Ludwig Triest

Permanent Genetic Resources added to Molecular Ecology Resources Database 1 December 2009-31 January 2010

This article documents the addition of 220 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Allanblackia floribunda, Amblyraja radiata, Bactrocera cucurbitae, Brachycaudus helichrysi, Calopogonium mucunoides, Dissodactylus primitivus, Elodea canadensis, Ephydatia fluviatilis, Galapaganus howdenae howdenae, Hoplostethus atlanticus, Ischnura elegans, Larimichthys polyactis, Opheodrys vernalis, Pelteobagrus fulvidraco, Phragmidium violaceum, Pistacia vera, and Thunnus thynnus. These loci were cross‐tested on the following species: Allanblackia gabonensis, Allanblackia stanerana, Neoceratitis cyanescens, Dacus ciliatus, Dacus demmerezi, Bactrocera zonata, Ceratitis capitata, Ceratitis rosa, Ceratits catoirii, Dacus punctatifrons, Ephydatia mülleri, Spongilla lacustris, Geodia cydonium, Axinella sp., Ischnura graellsii, Ischnura ramburii, Ischnura pumilio, Pistacia integerrima and Pistacia terebinthus.

Genetic consequences of habitat fragmentation in an agricultural landscape on the common Primula veris, and comparison with its rare congener, P. vulgaris

In Flanders (northern Belgium), the common (butlocally rare) P. veris and its rarerelative Primula vulgaris, twoself-incompatible perennials, mainly occur infragmented habitats of the intensively usedagricultural landscape. We investigated geneticvariation and structure for respectively 30 and27 allozyme loci in 24 and 41 populations ofP. veris and P. vulgaris, inrelation to species (local) abundance and topopulation size and plant density, and comparedpopulations of linear landscape elements(verges) with non verge populations. Therare P. vulgaris was geneticallydepauperate compared to the commoner P.veris. Both species showed a geographicalstructure of genetic variation and isolation bydistance. The scattered populations of P.veris from the Westhoek region (polderhabitats) were of smaller size, geneticallydepauperate and showed higher interpopulationdivergence compared to the Voeren region, wherepopulations are densely distributed andhabitats less fragmented. Verge and non vergepopulations showed similar populations sizesand levels of genetic variation. For bothspecies, small populations showed a loss ofgenetic variation, but still maintained highlevels of observed heterozygosity.Within-population plant density negativelyaffected allelic richness in P. veris.Ours results indicate that common species canalso be negatively affected by habitatfragmentation (perhaps even more so than rarespecies) and that verge populations can have ahigh potential conservation value. Both vergeand non-verge populations should be included inconservation efforts.

atetra, a new software program to analyse tetraploid microsatellite data: comparison with tetra and tetrasat

Despite the importance of tetraploid species, most population genetic studies deal with diploid ones because of difficulties in analysing codominant microsatellite data in tetraploid species. We developed a new software program—atetra—which combines both the rigorous method of enumeration for small data sets and Monte Carlo simulations for large ones. We discuss the added value of atetra by comparing its precision, stability and calculation time for different population sizes with those obtained from previous software programs tetrasat and tetra. The influence of the number of simulations on the calculation stability is also investigated. atetra and tetrasat proved to be more precise when compared with tetra, which, however, remains faster. atetra has the same precision than tetrasat, but is much faster, can handle an infinite number of partial heterozygotes and calculates more genetic variables. The more user‐friendly interface of atetra reduces possible mistakes.

Impact of three aquatic invasive species on native plants and macroinvertebrates in temperate ponds

Biological plant invasions pose a serious threat to native biodiversity and have received much attention, especially in terrestrial habitats. In freshwater ecosystems impacts of invasive plant species are less studied. We hypothesized an impact on organisms from the water column and from the sediment. We then assessed the impact of three aquatic invasive species on the plants and macroinvertebrates: Hydrocotyle ranunculoides, Ludwigia grandiflora and Myriophyllum aquaticum. Our research on 32 ponds in Belgium indicated that the reduction in the native plant species richness was a common pattern to invasion. However, the magnitude of impacts were species specific. A strong negative relationship to invasive species cover was found, with submerged vegetation the most vulnerable to the invasion. Invertebrate richness, diversity and abundance were measured in sediments of invaded and uninvaded ponds along a gradient of H. ranunculoides, L. grandiflora, and M. aquaticum species cover. We found a strong negative relationship between invasive species cover and invertebrate abundance, probably due to unsuitable conditions of the detritus for invertebrate colonization. Taxonomic compositions of aquatic invertebrate assemblages in invaded ponds differed from uninvaded ponds. Sensitive benthos, such as mayflies were completely absent in invaded ponds. The introduction of H. ranunculoides, L. grandiflora, and M. aquaticum in Belgian ponds has caused significant ecological alterations in the aquatic vegetation and the detritus community of ponds.

Comparative monitoring of diatoms, macroinvertebrates and macrophytes in the Woluwe River (Brussels, Belgium)

The river Woluwe in Brussels and Flanders (Belgium) is a small tributary of 15 km length that drains an area of 9400 ha in the Schelde river basin. The headwaters of the Woluwe are highly fragmented by diverse pond systems and are vaulted in the Brussels agglomeration. Hyporheic zones locally influence the water quality. The downstream stretch of the river receives sewage waters from households and industry. As the river Woluwe within a short distance represents a typical gradient from groundwater-fed sources in the forest towards severely polluted water, a comparative monitoring using diatoms, macroinvertebrates and macrophytes was done. The saprobic index based on diatoms, the Belgian Biotic Index (BBI) for macroinvertebrates and a macrophyte index based on the N-values of Ellenberg were used in this comparison and for estimating the correlation with the bimonthly measured chemical variables in 16 sampling stations. The diatom saprobic index and the macrophyte index were strongly correlated. Both groups showed strong correlations with phosphate, ammonium and chemical oxygen demand. The Belgian Biotic Index showed lower correlations with the nutrient variables, but was slightly better correlated to chemical oxygen demand, chloride and dissolved oxygen. None of the indices showed a correlation with nitrate. Local substrate or light conditions could interfere with the indicator system, especially for the macrophytes and occasionally for the macroinvertebrates. It was concluded that at least in this particular river system, the indices based on the primary producers were more indicative for the trophic status, whereas the BBI showed a broader relationship to the general degree of pollution. Therefore, these three indices are considered as complementary for monitoring the biological quality and the ecological status of a river system.

Genetic diversity in adult and seedling populations of Primula vulgaris in a fragmented agricultural landscape

Habitat fragmentation is known to generally reduce the size of plant populations and increase their isolation, leading to genetic erosion and increased between-population genetic differentiation. In Flanders (northern Belgium) Primula vulgaris is very rare and declining. Populations have incurred strong fragmentation for the last decades and are now restricted to a few highly fragmented areas in an intensively used agricultural landscape. Previous studies showed that small populations of this long-lived perennial herb still maintained high levels of genetic variation and low genetic differentiation. This pattern can either indicate recent gene flow or represent historical variation. Therefore, we used polymorphic microsatellite loci to investigate genetic variation and structure in adult (which may still reflect historical variation) and seedling (recent generation, thus affected by current processes) life stages. The recent generation (seedlings) showed a significant loss of observed heterozygosity (Ho) together with lower expected heterozygosity (He), a trend for higher inbreeding levels (FIS) and higher differentiation (FST) between populations compared to the adult generation. This might result from (1) a reduction in effective population size, (2) higher inbreeding levels in the seedlings, (3) a higher survival of heterozygotes over time due to a higher fitness of heterozygotes (heterosis) and/or a lower fitness of homozygotes (inbreeding depression), (4) overlapping generations in the adult life stage, or (5) a lack of establishment of new (inbred) adults from seedlings due to degraded habitat conditions. Combining restoration of both habitat quality and gene flow between populations may be indispensable to ensure a sustainable conservation of fragmented populations.

Fine-scale genetic structure of the common Primula Elatior (Primulaceae) at an early stage of population fragmentation

Many rare species are threatened by habitat fragmentation; however, less is known about effects of fragmentation on common species, despite their potential role in ecosystem productivity and functioning. We identified key factors and processes influencing gene flow in a large population of Primula elatior, a common distylous perennial herb, at an early stage of the fragmentation process, i.e., when fragmentation is taking place. Using 19 allozyme loci, we investigated genetic variation and fine-scale spatial genetic structure (SGS) at seedling and adult life stages in relation to fragmentation history (recent bottlenecks), selection, clonal propagation, sexual reproduction (seed and pollen dispersal, distyly), and patchy structure (patch size, plant density, and morph ratio). The main factors contributing to the strong SGS are seed and (to a lesser extent) pollen dispersal, through a spatial Wahlund effect and biparental inbreeding. Significant differences in allele frequencies between seedlings and adults indicate a temporal Wahlund effect. Patch plant density and biased morph ratio also affect the genetic patterns. Our results show that if P. elatior populations evolve into patchworks of small, isolated remnants, genetic erosion, reduced gene flow, and increased inbreeding can be expected, suggesting that such common plant species might require large population sizes to remain viable.

Restoration potential of biomanipulation for eutrophic peri-urban ponds: the role of zooplankton size and submerged macrophyte cover

Eight hypereutrophic phytoplankton dominated ponds from the Brussels Capital Region (Belgium) were biomanipulated (emptied with fish removal) to restore their ecological quality and reduce the risk of cyanobacterial bloom formation. Continuous monitoring of the ponds before and after the biomanipulation allowed the effects of the management intervention on different compartments of pond ecosystems (phytoplankton, zooplankton, submerged vegetation and nutrients) to be assessed. Fish removal resulted in a drastic reduction in phytoplankton biomass and a shift to the clear-water state in seven out of eight biomanipulated ponds. The reduction in phytoplankton biomass was associated with a marked increase in density and size of large cladocerans in six ponds and a restoration of submerged macrophytes in five ponds. The phytoplankton biomass in the ponds with extensive stands of submerged macrophytes was less affected by planktivorous fish recolonisation of some of the ponds later in the summer. The two non-vegetated ponds as well as one pond with sparse submerged vegetation showed a marked increase in phytoplankton biomass associated with the appearance of fish. Phytoplankton biomass increase coincided with the decrease in large Cladocera density and size. One pond lacking submerged macrophytes could maintain very low phytoplankton biomass owing to large Cladocera grazing alone. The results of this study confirmed the importance of large zooplankton grazing and revegetation with submerged macrophytes for the maintenance of the clear-water state and restoration success in hypereutrophic ponds. They also showed that large Cladocera size is more important than their number for efficient phytoplankton control and when cladocerans are large enough, they can considerably restrain phytoplankton growth, including bloom-forming cyanobacteria, even when submerged vegetation is not restored. The positive result of fish removal in seven out of eight biomanipulated ponds clearly indicated that such management intervention can be used, at least, for the short-term restoration of ecological water quality and prevention of noxious cyanobacterial bloom formation. The negative result of biomanipulation in one pond seems to be related to the pollution by sewage water.

The use of random amplified polymorphic DNA (RAPD) for hybrid detection in Scirpus from the river Schelde (Belgium)

Along the unique freshwater tidal zone of the river Schelde (Belgium), plants of Scirpus species occur primarily in small and fragmented populations. The majority of these are native Scirpus lacustris, S. pungens, S. triqueter, S. tabernaemontani and intermediate morphological forms. The distribution area of S. triqueter is even restricted to this tidal habitat. However, several cultivated S. tabernaemontani strains have recently been introduced. The latter species is often used to stabilize riverbanks. To determine the existing genetic diversity among these species, stems from plants from 44 different locations were subjected to random amplified polymorphic DNA (RAPD) analysis, using 22 decanucleotide primers. Data analysis of the amplified DNA fragments enabled us to unambiguously differentiate among these Scirpus taxa. Hybridization between S. triqueter and S. tabernaemontani was documented, and the studied hybrids were always genetically more similar to S. triqueter than to S. tabernaemontani. Among the introduced clones, at least two different origins could be distinguished. Several of the introduced S. tabernaemontani clones were clearly different from individuals of the same species that were native to this region. As there were different hybrid genotypes owing to different hybrid events or to introgression, this complex should be safeguarded from genetic pollution. New S. tabernaemontani genotypes with markers not previously observed in that hybrid complex of the unique tidal freshwater habitat should be avoided in replantation projects.

Urban impact on ecological integrity of nearby rivers in developing countries: the Borkena River in highland Ethiopia

Accelerated pollution and eutrophication of rivers and streams because of human activity are a concern throughout the world and severe in Africa where Ethiopia is case in point. The objective of this study was to assess the urban impact on the ecological integrity of the Borkena River at the eastern escarpment of the central Ethiopian highlands. The water quality status and macroinvertebrate distribution and diversity of the river were assessed during the dry and wet seasons. Diversity indices revealed that a severe decline in the ecological integrity of the Borkena River downstream of Dessie and within Kombolcha towns in terms of macroinvertebrate abundance and composition. Clustering and ordination analysis clearly separated reference sites from urban impacted sites. At the urban-impacted sites, dissolved oxygen was also depleted to 0.5 mg/l and BOD5 values were reached to a level of above 1,000 mg/l, with extremely low biological diversity of pollution-sensitive taxa. These patterns are the result of a combination of rampant dumping of untreated wastes exacerbated by geologic, topographic, climatic and land use factors.