopsides reported here for the first time in a plant species other than Arabidopsis thaliana,and a jasmonate-containing galactolipid. The production of these metabolites in response to endophyte presence, particularly in newly infected foliage, suggests that endophytes elicit similar chemical responses in plants to those usually produced following wounding, herbivory and pathogen invasion. Whether en-dophytes benefit their hosts may depend on a complex series of chemically mediated interactions between the plant, the endophyte, other microbial colonists and natural enemies

PRECOG: a tool for automated extraction and visualization of fitness components in microbial growth phenomics.

BACKGROUND: Phenomics is a field in functional genomics that records variation in organismal phenotypes in the genetic, epigenetic or environmental context at a massive scale. For microbes, the key phenotype is the growth in population size because it contains information that is directly linked to fitness. Due to technical innovations and extensive automation our capacity to record complex and dynamic microbial growth data is rapidly outpacing our capacity to dissect and visualize this data and extract the fitness components it contains, hampering progress in all fields of microbiology. RESULTS: To automate visualization, analysis and exploration of complex and highly resolved microbial growth data as well as standardized extraction of the fitness components it contains, we developed the software PRECOG (PREsentation and Characterization Of Growth-data). PRECOG allows the user to quality control, interact with and evaluate microbial growth data with ease, speed and accuracy, also in cases of non-standard growth dynamics. Quality indices filter high- from low-quality growth experiments, reducing false positives. The pre-processing filters in PRECOG are computationally inexpensive and yet functionally comparable to more complex neural network procedures. We provide examples where data calibration, project design and feature extraction methodologies have a clear impact on the estimated growth traits, emphasising the need for proper standardization in data analysis. CONCLUSIONS: PRECOG is a tool that streamlines growth data pre-processing, phenotypic trait extraction, visualization, distribution and the creation of vast and informative phenomics databases

Head-group acylation of monogalactosyldiacylglycerol is a common stress response, and the acyl-galactose acyl composition varies with the plant species and applied stress

Formation of galactose-acylated monogalactosyldiacylglycerols has been shown to be induced by leaf homogenization, mechanical wounding, avirulent bacterial infection, and thawing after snap-freezing. Here, lipidomic analysis using mass spectrometry showed that galactose-acylated monogalactosyldiacylglycerols, formed in wheat (Triticum aestivum) and tomato (Solanum lycopersicum) leaves upon wounding, have acyl-galactose profiles that differ from those of wounded Arabidopsis thaliana, indicating that different plant species accumulate different acyl-galactose components in response to the same stress. Additionally, the composition of the acyl-galactose component of Arabidopsis acMGDG depends on the stress treatment. After sub-lethal freezing treatment, acMGDG contained mainly non-oxidized fatty acids esterified to galactose, whereas mostly oxidized fatty acids accumulated on galactose after wounding or bacterial infection. Compositional data are consistent with acMGDG being formed in vivo by transacylation with fatty acids from digalactosyldiacylglycerols. Oxophytodienoic acid, an oxidized fatty acid, was more concentrated on the galactosyl ring of acylated monogalactosyldiacylglycerols than in galactolipids in general. Also, oxidized fatty acid-containing acylated monogalactosyldiacylglycerols increased cumulatively when wounded Arabidopsis leaves were wounded again. These findings suggest that, in Arabidopsis, the pool of galactose-acylated monogalactosyldiacylglycerols may serve to sequester oxidized fatty acids during stress responses

Cross‐contamination risks in sediment‐based resurrection studies of phytoplankton

Abstract Resurrection studies can answer some fundamental questions in aquatic ecology and evolutionary biology. For phytoplankton resting stages, longevity of thousands to millions of years has recently been reported. However, contamination during sediment sampling could distort these estimates, and this risk has not been systematically evaluated. Here we used 4.5 μm diameter microspheres to quantify contamination while reviving the resting stages of seven abundant estuarine diatom and cyanobacterial taxa. We observed a sharp decline in resting stages abundance from 106 (g wet sediment)−1 at the surface to < 0.8 (g wet sediment)−1 at 12.5 cm depth. Added microspheres (~ 4.5 × 107 cm−2) were translocated even deeper down the sediment and could well explain the vertical distributions and abundances of revived cells. Without this control, we could have claimed to have revived seven multi‐decades to centennial‐old taxa. Our findings suggest that improved contamination controls are needed for sediment core sampling of rare cells, microfossils, or DNA molecules

Cross-contamination risks in sediment-based resurrection studies of phytoplankton

Resurrection studies can answer some fundamental questions in aquatic ecology and evolutionary biology. For phytoplankton resting stages, longevity of thousands to millions of years has recently been reported. However, contamination during sediment sampling could distort these estimates, and this risk has not been systematically evaluated. Here we used 4.5 μm diameter microspheres to quantify contamination while reviving the resting stages of seven abundant estuarine diatom and cyanobacterial taxa. We observed a sharp decline in resting stages abundance from 106 (g wet sediment)−1 at the surface to < 0.8 (g wet sediment)−1 at 12.5 cm depth. Added microspheres (~ 4.5 × 107 cm−2) were translocated even deeper down the sediment and could well explain the vertical distributions and abundances of revived cells. Without this control, we could have claimed to have revived seven multi-decades to centennial-old taxa. Our findings suggest that improved contamination controls are needed for sediment core sampling of rare cells, microfossils, or DNA molecules

Concerted Evolution of Life Stage Performances Signals Recent Selection on Yeast Nitrogen Use

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