Effect of multiple-locus heterozybosity and salinity on clearance rate in a brackish-water clam,Rangia cuneata (Sowerby)

Previous studies of several species of marine bivalves and gastropods have reported a positive correlation between growth or size and level of multiple-locus heterozygosity. There is some evidence that the growth advantage of relatively heterozygous individuals is due to a lower rate of standard or routine metabolism, compared with more homozygous individuals, although heterozygosity-dependent differences in feeding rate may also be involved. The present study examined the relationship between clearance rate in three salinity treatments (5,15, and 25%.) and multiple-locus heterozygosity at nine polymorphic allozyme loci in the clam Rangia cuneata (Sowerby). Clearance rates were determined by disappearance of an algal suspension from a flowing-water system. Allozyme genotypes were determined using starch-gel electrophoresis. The polymorphic loci examined were those coding for a nonspecific esterase (Est), mannosephosphate isomerase (Mpi), leucine aminopeptidase (Lap), 6-phosphogluconate dehydrogenase (6-Pgd), phosphoglucose isomerase (Pgi), isocitrate dehydrogenase (Idh), malate dehydrogenase (Mdh), adenylate kinase (Adk), and phosphoglucomutase (Pgm). Weight-corrected clearance rates increased significantly (P < 0.05) with increasing multiple-locus heterozygosity and decreased significantly (P < 0.05) with increasing salinity. These data support the idea that heterozygosity-growth correlations may be due in part to differences in clearance rate. However, further study is needed to understand the exact physiological processes which relate heterozygosity and growth.     

Asymmetric gene flow and the evolutionary maintenance of genetic diversity in small,peripheral Atlantic salmon populations

Small populations may be expected to harbour less genetic variation than large populations, but the relation between census size (N), effective population size (N _e), and genetic diversity is not well understood. We compared microsatellite variation in four small peripheral Atlantic salmon populations from the Iberian peninsula and three larger populations from Scotland to test whether genetic diversity was related to population size. We also examined the historical decline of one Iberian population over a 50-year period using archival scales in order to test whether a marked reduction in abundance was accompanied by a decrease in genetic diversity. Estimates of effective population size (N _e) calculated by three temporal methods were consistently low in Iberian populations, ranging from 12 to 31 individuals per generation considering migration, and from 38 to 175 individuals per generation if they were regarded as closed populations. Corresponding N _e/N ratios varied from 0.02 to 0.04 assuming migration (mean=0.03) and from 0.04 to 0.18 (mean=0.10) assuming closed populations. Population bottlenecks, inferred from the excess of heterozygosity in relation to allelic diversity, were detected in all four Iberian populations, particularly in those year classes derived from a smaller number of returning adults. However, despite their small size and declining status, Iberian populations continue to display relatively high levels of heterozygosity and allelic richness, similar to those found in larger Scottish populations. Furthermore, in the R. Asón no evidence was found for a historical loss of genetic diversity despite a marked decline in abundance during the last five decades. Thus, our results point to two familiar paradigms in salmonid conservation: (1)␣endangered populations can maintain relatively high levels of genetic variation despite their small size, and (2) marked population declines may not necessarily result in a significant loss of genetic diversity. Although there are several explanations for such results, microsatellite data and physical tagging suggest that high levels of dispersal and asymmetric gene flow have probably helped to maintain genetic diversity in these peripheral populations, and thus to avoid the negative consequences of inbreeding.     

Enzymes of Heavy-Metal-Resistant and Non-Resistant Populations of Silene cucubalus and Their Interaction with Some Heavy Metals in vitro and in vivo

The in vitro and in vivo effects of copper, zinc, cadmium, nickel, cobalt, and manganese on nitrate reductase, malate dehydro-genase, isocitrate dehydrogenase, and glucose-6-phosphate dehydrogenase of zinc-, copper- and non-resistant populations of Silene cucubalus were investigated. During the in vitro experiments no resistant enzyme could be detected; enzymes of resistant and non-resistant ecotypes had a similar sensibility to all the metals. Nitrate reductase was the most sensitive enzyme. During the in vivo experiments remarkable differences were found. The nitrate reductase and the isocitrate dehydrogenase of the zinc-resistant population were activated when adding zinc to the culture medium, especially the nitrate reductase showed high activities at zinc concentrations where the nitrate reductase of the non-zinc-resistant populations was nearly completely inhibited. The zinc-resistant ecotype had a real need for zinc.     

Accumulation, subcellular localization and ecophysiological responses to copper stress in two Daucus carota L. populations

Copper accumulation, subcellular localization and ecophysiological responses to excess copper were investigated using pot culture experiments with two Daucus carota L. populations, from a copper mine and an uncontaminated field site, respectively. Significant differences of malondialdehyde (MDA) and hydrogen peroxide (H₂O₂) concentrations and antioxidant enzyme [superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX)] activities of leaves under Cu treatment were observed between the two populations. At high Cu concentrations (400 and 800 mg kg⁻¹), a significant increase in contents of MDA and H₂O₂ but a significant decrease in activities of SOD, CAT and APX were observed in uncontaminated population. Contrarily, the population from copper mine maintained a lower level of MDA and H₂O₂ but higher activities of SOD, CAT and APX. Copper accumulation in roots and shoots increased significantly with the increase of copper concentrations in soils in the two populations. No significant difference of the total Cu in roots and shoots was found between the two populations at same copper treatment. There were also no striking differences of cell wall-bound Cu and protoplasts Cu of leaves between the two populations. The difference was that Cu concentration in vacuoles of leaves was 1.5-fold higher in contaminated site (CS) population than in uncontaminated site population. Hence, more efficient vacuolar sequestration for Cu and maintaining high activities of SOD, CAT and APX in the CS population played an important role in maintaining high Cu tolerance.     

Genetic diversity of Mexican brook lamprey <Emphasis Type="Italic">Lampetra (Tetrapleurodon) geminis</Emphasis> (Alvarez del Villar, 1966)

Lampreys are the only surviving representatives of the oldest known vertebrates. The Mexican lamprey L. geminis (nonparasitic), is particularly interesting, because it is an endemic, biogeographical relict, and a threatened species. RAPD markers were used to describe genetic diversity in L. geminis A total of 77 specimens were collected from five populations, three in the R'o Grande de Morelia-Cuitzeo basin and two in the R'o Duero-Lerma-Chapala basin, Mexico. Eighty-eight RAPD markers were obtained from eight primers. Genetic diversity within each population was estimated using Shannon's index (S), heterozygosity (H) and gene diversity (h). These estimates revealed significant variation within populations, although a variance homogeneity test (HOMOVA) showed no significant differences among populations or between basins. Nei genetic distance values indicate a low genetic differentiation among populations. Analysis of molecular variance (AMOVA) indicates that most of the genetic diversity occurs within populations (91.4%), but that a statistically significant amount is found among populations (P0.001). Principal coordinates and cluster analyses of RAPD phenotypes show that specimens are not grouped by geographical origin. The genetic diversity found within L. geminispopulations may be explained by its breeding system and an overlapping of generations. The scarce genetic differentiation among populations is likely to the low rate of DNA change that characterizes the lamprey group.     

Selection affecting enzyme polymorphisms in laboratory populations of Drosophila melanogaster

Summary Gene frequencies at nine polymorphic enzyme loci were followed in 22 cage populations of D. melanogaster all founded from the same stock but subsequently maintained in different environments. The three factors distinguishing the different environments were temperature and the alcohol and yeast in the medium and for each factor each population experienced either a constant or variable condition. The variable conditions were either coarse-grained (period between changes longer than the generation time) or fine-grained (the period less than the generation time).Six polymorphisms were found to be unaffected by any experimental condition. However, the various constant temperatures tested differed in their effects on Alcohol dehydrogenase (Adh), Amylase (Amy) and Isocitrate dehydrogenase (Idh-NADP) gene frequencies. Furthermore, coarse-grained variation between high and low temperatures increased Amy and Idh-NADP heterozygosity and decreased Amy heterozygosity while fine-grained variation between such temperatures promoted Amy heterozygosity. In addition, joint coarse-grained variation for the type of alcohol and yeast promoted Amy and Idh-NADP heterozygosity as well as heterozygosity averaged over all nine loci.In general, the results suggested two major modifications to the hypothesis of a positive relationship between environmental and genetic variability. First, they indicated that the relationship is not always positive. It varies both between loci and between environments. Second however, they indicated that the relationship is more likely to be positive when the environmental variability is coarse- rather than fine-grained.     

Nineteen novel microsatellites in Chinese lake gudgeon Sarcocheilichthys sinensis Bleeker, 1971

Nineteen polymorphic microsatellites isolated from a partial genomic DNA library of Sarcocheilichthys sinensis enriched for CA repeats were developed. Variability was tested on 48 unrelated individuals collected from Jiangsu province, China. Observed heterozygosity ranged from 0.200 (Ss236) to 1.00 (Ss223, Ss224), whereas the expected heterozygosity ranged from 0.213 (Ss232) to 0.939 (Ss224). Shannon's information index ranged from 0.405 (Ss236) to 2.988 (Ss224). Sixteen of the 19 microsatellites conformed to Hardy–Weinberg equilibrium. These microsatellites can be used for the evaluation of genetic parameters in populations to supply valuable information for conservation management.     

Characterization of polymorphic microsatellite loci in the terrestrial isopod Armadillidium vulgare

The common pill bug, Armadillidium vulgare, is known to harbour two distinct strains of the feminizing proteobacteria Wolbachia. In order to study the effect of the presence of Wolbachia on the evolution of A. vulgare populations, we developed and characterized a set of nine polymorphic microsatellite loci from two microsatellite‐enriched genomic libraries. We screened 48 individuals from three French populations and found high genetic variation. Locus‐specific allelic diversity ranged from four to 28 and observed heterozygosity from 0.40 to 1.00, which indicates that these markers can be used to conduct population genetic studies in A. vulgare.     

GENETIC DIVERSITY AND CAPILLARIA HEPATICA (NEMATODA) PREVALENCE IN MICHIGAN DEER MOUSE POPULATIONS

There have been few field tests of the hypothesis that homozygous populations are prone to high levels of disease. I tested for a negative correlation between genetic diversity and parasitism by estimating the allozyme heterozygosity, population density, and proportion of individuals infected by Capillaria hepatica (Nematoda) in nine Michigan populations of deer mice (Peromyscus maniculatus). Parasite prevalence was correlated negatively with heterozygosity when the effects of density were held constant, but was not correlated with population density after controlling for the effects of genetic diversity. These data support the prediction that inbred populations will be more susceptible to parasite infestations.     

The effect of mating system on growth of Arabidopsis lyrata in response to inoculation with the biotrophic parasite Albugo candida

The effects of variation in host reproductive systems on response to pathogens are not well understood. We inoculated individuals from outcrossing and inbreeding populations of North American Arabidopsis lyrata with Albugo candida (white blister rust) to test the effect of mating system and heterozygosity on disease response. We observed three host infection phenotypes, classified as fully resistant, partially resistant and fully susceptible. Overall, inbreeding populations had more susceptible and fewer partially resistant individuals than outcrossing populations, but the highest proportion of resistant individuals was found in two of the inbreeding populations. Mating system did not affect relative growth rate of inoculated plants, but there were strong effects of population and infection phenotype. We conclude that mating system per se does not determine the resistance of natural A. lyrata populations to infection by Albugo, but that the increased variability in responses among inbreeding populations may be due to reduced effective population size.     

Disentangling the impact of demographic factors on population differentiation of an endangered freshwater crayfish (Austropotamobius pallipes) using population density and microsatellite data

1. Habitat fragmentation of stream ecosystems often results in decreased connectivity between populations and lower population sizes. Hence, understanding how habitat fragmentation affects genetic erosion is important for the preservation of freshwater biodiversity, in particular, as small populations suffer from loss of genetic diversity through genetic drift and loss of fitness because of inbreeding, increasing the risk of extinction. 2. Here, we assess the impact of demographic factors on population differentiation in the endangered freshwater crayfish Austropotamobius pallipes by analysing population genetic structure, estimating effective population sizes and comparing levels of polymorphism at five microsatellite loci with density estimates of 10 populations within a small French catchment that has become progressively confined to headwaters over the last six decades. 3. Levels of expected heterozygosity and allelic richness per population were relatively low (0.214–0.396 and 1.6–2.6, respectively). We found strong genetic differentiation between these geographically close populations (F_ST = 0.283), with weak statistical evidence for a pattern of isolation by distance. Estimates of effective population size were low (<150) in most populations, but potentially reached several thousands in three populations. 4. Population density and allelic richness were strongly positively correlated. A robust relationship between population density and heterozygosity values was also noted, but only after discarding two populations for which significant genetic signatures of a recent bottleneck were found; these two populations displayed high expected heterozygosity compared with a very low density. Populations with the highest densities of individuals had the highest effective population size estimates and vice versa. 5. Our results clearly show the importance of demographic factors and genetic drift on A. pallipes populations. Furthermore, analysis of genetic and population density data is a pragmatic and efficient approach to corroborate inferences from genetic data and can be particularly useful in the identification of populations experiencing a bottleneck and therefore in conservation genetics studies aiming at identifying priority populations for conservation.     

华北落叶松种子园控制授粉子代遗传多样性分析

以华北落叶松控制授粉群体的全部子代为材料,母本相同的个体视为同一家系,利用18对SSR分子标记对7个家系257个个体进行扩增,分析其遗传多样性及其遗传分化水平。结果表明:(1)18个SSR位点共检测到72个等位基因,平均等位基因数4个,有效等位基因数(Ne)为1.247~3.411。(2)7个家系的平均有效等位基因数为2.135个,观测杂合度(Ho)为0.518,期望杂合度(He)为0.502,Shannon信息指数0.846,其中55号家系遗传多样性水平最高,56号的遗传多样性水平最低。(3)遗传分化系数(GST)为0.113,各家系群体处于中等遗传分化水平,AMOVA分析结果显示82%的遗传变异存在于家系内,18%的遗传变异存在于家系间。(4)聚类分析结果表明,55号与59号家系的遗传距离最近,具有较近的亲缘关系,49号家系与其他家系的遗传距离最远。(5)结合遗传多样性及遗传分化水平结果,估算获得选择核心家系数及核心个体数,选择5个家系均可获得96%以上的遗传多样性,对于个体数较少的家系,选择15~20个个体;对于个体数较多的家系,选择35个个体,即可获得96%以上的遗传多样性。该研究结果对华北落叶松种子园育种群体的选择及其遗传多样性的保护具有重要意义。     

Transkingdom transfer of the phosphoglucose isomerase gene

Previous analysis of the gene encoding phosphoglucose isomerase (Pgi) suggests that this gene may have been transferred between a eukaryote and a bacterium. However, excluding the alternative hypothesis of ancient gene duplication has proven difficult because of both insufficient sampling of taxa and an earlier misidentification of a bacterialPgi sequence. This paper presents a phylogenetic analysis of published completePgi sequences together with analysis of new partialPgi sequences from six species of bacteria. The data identify a group of bacterialPgi sequences, including sequences fromEscherichia coli andHaemophilus influenzae, which are more closely related to eukaryoticPgi sequences than to other bacterial sequences. The topology of gene trees constructed using several different methods are all consistent with the hypothesis of lateral gene transfer andnot ancient gene duplication. Furthermore, an estimate of a molecular clock forPgi dates the divergence of theE. coli andH. influenzae sequences from the animal sequences to between 470 and 650 million years ago, well after other estimates of the divergence between eukaryotes and bacteria. This study provides the most convincing evidence to date of the transkingdom transfer of a nuclear gene.     

Association between host's genetic diversity and parasite burden in damselflies

Recent research indicates that low genetic variation in individuals can increase susceptibility to parasite infection, yet evidence from natural invertebrate populations remains scarce. Here, we studied the relationship between genetic heterozygosity, measured as AFLP‐based inbreeding coefficient f_AFLP, and gregarine parasite burden from eleven damselfly, Calopteryx splendens, populations. We found that in the studied populations, 5–92% of males were parasitized by endoparasitic gregarines (Apicomplexa: Actinocephalidae). Number of parasites ranged from none to 47 parasites per male, and parasites were highly aggregated in a few hosts. Mean individual f_AFLP did not differ between populations. Moreover, we found a positive association between individual's inbreeding coefficient and parasite burden. In other words, the more homozygous the individual, the more parasites it harbours. Thus, parasites are likely to pose strong selection pressure against inbreeding and homozygosity. Our results support the heterozygosity‐fitness correlation hypothesis, which suggests the importance of heterozygosity for an individual's pathogen resistance.     

Unexpectedly high genetic diversity and divergence among populations of the apomictic Neotropical tree Miconia albicans

• Since tropical trees often have long generation times and relatively small reproductive populations, breeding systems and genetic variation are important for population viability and have consequences for conservation. Miconia albicans is an obligate, diplosporous, apomictic species widespread in the Brazilian Cerrado, the savanna areas in central Brazil and elsewhere in the Neotropics. The genetic variability would be, theoretically, low within these male‐sterile and possibly clonal populations, although some variation would be expected due to recombination during restitutional meiosis.
• We used ISSR markers to assess genetic diversity of M. albicans and to compare with other tropical trees, including invasive species of Melastomataceae. A total of 120 individuals from six populations were analysed using ten ISSR primers, which produced 153 fully reproducible fragments.
• The populations of M. albicans presented mean Shannon's information index (I) of 0.244 and expected heterozygosity (H_e) of 0.168. Only two pairs of apparently clonal trees were identified, and genetic diversity was relatively high. A hierarchical amova for all ISSR datasets showed that 74% of the variance was found among populations, while only 26% of the variance was found within populations of this species. Multivariate and Bayesian analyses indicated marked separation between the studied populations.
• The genetic diversity generated by restitutional meiosis, polyploidy and possibly other genome changes may explain the morpho‐physiological plasticity and the ability of these plants to differentiate and occupy such a wide territory and different environmental conditions. Producing enormous amounts of bird‐dispersed fruits, M. albicans possess weedy potential that may rival other Melastomataceae alien invaders.

     

dbEST-derived SSR markers in sea urchin (Hemicentrotus pulcherrimus)

Sea urchin is an important model organism for evolutionary biology, embryology, and gene regulation study. We developed and evaluated simple sequence repeat (SSR) markers from expressed sequence tags (ESTs) of Strongylocentrotus purpuratus and Hemicentrotus pulcherrimus. Characteristics of nine EST-SSR loci were investigated using 41 Hemicentrotus pulcherrimus individuals. The number of alleles per locus ranged from two to five. The observed heterozygosity ranged from 0.122 to 0.7805, while the expected heterozygosity ranged from 0.4472 to 0.7696. These loci and markers will be useful for population genetics and systemic evolution among species of sea urchin.     

Residues of brodifacoum and other anticoagulant pesticides in target and non‐target species,Nelson Lakes National Park,New Zealand

Abstract

Cereal‐based bait containing 20 ppm brodifacoum was used in bait stations continuously from December 1997 to August 2000 in the Rotoiti Nature Recovery Project area, for controlling brushtail possum (Trichosurus vulpecula), ship rat (Rattus rattus), and house mouse (Mus musculus) populations. Concurrently (and before and after), baits containing brodifacoum, bromadiolone, flocoumafen, coumatetralyl, or warfarin were also used in St Arnaud village and on farms immediately adjacent to the project area. Brodifacoum residues were detected in the livers of 234 mammals from eight species, and two birds from two species captured alive, and a further seven birds from five species found dead in the project area (cause of death unknown). The highest concentration of brodifacoum residues in mammalian livers was recorded during the period brodifacoum was used in the project area. However, residues were present in the livers of some individuals at least 24 months after brodifacoum use in the project area stopped. These residues may have persisted in animals surviving brodifacoum use in the project area, and/or been transported into the area by animals moving to and from the adjacent village and farms, where brodifacoum use continued. Residues of flocoumafen, coumatetralyl, or warfarin, used only in the village and on farms, were also detected in the livers of animals captured up to at least 8 km from the nearest source. The results concur with other studies cautioning against indiscriminate or prolonged use of persistent anticoagulants for vertebrate pest control. However, the risks from such pesticides must be balanced against the demonstrable benefits of reducing pest abundance.     

Microsatellite loci for tucumã of Amazonas (Astrocaryum aculeatum) and amplification in other Arecaceae

• Premise of the study: Microsatellite loci were developed for tucumã of Amazonas (Astrocaryum aculeatum), and cross-species amplification was performed in six other Arecaceae, to investigate genetic diversity and population structure and to provide support for natural populations management. • Methods and Results: Fourteen microsatellite loci were isolated from a microsatellite-enriched genomic library and used to characterize two wild populations of tucumã of Amazonas (Manaus and Manicoré cities). The investigated loci displayed high polymorphism for both A. aculeatum populations, with a mean observed heterozygosity of 0.498. Amplification rates ranging from 50% to 93% were found for four Astrocaryum species and two additional species of Arecaceae. • Conclusions: The information derived from the microsatellite markers developed here provides significant gains in conserved allelic richness and supports the implementation of several molecular breeding strategies for the Amazonian tucumã.     

Regulation of gemma formation in the copper moss <Emphasis Type="Italic">Scopelophila cataractae</Emphasis> by environmental copper concentrations

Considerable attention has recently been focused on the use of hyperaccumulator plants for the phytoremediation of soils contaminated with heavy metals. The moss, Scopelophila cataractae (Mitt.) Broth., is a typical hyperaccumulator that is usually observed only in copper-rich environments and which accumulates high concentrations of copper in its tissues. However, many of the physiological processes and mechanisms for metal hyperaccumulation in S. cataractae remain unknown. To address this issue, we examined the mechanisms regulating gemma formation, which is considered the main strategy by which S. cataractae relocates to new copper-rich areas. From this study we found that treatment of S. cataractae with high concentrations of copper suppressed gemma formation but promoted protonemal growth. The suppressive effect was not observed by treatment with heavy metals other than copper. These results suggest the importance of copper-sensitive asexual reproduction in the unique life strategy of the copper moss, S. cataractae.     

Isolation of microsatellite markers for Contarinia nasturtii,a European pest invading the New World

Microsatellite markers were developed for epidemiological studies on Contarinia nasturtii (Diptera, Cecidomyiidae), a native European pest that was introduced to the New World in 1996. Nine loci were found to be polymorphic and suitable for the analysis of 56 male individuals that were collected using pheromone traps. These markers all conform to Hardy–Weinberg expectations in at least one of the two tested populations, and carry an average number of 11 alleles among populations and observed levels of heterozygosity ranging from 0.32 to 0.86. Primers for all markers also successfully amplified fragments from Contarinia pisi and Contarinia tritici.