Natural populations of the Closterium ehrenbergii (Desmidiales, Chlorophyta) species complex in Nepal

Several natural populations of the Closterium ehrenbergii Meneghini ex Ralfs species complex were collected in Nepal, in October–December 1982. Water temperature and pH were also recorded. Clonal isolates from these populations were identified to one of four mating groups (H, I, J and M) by test crossing with standard mating-type strains of known mating groups. Groups H and M have smooth walled zygospores, while Groups I and J have scrobiculated zygospore walls. Several undetermined isolates were found in some population samples. In contrast to the previously reported population samples from Nepal, especially from dried soil samples, some of these populations appeared to be rather heavily loaded with mutations that are deleterious to the sexual cycle (i.e. sexual compatibility, zygospore formation and germination). By genetic analysis, a zygote maturation-defective mutation (zym) was detected. One reason for such a heavy genetic load was suggested to be that most population samples had been maintained exclusively by asexual reproduction for a long period in large lakes and nearby ponds, or left-over vegetative populations in paddy fields after other members entered into dormancy through sexual reproduction. The significance of studying such mutations at sexual gene loci is discussed in the light of speciation problems in microalgae.     

Morphological and cytogenetic characteristics of intergroup hybrids between closely related mating groups of the Closterium ehrenbergii species complex (Desmidiales, Chlorophyta)

Five F₁ hybrid strains were established from rare survivors in intergroup crosses between three closely related mating groups (A, B and H) of the Closterium ehrenbergii Meneghini ex Ralfs species complex. Cell sizes of these five strains studied under our standard culture conditions were compared to those of their parental stains and also to the total range of cell-size variation in each mating group. All five F₁ strains were larger in mean cell width than their parental strains. In cell length, three of them were larger than, one was the same as, and the other was intermediate between their parental strains. Their cell sizes were always larger than the range of their respective smaller parental mating group and three of them were larger than the range of their respective larger parental mating group.     

Pattern of genetic differentiation in the Maculinea alcon species group (Lepidoptera, Lycaenidae) in Central Europe

The taxonomic status of Alcon Blues living in Central and Western Europe (conventionally: Maculinea alcon and Maculinea rebeli ) is quite confused. Some authors distinguish them as separate species of the M. alcon species group, while others consider them as subspecies or simply ecological forms. Our aim was to study the pattern of genetic differentiation among several populations of these taxa with reference to their taxonomic position. Imagoes were collected from 27 localities in Central Europe between 1999 and 2003. Samples originated from four regions: northern Hungary, central Hungary, Romania: Transylvania and Slovenia. Enzyme polymorphism was analysed using polyacrylamide gel electrophoresis. In all samples 16 enzyme loci were studied. In the analysis of the data, F -statistics was computed and the total genetic variation was partitioned into within and between population components. Nei's genetic distances were calculated and UPGMA dendrogram was constructed on the basis of the distance matrix. Hierarchical F -statistics and amova was computed to study the pattern of genetic differentiation among the samples. Principal component analysis (PCA) was also carried out using the allele frequencies of the samples. The results of all analyses indicated a high level of differentiation among the samples. The differences among the samples collected in different years from the same population accounted for a considerable amount of the between sample variation. The distribution of the between sample variation did not exhibit a species pattern. The results of amova and PCA indicated that the geographic pattern was slightly more expressed in the between sample variation compared with the species pattern of it. These results seem to contradict with the conventional taxonomical subdivision.     

High genetic diversity and no inbreeding in the endangered copper redhorse, Moxostoma hubbsi (Catostomidae, Pisces): the positive sides of a long generation time

The evolutionary potential of a species is determined by its genetic diversity. Thus, management plans should integrate genetic concerns into active conservation efforts. The copper redhorse (Moxostoma hubbsi) is an endangered species, with an endemic distribution limited to the Richelieu River and a short section of the St Lawrence River in Quebec, Canada. The population, gradually fragmented since 1849, is characterized by a decline in population size and a lack of recruitment. A total of 269 samples were collected between 1984 and 2004 and genotyped using 22 microsatellite loci, which indicated that these fish comprise a single population, with a global F(ST) value of only 0.0038. Despite a small census size ( approximately 500), a high degree of genetic diversity was observed compared to common values for freshwater fishes (average number of 12.5 alleles/locus and average HO of 0.77 +/- 0.08). No difference was observed between expected and observed pairwise values of relatedness (r(xy): -0.00013 +/- 0.11737), suggesting an outbred population. Long-term Ne was estimated at 4476 whereas contemporary Ne values ranged from 107 to 568, suggesting a pronounced yet gradual demographic decline of the population, as no bottleneck could be detected for the recent past. By means of simulations, we estimated Ne would need to remain at more than approximately 400 to retain 90% of the genetic diversity over 100 years. Overall, these observations corroborate other recent empirical studies confirming that long generation times may act as a buffering effect contributing to a reduction in the pace of genetic diversity erosion in threatened species.     

Identification of a unicellular,non-pigmented alga that mediates growth inhibition in anuran tadpoles: a new species of the genus Prototheca (Chlorophyceae: Chlorococcales)

A non-pigmented, unicellular alga isolated from the faeces of British anuran tadpoles and which is associated with growth inhibition in these tadpoles, was described and identified using cytological, ultrastructural, nutrient assimilation and immunological studies. The alga possessed all the distinctive morphological features of the genus Prototheca, it grew weakly on Prototheca Isolation Medium (PIM), it required thiamine for continued growth and replication, and it could assimilate the five major substrates used to speciate the protothecans. All of these characteristics, together with previous nucleic acid hybridisation studies, indicated that the microorganism belonged to the genus Prototheca. There are currently five species recognised as valid (Pore, 1985 & 1986): Prototheca zopfii Kruger, 1884, P. wickerhamii Tubaki & Soneda, 1959, P. moriformis Kruger, 1884, P. stagnora Cooke, 1968 and P. ulmea Pore, 1986.The immunology showed that the new species was related to two of the protothecans, but overall it showed that the alga was antigenically distinct from the other protothecans tested in the immunoassay. This, together with its inability to grow strongly on PIM, its ability to assimilate a wide rage of carbon substrates and its ability to mediate growth inhibition in anuran tadpoles, indicated a new species of Prototheca. We therefore propose the name Prototheca richardsi sp. n.     

Isozyme analysis of Galaxias species (Teleostei: Galaxiidae) from the Taieri River, South Island, New Zealand: a species complex revealed

We examined genetic differentiation among 23 samples of non-migratory river galaxias from 17 streams in the Taieri River system, South Island, New Zealand. Four major genetic types were found, two of which occur in narrow sympatry in one location. These were compared with topotypical material representing Galaxias anomalus from the Clutha system (Otago) and G. vulgaris from the Waimakariri system (Canterbury) in order to establish identity. Morphological examination of these four major genetic types revealed consistent concomitant differences. The results suggest that there are at least three species of river galaxias in the Taieri system: G. anomalus, G. vulgaris and at least one previously undescribed species. We propose that the genetic structuring and subsequent speciation of this group has been promoted by the absence of the marine juvenile phase that is found in five other members of the genus native to New Zealand. This structuring may be exacerbated by population fragmentation over the last century owing to the negative influence of introduced trout. The phylogenetic diversity within the river system mirrors the diverse flora and invertebrate fauna of the region, and has conservation implications that parallel those resulting from our improved knowledge of the New Zealand herpetofauna through the application of genetic analysis.     

Temporal environmental change, clonal physiology and the genetic structure of a Daphnia assemblage (D. galeata–hyalina hybrid species complex)

1. In a combined field and laboratory study, seasonal relationships between water temperature and oxygen content, genetic structure (composition of MultiLocus Genotypes, MLGs) of a Daphnia assemblage (D. galeata–hyalina hybrid species complex), and the physiological properties of clones of frequent MLGs were studied. In accordance with the oxygen‐limited thermal tolerance hypothesis, essential physiological variables of oxygen transport and supply were measured within the tolerable temperature range. 2. A few MLGs (types T1–T4) were frequent during early spring and late autumn at surface temperatures below 10 °C. Clones of T1–T4 showed a low tolerance towards higher temperatures (above 20 °C) and a high phenotypic plasticity under thermal acclimation in comparison to clones derived from frequent MLGs from later seasons, and stored high–medium quantities of carbohydrates at 12 and 18 °C. 3. Another MLG (T6) succeeded the MLGs T1–T4. T6 was frequent over most of the year at temperatures above 10 °C and below 20 °C. A clone derived from T6 exhibited a high tolerance towards warm temperatures and a more restricted phenotypic plasticity. It stored high–medium quantities of carbohydrates at 12, 18 and 24 °C and showed a high capacity for acclimatory adjustments based on haemoglobin expression. 4. During the summer period at temperatures ≥20 °C, the MLG T6 was found mainly near to the thermocline, where temperature and oxygen content were distinctly lower, and to a lesser extent in surface water. At the surface, another MLG (T19) was predominant during this period. A clone of this MLG showed a very high tolerance towards warm temperatures, minimal phenotypic plasticity, low carbohydrate stores and a high capacity for circulatory adjustments to improve oxygen transport at higher temperatures. 5. This study provides evidence for connections between the spatio‐temporal genetic heterogeneity of a Daphnia assemblage and the seasonal changes of water temperature and oxygen content. The data also suggest that not only the actual temperature but also the dynamics of temperature change may influence the genetic structure of Daphnia populations and assemblages.     

Data on molecular taxonomy and genetic diversification of the European Hermit beetles, a species complex of endangered insects (Coleoptera: Scarabaeidae, Cetoniinae, Osmoderma)

A molecular analysis was carried out on the European hermit beetles (the Osmoderma eremita species complex) to explore their genetic diversification and the robustness of previous morphologically based taxonomic arrangements. Complete sequences of mtDNA cytochrome C oxidase I gene were obtained from 26 individuals. Mean levels of interspecific sequence divergence ranged from 0.044 to 0.186. The results indicate a clear-cut distinction between two clades. The first one includes the W-European O. eremita Scopoli 1763 ; and the two Italian endemic taxa Osmoderma italicum Sparacio, 2000 and Osmoderma cristinae Sparacio, 1994 ; from southern peninsular Italy and Sicily, respectively. The second one includes the widespread E-European Osmoderma barnabita ; and the southern Balcanic Osmoderma lassallei Baraud and Tauzin, 1991 from Greece and European Turkey. Within the two clades, molecular and morphological data well support a specific rank for O. lassallei and O. barnabita on one side, and for O. eremita and O. cristinae on the other side, while the taxonomic position of O. italicum , more closely related to O. eremita , is still uncertain. Current geographical distribution, interspecific genetic diversification, and very low levels of intraspecific genetic divergence in western European populations of O. eremita sensu stricto are hypothesized to be the result of multiple speciation events (mainly occurred in refugial forest areas of the Italian and Balkan peninsulas and Sicily before and during the Pleistocene glacial peaks), followed by fast post-glacial northward and westward expansion of some species.     

A study of genetic distances and variability in several species pf the gemis Ctenomys (Rodentia: Octodontidae) with special references to a probable causal role of chromosomes in speciation

Genetic variability measured by allozymic electrophoresis has been studied in several species of the subterranean rodent genus Ctenomys (Octodontidae). The study was carried out with the main purpose of analysing a special group known as the 'Corrientes group' which inhabits the province of Corrientes in Argentina. The members of the group are, with high probability, isolated reproductively due to their karyotypic differences diploid numbers are between 42 and 70). To evaluate whether the chromosomal differences were the causes of speciation or if they arose a posteriori, we compared the level of the genetic distance among taxa within the orrientes group with all those measured between intraspecific populations in Ctenomys . The results indicate that the distances among the populations of the Corrientes group are at or below the level of those measured among traspecific populations in Ctenomys . Thus, it is unlikely that genetic differentiation triggered speciation. It is shown that this low level of genetic differentiation is not in contradiction with the high levels expected for species originating from one of the possible mechanisms of chromosomal speciation named, chromosomal transilience. Although for geographical reasons it seems obviousthat gene flow is precluded among the members of the Corrientes group, the Slatkin method for estimating Nm values was also used. Because high values of Nm exist but no isolation by distance could be detected, it is suggested that reproductive and geographical isolation are very ecent. Finally, analysis of population variability suggests that the high levels of heterozygosity observed (a) can be explained by the population structure, and (b) are within the range of expected values if bottlenecks have occurred in the recent history of the Corrientes group. As a general conclusion, the results indicate that in the Corrientes group the genetic data support a putative causal role for chromosomes in speciation.