Damien M. de Vienne

Speciation in fungi.

In this review on fungal speciation, we first contrast the issues of species definition and species criteria and show that by distinguishing the two concepts the approaches to studying the speciation can be clarified. We then review recent developments in the understanding of modes of speciation in fungi. Allopatric speciation raises no theoretical problem and numerous fungal examples exist from nature. We explain the theoretical difficulties raised by sympatric speciation, review the most recent models, and provide some natural examples consistent with speciation in sympatry. We describe the nature of prezygotic and postzygotic reproductive isolation in fungi and examine their evolution as functions of temporal and of the geographical distributions. We then review the theory and evidence for roles of cospeciation, host shifts, hybridization, karyotypic rearrangement, and epigenetic mechanisms in fungal speciation. Finally, we review the available data on the genetics of speciation in fungi and address the issue of speciation in asexual species.

Restriction fragment length polymorphism analysis of loci associated with disease resistance genes and developmental traits in Pisum sativum L.

An F2 population of pea (Pisum sativum L.) consisting of 174 plants was analysed by restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) techniques. Ascochyta pisi race C resistance, plant height, flowering earliness and number of nodes were measured in order to map the genes responsible for their variation. We have constructed a partial linkage map including 3 morphological character genes, 4 disease resistance genes, 56 RFLP loci, 4 microsatellite loci and 2 RAPD loci. Molecular markers linked to each resistance gene were found: Fusarium wilt (6 cM from Fw), powdery mildew (11 cM from er) and pea common Mosaic virus (15 cM from mo). QTLs (quantitative traits loci) for Ascochyta pisi race C resistance were mapped, with most of the variation explained by only three chromosomal regions. The QTL with the largest effect, on chromosome 4, was also mapped using a qualitative, Mendelian approach. Another QTL displayed a transgressive segregation, i.e. the parental line that was susceptible to Ascochyta blight had a resistance allele at this QTL. Analysis of correlations between developmental traits in terms of QTL effects and positions suggested a common genetic control of the number of nodes and earliness, and a loose relationship between these traits and height.

A congruence index for testing topological similarity between trees

MOTIVATION Phylogenetic trees are omnipresent in evolutionary biology and the comparison of trees plays a central role there. Tree congruence statistics are based on the null hypothesis that two given trees are not more congruent (topologically similar) than expected by chance. Usually, one searches for the most parsimonious evolutionary scenario relating two trees and then one tests the null hypothesis by generating a high number of random trees and comparing these to the one between the observed trees. However, this approach requires a lot of computational work (human and machine) and the results depend on the evolutionary assumptions made. RESULTS We propose an index, I(cong), for testing the topological congruence between trees with any number of leaves, based on maximum agreement subtrees (MAST). This index is straightforward, simple to use, does not rely on parametrizing the likelihood of evolutionary events, and provides an associated confidence level. AVAILABILITY A web site has been created that allows rapid and easy online computation of this index and of the associated P-value at http://www.ese.u-psud.fr/bases/upresa/pages/devienne/index.html

Genetic dissection of the relationship between carbon metabolism and early growth in maize, with emphasis on key-enzyme loci

The determinism of carbon metabolism traits during early growth in maize has been investigated using a marker-based quantitative genetics approach. In addition to growth traits, concentration of carbohydrates and activity of four key enzymes of their metabolism (sucrose phosphate synthase, ADP-glucose pyrophosphorylase, invertases and sucrose synthase) have been measured in leaves of individuals of a recombinant inbred line population. Using more than 100 RFLP markers, quantitative trait loci (QTLs) were mapped for each biochemical and developmental trait. Causal relationships, suggested by previous physiological studies, were reinforced by common locations of QTLs for different traits. Thus, the strong correlation between growth rate and invertase activity, which may reflect sink organ strength, could be explained to a large extent by a single region of chromosome 8. Moreover, some of the structural genes of the enzymes mapped to regions with QTLs affecting the activity of the encoded enzyme and/or concentration of its product, and sometimes growth traits. These results emphasize the possible role of the polymorphism of key-enzyme genes in physiological processes, and hence in maize growth.

Fungal evolutionary genomics provides insight into the mechanisms of adaptive divergence in eukaryotes

Fungi are ideal model organisms for dissecting the genomic bases of adaptive divergence in eukaryotes. They have simple morphologies and small genomes, occupy contrasting, well‐identified ecological niches and tend to have short generation times, and many are amenable to experimental approaches. Fungi also display diverse lifestyles, from saprotrophs to pathogens or mutualists, and they play extremely important roles in both ecosystems and human activities, as wood decayers, mycorrhizal fungi, lichens, endophytes, plant and animal pathogens, and in fermentation or drug production. We review here recent insights into the patterns and mechanisms of adaptive divergence in fungi, including sources of divergence, genomic variation and, ultimately, speciation. We outline the various ecological sources of divergent selection and genomic changes, showing that gene loss and changes in gene expression and in genomic architecture are important adaptation processes, in addition to the more widely recognized processes of amino acid substitution and gene duplication. We also review recent findings regarding the interspecific acquisition of genomic variation and suggesting an important role for introgression, hybridization and horizontal gene transfers (HGTs). We show that transposable elements can mediate several of these genomic changes, thus constituting important factors for adaptation. Finally, we review the consequences of divergent selection in terms of speciation, arguing that genetic incompatibilities may not be as widespread as generally thought and that pleiotropy between adaptation and reproductive isolation is an important route of speciation in fungal pathogens.

Association of protein amount polymorphism (PAP) among maize lines with performances of their hybrids

SummaryIt has been suggested that molecular foundations of phenotypic diversity reside in the variability of genome expression. This variability can be appraised through the polymorphism of individual protein amounts (PAP: protein amount polymorphism). Eight maize inbred lines and ten of their single-cross hybrids were analyzed by two-dimensional polyacrylamide gel electrophoresis in order to examine the potential of PAP for predicting hybrid vigor. The 28 possible pairs of lines were characterized for: (i) the number, H of expected heterozygous structural loci in their hybrid, in the sample of loci revealed by 2D-PAGE; (ii) four distance indices based on PAP; (iii) the hybrid values for five agromorphological characters measured in four different year/locations. For the subset of ten hybrids analyzed by 2D-PAGE, the number of cases of nonadditive inheritance (NA) was also counted. Whereas H appeared to be related neither to the PAP indices, nor to NA, nor to hybrid performances, PAP indices were correlated to NA, and both were positively associated to hybrid performances. The possibility that PAP is responsible for quantitative trait variation is discussed. This could result in the definition of biological predictors of heterosis.

Phylogenetic determinants of potential host shifts in fungal pathogens

Understanding what determines the host range of pathogens and the potential for host shifts is of critical importance to controlling their introductions into new environments. The phylogeny of the hosts has been shown to be important: pathogens are more likely to be infectious on hosts closely related to their host‐of‐origin because of the similar host environments that is shared by descent. The importance of pathogen phylogenies for predicting host range has never been investigated, although a pathogen should also be able to exploit a new host that its close relative can infect. We performed cross‐inoculations using a plant–fungal association and showed that both host and pathogen phylogenies were significant predictors of host range, with at least partly independent effects. Furthermore, we showed that some pathogens were better at infecting novel hosts. Our results should have implications in the context of biological invasions and emergences of new diseases due to globalization.

Quantification of dominance for proteins pleiotropically affected by opaque-2 in maize

The maize opaque-2 (o2) mutation is known to have numerous pleiotropic effects. Using two different genetic backgrounds, endosperm proteins from isogenic lines of the o2 locus and their reciprocal hybrids were compared by high-resolution two-dimensional electrophoresis (2-D PAGE), and the 2-D gels stained with Coomassie blue or silver were analysed using a computer-assisted system for quantification of polypeptide spots. The aim of this work was (i) to identify polypeptides, the amount of which is affected by the single gene substitution, and (ii) to assess the dominance relations for every affected polypeptide. The triploid nature of the endosperm allowed the effects of 0, 1, 2 and 3 doses of the wild-type O2 allele to be examined. When using a basic pH range in the first dimension (IEF), we observed the known effects of o2 on prolamin storage proteins, mostly being 22 kDa, zeins. In a pH 5-7 IEF range, we detected 14 polypeptides whose amount is affected by the o2 mutation in both backgrounds. Eight of them were more abundant in the wild type than in the mutant and six were more abundant in the mutant than in the wild type. Dominance of the wild type allele was found for the amounts of 10 polypeptides, while the amounts of two of them displayed additive inheritance. This is an example where pleiotropically related characters display different inheritance, which could generate heterozygous phenotypes with original properties as compared to their homozygous parents. Because O2 belongs to the basic region-leucine zipper family of transcription activators, our results show that 2-D PAGE of isogenic lines represents a straightforward method to identify gene products affected by a transcription factor, whether they are products of its target genes, or products indirectly affected by far-reaching effects. This is a prerequisite to understanding the multiple molecular and physiological consequences of the action of such factors.

Is the polymorphism of protein amounts related to phenotypic variability? A comparison of two-dimensional electrophoresis data with morphological traits in maize

SummaryThe hypothesis that the quantitative variations in gene product levels could be a more important basis for morphological and adaptative change than the classical qualitative variability revealed by electrophoretic techniques was studied by comparing five maize lines from three sets of variables: (i) qualitative variations of proteins (presence/absence) revealed by two-dimensional polyacrylamide gel electrophoresis (2D PAGE), at a physiological seedling stage; (ii) quantitative variations in proteins (spots more or less intense) revealed by 2D PAGE, at the same physiological stage; (iii) general combining abilities of fourteen heritable, morphological or agronomical characters measured at various juvenile and adult stages. Distances between lines were defined, based on qualitative and quantitative variations of proteins. These distances do not appear to be correlated and do not give the same patterns of divergence between lines, as shown by principal coordinate analyses. Mahalanobis distances computed from the general combining abilities of the morphological characters are significantly correlated (r=0.75) to quantitative but not to qualitative distances. The comparison of the first planes of the principal coordinate analyses performed on the three kinds of distances clearly confirms this finding. Our results are discussed in connection with the possible genetic meaning of the two molecular distances and with the hypothesis that regulatory processes are primarily implicated in morphological variation.

Two-dimensional electrophoresis of proteins as a source of monogenic and codominant markers for population genetics and mapping the expressed genome

The positional polymorphism of polypeptides revealed using two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) was analysed in segregating families of four plant species, a gymnosperm (maritime pine), and three angiosperms (maize, barley and pea). All of the 170 variations scored had monogenic and codominant inheritance, making 2-D PAGE a quite abundant and cheap source of good-quality genetic markers. Genetic mapping showed that the loci involved are well distributed on the chromosomes. In particular, the construction of a composite map of the maize genome including 253 markers revealed that the protein loci are interspersed with the RFLP loci, and provide in some instances markers for chromosomal regions previously lacking molecular markers. In the context of the genome mapping projects, such markers are physiologically relevant in that they reveal loci whose transcripts are translated in the organ analysed.