Resource-niche complementarity and autotrophic compensation determines ecosystem-level responses to increased cladoceran species richness

This study examines the relationship between cladoceran species richness and ecosystem functioning. I conducted an experiment in which four cladocerans, Daphnia. magna, D. longispina, D. pulex and Chydorus sphaericus, were cultured in microcosms using different species combinations and levels of species richness. The results demonstrate that even within this closely related group of organisms the effects on ecosystem-level variables, such as total algae and zooplankton biomass, per capita productivity, and nutrient concentrations, as well as phytoplankton community structure, were highly variable between different combinations of these species. Since only four species where involved in this study, species-specific effects dominated the general relationship between species richness and ecosystem functioning. Particular combinations of species resulted in effects that indicated more efficient grazing. These effects, which were most pronounced in combinations including both D. magna and C. sphaericus, were manifested as an indirect effect as the prey community shifted towards grazing-resistant species. As a result, the productivity of the prey community decreased, because phytoplankton species with lower per capita productivity became more dominant. I suggest that the primary mechanism that caused this significant effect was complementarity in prey-size use of D. magna and C. sphaericus. In terms of prey-size range, D. pulex and D. longispina were redundant when D. magna was present and were quickly out-competed by the latter despite higher per capita filtering efficiency. The results show that different mechanisms are important for different combinations of species. Furthermore, the ability of the prey community to respond to changes of consumer species composition is an important factor in experiments in which consumer species richness is experimentally manipulated. Received: 2 November 1998 / Accepted: 8 September 1999     

Impacts of tree species diversity on litter decomposition in northern temperate forests of Wisconsin, USA: a multi-site experiment along a latitudinal gradient

Over the past decade an increasing amount of research has sought to understand how the diversity of species in an ecosystem can influence fluxes of biologically important materials, such as the decomposition of organic matter and recycling of nutrients. Generalities among studies have remained elusive, perhaps because experimental manipulations have been performed at relatively small spatial scales where site-specific variation generates patterns that appear idiosyncratic. One approach for seeking generality is to perform parallel experiments at different sites using an identical species pool. Here we report results from a study where we manipulated the diversity of leaf litter from the same six dominant tree species in the litter layer of three forested ecosystems. These ecosystems spanned a 300 km latitudinal transect in Wisconsin, USA, and were characterized by a large gradient in temperature and moisture, and thus, rates of decomposition. After allowing combinations of one, two, four, and six species of leaf litter to decompose for 1 year, we found that increasing leaf litter richness led to slower rates of decomposition and higher fractions of nitrogen lost from litter. Across all sites, climate and initial litter chemistry explained more of the variation in decomposition rates than did litter richness. Effects of leaf litter diversity were non-additive, meaning they were greater than expected from the impacts of individual species, and appeared to be strongly influenced by the presence/absence of just 1–2 species (Tilia americana and Acer saccharum). The rate of decomposition of these two species was highly site-specific, which led to strong negative effects of litter richness only being observed at the southernmost sites where T. americana and A. saccharum decomposed more quickly. In contrast, litter diversity increased nitrogen loss at the northernmost sites where decomposition of T. americana was notably slowed. Our study shows that species diversity affected at least one of the two litter processes at each site along this 300-km gradient, but the exact nature of these effects were spatially variable because the performance of individual species changed across the heterogeneous landscape.     

Secondary succession in abandoned fields of dry tropical Northern Cameroon

Abstract. This study evaluates the processes of recolonization of abandoned fields by native vegetation under conditions of intensive human activity (fire, intensive grazing, firewood cutting) in a semi-arid tropical region savanna of northern Cameroon. Secondary plant succession was studied in two series of formerly cultivated fields 1–35 yr after the beginning of the fallow period. Floristic changes and the dynamics of woody plant populations were compared between areas with vertisols (clay texture) and sandy soils, as a function of length of fallows. Vegetation changed continuously during the 35 years following field abandonment. However, a very abrupt break occurred between 6 and 10 yr, due to increasingly intense human pressure during this period. Up to that point, ecological models and mechanisms of succession presented in the literature are more or less confirmed by our results. Usually, secondary succession is blocked at a stage of wooded grassland as a result of human activities.     

Resistance to rust fungi in Lolium perenne depends on within-species variation and performance of the host species in grasslands of different plant diversity

The hypothesis that plant species diversity and genetic variation of the host species decrease the severity of plant diseases is supported by studies of agricultural systems, but experimental evidence from more complex systems is scarce. In an experiment with grassland communities of varying species richness (1, 2, 4, 8, 16, and 60 species) and functional group richness (1, 2, 3, and 4 functional groups), we used different cultivars of Lolium perenne (perennial ryegrass) to study effects of biodiversity and cultivar identity on the occurrence and severity of foliar fungal diseases caused by Puccinia coronata (crown rust) and P. graminis (stem rust). Cultivar monocultures of perennial ryegrass revealed strong differences in pathogen susceptibility among these cultivars. Disease intensity caused by both rust fungi decreased significantly with growing species richness of species mixtures. The response to the diversity gradient was related to the decreased density and size of the host individuals with increasing species richness. The occurrence of other grass species known to be possible hosts of the pathogens in the experimental mixtures did not promote disease intensity in L. perenne, indicating that there was a high host specificity of pathogen strains. Differences in pathogen susceptibility among perennial ryegrass cultivars persisted independent of diversity treatment, host density and host individual size, but resulted in a cultivar-specific pattern of changes in pathogen infestation across the species-richness gradient. Our study provided evidence that within-species variation in pathogen susceptibility and competitive interactions of the host species with the environment, as caused by species diversity treatments, are key determinants of the occurrence and severity of fungal diseases. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Habitat selection as a hierarchical spatial process: the green woodpecker at the northern edge of its distribution range

Habitat selection can be envisaged as a hierarchical spatial process, from choice of home range to choice of dietary item. The green woodpecker (Picus viridis) is described as being closely bound to cultivated land and deciduous forests, mainly due to its summer diet composed of ants (Hymenoptera: Formicidae) found on meadows and pastures. To explore possible responses of this woodpecker to recent changes in land use practice, we studied home ranges, feeding habitats and food selection of a marginal population (four radio-marked males and five females) in a 30,000-ha conifer-dominated landscape at the northern edge of its distribution range in south-central Scandinavia. We asked: (1) Is the green woodpecker confined to areas with cultivated land and deciduous forest? (2) If so, are important food items (ants) particularly abundant or exclusively found there? (3) Can clearcuts and young plantations substitute for cultivated land as feeding habitat? Home ranges (mean=100 ha) were invariably confined to the parts of the landscape that contained cultivated land (<1% of the total area). In summer, birds preferred to feed in cultivated land, presumably due to a higher overall biomass of ants compared to forest habitats. They avoided clearcuts, but preyed extensively upon soil-dwelling ants in young conifer stands (16–30 years old). We failed to find preferences for particular ant groups (Lasius niger and L. flavus) associated with cultivated land. The principal summer food was Serviformica, an ant group that was equally abundant in cultivated land and forest habitat. A positive correlation between ant body mass and a preference index suggests that the birds selected the larger ant species independent of habitat type. In winter, birds fed exclusively on mound-building Formica rufa-ants in closed-canopy, older forest stands. Our results indicate that the green woodpecker successfully utilizes young conifer plantations as feeding habitat. At a larger scale, we hypothesize that green woodpecker populations fail to establish in managed forest tracts, not because of food shortage, but because the landscapes lack cultivated land serving as a key stimulus encouraging individuals to settle.     

Crop-weed interactions in the Beta vulgaris complex at a local scale: allelic diversity and gene flow within sugar beet fields

Crop-wild hybrids and weed beets are the main source of agronomic concern for sugar beet production all over Europe. In order to understand the dynamics of crop-wild interactions and the evolution of weediness in Beta vulgaris, we investigated genetic features of bolting individuals occurring at a local scale, i.e. within two sugar beet fields of the French northern area of sugar beet production. By analysing ploidy level, mitochondrial DNA and microsatellite polymorphism, the genetic diversity and the genetic relationships among three different classes of individuals (variety, in-row and out-row weed-beets) from a given field were examined. Such genetic analyses provide a unique opportunity to obtain evidence for the weeds origin and the evolutionary hypotheses previously stated. All the individuals shared in common the Svulg mitochondrial haplotype, and thus a common maternal origin. Conversely, the large genetic diversity at microsatellite loci highlighted the large diversity of the pollinator plants (cultivated and wild plants) during the-seed production process, as well as during the further evolution of weed beets in the sugar production area. Received: 23 April 2001 / Accepted: 15 June 2001     

A novel comparative research platform designed to determine the functional significance of tree species diversity in European forests

One of the current advances in functional biodiversity research is the move away from short-lived test systems towards the exploration of diversity-ecosystem functioning relationships in structurally more complex ecosystems. In forests, assumptions about the functional significance of tree species diversity have only recently produced a new generation of research on ecosystem processes and services. Novel experimental designs have now replaced traditional forestry trials, but these comparatively young experimental plots suffer from specific difficulties that are mainly related to the tree size and longevity. Tree species diversity experiments therefore need to be complemented with comparative observational studies in existing forests. Here we present the design and implementation of a new network of forest plots along tree species diversity gradients in six major European forest types: the FunDivEUROPE Exploratory Platform. Based on a review of the deficiencies of existing observational approaches and of unresolved research questions and hypotheses, we discuss the fundamental criteria that shaped the design of our platform. Key features include the extent of the species diversity gradient with mixtures up to five species, strict avoidance of a dilution gradient, special attention to community evenness and minimal covariation with other environmental factors. The new European research platform permits the most comprehensive assessment of tree species diversity effects on forest ecosystem functioning to date since it offers a common set of research plots to groups of researchers from very different disciplines and uses the same methodological approach in contrasting forest types along an extensive environmental gradient.     

Improving the assessment of species compositional dissimilarity in a priori ecological classifications: evaluating map scale,sampling intensity and improvement in a hierarchical classification

Question: Can species compositional dissimilarity analyses be used to assess and improve the representation of biodiversity patterns in a priori ecological classifications? Location: The case study examined the northern‐half of the South‐east Queensland Bioregion, eastern Australia. Methods: Site‐based floristic presence–absence data were used to construct species dissimilarity matrices (Kulczynski metric) for three levels of Queensland's bioregional hierarchy – subregions (1:500 000 scale), land zones (1:250 000 scale) and regional ecosystems (1:100 000 scale). Within‐ and between‐class dissimilarities were compiled for each level to elucidate species compositional patterns. Randomized subsampling was used to determine the minimum site sampling intensity for each hierarchy level, and the effects of lumping and splitting illustrated for several classes. Results: Consistent dissimilarity estimates were obtained with five or more sites per regional ecosystem, 10 or more sites per land zone, and more than 15 sites per subregion. On average, subregions represented 4% dissimilarity in floristic composition, land zones approximately 10%, and regional ecosystems over 19%. Splitting classes with a low dissimilarity increased dissimilarity levels closer to average, while merging ecologically similar classes with high dissimilarities reduced dissimilarity levels closer to average levels. Conclusions: This approach demonstrates a robust and repeatable means of analysing species compositional dissimilarity, determining site sampling requirements for classifications and guiding decisions about ‘lumping’ or ‘splitting’ of classes. This will allow more informed decisions on selecting and improving classifications and map scales in an ecologically and statistically robust manner.     

The mycorrhizal colonization of six wetland plant species at sites differing in land use history

 Five wetland prairie sites and six native plant species in western Oregon were examined to determine patterns of vesicular-arbuscular mycorrhizal fungal (VAMF) colonization. The sites differed in type and intensity of past land use. VAMF colonization was tested in situ on seedlings from both field-sown seeds and from transplants. Colonization was measured as the percentage of root length with arbuscles or vesicles. All species (Deschampsia cespitosa, Downingia elegans, Eriophyllum lanatum, Hordeum brachyantherum, Microseris laciniata, and Plagiobothrys figuratus) became colonized by VAMF during the study. This is the first report of mycorrhizal colonization of these important native species. All sites supported mycorrhizal colonization of some of the experimental species. Average VAMF colonization ranged from 58% to 92% but was unrelated to subjective rankings of land use intensity. These results suggest that VAMF inoculum at all sites was sufficient to support revegetation by at least some species of native plants. Accepted: 16 September 1999     

Genetic diversity and classification of cyanobacteria in different Azolla species by the use of PCR fingerprinting

Symbiotically associated cyanobacteria from 18 accessions within all known species in the genus Azolla were examined and classified by the use of polymerase chain reaction (PCR)-fingerprinting. A repetitive sequence specific for cyanobacteria, the short tandemly repeated repetitive (STRR) sequence, was used as a primer in the reaction. Cyanobacterial filaments isolated directly from the Azolla leaf cavity or contained within homogenised symbiotic Azolla tissue were used as templates. Based on the fingerprint pattern, distinct differences were demonstrated between cyanobacteria isolated from the Euazolla and Rhizosperma sections. In addition, individual fingerprints were obtained from all cyanobacteria isolated from the different Azolla species. The fingerprints were used to generate a phylogenetic tree. Three clusters were distinguished: one contained the four isolates from the section Euazolla, a second the isolate from Azolla filiculoides, and a third the three isolates from the section Rhizosperma. By the use of STRR-PCR fingerprinting, new data on the taxonomy of cyanobacteria in Azolla were obtained, which have been difficult to generate by other classification methods. PCR-fingerprinting may, therefore, be a valuable tool for diversity and classification studies of symbiotic cyanobateria from Azolla and, as co-evolution between the cyanobacteria and its corresponding host exists the method may also be useful for the taxonomy of Azolla. Received: 19 December 1998 / Accepted: 31 May 1999     

Use of microsatellites to evaluate genetic diversity and species relationships in the genus Lycopersicon

In order to determine how informative a set of microsatellites from tomato is across the genus Lycopersicon, 17 microsatellite loci, derived from regions in and around genes, were tested on 31 accessions comprising the nine species of the genus. The microsatellite polymorphisms were used to estimate the distribution of diversity throughout the genus and to evaluate the efficacy of microsatellites for establishing species relationships in comparison with existing phylogeny reconstructions. Gene diversity and genetic distances were calculated. A high level of polymorphism was found, as well as a large number of alleles unique for species. The level of polymorphism detected with the microsatellite loci within and among species was highly correlated with the respective mating systems, cross-pollinating species having a significantly higher gene diversity compared to self-pollinating species. In general, microsatellite-based trees were consistent with a published RFLP-based dendrogram as well as with a published classification based on morphology and the mating system. A tree constructed with low-polymorphic loci (gene diversity <0.245) was shown to represent a more-reliable topology than a tree constructed with more-highly polymorphic loci. Received: 19 February 2001 / Accepted: 26 March 2001     

Plant species diversity in abandoned coppice forests in a temperate deciduous forest area of central Japan

We investigated plant species diversity as it related to stand structure and landscape parameters in abandoned coppice forests in a temperate, deciduous forest area of central Japan, where Fagus crenata was originally dominant. The species occurring in the study plots were classified into habitat types based on a statistical analysis of their occurrence bias in particular habitats (e.g., primary forest, coniferous plantation) in the landscape studied. The relationships between stand structure, which reflected the gradient of management, and forest floor plant species diversity (H and J) and richness (number of species per unit area) were not significant. However, these factors did influence the forest floor plant composition of the different types of habitat. According to the multiple regression analysis, species diversity and the richness of forest floor plants was affected by landscape parameters rather than by stand structure. For trees, species richness was mainly affected by the relative dominance of F. crenata, which is one of the stand structure parameters that decreases with intensive management. This is probably because many of the tree species that are characteristic of coppice forests increase after F. crenata have been eliminated by management; these species are not dominant in the original forest, where they are suppressed by F. crenata, the shade-tolerant dominant species. The species diversity (H and J) of trees was positively correlated with some landscape parameters, including the road density around the study plot, which may be associated with the intensity of management activity. The number of disturbance-tolerant species increased with increasing road density. Stand structure mainly affected disturbance-intolerant forest floor plant species and disturbance-tolerant tree species. Thus, the species diversity responses differed between forest floor plants and trees. The impact of forest management on species diversity was more prominent for forest floor plants.     

Ants on swollen-thorn acacias: species coexistence in a simple system

 On the black cotton soils of the Laikipia ecosystem in Kenya, two swollen-thorn acacia species support nine ant species, four of which are apparently obligate plant-ants. Among the ants, there are five species of Crematogaster, two species of Camponotus, and one each of Tetraponera and Lepisota. Acacia drepanolobium is host to four ant species that are both common and mutually exclusive. These four ant species, and an additional non-exclusive ant species, tend to occur on trees of different sizes, implying a succession of ant occupants. Nonetheless, all four exclusive species occur in substantial proportions on trees of intermediate size. There is direct evidence that an early successional ant species (Tetraponera penzigi) is actively evicted by two late successional ant species in the genus Crematogaster. There was also some evidence of height differentiation among ant species resident on A. seyal. Different acacia-ant species had different direct effects on A. drepanolobium. Extrafloral nectaries were eaten and destroyed only on trees inhabited by Tetraponera. Axillary shoots were eaten only on trees inhabited by C. nigriceps (potentially another early successional ant). This was associated with more new terminal shoots and healthier leaves than other trees, but also the virtual elimination of flowering and fruiting. Different resident acacia-ant species also had characteristic relationships with other insects. Among the four mutually exclusive ant species, only Crematogaster sjostedti was associated with two species of Camponotus, at least one of which (C. rufoglaucus) appears to be a foraging non-resident. A. drepanolobium trees occupied by C. sjostedti were also far more heavily infested with leaf galls than were trees occupied by other ant species. A. drepanolobium trees occupied by C. mimosae and C. sjostedti uniquely had tended adult scale insects. This diversity of ant inhabitants, and their strikingly different relationships with their hosts and other insect species, are examples of coexisting diversity on an apparently uniform resource. Received: 13 November 1995 / Accepted: 16 May 1996     

Distribution of genetic diversity in Pinus pinaster Ait. as revealed by chloroplast microsatellites

 Using nine chloroplast simple sequence.repeats (cpSSRs) markers, we evaluated haplotypic variation within and among natural populations of Maritime pine (Pinus pinaster Ait.) in order to shed light on the history of this species. Seven out of the nine cpSSRs analysed were polymorphic, giving a total of 24 different variants. The 24 variants combined in 34 different haplotypes. The populations which generally showed the lowest level of haplotypic diversity are those located in Portugal. The Landes (France) and Pantelleria (Italy) populations represent the two main reservoirs of haplotypic diversity. The proportion of genetic differentiation among populations, estimated using R_st, which is a measure based upon a strict stepwise mutation model, was 0.235. The high level of differentiation was also confirmed by the AMOVA analysis (Φ_ST=0.254, P<0.001). Four main groups of populations were identified on the basis of Principal Component Analysis, with the differences being statistically significant (Φ_CT=0.299, P<0.001). Based on our results the presence of refugia located in the South of Portugal, previously proposed for this species, may be excluded, and a different possible recolonization process of Maritime pine in the post-glacial period has been proposed. Populations from North Africa and France might have represented a starting point of the recolonization process of Portugal and of the Italian part of the natural range, respectively. This hypothesis seems to be confirmed by the analysis of the distribution of the pairwise differences among individuals within populations: Landes and Pantelleria populations showed a bimodal distribution, as would be expected for ancient gene pools. Received: 5 November 1997 / Accepted: 5 January 1998     

Refuge use by fish as a function of body weight changes

Refuge use provides a good model for the study of trade-offs between the benefits of predator avoidance and the costs of lost feeding opportunities. We manipulated the latter costs by subjecting similar-sized three-spine sticklebacks to 2 days of food deprivation followed by a 2-day re-feeding period and recorded associated changes in body weight and refuge use. Food deprivation resulted in a decrease and re-feeding in an increase in the duration of refuge use by fish. Emergence times of fish from the refuge were extremely variable (with a ratio of 1:127 between the shortest and the longest ones) but individual ranks were highly consistent between different days of testing, suggesting that emergence times were individually characteristic. Percentage weight change of fish in response to the experimental treatments also showed a high level of inter-individual variation ranging from 0–17%. A significant positive correlation was found between the percentage weight lost and the percentage decrease in emergence time from a refuge after food deprivation and similarly between the percentage weight gained and the percentage increase in refuge use after re-feeding. The relationship between energy turnover and behavioural strategies is discussed. Received: 13 December 1998 / Received in revised form: 2 May 1999 / Accepted: 7 June 1999     

Limb development in a primitive crustacean,Triops longicaudatus: subdivision of the early limb bud gives rise to multibranched limbs

 Recent advances in developmental genetics of Drosophila have uncovered some of the key molecules involved in the positioning and outgrowth of the leg primordia. Although expression patterns of these molecules have been analyzed in several arthropod species, broad comparisons of mechanisms of limb development among arthropods remain somewhat speculative since no detailed studies of limb development exist for crustaceans, the postulated sister group of insects. As a basis for such comparisons, we analysed limb development in a primitive branchiopod crustacean, Triops longicaudatus. Adults have a series of similar limbs with eight branches or lobes that project from the main shaft. Phalloidin staining of developing limbs buds shows the distal epithelial ridge of the early limb bud exhibits eight folds that extend in a dorsal ventral (D/V) arc across the body. These initial folds subsequently form the eight lobes of the adult limb. This study demonstrates that, in a primitive crustacean, branched limbs do not arise via sequential splitting. Current models of limb development based on Drosophila do not provide a mechanism for establishing eight branches along the D/V axis of a segment. Although the events that position limbs on a body segment appear to be conserved between insects and crustaceans, mechanisms of limb branching may not. Received: 28 February 1996/Accepted: 24 June 1996     

Breakdown of self-incompatibility in a natural population of Petunia axillaris (Solanaceae) in Uruguay containing both self-incompatible and self-compatible plants

 Many members of the Solanaceae display a type of gametophytic self-incompatibility which is controlled by a single multiallelic locus, called the S-locus. From our previous survey of more than 100 natural populations of Petunia axillaris (a solanaceous species) in Uruguay, we had found that the majority of the populations of subspecies axillaris were comprised of virtually all self-incompatible individuals. The rest were ”mixed populations” which contained mostly self-incompatible and some self-compatible individuals. In this study, we examined the self-incompatibility behavior and determined the S-genotypes of 33 plants raised from seeds obtained from one such mixed population, designated U1. We found that 30 of the 33 plants (designated U1–1 through U1–33) were self-incompatible and a total of 18 different S-alleles were represented. To determine the S-genotypes of the three self-compatible plants (U1–2, U1–16, and U1–22) and the possible causes for the breakdown of their self-incompatibility, we carried out reciprocal crosses between each of them and each of the 18 S-homozygotes (S ₁ S ₁ through S ₁₈ S ₁₈ ) obtained from bud-selfed progeny of 14 of the 30 self-incompatible plants. For U1–2 and U1–16, we also carried out additional crosses with U1–25 (with S ₁ S ₁₃ genotype) and an S ₁₃ S ₁₅ plant (obtained from a cross between an S ₁₃ -homozygote and an S ₁₅ -homozygote), respectively. Based on all the pollination results and analysis of the production of S-RNases, products of S-alleles in the pistil, we determined the S-genotypes of U1–2, U1–16, and U1–22, and propose that the breakdown of self-incompatibility in these three plants is caused by suppression of the production of S₁₃-RNase from the S ₁₃ -allele they all carry. We have termed this phenomenon ”stylar-part suppression of an S-allele” or SPS. Received: 25 September 1998 / Revision accepted: 22 December 1998     

Relation of water transport to leaf gas exchange properties in three mangrove species

Mangrove species more tolerant to salinity may function with less efficient water transport, which may be related to more conservative water use. To test the hypothesis, we investigate the gas exchange and hydraulic properties of three mangrove species: Rhizophora mangle L., Laguncularia racemosa Gaert and Avicennia germinans (L.)L. Experiments were performed with adult plants growing naturally in the field under a salinity of 35‰. Gas exchange parameters showed that A. germinans had significantly higher photosynthetic rates, and lower stomatal conductance and transpiration rates, compared to the other two mangroves. In concert with this, instantaneous water use efficiency was significantly high in A. germinans, intermediate in L. racemosa and lowest in R. mangle. The hydraulic parameters of the three mangrove species were in the lowest end of the range reported for tropical trees. However, the three mangrove species exhibited measurable differences in hydraulic parameters related to the control of water requirements for maintenance of carbon gain. L. racemosa and A. germinans showed less efficient water transport at shoot level but were the more efficient species in water use at the leaf level in comparison to R. mangle. Received: 7 April 1999 / Accepted: 25 July 1999     

Evolution of a new nonclassical MHC class I locus in two Old World primate species

 HLA-G is a nonclassical major histocompatibility complex (MHC) class I molecule that is expressed only in the human placenta, suggesting that it plays an important role at the fetal-maternal interface. In rhesus monkeys, which have similar placentation to humans, the HLA-G orthologue is a pseudogene. However, rhesus monkeys express a novel placental MHC class I molecule, Mamu-AG, which has HLA-G-like characteristics. Phylogenetic analysis of AG alleles in two Old World primate species, the baboon and the rhesus macaque, revealed limited diversity characteristic of a nonclassical MHC class I locus. Gene trees constructed using classical and nonclassical primate MHC class I alleles demonstrated that the AG locus was most closely related to the classical A locus. Interestingly, gene tree analyses suggested that the AG alleles were most closely related to a subset of A alleles which are the products of an ancestral interlocus recombination event between the A and B loci. Calculation of the rates of synonymous and nonsynonymous substitution at the AG locus revealed that positive selection was not acting on the codons encoding the peptide binding region. In exon 4, however, the rate of nonsynonymous substitution was significantly lower than the rate of synonymous substitution, suggesting that negative selection was acting on these codons. Received: 22 April 1998 / Revised: 15 July 1998     

Sequencing of the internal transcribed spacer region ITS1 as a molecular tool detecting variation in the Stylosanthes guianensis species complex

 The internal transcribed spacer (ITS) regions 1 and 2 of the ribosomal DNA from Stylosanthes guianensis CIAT 1283 and cv ‘Schofield’ were amplified by polymerase chain reaction using conserved ITS primers from the 18S, 5.8S and 26S ribosomal genes flanking those regions. The entire region of 683 bp long was cloned, and seven clones were sequenced. Comparison of the ITS spacer regions with published DNA sequences of other plant species revealed limited homology only; this was in contrast to their comparison with the 5.8S rDNA sequences. The ITS1 region of 45 S. guianensis accessions was amplified by PCR and sequenced on both strands using the conserved primers ITS2-ITS5. These sequences, ranging from 201 to 204 bp, were aligned to each other to assess intra-specific polymorphism. Within the S. guianensis (Aubl.) Sw. species complex, 11 DNA sequence types could be distinguished based on an insertion/deletion (indel) event and 15 single base-pair substitutions. In 1 of the S. guianensis types, two kinds of ITS1 sequence were observed in each individual, reminiscent of an incomplete homogenization of the repeat structure in this type. Polymorphisms in the sequence of the ITS1 region were used to define molecular markers for S. guianensis on the basis of PCR-restriction fragment length polymorphism and selective PCR. Received: 24 June 1997 / Accepted: 31 October 1997