Search

Understanding the responses of insect herbivores to plant chemical defences is pivotal for the management of crops and pests. However, the mechanisms of interaction are not entirely understood. In this study, we compared the whole transcriptome gene expression of the aphid Macrosiphum euphorbiae grown on two different varieties of tomato that differ in their inducible chemical defences. We used two isogenic lines of tomato with a shared genetic background that only differ in the presence of type IV glandular trichomes and their associated acylsucrose excretions. This works also reports a de novo transcriptome of the aphid M. euphorbiae. Subsequently, we identified a unique and distinct gene expression profile for the first time corresponding to aphid´s exposure to type IV glandular trichomes and acylsugars. The analysis of the aphid transcriptome shows that tomato glandular trichomes and their associated secretions are highly efficient in triggering stress-related responses in the aphid, and demonstrating that their role in plant defence goes beyond the physical impediment of herbivore activity. Some of the differentially expressed genes were associated with carbohydrate, lipid and xenobiotic metabolisms, immune system, oxidative stress response and hormone biosynthesis pathways. Also, the observed responses are compatible with a starvation syndrome. The transcriptome analysis puts forward a wide range of genes involved in the synthesis and regulation of detoxification enzymes that reveal important underlying mechanisms in the interaction of the aphid with its host plant and provides a valuable genomic resource for future study of biological processes at the molecular level using this aphid.

Photosystem I (PS I) from the primitive cyanobacterium Gloeobacter violaceus has been purified and characterised. Despite the fact that the isolated complexes have the same subunit composition as complexes from other cyanobacteria, the amplitude of flash-induced absorption difference spectra indicates a much bigger antenna size with about 150 chlorophylls per P700 as opposed to the usual 90. Image analysis of the PS I preparation from Gloeobacter reveals that the PS I particles exist both in a trimeric and in a monomeric form and that their size and shape closely resembles other cyanobacterial PS I particles. However, the complexes exhibit a higher molecular weight as could be shown by gel filtration. The preparation contains novel polypeptides not related to known Photosystem I subunits. The N-terminal sequence of one of those polypeptides has been determined and reveals no homology to known or hypothetical proteins. Immunoblotting shows a cross-reaction of three of the polypeptide bands with an antibody raised against the major LHC from the diatom Cyclotella cryptica. Electron microscopy reveals a novel T-shaped complexwhich has never been observed in any other cyanobacterial PS I preparation. 77 K spectra of purified PS I show an extreme blue-shift of the fluorescence emission, indicating an unusual organisation of the PS I antenna system in Gloeobacter.

Two cell types of the same clone of Phaeocystis globosa, solitary non-flagellate cells and flagellates, were grown in batch cultures under identical conditions. The non-flagellate cells had a shorter lag phase (1.4 versus 2.8 days) and a higher growth rate (0.72 versus 0.65 day(-1)) than flagellate cells. The flagellates had a longer stationary phase (15.6 versus 9.5 days) and a lower death rate (0.07 versus 0.52 day(-1)) than non-flagellate cells. All differences were statistically significant. Biomass yield did not differ between the two cell types. The short lag phase and high growth rate of non-flagellate cells corresponds to field observations of rapidly developing non-flagellate Phaeocystis blooms that are typically observed in nutrient-rich environments such as temperate seas in spring. The flagellate cell type, with its longer stationary phase and lower death rate than non-flagellate cells, is better equipped for survival in oligotrophic environments. This explains why the flagellates of Phaeocystis are abundant after the spring phytoplankton bloom in temperate seas and in other nutrient-poor environments such as the open ocean.

A strain of Phaeocystis sp., isolated in the Southern Ocean, was cultured under iron- and light-limited conditions. The cellular content of chlorophyll a and accessory light-harvesting (LH) pigments increased under low light intensities. Iron limitation resulted in a decrease of all light-harvesting pigments. However, this decrease was greatly compensated for by a decrease in cell volume. Cellular concentrations of the LH pigments were similar for both iron-replete and iron-deplete cells. Concentrations of chlorophyll a were affected only under low light conditions, wherein concentrations were suppressed by iron limitation. Ratios of the LH pigments to chlorophyll a were highest for iron-deplete cells under both light conditions. The photoprotective cycle of diato/diadinoxanthin was activated under high light conditions, and enhanced by iron stress. The ratio of diatoxanthin to diadinoxanthin was highest under high light, low iron conditions. Iron limitation induced synthesis of 19'-hexanoyloxyfucoxanthin and 19'-butanoyloxyfucoxanthin at the cost of fucoxanthin. Fucoxanthin formed the main carotenoid in iron-replete Phaeocystis cells, whereas for iron-deplete cells 19'-hexanoyloxyfucoxanthin was found to be the main carotenoid. This shift in carotenoid composition is of importance in view of the marker function of both pigments, especially in areas where Phaeocystis sp. and diatoms occur simultaneously. A hypothesis is presented to explain the transformation of fucoxanthin into 19'-hexanoyloxyfucoxanthin and 19'-butanoyloxyfucoxanthin, referring to their roles as a light-harvesting pigment.

Clostridium acetobutylicum was metabolically engineered to produce a biofuel consisting of an isopropanol/butanol/ethanol mixture. For this purpose, different synthetic isopropanol operons were constructed and introduced on plasmids in a butyrate minus mutant strain (C acetobutylicum ATCC 824 Delta cac15 Delta upp Delta buk). The best strain expressing the isopropanol operon from the thl promoter was selected from batch experiments at pH 5. By further optimizing the pH of the culture, a biofuel mixture with almost no by-products was produced at a titer, a yield and productivity never reached before, opening the opportunities to develop an industrial process for alternative biofuels with Clostridia! species. Furthermore, by performing in vivo and in vitro flux analysis of the synthetic isopropanol pathway, this flux was identified to be limited by the [acetate](int) and the high Km of CoA-transferase for acetate. Decreasing the Km of this enzyme using a protein engineering approach would be a good target for improving isopropanol production and avoiding acetate accumulation in the culture medium. (C) 2013 Elsevier Inc. All rights reserved.

Streptococcus uberis is a highly prevalent causative agent of bovine mastitis, which leads to large economic losses in the dairy industry. The aim of this study was to examine the host response during acute inflammation after experimental challenge with capsulated Strep. uberis. Gene expression in response to Strep. uberis was compared between infected and control quarters in 3 animals. All quarters (n=16) were sampled at 16 different locations. Microarray data showed that 239 genes were differentially expressed between infected and control quarters. No differences in gene expression were observed between the different locations. Microarray data were confirmed for several genes using quantitative PCR analysis. Genes differentially expressed due to early Strep. uberis mastitis represented several stages of the process of infection: (1) pathogen recognition; (2) chemoattraction of neutrophils; (3) tissue repair mechanisms; and (4) bactericidal activity. Three different pathogen recognition genes were induced: ficolins, lipopolysaccharide binding protein, and toll-like receptor 2. Calgranulins were found to be the most strongly upregulated genes during early inflammation. By histology and immunohistochemistry, we demonstrated that changes in gene expression in response to Strep. uberis were induced both in infiltrating somatic milk cells and in mammary epithelial cells, demonstrating that the latter cell type plays a role in milk production as well as immune responsiveness. Given the rapid development of inflammation or mastitis after infection, early diagnosis of (Strep. uberis) mastitis is required for prevention of disease and spread of the pathogen. Insight into host responses could help to design immunomodulatory therapies to dampen inflammation after (early) diagnosis of Strep. uberis mastitis. Future research should focus on development of these early diagnostics and immunomodulatory components for mastitis treatment.

Although increasing concentrations of atmospheric CO2 are predicted to have substantial impacts on plant growth and functioning of ecosystems, there is insufficient understanding of the responses of belowground processes to such increases. We investigated the effects of different dark septate root endophytic (DSE) fungi on growth and nutrient acquisition by Pinus sylvestris seedlings under conditions of N limitation and at ambient and elevated CO2 (350 or 700 μ1 CO2 l−1). Each seedling was inoculated with one of the following species: Phialocephala fortinii (two strains), Cadophora finlandica, Chloridium paucisporum, Scytalidium vaccinii, Meliniomyces variabilis and M. vraolstadiae. The trial lasted 125 days. During the final 27 days, the seedlings were labeled with 14CO2 and 15NH4+. We measured extraradical hyphal length, internal colonization, plant biomass, 14C allocation, and plant N and 15N content. Under elevated CO2, the biomass of seedlings inoculated with DSE fungi was on average 17% higher than in control seedlings. Simultaneously, below-ground respiration doubled or trebled, and as a consequence carbon use efficiency by the DSE fungi significantly decreased. Shoot N concentration decreased on average by 57% under elevated CO2 and was lowest in seedlings inoculated with S. vaccinii. Carbon gain by the seedlings despite reduced shoot N concentration indicates that DSE fungi increase plant nutrient use efficiency and are therefore more beneficial to the plant under elevated CO2.

Photosystem I (PS I) from the primitive cyanobacterium Gloeobacter violaceus has been purified and characterised. Despite the fact that the isolated complexes have the same subunit composition as complexes from other cyanobacteria, the amplitude of flash-induced absorption difference spectra indicates a much bigger antenna size with about 150 chlorophylls per P700 as opposed to the usual 90. Image analysis of the PS I preparation from Gloeobacter reveals that the PS I particles exist both in a trimeric and in a monomeric form and that their size and shape closely resembles other cyanobacterial PS I particles. However, the complexes exhibit a higher molecular weight as could be shown by gel filtration. The preparation contains novel polypeptides not related to known Photosystem I subunits. The N-terminal sequence of one of those polypeptides has been determined and reveals no homology to known or hypothetical proteins. Immunoblotting shows a cross-reaction of three of the polypeptide bands with an antibody raised against the major LHC from the diatom Cyclotella cryptica. Electron microscopy reveals a novel T-shaped complexwhich has never been observed in any other cyanobacterial PS I preparation. 77 K spectra of purified PS I show an extreme blue-shift of the fluorescence emission, indicating an unusual organisation of the PS I antenna system in Gloeobacter.

Two cell types of the same clone of Phaeocystis globosa, solitary non-flagellate cells and flagellates, were grown in batch cultures under identical conditions. The non-flagellate cells had a shorter lag phase (1.4 versus 2.8 days) and a higher growth rate (0.72 versus 0.65 day(-1)) than flagellate cells. The flagellates had a longer stationary phase (15.6 versus 9.5 days) and a lower death rate (0.07 versus 0.52 day(-1)) than non-flagellate cells. All differences were statistically significant. Biomass yield did not differ between the two cell types. The short lag phase and high growth rate of non-flagellate cells corresponds to field observations of rapidly developing non-flagellate Phaeocystis blooms that are typically observed in nutrient-rich environments such as temperate seas in spring. The flagellate cell type, with its longer stationary phase and lower death rate than non-flagellate cells, is better equipped for survival in oligotrophic environments. This explains why the flagellates of Phaeocystis are abundant after the spring phytoplankton bloom in temperate seas and in other nutrient-poor environments such as the open ocean.

Two cell types of the same clone of Phaeocystis globosa, solitary non-flagellate cells and flagellates, were grown in batch cultures under identical conditions. The non-flagellate cells had a shorter lag phase (1.4 versus 2.8 days) and a higher growth rate (0.72 versus 0.65 day(-1)) than flagellate cells. The flagellates had a longer stationary phase (15.6 versus 9.5 days) and a lower death rate (0.07 versus 0.52 day(-1)) than non-flagellate cells. All differences were statistically significant. Biomass yield did not differ between the two cell types. The short lag phase and high growth rate of non-flagellate cells corresponds to field observations of rapidly developing non-flagellate Phaeocystis blooms that are typically observed in nutrient-rich environments such as temperate seas in spring. The flagellate cell type, with its longer stationary phase and lower death rate than non-flagellate cells, is better equipped for survival in oligotrophic environments. This explains why the flagellates of Phaeocystis are abundant after the spring phytoplankton bloom in temperate seas and in other nutrient-poor environments such as the open ocean.

A strain of Phaeocystis sp., isolated in the Southern Ocean, was cultured under iron- and light-limited conditions. The cellular content of chlorophyll a and accessory light-harvesting (LH) pigments increased under low light intensities. Iron limitation resulted in a decrease of all light-harvesting pigments. However, this decrease was greatly compensated for by a decrease in cell volume. Cellular concentrations of the LH pigments were similar for both iron-replete and iron-deplete cells. Concentrations of chlorophyll a were affected only under low light conditions, wherein concentrations were suppressed by iron limitation. Ratios of the LH pigments to chlorophyll a were highest for iron-deplete cells under both light conditions. The photoprotective cycle of diato/diadinoxanthin was activated under high light conditions, and enhanced by iron stress. The ratio of diatoxanthin to diadinoxanthin was highest under high light, low iron conditions. Iron limitation induced synthesis of 19'-hexanoyloxyfucoxanthin and 19'-butanoyloxyfucoxanthin at the cost of fucoxanthin. Fucoxanthin formed the main carotenoid in iron-replete Phaeocystis cells, whereas for iron-deplete cells 19'-hexanoyloxyfucoxanthin was found to be the main carotenoid. This shift in carotenoid composition is of importance in view of the marker function of both pigments, especially in areas where Phaeocystis sp. and diatoms occur simultaneously. A hypothesis is presented to explain the transformation of fucoxanthin into 19'-hexanoyloxyfucoxanthin and 19'-butanoyloxyfucoxanthin, referring to their roles as a light-harvesting pigment.