Cyclooxygenases and lipoxygenases are used by the fungus Podospora anserina to repel nematodes

Oxylipins are secondary messengers used universally in the living world for communication and defense. The paradigm is that they are produced enzymatically for the eicosanoids and non-enzymatically for the isoprostanoids. They are supposed to be degraded into volatile organic compounds (VOCs) and to participate in aroma production. Some such chemicals composed of eight carbons are also envisoned as alternatives to fossil fuels. In fungi, oxylipins have been mostly studied in Aspergilli and shown to be involved in signalling asexual versus sexual development, mycotoxin production and interaction with the host for pathogenic species. Through targeted gene deletions of genes encoding oxylipin-producing enzymes and chemical analysis of oxylipins and volatile organic compounds, we show that in the distantly-related ascomycete Podospora anserina, isoprostanoids are likely produced enzymatically. We show the disappearance in the mutants lacking lipoxygenases and cyclooxygenases of the production of 10-hydroxy-octadecadienoic acid and that of 1-octen-3-ol, a common volatile compound. Importantly, this was correlated with the inability of the mutants to repel nematodes as efficiently as the wild type. Overall, our data show that in this fungus, oxylipins are not involved in signalling development but may rather be used directly or as precursors in the production of odors against potential agressors.

Spatial genetic structure of <Emphasis Type="Italic">Lissotriton helveticus L</Emphasis>. following the restoration of a forest ponds network

Preserving amphibian genetic diversity through ecological restoration and conservation actions is a major challenge since their populations are declining worldwide. We studied the genetic diversity and spatial genetic structure of the palmate newt (Lissotriton helveticus) 2 years after the restoration of a pond network in northwestern France with the aim of reconstructing fine-scale genetic structure and patterns of colonization. We sampled newts from 29 forest ponds including both restored and non-degraded reference ponds, and genotyped 391 individuals at 12 microsatellite loci. We used two Bayesian clustering methods to spatially delineate genetic clusters, and we also detected potential recent migrants within the network. All ponds showed low levels of observed heterozygosity (Ho?=?0.534) and a mean F _IS of 0.251, possibly indicating a Wahlund or bottleneck effect. Pairwise F _ST suggested limited evidence of genetic differentiation among ponds. Within the pond network, we identified 3 to 4 genetic clusters. Combined with the detection of migrants, the results suggest an increase in gene flow within the restored pond network and that a high number of migrants came from the reference ponds. Our findings indicate an unexpected high dispersal ability for this small-bodied species. Overall, the absence of population structure represents a positive beginning for the restoration project. It also emphasizes the importance of spatial design in restoring a pond network and that such genetic data and methods should be used to monitor amphibians in restored habitats.     

Plant species diversity and polyploidy in islands of natural vegetation isolated in extensive cultivated lands

Natural polyploidy is often related to a longer life span, vegetative reproduction and higher competitive ability. In this paper, we test the possibility that these characteristics may favour the survival of polyploid taxa under conditions of long-term habitat fragmentation. In islands of natural vegetation isolated in extensive vineyards located in the South of France and in a large neighbouring area of natural vegetation, plant species richness and the relative abundance of polyploid taxa were assessed according to island size, isolation and vegetation structure. High species richness was observed, with numerous species restricted to the islands, suggesting that these may constitute refugia. However, species richness was not related to island size or to degree of isolation except for the flora of the woody areas. A very positive effect of area fragmentation on plant richness was observed, which is probably attributable to relatively low species overlap among the islands. Particularly high species richness was observed in open areas, provided that these were not extensively colonized by shrubs which seem to be responsible for local extinction of many annual taxa. Polyploids, which comprised mostly perennial herbs and woody species, were predominant in all the islands and in the large reference area. In open habitats invaded by shrubby species, a higher relative frequency of polyploids was observed in islands than in the reference area. Moreover, polyploid taxa were present in a larger number of islands than the diploid taxa, which were often restricted to a single island, suggesting that, after a long period of isolation, the polyploids may still have a lower probability of extinction. Evidence was obtained from vegetation structure analysis that diploid and polyploid annual herbs were restricted to open habitats and were both eliminated by shrubby species. Conversely, the diploid perennial herbs were also significantly affected by shrub colonization whereas the polyploids were mostly present in shrubby areas. This suggests that the higher competitive ability of polyploid perennial herbs may constitute a critical factor responsible for their wider distribution over the islands. We report the implications of our findings on conservation strategies, more particularly for a Mediterranean flora.     

Phylogenetic relationships and gene flow between sympatric diploid and tetraploid plants ofDactylis glomerata (Gramineae)

Phylogenetic relationships between sympatric, morphologically indistinguishable diploid and tetraploid plants ofDactylis glomerata L. (Gramineae) in Galicia (Spain) were assessed using allozyme markers for 6 distinct systems. The study exploited recent introduction in Galicia and subsequent hybridization of an alien 4xDactylis subspecies possessing distinct allozymes from those of all the native plants. Opportunities for gene exchanges between the ploidies were estimated from in situ observations of flowering, examination of progenies in 2x/4x natural and experimental crosses, and enzyme analyses. Results show a high genetic similarity between the Galician diploids and tetraploids, which possess peculiar alleles in common. Although the ploidy levels usually have distinct flowering periods, interploidal crosses do occasionally occur. Gene flow is likely much more important from the diploid to the tetraploid level. A good genetic intermixing occurs between the Galician and the alien tetraploid entities which have simultaneous flowering. Autopolyploidization of the diploids followed by various rates of hybridization is proposed as one very probable origin of natural tetraploids inDactylis.     

Effect of water stress on the lipid and fatty acid composition of cotton (Gossypium hirsutum) chloroplasts

In order to investigate the effects of water stress on the photosynthetic compartment, analysis of chloroplast polar lipids and their fatty acid composition was made. Cotton ( Gossypium hirsutum L. cv. Reba) plants were submitted to water stress by witholding irrigation. Chloroplasts were purified on a Percoll gradient and three fractions were collected: intact, apparently intact and broken chloroplasts. The percentage of broken chloroplasts increased with a decrease in leaf water potential, indicating a greater weakness of the membrane. Galactolipid content (expressed as mg lipid per mg chlorophyll), particularly digalactosyldiglyceride, decreased with decreasing water potential. Phospholipid content decreased in the broken chloroplast fraction. The fatty acid composition of chloroplasts was also affected. The perecentage of linolenic acid (18:3), the major fatty acid of thylakoids, decreased, whereas that of linoleic acid(18:2) and oliec acid (18:1) increased. An accumulation of fatty acids having less than 16 carbon atoms was also observed. These changes in the lipid and fatty acid composition of cotton chloroplasts under water stress might affect the properties of the thylakoid membrane and thereby the photosynthetic activity and the compartmentation of the leaf cells.     

Involvement of intestinal epithelial cells in dendritic cell recruitment during C. parvum infection

Dendritic cells (DCs) play a key role in activating and orientating immune responses. Little is currently known about DC recruitment during Cryptosporidium parvum infection. In the intestine, epithelial cells act as sensors, providing the first signals in response to infection by enteric pathogens. We analyzed the contribution of these cells to the recruitment of DCs during cryptosporidiosis. We found that intestinal epithelial cells produced a broad range of DC-attracting chemokines in vitro in response to C. parvum infection. The supernatant of the infected cells induced the migration of both bone marrow-derived DCs (BMDC) and the SRDC lymphoid dendritic cell line. Chemokine neutralization abolished DC migration in these assays. We next analyzed chemokine mRNA expression in the mucosa of C. parvum-infected neonatal mice and recruitment of the various subsets of DCs. Myeloid (CD11c+ CD11b+) and double-negative DCs (CD11c+ CD11b- CD8alpha-) were the main subsets recruited in the ileum during C. parvum infection, via a mechanism involving IFNgamma. DCs were also recruited and activated in the draining lymph nodes during C. parvum infection, as shown by the upregulation of expression of MHC II and of the costimulation molecules CD40 and CD86.     

Genetic variation and evolutionary history of holly oak: a circum-Mediterranean species-complex [Quercus coccifera L./Q. calliprinos (Webb) Holmboe, Fagaceae]

Holly oak is the only evergreen oak possessing a circum-Mediterranean range; it has two predominant morphological forms, calliprinos and coccifera, described in the eastern and western Mediterranean Basin respectively. The concordance of allozyme and morphotype variation was analysed in the whole holly oak range, and the most plausible historical events responsible for the current geographic pattern of genetic variation were investigated. Individuals from 24 populations were scored for allozyme variation at seven polymorphic loci. Multi-locus genotypes were analysed by using a correspondence analysis (CA) and a Bayesian clustering approach. The relative positions of the populations were obtained from multi-dimensional scaling coupled with UPGMA treatment. A continuous genotype distribution was observed in the CA, and two groups were identified using the Bayesian approach. With a 0.95 threshold, 66 and 69% of the individuals showing the calliprinos and the coccifera morphotypes respectively were assigned to the corresponding genetic groups, which differed by private alleles. As compared to coccifera, the genetic group calliprinos was characterized by higher allelic richness and a strong geographical genetic structure. In agreement with fossil records, the most parsimonious explanation for lack of geographical structure in coccifera is a substantial regression of holly oak in the western Mediterranean Basin during the glaciations and a fast westward post-glacial expansion of coccifera populations, probably from Greece. Two population groups were obtained from the scaling/UPGMA treatment. One included all of the calliprinos populations and a Greek coccifera population, suggesting that the two morphotypes are closely related genetically and constitute two components of the same species.     

ASSOCIATION BETWEEN QUANTITATIVE TRAITS AND ALLOZYME HETEROZYGOSITY IN A TETRASOMIC SPECIES: DACTYLIS GLOMERATA

Tetraploid individuals of orchardgrass (Dactylis glomerata L. subsp. hispanica Roth.) sampled from a natural population were used to evaluate the correlation between both single and multilocus heterozygosity at 7 enzyme loci, and several quantitative traits (plant size at time of collection, leaf weight, and panicle number in experimental trials). Four hundred and forty-eight plants were studied at the 7 loci and 288 of these individuals were scored for an additional eighth locus. Five genotype classes (monogenic, simplex, and duplex digenic, trigenic, and tetragenic) were distinguished according to their heterozygosity level. Multilocus heterozygosity showed a significant positive correlation with both leaf and panicle yield in experimental conditions, but not with original plant size, which was found to be markedly influenced by environmental microheterogeneity. Multilocus heterozygosity, estimated from both the number of heterozygous loci and the number of distinct alleles per locus, had a significant influence on plant performance. Individual locus effects were positive and significant at two loci (GOT1 and PX1). Panicle number increased regularly with heterozygosity level (from monogenics to tetragenics) at the GOT1 locus, as did leaf weight and panicle number at the PX1 locus. Such variation would be predicted by overdominance at these loci or at linked loci. Significant relationships between leaf yield and heterozygosity level at the GOT1 locus distinguished the homozygotes from the heterozygotes (of any class) and was thus more consistent with inbreeding effects. No significant differences were observed among the five genotype classes for any quantitative trait at the six remaining loci. At both the GOT1 and PX1 loci, heterozygosity had a significant independent effect on leaf weight and panicle number even when the correlation between these traits was removed by analysis of covariance.     

Prey selection by Dicyphus hesperus of infected or parasitized greenhouse whitefly

The development of effective biological control programs in which predators are integrated with other natural enemies such as parasitoids and entomopathogens requires an understanding of their interactions. In this study we examined the extent to which the omnivorous mirid bug Dicyphus hesperus, an effective biological control agent of greenhouse whiteflies, accepts prey that are either parasitized by the specialist whitefly parasitoid, Encarsia formosa or infected by the generalist fungal entomopathogen, Beauveria bassiana. In non-choice laboratory experiments, we measured how parasitism and infection of greenhouse whitefly, Trialeurodes vaporariorum, as related to parasitoid age and course of fungal infection, might alter the probability of feeding by second instar or adult female predators. Predation by D. hesperus was similar for parasitized and unparasitized whiteflies, regardless of parasitoid age. However, predators generally avoided feeding on infected whiteflies, particularly when infection was manifested through the presence of oosporein or hyphae on the surface of prey.