Effect of cultivating croplands and grazing in arid grassland habitats on the conservation of melitaeine butterflies in a mountainous area in Northern China

In the study area (Yanjiaping Village, Hebei Province, China), grazing extensity varies at different locations, small and discontinuous croplands are imbedded in some arid grassland, which are habitats for the melitaeine butterflies, Euphydryas aurinia and Melitaea phoebe. These two species of butterfilies coexist in this area, in which grazing and cultivation are the main disturbances. Grazing and cultivation have a reciprocal effect on E. aurinia, rather than M. phoebe. We observed that E. aurinia preferred to occupy patches with moderate grazing and imbedded with small and discontinuous croplands, where E. aurinia also has high population density. The percentage of E. aurinia larval groups in the ribbings was significantly higher than that of M. phoebe, whereas larvae of both species tended to increase in recent years. Our data also showed that the population density and the patch occupancy rate of both E. aurinia and M. phoebe were the highest under moderate grazing. It indicates that cultivation of small and discontinuous croplands within the patch has a significant effect on the population density of both species of melitaeine butterflies. Thus, to artificially create or maintain semi-natural habitats, complemented by moderate grazing, might be an ecological strategy to conserve melitaeine butterflies effectively. Considering the distinct impacts of cultivation and grazing on the population distribution and dynamics of the two different species, human disturbance in the mountainous area might be strategically involved in proposing conservation plans for the target species in the future.     

Genetic differentiation of theErigeron species (Asteraceae) in the Alps: A case of unusual allozymic uniformity

Allozyme variation was studied in all nine diploidErigeron species known from the Alps:E. alpinus, E. neglectus, E. polymorphus, E. candidus, E. uniflorus, E. atticus, E. gaudinii, E. acer, andE. angulosus. A total of 248 individuals from 24 natural populations was investigated using starch gel electrophoresis. Seven enzymes and 13 loci were assessed. Genetic variation within populations was low with the proportion of polymorphic loci ranging from 0.0–0.385, and average number of alleles per polymorphic locus from 2.0–2.5. In general, 70–100% of the genetic variation was attributed to between population differences. Mean genetic identities for pair-wise comparisons of populations averaged 0.893 within species, and 0.890 among species. Interspecific genetic variation of populations usually did not exceed intraspecific variation. It was concluded that theErigeron species from the Alps may have arisen by recent speciation probably during the epoches of glaciation. Morphological and ecological differences between species seem to be based on few gene loci.     

Microsatellite variation and differentiation among local populations of Castanopsis species in Japan

Microsatellite variations in Castanopsis species in Japan were examined to clarify the genetic relationships among 25 local populations according to the difference in the number of layers of adaxial epidermis in the leaves. Six microsatellite loci were assayed for 629 seedlings from the populations, and these seedlings were classified into five types according to the state of the leaf epidermis. Remarkable differences in the allele frequency of the six microsatellite loci were observed among these local populations. The coefficients of genetic differentiation, R_ST, of each locus ranged from 0.209 to 0.388. An unweighted pair-group method (UPGMA) phenogram constructed on the population pairwise R_ST over the loci revealed three clusters (A–C), and six sub-clusters. These clusters reflected the differences in the occurrence frequency of seedlings in each epidermis type within a population. Our findings suggest that clusters A and C are the local populations dominated by Castanopsis sieboldii and Castanopsis cuspidata, respectively, while local populations of cluster B are composed of the two Castanopsis species and/or include many individuals derived by hybridization. The six sub-clusters were found to reflect the geographic relationship among the populations, suggesting a different process for geographic population dynamics during the postglacial period.     

Extinction,recolonization, and the genetic structure of tidepool copepod populations

Summary Extinction and recolonization in metapopulations may either increase or decrease genetic differentiation among populations, but recent genetic models predict increased differentiation under most circumstances of recolonization. I examine this prediction empirically using tidepool populations of the marine tidepool copepodTigriopus californicus. The probability of extinction of tidepool populations was sufficiently high to invoke the model's predictions, but varied among populations. Nearly 75% of colonizing groups consisted of 10 or fewer individuals. The genetic effective size of colonizing groups might be as high as 18, depending on assumptions, but colonists probably originated from a subset of local populations. In contrast to my predictions, genetic differentiation was smaller among younger tidepool populations than among older populations on each of three rock outcrops, suggesting that genetic differentiation was reduced by metapopulation dynamics. The discrepancy between the prediction and the results may be explained by the unmet assumptions of classical metapopulation structure underlying the genetic models.     

Assessing genetic diversity of Elymus sibiricus (Poaceae: Triticeae) populations from Qinghai-Tibet Plateau by ISSR markers

Inter-simple sequence repeats (ISSR) markers were used to assess the genetic diversity and population structure in eight populations of Elymus sibiricus L. from the southeast of Qinghai-Tibet Plateau of China. Of the 100 primers screened, 13 produced highly reproducible ISSR bands. Using these primers, 193 discernible DNA fragments were generated with 149 (77.2%) being polymorphic, indicating considerable genetic variation at the species level. In contrast, there were relatively low levels of polymorphism at the population level with the percentage of polymorphic bands (PPB) ranging from 44.04 to 54.92%. The mean gene diversity (H_E) was estimated to be 0.181 within populations (range 0.164–0.200), and 0.274 at the species level. A high level of genetic differentiation among populations was detected based on Nei's genetic diversity analysis (33.1%), Shannon's index analysis (34.5%), Bayesian method (33.2%) and AMOVA analysis (42.5%). No significant statistical differences (analysis of molecular variance [AMOVA], P = 0.08) in ISSR variation were found between the sample collection regions. However, among populations (42.5% of the variance) and within populations (57.5% of the variance), there were significant differences (P < 0.001). Populations shared high levels of genetic identity. This pattern of genetic variation was different from that for most of inbreeding Triticeae species reported. In addition, a geographical pattern of population differentiation, where the populations from south and north of sampling sites were clearly separated from each other, was revealed by both the cluster and principal coordinates analyses. Generally, the result of this study indicates that E. sibiricus contains high molecular variation in its populations. The implications of these results for the conservation of the species are discussed.     

Low genetic diversity and significant population structuring in the relict Amentotaxus argotaenia complex (Taxaceae) based on ISSR fingerprinting

Amentotaxus, a genus of the Taxaceae, represents an ancient lineage that has long existed in Eurasia. All Amentotaxus species experienced frequent population expansion and contraction over periodical glaciations in Tertiary and Quaternary. Among them, Amentotaxus argotaenia complex consists of three morphologically alike species, A. argotaenia, Amentotaxus yunnanensis, and Amentotaxus formosana. This complex is distributed in the subtropical region of mainland China and Taiwan where many Pleistocene refugia have been documented. In this study, genetic diversity and population structuring within and between species were investigated based on the inter-simple sequence repeats (ISSR) fingerprinting. Mean genetic diversity within populations was estimated in three ways: (1) the percentage of polymorphic loci out of all loci (P) (2) Neis unbiased expected heterozygosity (He), and (3) Shannons index of phenotypic diversity. For a total of 310 individuals of 15 populations sampled from the three species, low levels of ISSR genetic variation within populations were detected, with P=4.66–16.58%, He=0.0176–0.0645 and Hpop=0.0263–0.0939, agreeing with their seriously threatened status. AMOVA analyses revealed that the differences between species only accounted for 27.38% of the total variation, whereas differences among populations and within populations were 57.70 and 14.92%, respectively, indicating substantial isolation between the patch-like populations. A neighbor-joining tree identified a close affinity between A. yunnanensis and A. formosana. Genetic drift due to small population size, plus limited current gene flow, resulted in significant genetic structuring. Low levels of intrapopulational genetic variation and considerable interpopulational divergence were also attributable to demographic bottlenecks during and/or after the Pleistocene glaciations.     

Genetic diversity and structure of the endemic Caesalpinia hintonii complex (Leguminosae: Caesalpinioideae) in Mexico

The Caesalpinia hintonii complex is formed by five endemic species (C. hintonii, C. laxa, C. macvaughii, C. melanadenia and C. epifanioi) occurring in central Mexico. This species complex is under incipient genetic divergence as by-product of local adaptations in reproductive and morphological traits to different habitats. We estimate the genetic variation and structure of populations of this species complex to assess the extent of genetic differentiation among populations and related species along its geographic distribution. Estimations of genetic diversity and structure were done based on ten enzymes and 18 loci. Mean number of alleles per locus ranged from 1.5 to 1.9. Polymorphic loci ranged from 42.1 to 68.4. Observed (H_o: range 0.191–0.275) and expected (H_e: range 0.205–0.317) heterozygosities in this complex were higher compared with other endemic and legume species. Neis genetic diversity estimates showed that most genetic variation was found within (H_S = 0.325) rather than among populations (D_ST = 0.085). Populations of the species C. hintonii showed a considerable genetic differentiation (F_ST = 0.207). The results of genetic diversity and structure within and among populations are in accord with the great morphological differentiation described for this species complex.     

Analysis of genetic diversity in the endangered Hucho taimen from China

Hucho taimen are listed as endangered in China. The population size has declined recently, prompting an increase in the level of listing from grade three in 2002 to grade five in 2006. We analyzed the genetic diversity of wild populations using 17 microsatellite markers to establish a scientific basis for conservation of this species. We collected tissue samples from four populations in the Heilongjiang River basin: Huma River (HM), Hutou (HT), Haiqing (HQ), and Zhuaji (ZJ). A total of 21 loci were amplified, 18 of which were polymorphic. The number of alleles per locus ranged from 2 to 9 (mean: 4.1905). There were 13 highly polymorphic loci and 5 moderately polymorphic loci. Analysis of five genetic diversity parameters (Na, Ne, Ho, He, and PIC) suggested moderate levels of diversity within the populations. The populations were ranked HT > HQ > ZJ > HM, but the differences in diversity were not statistically significant (P > 0.05). A comparison of variation among all four populations suggested Hardy–Weinberg disequilibrium at 20% of the loci. Genetic differentiation (Fst) was 0.0644 and the gene flow among populations was estimated at 3.36 individuals per generation. The majority of diversity (93.88%) occurred among individuals within a population. In contrast, relatively little (6.12%) of the genetic diversity was distributed between the populations. An analysis of genetic differentiation and genetic distance between pairs of populations revealed that both parameters were higher in comparisons of the HM population to the HT, HQ, and ZJ populations than among the three latter populations. This suggests that the HM population has a distinct genetic structure. We hypothesize that habitat degradation and excessive fishing, not low genetic diversity, has caused the decline in H. taimen populations. However, this species should be protected from further declines in genetic diversity.     

Genetic diversity and environmental associations of wild barley,Hordeum spontaneum (Poaceae), in Iran

Genetic diversity and structure of populations of the wild progenitor of barleyHordeum spontaneum in Iran was studied by electrophoretically discernible allozymic variation in proteins encoded by 30 gene loci in 509 individuals representing 13 populations of wild barley. The results indicate that: a)Hordeum spontaneum in Iran is extremely rich genetically but, because of predominant self-pollination, the variation is carried primarily by different homozygotes in the population. Thus, genetic indices of polymorphismP-1% = 0.375, range = 0.267–0.500, and of genetic diversity,He = 0.134, range = 0.069–0.198, are very high. b) Genetic differentiation of populations includes clinal, regional and local patterns, sometimes displaying sharp geographic differentiation over short distances. The average relative differentiation among populations isGst = 0.28, range = 0.02–0.61. c) A substantial portion of the patterns of allozyme variation in the wild gene pool is significanctly correlated with the environment and is predictable ecologically, chiefly by combinations of temperature and humidity variables. d) The natural populations studied, on the average, are more variable than two composite crosses, and more variable than indigenous land races of cultivated barely,Hordeum vulgare, in Iran. — The spatial patterns and environmental correlates and predictors of genetic variation ofH. spontaneum in Iran indicate that genetic variation in wild barley populations is not only rich but also at least partly adaptive. Therefore, a much fuller exploitation of these genetic resources by breeding for disease resistance and economically important agronomic traits is warranted.     

Comparison of genetic population structure of the large blue butterflies Maculinea nausithous and M. teleius

We investigated the genetic population structure of two rare myrmecophilous lycaenid butterflies, Maculinea nausithous and M. teleius, which often live sympatrically and have similar biology. In Europe, both species occur in highly fragmented populations and are vulnerable to local extinction. The proportion of variable allozyme loci, average heterozygosity and genetic differentiation among populations was higher in M. nausithous than in sympatrically living M. teleius populations. We hypothesise that the differences in heterozygosity are mainly due to the known higher efficiency of typical host ant nests in rearing M. nausithous pupae compared to M. teleius pupae. This implies a larger probability of larval survival in M. nausithous, which buffers populations against environmental and demographic stochasticity. In contrast, the lower carrying capacity of ant nests in rearing M. teleius pupae requires higher nest-densities and makes M. teleius populations more prone to losing genetic variation through drift if this condition is not fulfilled. The single investigated Russian population of M. teleius showed much higher levels of heterozygosity than any of the Polish populations, suggesting a more viable and still intact metapopulation structure.     

An exploratory analysis of geographic genetic variation in southern African nyala (Tragelaphus angasii)

We report patterns of genetic variation based on microsatellite, allozyme and mitochondrial control region markers in nyala from geographic locations sampled in South Africa, Mozambique, Malawi and Zimbabwe. Highly significant differences were observed among allele frequencies at three microsatellite loci between populations from KwaZulu-Natal, Limpopo and Malawi, with the Malawi and KwaZulu-Natal groupings showing the highest differentiation (R_ST=0.377). Allozyme frequencies showed minor, non-statistically significant regional differences among the South African populations, with maximum F_ST values of 0.048–0.067. Mitochondrial DNA analyses indicated a unique haplotype in each location sampled. Since none of these indices of population differentiation showed significant correlation to absolute geographic distance, we conclude that geographic variation in this species is probably a function of a distribution pattern stemming from habitat specificity. It is suggested that translocations among geographically distant regional populations be discouraged at present, pending a more elaborate investigation. Transfer of native individuals among local populations may, however, be required for minimizing the likelihood of inbreeding depression developing in small captive populations.     

同域分布垂穗披碱草和达乌力披碱草遗传多样性和遗传结构分析

披碱草属(Elymus L.)是禾本科(Poaceae)小麦族(Triticeae)中的一个多年生属,该属在青藏高原地区有广泛分布,多数物种是草原和草甸的组成成分,许多种类为品质优良的牧草.垂穗披碱草(E.nutans)和达乌力披碱草(E.dahuricus)同为禾本科小麦族披碱草属异源六倍体物种,染色体组组成皆为St...     

Genetic Variation Within and Among Populations of Rhodiola alsia (Crassulaceae) Native to the Tibetan Plateau as Detected by ISSR Markers

Genetic variation of 10 Rhodiola alsia (Crassulaceae) populations from the Qinghai–Tibet Plateau of China was investigated using intersimple sequence repeat (ISSR) markers. R. alsia is an endemic species of the Qinghai–Tibet Plateau. Of the 100 primers screened, 13 were highly polymorphic. Using these primers, 140 discernible DNA fragments were generated with 112 (80%) being polymorphic, indicating pronounced genetic variation at the species level. Also there were high levels of polymorphism at the population level with the percentage of polymorphic bands (PPB) ranging from 63.4 to 88.6%. Analysis of molecular variance (AMOVA) showed that the genetic variation was mainly found among populations (70.3%) and variance within populations was 29.7%. The main factors responsible for the high level of differentiation among populations are probably the isolation from other populations and clonal propagation of this species. Occasional sexual reproduction might occur in order to maintain high levels of variation within populations. Environmental conditions could also influence population genetic structure as they occur in severe habitats. The strong genetic differentiation among populations in our study indicates that the conservation of genetic variability in R. alsia requires maintenance of as many populations as possible.     

Genetic differentiation among populations ofArabis serrata (Brassicaceae) along its geographic distribution

Levels and distribution of genetic variation were investigated in ten populations of the perennialArabis serrata distributed along a latitudinal gradient throughout Japan. Populations of this endemic species occupy predominantly three types of habitats: limestone and derived soils, volcanic and disturbed sites. Previous studies showed that plants ofA. serrata are differentiated in morphological and ecological traits under both natural conditions and common garden experiments suggesting genetic differentiation among populations. To assess the degree of genetic differentiation under different habitats ofA. serrata populations, we analyzed the isozyme genetic structure. Ten populations, located in mountains of central and northern Honshu and Hokkaido, were analyzed by starch gel enzyme electrophoresis. Fourteen loci of eight enzymes were resolved and six loci were monomorphic for all the populations. Populations sampled maintain low levels of genetic variation (P=0.16;H=0.05;A=1.16) compared to that maintained by other outcrossing seed plants. In some cases, few or no heterozygous individuals were detected, and consequently, mean observed heterozygosity was zero or near zero. Six (29%) of the fixation indices,F, estimated deviated significantly from Hardy Weinberg genotypic expectations indicating a deficiency of heterozygotes in most of the cases. The mean genetic identity (Nei'sI) between population pairs was 0.852 and indicates a moderate level of genetic differentiation among populations.Arabis serrata has most of its genetic variation distributed among rather than within populations. The among-population component of the total genetic diversity (G _ST mean value) was 0.416, indicating genetic differentiation between populations. There groups of populations were recognized in an unrooted tree generated by the Neighbor-Joining method. These results suggest groups of populations differentiated regionally. Estimates of interpopulational gene flow (Nm) were very variable (range 0.049–3.718) with a meanNm=1.203 for all populations.     

Surprising diversity in the Pannonian populations of Marsh Fritillary (Euphydryas aurinia,Lepidoptera: Nymphalidae): Morphometric and molecular aspects

Since genetic variation is the basis of evolutionary potential of a species, its structure needs to be understood. Thus, the aim of this study was to analyze and contrast the structure of genetic and phenotypic variation in the Euphydryas aurinia populations of southeastern central Europe. Genetic variation was studied by two types of molecular genetic markers: mtDNA COI sequences and allozymes. As the great hiatus in the European distribution of E. aurinia is located in the central part of the Carpathian Basin, we expected that the populations East and West to this gap would be highly differentiated. Populations of Central Transdanubia actually represent the easternmost margin of the West European distribution of E. aurinia. In view of the peripheral position of these populations, we supposed to find some genetic sign of local adaptation, as a consequence of diversifying selection and an increased level of fluctuating asymmetry as a result of environmental stress. The analyses of the molecular genetic markers revealed a basic East–West differentiation among the populations of southeastern central Europe which was further structured in the western part of the study area. The results suggested that the genetic differentiation between the two western regions is probably the consequence of diversifying selection. The pattern of phenotypic differentiation among the western populations, however, was different. A geographic cline was revealed (decreasing wing size) toward the eastern margin of the distribution in parallel with increasing fluctuating asymmetry. The conservation inferences of the results are considered.     

Combining larval habitat quality and metapopulation structure – the key for successful management of pre-alpine Euphydryas aurinia colonies

This study aims to analyse larval habitat preferences and landscape level population structure of the threatened Marsh Fritillary butterfly, Euphydryas aurinia, and discusses implications for the conservation and management of this strongly declining species in central Europe. Whereas current management strategies are mainly based on studies of habitat requirements of adult individuals, we intend to emphasise larval habitat quality and population processes at the landscape level as additional key factors. Microhabitat preference analysis of egg-laying females showed that eggs were predominantly laid on prominent large-sized host plant individuals. Additionally, when Succisa pratensis was used as a host plant (as opposed to Gentiana asclepiadea), host individuals in open vegetation structure were preferred. Optimal oviposition conditions were present in recently abandoned calcareous fen meadows and at the edges of such meadows currently in use. A two-year patch-occupancy study in the northern pre-alpine region of south-west Germany indicated that E. aurinia lives in a metapopulation. In a logistic-regression model, patch size, isolation, and habitat quality explained 82% of the observed patch-occupancy pattern in 2001. Our data suggest that a suitable conservation strategy must incorporate both the conservation of a network of suitable habitat patches, and efforts to maximise local habitat quality by ensuring that host plants can grow to a large size and are surrounded by sparse and low vegetation cover.     

Genetic diversity in the rare orchid Gymnadenia odoratissima and a comparison with the more common congener, G. conopsea

Patterns and levels of genetic diversity mayhave significant influence on the long termpersistence of local populations and revealingsuch information is important in protectingrare species. In this study we investigated thegenetic pattern in five microsatellite lociwithin five Swedish populations of the rareorchid species Gymnadenia odoratissima. Thegeographic distribution of G. odoratissima isrestricted to Europe and in Scandinavia it isonly found in three provinces in southernSweden; Östergötland,Västergötland and on the island ofGotland.Compared with the more widespread congener G.conopsea our results indicate lower levels ofgenetic variation within and higher degrees ofgenetic differentiation among populations ofG. odoratissima (H_EL = 0.6–0.8 in G. conopseaand 0.3–0.7 in G. odoratissima; F_ST over allpopulations = 0.06 in G. conopsea and 0.19 inG. odoratissima). Also, we found a cleardistinction among mainland and islandpopulations of G. odoratissima wherepopulations on the island of Gotland seem toexhibit higher levels of gene flow andintragenetic variation, probably as a result ofa larger number of existing populations.Future conservation of this species shouldfocus on facilitation on colonisation events,especially on the mainland, and preservation ofthe genetically more variable Gotlandpopulations.     

Genetic diversity of fragmented populations of<Emphasis Type="Italic"> Polygala reinii</Emphasis> (Polygalaceae), a perennial herb endemic to Japan

Genetic variation at 10 allozyme loci was analyzed in 14 populations of Polygala reinii (Polygalaceae), a perennial herb endemic to central Honshu, Japan, with a fragmented geographical distribution. The levels of genetic variation within species (P=80.0, A=3.10, H_E=0.303) and within populations (P=42.1, A=1.61, H_E=0.163) were considerably higher than the mean for other endemic plants or short-lived perennial herbs. Genetic differentiation among populations was also high (G_ST=0.404). The genetic distance phenogram tended to show a clustering of the populations reflecting the fragmentation of the species range. A principal component analysis revealed the same tendency, as well as three groupings of populations in the Tokai district, on the Kii Peninsula and in the northern Kinki district. A negative correlation was obtained between the levels of gene flow and geographical distance among the populations (r=–0.745, P<0.0001). These results indicated limited gene flow among populations in P. reinii, presumably due to the geographical isolation accompanying the fragmented distribution. On the other hand, the geographical differentiation between the Japan Sea and Pacific Ocean sides was found in P. reinii, suggesting the influence of postglacial migration on the establishment of the genetic structure of this species.     

The fragmented population structure of the Sardinian chalk hill blue butterfly (Lepidoptera, Lycaenidae)

The endemic Sardinian chalk hill blue butterfly, Polyommatus coridon gennargenti, is considered vulnerable to extinction because of its low genetic variation and restricted distribution. The species also has a fragmented distribution, which follows the patchy distribution pattern of its larval host-plant. A preliminary investigation of the population structure of P. coridon gennargenti was carried out on a small network of four local populations by means of capture–recapture methods. Estimated population sizes and movement rates among the four adjacent local populations suggest that this taxon has a metapopulation structure composed of loosely connected small local populations. Natural fragmentation, isolation, and traditional land use contribute to the vulnerability of P. coridon gennargenti to extinction. Low effective population sizes and restricted movement between habitat patches lead to inbreeding and an increased vulnerability to extinction of this island population.     

A genetic comparison of two species of clam shrimp in the genus Eulimnadia: An electrophoretic approach

Herein we report the first genetic comparison among species in the genus Eulimnadia. Multilocus genotypic patterns (using six allozyme loci) were compared for a total of 2277 clam shrimp from nine populations from Arizona and New Mexico. Seven of these populations were morphologically typed as Eulimnadia texana Packard and two were typed as Eulimnadia diversa Mattox. All populations were hermaphrodite-biased, and highly inbred (inbreeding coefficients ranging from 0.33 to 0.98). Genetic distances showed the two species to be within the range described for other arthropods. One of the two E. diversa populations appeared to be a hybrid between E. texana and E. diversa, showing electrophoretic patterns similar to both species, although morphologically, they were typed as E. diversa. A phenogram (generated using coancestry distances and a neighbor joining algorithm) placed this hybrid population half-way between these two species, and a breakdown of individuals within this hybrid population (based on allozyme scores) indicated individuals very similar to the second E. diversa population, and two groups of apparent hybrid individuals. Therefore, the distinction between these two species is questionable due to their apparent hybridization in this area of Arizona. Genetic population structuring was noted among the seven E. texana pools. Estimated migration rates were less than one migrant per generation. Even in the geographically close pools in New Mexico, which were separated by only hundreds of meters, significant sub-structuring was noted, and estimates of migration rate were less than two migrants per generation.