DNA microarray-based genotyping of Chlamydophila psittaci strains from culture and clinical samples

The avian and human pathogen Chlamydophila (C.) psittaci represents a genetically heterogeneous species. To facilitate epidemiological surveys, more rapid yet highly specific molecular tests are needed. Currently used typing methods, i.e. serotyping and PCR-RFLP, have only limited sensitivity and are incapable of covering the wide spectrum of naturally occurring types of C. psittaci strains. In the present study, a new DNA microarray assay based on the ArrayTube (AT) technology was used to genotype C. psittaci in 98 isolates and 23 clinical tissue samples. The present array carries 35 oligonucleotide probes derived from variable domains 2 and 4 of the ompA gene. The assay proved highly sensitive, allowing correct genotyping of DNA from 2 inclusion-forming units. The results of DNA microarray genotyping of cultured strains proved highly concordant with the data from PCR-RFLP typing and serotyping. Sequencing of the ompA gene served as the reference test to verify the accuracy of AT genotyping results. In 15 instances (15.3%), strains were successfully typed by the AT assay, while serotyping and/or PCR-RFLP genotyping failed to produce unambiguous results. Eleven of these samples were ompA sequenced to confirm the AT findings. In addition to the currently accepted nine ompA genotypes, the microarray test was shown to recognise new provisional genotypes, such as Mat116 and YP84. In conclusion, the new AT assay proved to be suitable for rapid, sensitive and reproducible genotyping of C. psittaci strains and can be recommended for routine diagnosis.     

Genes for race-specific resistance against blackleg disease in Brassica napus L.

Specificity of interaction at the cotyledon stage was recently demonstrated between the blackleg pathogen, Leptosphaeria maculans, and Brassica napus. Three pathogenicity groups were distinguished, PG2 avirulent towards ‘Quinta’ and ‘Glacier’, PG3 avirulent towards ‘Quinta’, and PG4 virulent on the two cultivars. The genetic control of the interactions was investigated on both the pathogen and the plant. Tetrad analysis was performed following PG3 × PG4 and PG2 × PG4 crosses.‘Quinta’ and ‘Glacier’ were crossed with the susceptible winter oilseed rape cultivar ‘Score’. The analysis of F₁, F₂ and testcross populations suggested that the incompatible interaction between ‘Quinta’ and PG3 isolates is conditioned by the presence of the dominant single resistance allele Rlml in ‘Quinta’ and the matching avirulence gene AvrLml in L. maculans. Race-specific resistance of ‘Glacier’ to PG2 isolates was conditioned by the matching gene pair Rlm2/AvrLm2. Finally, the data suggest that two avirulence genes matching two dominant loci control the ‘Quinta’-PG2 interaction. The consequences of the occurrence of race-specific resistance in B. napus are discussed with respect to future breeding for blackleg resistance.     

Identification of RAPD markers linked to linolenic acid genes in rapessed

Summary The inheritance of the low linolenic acid content (derivated from mutant lines) in rapeseed was investigated. Molecular techniques of gene mapping through RAPD markers were applied on a microspore-derived progeny from a high × low linolenic acid F₁ hybrid. Bulked segregant analysis made it possible to test rapidly number of RAPD primers. Two linkage groups of 6 markers (72.7 cM and 75.6 cM) were determined. Each corresponded to a major QTL which explained 24% and 30.7% of the total phenotypic variation of the linolenic acid content. It was confirmed that two independant mutations were implied in the low linonenic acid content.     

Response to Chemical Alarm Cues under Weakly Acidic Conditions: A Graded Loss of Antipredator Behaviour in Juvenile Rainbow Trout

A wide variety of aquatic organisms, including juvenile salmonids, assess local predation risks using chemosensory cues. Such chemical cues are typically released from injured conspecifics and their detection may lead to species-typical antipredator behaviour, increasing the probability of prey to survive during predator encounters. Studies have demonstrated however, that under weak acidification (pH ~6.0), the response towards these chemical alarm cues is impaired. However, it remains unknown if the loss of response is graded (i.e., the behavioural response decreases with a reduction in pH) or if there is a threshold pH at which prey can no longer detect the alarm cues. We conducted two laboratory experiments to examine the effects of a graded reduction in pH on the behavioural response of juvenile rainbow trout to conspecific chemical alarm cues. The results of our first experiment suggest that at pH 6.6 and above, the alarm cues elicited a strong antipredator response, while alarm cues buffered to pH 6.2 did not (i.e. not different from distilled water). However, alarm cues buffered to pH 6.4 elicited a weak response, suggesting a graded response. We directly tested this in our second experiment using a repeated measures design. The response to alarm cues at varying pH levels did indeed follow a graded loss of function. Together, our results suggest that juvenile rainbow trout exhibit a reduction in the response to conspecific alarm cues proportional to ambient acidity and that the response to these critically important cues is lost at pH below 6.4. As the detection and response to these chemical alarm cues have been shown to confer direct survival benefit to individuals, these results are therefore presented in relation to possible sub-lethal effects of anthropogenic acidification to freshwater fish.     

Synthesis and sensorial properties of 2-alkylalk-2-enals and 3-(acetylthio)-2-alkyl alkanals

Parallel synthesis was applied to prepare a series of 3-(acetylthio)-2-alkyl alkanals by Michael addition of thioacetic acid under alkaline conditions to alpha,beta-unsaturated 2-alkyl-substituted aldehydes, which were obtained by aldol condensation of the corresponding primary aldehydes as starting materials. The target compounds were characterized in terms of GC, MS, and NMR data. The sensory properties of the odorants, such as odor quality and odor detection threshold value, were determined with a trained panel. Structure-activity relationships are discussed, suggesting that the 1,3-oxygen-sulfur functionality, required for the "olfactophore" of tropical/vegetable notes, can further be extended to the acetylthio derivatives.     

Litter decomposition in peatlands is promoted by mixed plants

Purpose

The carbon sink function of peatlands is primarily driven by a higher production than decomposition of the litter Sphagnum mosses. The observed increase of vascular plants in peatlands could alter the decomposition rate and the carbon (C) cycle through a litter mixing effect, which is still poorly studied. Here, we examine the litter mixing effect of a peat moss (Sphagnum fallax) and two vascular plants (Pinus uncinata and Eriophorum vaginatum) in the field and laboratory-based experiment.

Materials and methods

During the laboratory incubation, mass loss, CO₂ production, and dissolved organic carbon concentration were periodically monitored during 51 days. The collected data were then processed in a C dynamics model. The calculated enzymatic activity was correlated to the measured β-glucosidase activity in the litter. In the field experiment, mass loss and CO₂ production from litter bags were annually measured for 3 years.

Results and discussion

Both laboratory and field experiments clearly show that the litter mixture, i.e., Sphagnum-Pinus-Eriophorum, had a synergistic effect on decomposition by enhancing the mass loss. Such enhanced mass loss increased the water extractable C and CO₂ production in the litter mixture during the laboratory experiment. The synergistic effect was mainly controlled by the Sphagnum-Eriophorum mixture that significantly enhanced both mass loss and CO₂ production. Although the β-glucosidase activity is often considered as a major driver of decomposition, mixing the litters did not cause any increase of the activity of this exo-enzyme in the laboratory experiment suggesting that other enzymes can play an important role in the observed effect.

Conclusions

Mixing litters of graminoid and Sphagnum species led to a synergistic effect on litter decomposition. In a context of vegetation dynamics in response to environmental change, such a mixing effect could alter the C dynamics at a larger scale. Identifying the key mechanisms responsible for the synergistic effect on litter decomposition, with a specific focus on the enzymatic activities, is crucial to better predict the capacity of peatlands to act as C sinks.

     

Identification of Maintainer Genes in Brassica napus L. for the Male-Sterility-Inducing Cytoplasm of Diplotaxis muralis L.

Male fertility of F¹ interspecific hybrid plants derived from crosses between cytoplasmic male-sterile Brassica campestris in Diplotaxis muralis cytoplasm and 147 B. napus cultivars was Investigated. F¹, plants obtained, from crosses with the B. napus cultivars‘Mangum’and‘Hinchu’were male-sterile while F¹ plants derived from all other crosses were male-fertile. This indicated that these two cultivars carried maintainer genes far the male-sterility-inducing cytoplasm of D. muralis. Sterility was stable In plants derived from backcrosses of male-sterile F; plants with‘Mangun and‘Hinchu’but the seed set of backcross plants was low. With restorer genes readily available in B. napus, these findings could lead to the development of a new cytoplasmic male sterility system for the breeding of B. napus hybrid cultivars.     

Spawning areas and migration patterns in the early life history of Squalius cephalus (Linnaeus, 1758): Use of otolith microchemistry for conservation and sustainable management

1. In the context of the River Rhône restoration programme, the objective of this study was to assess the dispersal and population connectivity of the European chub, Squalius cephalus (Linnaeus, 1758) in a section of the natural, free-flowing part of the lower River Rhône.
2. The elemental water signatures for Sr:Ca and Ba:Ca were measured at seven stations within the river section, including its tributaries and backwaters, to determine whether they could be differentiated by microchemistry. From August to October the signatures differed among three sectors of the study reach: the Rhône; the Ardèche; and the backwaters.
3. The elemental signatures of the otoliths of 178 young-of-the-year (YOY) chub from the stations were measured to assess spawning areas and identify early migratory life histories. Analyses of otolith microchemistry identified the natal origin of 95% of the fish. Sr:Ca signatures of otoliths showed early downstream migration for 30.4% of the YOY chub; however, 70% of the fish recruited relatively close to their spawning origin suggesting a potential inshore retention of fish.
4. The results showed (i) a non-negligible contribution of the tributaries and backwaters in the recruitment of fish into the main channel; and (ii) the ability of chub to migrate over several kilometres during their first few months of life.
5. This study shows that the combination of water and otolith microchemistry analyses of fish growth increases the understanding of fish life history. Moreover, the study revealed that half of the YOY fish from this river section came from the tributaries and the other half came from reproduction in the River Rhône (despite its high anthropization).
6. These methods offer promising future applications of otolith microchemistry for river management in the context of ecological rehabilitation, especially to assess the effectiveness of measures applied in the restoration of floodplain connectivity for riverine fish population conservation.