Stability of expression of reference genes in porcine peripheral blood mononuclear and dendritic cells

Real-time quantitative PCR (RT-qPCR) is a critical tool used to evaluate changes in gene expression. The precision of this tool is reliant upon the selection of reference genes whose expression remains unaltered in culture conditions and following stimulation. Stably expressed reference genes are used to normalize data so observed changes in expression are not due to artifacts but rather reflect physiological changes. In this study, we examined the expression stability of the porcine genes glyceraldehyde 3-phosphate dehydrogenase (GAPDH), succinate dehydrogenase complex subunit A (SDHA), eukaryotic elongation factor 1 gamma-like protein (eEF1), ribosomal protein L19 (RPL19), beta-actin (ACTB) and ATP synthase mitochondrial F0 complex (ATP5G1) in peripheral blood mononuclear cells (PBMCs), monocytes, monocyte-derived dendritic cells (MoDCs), blood isolated dendritic cells (BDCs) and T cells with or without stimulation with lipolysaccharide (LPS). An M value was used as a measure of gene stability as determined using geNORM software. Recommendations for the use of reference genes include using GAPDH and B-actin in PBMCs: RPL19 and SDHA in T cells; RPL19 and B-actin in monocytes; RPL-19 and SDHA in BDCs: and RPL-19 and ATP5GA in MoDCs.     

Biochemical composition is not the main factor influencing variability in carbon isotope composition of tree rings

From June to December, we determined the effects of variations in biochemical composition on delta(13)C of tree rings of 2-year-old oaks (Quercus petraea (Matt.) Liebl.) growing under semi-natural conditions, and the dependence of these effects of water stress during the growth season. Percent abundance, carbon concentration and delta(13)C were measured in holocellulose, lignin, extractive-free wood, starch and a water-soluble fraction. Relative concentrations of lignin and holocellulose in the extractive-free wood varied little during the season or in response to water stress, indicating that these compounds could not quantitatively explain the variations in whole-wood delta(13)C. Among all sampled tree rings, the relative concentration of each structural compound (holocellulose and lignin) accounted for less than 5% of the delta(13)C variability in whole wood. Variations in holocellulose and extractive-free wood delta(13)C between tree rings were almost identical (r > 0.95), whereas variations in lignin delta(13)C were less well correlated to these compounds. Whole-wood delta(13)C had a slightly altered isotopic signal compared with that of the structural compounds because of the presence of the extractive component. These results showed that variations in lignin delta(13)C and lignin concentration have little influence on extractive-free wood delta(13)C and whole-wood delta(13)C. Rather, holocellulose influences delta(13)C the most. Thus, we confirmed that, for climatic reconstruction from tree rings, removal of extractives by soxhlet is generally sufficient and sometimes unnecessary. Our findings also indicate that, in the case of rapid and severe water stress, the structural component did not accurately record the associated increase in delta(13)C because of dilution with previously formed organic matter and cessation of trunk growth. The effect of drought on carbon isotope ratios was more pronounced in the extractive compounds, making them good water stress indicators but only on a scale of days to months.     

The consequences of late outbreaks of the aphid Aphis gossypii in cotton growing in central Africa: Towards a possible method for the prevention of cotton stickiness

'Sticky cotton' causes substantial economic loss in central Africa. The phenomenon is the result of honeydew excreted by sapsucking insects, mainly the aphid Aphis gossypii Glover. The chemical protection currently available to farmers does not limit this type of damage effectively. From exhaustive counting of infested leaves and quantitative measurement of stickiness with a thermodetector, a positive relationship can be established between the seriousness of fibre stickiness and aphid outbreaks at the end of the season. A control method involving the topping of plants after boll opening reduces the number of leaves on which aphids feed and thus the abundance of the aphids.     

Etapes du developpement embryonnaire et larvaire du sar, Diplodus sargus L., en elevageStages of the embryonic and larval development of the white sea bream, Diplodus sargus L., in rearing conditions

Eggs of white sea bream (Diplodus sargus), taken from wild parents, were incubated and, after hatching, produced larvae which were then reared up to the fry stage. The different stages of development during the embryonic and larval periods are described. Some characteristics of the eggs (buoyancy, duration of incubation, critical periods) and of the larvae (metrical characters) are reported.     

Low genetic diversity and strong geographic structure in introduced populations of the Eucalyptus foliar pathogen Teratosphaeria destructans

The aggressive Eucalyptus leaf pathogen, Teratosphaeria destructans, causes widespread damage in tropical and subtropical Eucalyptus-growing regions of Indonesia, China, Thailand, East Timor, Vietnam, Lao, and South Africa. Little is known regarding the origin, pathways of dispersal, or reproductive biology of this pathogen. The aim of this study was to investigate the genetic structure of a global collection of T. destructans isolates. This was achieved by developing and using polymorphic microsatellite markers. Low genotypic diversity and a limited number of private alleles were found in all investigated populations, with the highest maximum diversity of 10.7% in isolates from South Sumatra. This supports the hypothesis that T. destructans was introduced to these regions. High levels of clonality were common in all populations, especially in isolates sampled from the recent disease outbreak in South Africa, which were all identical. The global collection of isolates grouped into three distinct clusters, corresponding largely to their sampled regions. Low levels of genotypic diversity, high levels of clonality, and strong geographic structure suggest independent introductions into all the sampled areas from an unknown source. The results imply that strict biosecurity measures are needed to avoid introductions of additional genotypes in these areas.     

Soil‐carbon preservation through habitat constraints and biological limitations on decomposer activity

We review recent experimental results on the role of soil biota in stabilizing or destabilizing soil organic matter (SOM). Specifically, we analyze how the differential substrate utilization of the various decomposer organisms contributes to a decorrelation of chemical stability, residence time, and carbon (C) age of organic substrates. Along soil depth profiles, a mismatch of C allocation and abundance of decomposer organisms is consistently observed, revealing that a relevant proportion of soil C is not subjected to efficient decomposition. Results from recent field and laboratory experiments suggest that (1) bacterial utilization of labile carbon compounds is limited by short‐distance transport processes and, therefore, can take place deep in the soil under conditions of effective local diffusion or convection. In contrast, (2) fungal utilization of phenolic substrates, including lignin, appears to be restricted to the upper soil layer due to the requirement for oxygen of the enzymatic reaction involved. (3) Carbon of any age is utilized by soil microorganisms, and microbial C is recycled in the microbial food web. Due to stoichiometric requirements of their metabolism, (4) soil animals tend to reduce the C concentration of SOM disproportionally, until it reaches a threshold level. The reviewed investigations provide new and quantitative evidence that different soil C pools underlie divergent biological constraints of decomposition. The specialization of decomposers towards different substrates and microhabitats leads to a relatively longer persistence of virtually all kinds of organic substrates in the nonpreferred soil spaces. We therefore propose to direct future research explicitly towards such biologically nonpreferred areas where decomposition rates are slow, or where decomposition is frequently interrupted, in order to assess the potential for long‐term preservation of C in the soil.     

Uptake and metabolic fate of [14C]-2,4-dichlorophenol and [14C]-2,4-dichloroaniline in wheat (Triticum aestivum) and soybean (Glycine max)

The uptake and metabolism of [14C]-2,4-dichlorophenol (DCP) and [14C]-2,4-dichloroaniline (DCA) were investigated in wheat and soybean. Seeds were exposed to a nutrient solution containing 50 microM of one of two radiolabeled compounds, and plant organs were harvested separately after 18 days of growth. In wheat, uptake of [14C]-2,4-DCP was 16.67 +/- 2.65 and 15.50 +/- 2.60% of [14C]-2,4-DCA. In soybean, uptake of [14C]-2,4-DCP was significantly higher than [14C]-2,4-DCA uptake, 38.39 +/- 2.56 and 18.98 +/- 1.64%, respectively. In the case of [14C]-2,4-DCP, the radioactivity absorbed by both species was found mainly associated with roots, whereas [14C]-2,4-DCA and related metabolites were associated with aerial parts, especially in soybean. In wheat, nonextractable residues represented 7.8 and 8.7% of the applied radioactivity in the case of [14C]-2,4-DCP and [14C]-2,4-DCA, respectively. In soybean, nonextractable residues amounted to 11.8 and 5.8% of the total radioactivity for [14C]-2,4-DCP and [14C]-2,4-DCA, respectively. In wheat, nonextractable residues were nearly equivalent to extractable residues for [14C]-2,4-DCP, whereas they were greater for [14C]-2,4-DCA. In soybean, the amount of extractable residues was significantly greater for both chemicals. However, in both species, nonextractable residues were mainly associated with roots. Isolation of soluble residues was next undertaken using excised shoots (wheat) or excised fully expanded leaves including petioles (soybean). Identification of metabolite structures was made by comparison with authentic standards, by enzymatic hydrolyses, and by electrospray ionization-mass spectrometric analyses. Both plant species shared a common metabolism for [14C]-2,4-DCP and [14C]-2,4-DCA since the malonylated glucoside conjugates were found as the final major metabolites.     

Assessment of earthworm contribution to soil hydrology: a laboratory method to measure water diffusion through burrow walls

The capacity for water diffusion in burrow walls (i.e. the coefficient of sorptivity) either burrowed by Lumbricus terrestris (T-Worm) or artificially created (T-Artificial) was studied through an experimental design in a 2D terrarium. In addition, the soil density of earthworm casts, burrow walls (0–3 mm around the burrow) and the surrounding soil (>3 mm) were measured using the method of petroleum immersion. This study demonstrated that the quantity of water which transits through burrows of L. terrestris in the soil matrix was lower than that transited through soil fractures, due to a reduction of soil porosity in burrow walls (compaction: cast > worms burrow walls > surrounding soil > artificial burrow walls). Earthworm behaviour, in particular burrow reuse with associated cast pressing on walls, could explain the larger burrow wall compaction in earthworm burrows. If water diffusion was lower through the compacted burrows, burrow reuse by the worms makes them more stable (worms would maintain the structure over years) than unused burrows. The present experimental design could be used to test and measure the specific differences between earthworm species in their contributions to water diffusion. Probably, these contributions depend on the presumed related-species behaviours which would determine the degree of burrow wall compaction.