Two highly validated multiplexes (12-plex and 8-plex) for species delimitation and parentage analysis in oaks (Quercus spp.)

Multiplex PCR is a fast and cost‐effective technique allowing increased genotyping throughput of microsatellites. We developed two multiplexes for Quercus petraea and Q. robur, a 12‐plex of EST‐SSRs (eSSRs) and an 8‐plex of genomic SSRs (gSSRs). We studied the origin of allele calling errors at the human reader and software levels. We showed that the robustness of allele identification can be improved by binning on raw peak sizes prior to genetic data analysis. We checked through simulation the power of these markers for species delimitation and hybrid detection. The resolution achieved with all 20 markers was greatly improved compared to that of previous studies based on a subset of the markers. Preliminary PCR tests suggest that these multiplexes might be useful to study other oak species as well. The strategy used for multiplex microsatellite development (from PCR conditions to the definition of allele calling rules) should be broadly applicable.     

Blind population genetics survey of tropical rainforest trees

Rainforest tree species can be difficult to identify outside of their period of reproduction. Vascular tissues from Carapa spp. individuals were collected during a short field trip in French Guiana and analysed in the laboratory with nuclear and chloroplast markers. Using a Bayesian approach, > 90% of the samples could be assigned to one of two distinct clusters corresponding to previously described species, making it possible to estimate the genetic structure of each species and to identify cases of introgression. We argue that this blind procedure represents a first-choice rather than a fallback option whenever related taxa are investigated.     

Species composition and relative abundance of copepods in Chilean fjords

We studied the species composition of copepods in the fjordregion, Isla Madre de Dios, Chile during October 1979. Fifteennew records for the fjords are cited. We found the antarcticcopepod Paraeuchaeta antarctica which is consistent with earlyrecords of other antarctic zooplankters in the area. The analysisof relative abundances shows that changes in dominance occuronly among four most abundant species of copepods.     

Development of a Chinese-Indian hybrid (Chindian) rhesus macaque colony at the California National Primate Research Center by introgression

Background  Fullbred Chinese and Indian rhesus macaques represent genetically distinct populations. The California National Primate Research Center introduced Chinese founders into its Indian-derived rhesus colony in response to the 1978 Indian embargo on exportation of animals for research and the concern that loss of genetic variation in the closed colony would hamper research efforts. The resulting hybrid rhesus now number well over a thousand animals and represent a growing proportion of the animals in the colony.
Methods  We characterized the population genetic structure of the hybrid colony and compared it with that of their pure Indian and Chinese progenitors.
Results  The hybrid population contains higher genetic diversity and linkage disequilibrium than their full Indian progenitors and represents a resource with unique research applications.
Conclusions  The genetic diversity of the hybrids indicates that the strategy to introduce novel genes into the colony by hybridizing Chinese founders and their hybrid offspring with Indian-derived animals was successful.     

More introgression with less gene flow: chloroplast vs. mitochondrial DNA in the Picea asperata complex in China, and comparison with other Conifers

Recent work has suggested that rates of introgression should be inversely related to levels of gene flow because introgressed populations cannot be 'rescued' by intraspecific gene flow if it is too low. Mitochondrial and chloroplast DNA (mtDNA and cpDNA) experience very different levels of gene flow in conifers due to their contrasted maternal and paternal modes of transmission, hence the prediction that mtDNA should introgress more readily than cpDNA in this group. Here, we use sequence data from both mtDNA and cpDNA to test this hypothesis in a group of closely related spruces species, the Picea asperata complex from China. Nine mitochondrial and nine chloroplast haplotypes were recovered from 459 individuals in 46 natural populations belonging to five species of the Picea asperata complex. Low variation was found in the two mtDNA introns along with a high level of differentiation among populations ( G _ST = 0.90). In contrast, we detected higher variation and lower differentiation among populations at cpDNA markers ( G _ST = 0.56), a trend shared by most conifer species studied so far. We found that cpDNA variation, although far from being fully diagnostic, is more species-specific than mtDNA variation: four groups of populations were identified using cpDNA markers, all of them related to species or groups of species, whereas for mtDNA, geographical variation prevails over species differentiation. The literature suggests that mtDNA haplotypes are often shared among related conifer species, whereas cpDNA haplotypes are more species-specific. Hence, increased intraspecific gene flow appears to decrease differentiation within species but not among species.     

Species Delimitation--a Geneious plugin for the exploration of species boundaries

Species Delimitation is a plugin to the Geneious software to support the exploration of species boundaries in a gene tree. The user assigns taxa to putative species and the plugin computes statistics relating to the probability of the observed monophyly or exclusivity having occurred by chance in a coalescent process. It also assesses the within and between species genetic distances to infer the probability with which members of a putative species might be identified successfully with tree-based methods.     

Positive relationships between genetic diversity and abundance in fishes

Molecular markers, such as mitochondrial DNA and microsatellite loci, are widely studied to assess population genetics and phylogeography; however, the selective neutrality of these markers is increasingly being questioned. Given the importance of molecular markers in fisheries science and conservation, we evaluated the neutrality of both mtDNA and microsatellite loci through their associations with population size. We surveyed mtDNA and microsatellite data from the primary literature and determined whether genetic diversity increased with abundance across a total of 105 marine and freshwater fishes, with both global fisheries catch data and body size as proxies for abundance (with an additional 57 species for which only body size data were assessed). We found that microsatellite data generally yielded higher associations with abundance than mtDNA data, and within mtDNA analyses, number of haplotypes and haplotype diversity were more strongly associated with abundance than nucleotide diversity, particularly for freshwater fishes. We compared genetic diversity between freshwater and marine fishes and found that marine fishes had higher values of all measures of genetic diversity than freshwater fishes. Results for both mtDNA and microsatellites generally conformed to neutral expectations, although weaker relationships were often found between mtDNA nucleotide diversity and ‘abundance’ compared to any other genetic statistic. We speculate that this is because of historical events unrelated to natural selection, although a role for selection cannot be ruled out.     

A survey of the relative abundance of specific groups of cellulose degrading bacteria in anaerobic environments using fluorescence in situ hybridization

AIMS: The utility of fluorescence in situ hybridization (FISH) for detecting uncultured micro-organisms in environmental samples has been shown in numerous habitats. In this study a suite of three FISH probes for cellulolytic bacteria is described and their efficacy is demonstrated by quantifying the relative abundance of the target micro-organisms in a range of industrial biomass samples. METHODS AND RESULTS: The probes were designed from data derived from an artificial landfill leachate reactor study and 16S rRNA gene databases. The original biomass sample proved to be well described by the three probes targeting a total of 51% of the bacterial (EUBMIX targeted) cells in quantitative FISH experiments. CONCLUSIONS: Three probes were developed and applied to samples from a range of industrial digesters. The CSTG1244 probe, specific for organisms closely related to Clostridium stercorarium, were observed in the widest range of samples (7 of the 19 samples tested). The CTH216a FISH probe, specific for organisms closely related to Clostridium thermocellum, described the highest proportion of the bacterial population within any one sample (46% in an anaerobically digested sludge sample). Finally, the BCE216a probe, specific for organisms closely related to Bacteroides cellulosolvens, achieved the lowest level of hybridisation of the three probes tested. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates that the three groups of anaerobic cellulolytic micro-organisms were present in different bioreactors but at variable abundances ranging from low (where other organisms would have been responsible for cellulolysis) to high. We showed the potential of using group specific FISH probes and quantitative FISH in environmental studies. The utility of using newly designed FISH probes was demonstrated by their ability to detect and quantify the target bacterial groups in samples from a range of industrial wastewater digesters.     

Statistical mechanics of relative species abundance

Statistical mechanics of relative species abundance (RSA) patterns in biological networks is presented. The theory is based on multispecies replicator dynamics equivalent to the Lotka–Volterra equation, with diverse interspecies interactions. Various RSA patterns observed in nature are derived from a single parameter related to productivity or maturity of a community. The abundance distribution is formed like a widely observed left-skewed lognormal distribution. It is also found that the “canonical hypothesis” is supported in some parameter region where the typical RSA patterns are observed. As the model has a general form, the result can be applied to similar patterns in other complex biological networks, e.g. gene expression.     

北京东灵山辽东栎(Quercus liaotungensis)林沿海拔梯度的物种多度分布

多度格局对理解群落结构具有重要意义。选用5个描述种-多度关系的生态位模型(断棍模型BSM、生态位优先占领模型NPM、生态位重叠模型ONM、随机分配模型RAM、优势优先模型DPM),分乔、灌、草3层分别对北京东灵山辽东栎林进行了研究。结果表明:在乔木层,沿海拔梯度从低海拔到高海拔,能很好地反映物种多度格局变异的模型有由RAM、NPM向DPM过渡的趋势;NPM和BSM均能较好地模拟灌木层的绝大多数海拔段,但NPM的效果更好;草本层以BSM的模拟效果最好。生态位模型可以反映出辽东栎林乔木层物种多度分布沿海拔存在的明显变异,而灌木层和草本层物种多度分布沿海拔梯度无明显变化或很难由生态位模型反映出来,是否引入新的模型方法,如中性模型、近中性模型,还有待于进一步研究。     

Species delimitation in the genusLipomyces by nuclear genome comparison

Species delimitation inLipomyces was attempted by nuclear genome comparison in conjuction with the re-evaluation of 48 physiological characters of 65 strains.High intraspecific (>75%) and low interspecific (<28%) similarity values established thatL. japonicus, L. lipofer andL. tetrasporus are genetically isolated, and also distinct fromL. kononenkoae andL. starkeyi.Ambiguous similarity values were obtained withL. kononenkoae andL. starkeyi. Strains previously assigned toL. kononenkoae constitute two related clusters. While similarity values within each cluster range from 76–99%, representatives of the two clusters reassociate for only 47%. Since these clusters are differentiated by their ecologically relevant maximum growth temperature,L. kononenkoae is subdivided. Strains previously assigned toL. starkeyi resolve into four closely related clusters. While similarity values within each cluster range from 78–100%, representatives of the four clusters reassociate for only 59–69%. Since these four clusters are poorly differentiated, the subdivision ofL. starkeyi does not appear possible without recourse to other criteria.Four unassigned strains constitute a further two clusters. Reassociation within these clusters is of the order of 91–100%, while reassociation between them occurs only at 59%. Reassociation of representatives of these clusters with those of theL. kononenkoae andL. starkeyi complexes is around 40% and 31%, respectively. These two clusters consequently appear to be intermediate betweenL. kononenkoae andL. starkeyi, and will, as such, have to be considered in any delimitation of these two species. A key to the taxa ofLipomyces and related genera of the Lipomycetaceae is given.     

Increased inbreeding and inter-species gene flow in remnant populations of the rare <Emphasis Type="Italic">Eucalyptus benthamii</Emphasis>

Eucalyptus benthamii Maiden & Cambage is a forest tree of interest for conservation and plantation forestry. It is vulnerable to extinction, occurring on the alluvial floodplains of the Nepean River and its tributaries, south-west of Sydney, Australia. These floodplains were largely cleared of native vegetation for agriculture by the mid-1800s. Flooding of the Cox Valley for Sydneys water supply further decreased the species distribution. The species is now confined to one population of approximately 6500 trees in the Kedumba valley and three remnant populations on the Nepean River at Bents Basin (about 300 trees), Wallacia (nine trees) and Camden (about 30 trees). Genetic analysis of the four populations using microsatellite markers revealed significant divergence among all populations, despite the Bents Basin, Wallacia and Camden remnants being separated by distances of only a few kilometres. Trees in these populations have been estimated to range from 35 to 200 years old, suggesting genetic divergence among populations occurred prior to land clearing. To investigate the impact of fragmentation on the next generation, outcrossing rates were estimated from 41 families. While no direct relationship was found between population size and outcrossing rates, fragmentation and the isolation of trees appears to have resulted in higher levels of selfing and biparental inbreeding in seed collected from the Camden and Wallacia remnants. There was also evidence from seedling morphology that inter-species gene flow increased with fragmentation since 20% of the progeny from Camden and 30% of the progeny from Wallacia were hybrids. Seed viability and germination rates were significantly lower in the remnant populations, reducing their value as seed sources for regeneration and plantation forestry. To maintain the genetic integrity of the remnant populations, germplasm should be sourced from the local area. Outcrossed, non-hybrid seed could be produced by controlled pollination in ex-situ conservation stands or by using seedling morphology and microsatellites to screen seedlings from the remnant populations.     

Conservation of (GA)n microsatellite loci between Quercus species

Microsatellite (simple sequence repeat, SSR) amplification was performed in eight different members of the Fagaceae family by using sets of primers developed from sessile oak, Quercus petraea . In total, 136 cases of heterologous amplification were carried out, and 66% resulted in interpretable amplification products. From these, 12 PCR amplification products were sequenced and all 12 contained a sequence homologous to the original locus from Q. petraea . Although SSR primers worked even across different genera, with increasing evolutionary distance there was a clear tendency for decreasing ability to successfully amplify loci and a decreasing proportion of polymorphism amongst those markers which could be amplified. Two of the loci, ssrQpZAG46 and ssrQpZAG110, were polymorphic in all Quercus species tested. Only at one locus, ssrQpZAG58, a specific PCR product could be amplified in all species analysed. For four loci found in two species, we observed significant interspecies differences in the size range of the amplified alleles. Sequence analysis of two alleles showed that the size differences are not only due to variations in the number of (GA) repeats but also to an insertion of approximately 80 nucleotides in the flanking region. Our findings prove the usefulness of SSR markers within and amongst closely related genera of plants.     

Using chloroplast DNA to trace postglacial migration routes of oaks into Britain

Postglacial migration is a major factor responsible for the patterns of genetic variation we see in natural populations. Fossil pollen data indicate that early postglacial colonists such as oak, were able to take both western and eastern migration routes into Britain. Analysis at a finer level is now permitted by the use of modern molecular techniques. A 13-bp duplication in the chloroplast tRNA^Leul intron occurs in natural populations of East Anglian oaks, but is not found in other parts of Britain or from mainland Europe. The distribution of this marker suggests that the mutation occurred either in southern England, or during migration from the mainland, and became fixed in a source population from which East Anglia was colonized. Planting of non-native trees for roadside boundaries and in the grounds of old houses and estates, explains the absence of the marker from some East Anglian oaks.     

Even more oak species in Mexico? Genetic structure and morphological differentiation support the presence of at least two specific entities within Quercus laeta

Differentiation among populations, sometimes despite ongoing gene exchange, is a key step in speciation. Therefore, comparison of intra- and interspecific differentiation patterns is of great significance to understanding speciation. The genus Quercus is an interesting system to test speciation models in the presence of gene flow, due to its weak interspecific reproductive barriers. The aim of the present study was to characterize the degree and pattern of morphological and genetic differentiation among different morphotypes in the white oak Quercus laeta, some corresponding to the previously described species Quercus centralis, Q. laeta, Quercus prinopsis, and Quercus transmontana, as well as geographically structured variation within Q. transmontana not described previously. Our goal was to evaluate if some of these can be considered distinct specific entities or are rather part of a continuum of variation. Nine microsatellite loci and two intergenic regions of chloroplast DNA were analyzed. Morphological differences were evaluated using geometric morphometrics. Chloroplast DNA showed low differentiation, suggesting introgression or sharing of ancestral haplotypes among the Q. laeta morphotypes. Nuclear microsatellites indicated differentiation into two distinct main genetic groups, which were congruent with morphological differentiation. In conclusion, nuclear markers and morphological variations suggest the existence of at least two different entities within Q. laeta.     

Characterization of polymorphic microsatellite loci in the endangered Miho spine loach (Iksookimia choii) and cross-species amplification within the Cobitidae family

The Miho spine loach (Iksookimia choii) is an endangered freshwater fish endemic to Korea. To aid a conservation programme for this species, 20 polymorphic microsatellite loci were isolated and characterized in 67 individuals from three populations. The number of alleles per locus ranged from 8 to 33. Within populations, the observed and expected heterozygosities ranged from 0.59 to 1.00 and from 0.56 to 0.96, respectively, indicating its usefulness in population genetics studies. Cross-species amplification of the loci was tested in six species of the family Cobitidae, and 18 loci were amplified in two or more related species.     

Temporal mortality pattern of pedunculate oaks in southern Sweden

Decline of the oak forests decline in southern Sweden has been reported for more than two decades. Little empirical data exists, however, to study the temporal pattern of the phenomenon in detail. In this study we quantified the temporal pattern of non-windfall oak mortality by analyzing the dataset of 44 dendrochronologically dated dead pedunculate oak (Quercus robur L.) trees. We compared tree-ring chronologies from recently dead and living trees from the same sites (number of sites=13) located in the nemoral and boreo-nemoral zones in southern Sweden. For each dead tree, tree-ring chronologies were analyzed for the presence of pre-death growth depressions. A growth depression was defined as a period (of one or more years) when growth remained below the 5%, 7%, or 10% quantiles of the ring-width distribution obtained from living trees for a particular year and site.

The most recent peak in oak mortality occurred around the year 2000. Growth depressions were recorded in 80% (n=35) of all dead oaks and were most prominent during the 1990s. While some oaks showed an obvious reduction in growth over several decades, 51% of the dead trees had growth depression for at least 4 years prior to death. Although diameter growth rate differed between living and recently dead trees for at least 30 years, this difference started to amplify in late 1980s–early 1990s. Presence of pre-death growth depression in tree-ring chronologies implies that (a) non-windfall mortality of oak is a decade-long process and (b) the actual death events might be lagging behind the timing of the mortality-inducing factors. ANOVA revealed significant differences in tree responses to the drought year 1992. The cumulative growth increment ratio between 1992–1994 and 1989–1991, was higher in living trees than in those that had recently died. We suggest that the spring and summer drought of 1992 resulted in the mortality of oaks that was observed in southern Sweden at the end of the 20th century. If this time lag exists, it may complicate analyses of decline-related factors and the choice of appropriate actions by forest managers. We conclude that studies of oak decline may benefit from widening the time perspective to include several decades preceding the sampling year.     

Invasive species can't cover their tracks: using microsatellites to assist management of starling (Sturnus vulgaris) populations in Western Australia

Invasive species are known to cause environmental and economic damage, requiring management by control agencies worldwide. These species often become well established in new environments long before their detection, resulting in a lack of knowledge regarding their history and dynamics. When new invasions are discovered, information regarding the source and pathway of the invasion, and the degree of connectivity with other populations can greatly benefit management strategies. Here we use invasive common starling ( Sturnus vulgaris ) populations from Australia to demonstrate that genetic techniques can provide this information to aid management, even when applied to highly vagile species over continental scales. Analysis of data from 11 microsatellites in 662 individuals sampled at 17 localities across their introduced range in Australia revealed four populations. One population consisted of all sampling sites from the expansion front in Western Australia, where control efforts are focused. Despite evidence of genetic exchange over both contemporary and historical timescales, gene flow is low between this population and all three more easterly populations. This suggests that localized control of starlings on the expansion front may be an achievable goal and the long-standing practice of targeting select proximal eastern source populations may be ineffective on its own. However, even with low levels of gene flow, successful control of starlings on the expansion front will require vigilance, and genetic monitoring of this population can provide essential information to managers. The techniques used here are broadly applicable to invasive populations worldwide.