Systematics and gene geography of Juniperus communis L. inferred from isoenzyme data

Using isoenzyme analysis, 35 populations of Juniperus communis L. from various parts of the Russian species range and by one population from Sweden and Alaska were studied. The total sample size was 1200 plants. As a result, the existence of J. communis var. oblonga in North Caucasus and J. communis var. depressa in North America was confirmed, but genetic differences between J. communis var. communis and J. communis var. saxatilis were not detected in the main part of the Russian species range (European part of Russia, Ural, Siberia). These populations proved to be genetically uniform with the same predominant allelic frequencies, which may evidence recent settling of this species from one of Central or East European refugium. J. communis var. saxatilis from northeastern Russia inhabiting the region behind Verkhoyansk mountain and Russian Far East showed considerable differentiation in frequencies of alleles at three loci and geographical subdivision. These populations also exhibit high intrapopulation variation. This can be connected with the refugium in this territory. The origin of this group is probably connected with migrations from Central Asia (Tibet) in the direction to northeastern Russia along mountains connecting Central and North Asia. It is also assumed that migrations of this species previously proceeded across the Beringian land bridge.     

HS‐SPME/GC Analysis Reveals the Population Variability of Terpene Contents in Juniperus communis Needles

Juniperus communis var. communis L. is an aromatic plant – typical boreal element of flora. In the extensive literature concerning J. communis, there is much data on the composition and the content of essential oil of needles and coneberries, but a detailed analysis of terpene distribution within and between populations is missing. A representative pool of 74 J. communis individuals originating from ten populations of Northern Poland was investigated in order to evaluate the intra‐ and interpopulational variability of the terpene pattern. Headspace solid‐phase microextraction (HS‐SPME) coupled with GC/MS and GC/FID was applied in achiral and enantioselective analysis. The majority of the samples (85%), despite different origin, were similar in the terpene pattern. High diversity of terpenes was observed within the populations and low diversity between them. High variation of enantiomeric composition was in accordance with large variation of individual compounds in general (achiral analysis). J. communis samples from Northern Poland could be distinguished by the α‐pinene/sabinene ratio, and they were divided into three chemical races.     

Biogeographic Variation of Foliar n‐Alkanes of Juniperus communis var. saxatilis Pallas from the Balkans

The composition of the epicuticular n‐alkanes isolated from the leaves of ten populations of Juniperus communis L. var. saxatilis Pallas from central (continental) and western (coastal) areas of the Balkan Peninsula was characterized by GC‐FID and GC/MS analyses. In the leaf waxes, 14 n‐alkane homologues with chain‐lengths ranging from C₂₂ to C₃₅ were identified. All samples were dominated by n‐tritriacontane (C₃₃), but differences in two other dominant n‐alkanes allowed separating the coastal from the continental populations. Several statistical methods (ANOVA, principal component, discriminant, and cluster analyses as well as the Mantel test) were deployed to analyze the diversity and variability of the epicuticular‐leaf‐n‐alkane patterns of the ten natural populations of J. communis var. saxatilis and their relation to different geographic and bioclimatic parameters. Cluster analysis showed a high correlation of the leaf‐n‐alkane patterns with the geographical distribution of the investigated samples, differentiating the coastal from the continental populations of this taxon. Several bioclimatic parameters related to aridity were highly correlated with this differentiation.     

Allozyme and shell variation in two marine snails (Littorina, Prosobranchia) with different dispersal abilities

As adults Littorina littorea and L. saxatilis have overlapping distributions, both on the shore and on a geographical scale, this implies that they are affected by similar selective forces to a large extent. The two species have, however, different reproductive strategies. Littorina littorea has a planktonic larval stage lasting 4–6 weeks, whereas L. saxatilis has direct development. This influences dispersal rates of the two species, and gene flow and effective population size are both assumed to be much smaller in L. saxatilis than in L. littorea. Intraspeciflc variation in morphological and allozyme characters differs between the two species. Littorina littorea is homogeneous for both types of characters, both within and between populations, while populations of L. saxatilis may show pronounced differences in shell characters with micro-environment changes, and allozyme variation is markedly higher in this species too. The polymorphism in shell characters of L. saxatilis and the monomorphism of L. littorea, may be expected from theory as a consequence of the different dispersal strategies. Also the lack of divergence between geographically separate populations of L. littorea, but the high degree of between population differentiation in L. saxatilis, could be attributed to the high and low dispersal rates, respectively. However, the larger allozyme variation within subpopulations of L. saxatilis compared to that of L. littorea, is surprising as the former species has a smaller effective population size than the latter. Furthermore, there is no correlation between dispersal ability and overall allozyme heterozygosity, when 10 additional species of Littorina are included in the comparison. It is suggested that different evolutionary backgrounds of different species will possibly explain this, as neither selectionist nor neutralist models alone account for the observation. That is, an earlier passage through a population bottleneck may, for example, influence the genetic variation for a long period of time.     

Using genetic and morphological analysis to distinguish endangered taxa from their hybrids with the cultivated exotic pest plant <Emphasis Type="Italic">Lantana strigocamara</Emphasis> (syn: <Emphasis Type="Italic">Lantana camara</Emphasis>)

Because highly invasive species can rapidly assimilate rare taxa, we questioned whether two Florida endangered Lantana depressa varieties existed 21 years after Sanders documented their widespread hybridization with exotic Lantana strigocamara, and whether morphological traits could accurately discriminate genetic individuals. Stepwise discriminant analysis of morphological characters discriminated the three taxa, correctly classifying 98, 91, 89% of L. strigocamara, L. depressa var. depressa, and var. floridana. Hybrids blurred taxonomic distinctions of varieties and reduced classification accuracy by 7–17%. Species-specific Random Fragment Length Polymorphism (RFLP-PCR) confirmed hybridization has occurred. Intersimple Sequence Repeat (ISSR) fingerprints analyzed with STRUCTURE identified three groups indicating introgression. Morphological traits significantly, but weakly correlated with q ratios (P = 0.0001; r ² = 0.45). Although L. strigocamara introgression is widespread and ongoing, wild populations contain individuals that are predominantly L. depressa genome, supporting actions to remove adventive L. strigocamara, prevent its sale, and promote sales of genetically confirmed natives.     

Assessment of genetic variation in Sicilian Helichrysum (Asteraceae) and implication to germplasm conservation

Abstract

Some taxa of Sicilian Helichrysum related to sect. Stoeachadina have an uncertain systematic position; moreover, they are rare and endangered. In order to preserve these taxa in a genebank, a strategy to keep their genetic variation as high as possible is required. The high informative content of AFLP markers was used to assess genetic variations within and between populations and their relationships. The results show that all populations are genetically similar, even though there are two distinct groups: the first is the population referred to H. pendulum, and the second is represented by other populations belonging to several different specific and intraspecific taxa. The H. pendulum population is homogeneous, and characterised by the lowest genetic variation among all the populations considered. The second group shows a greater variation as evidenced by the presence of three subgroups: (i) populations referred to H. panormitanum var. panormitanum and H. panormitanum var. stramineum, (ii) populations referred to H. errerae var. errerae and H. errerae var. messerii, (iii) a population referred to H. hyblaeum, which, despite morphological and chorological differences, is similar to the other populations of the second group from a genetic stand point. Each population of the above-mentioned taxa has local fragment markers whose frequency is useful to determine how many plants should be sampled in order to collect the seeds necessary to preserve the population's genetic diversity in a genebank.     

Rapid colonization of Belgian breakwaters by the direct developer,Littorina saxatilis (Olivi) (Prosobranchia,Mollusca)

The Belgian coast has no natural rocky sites but a number of man-made constructions are colonized by rocky shore organisms. The rough periwinkle, Littorina saxatilis (Olivi), lacks a planktonic larval stage but is found on most breakwaters along the Belgian coast, a few built as recently as 1986. This indicates a good potential of dispersal along this sandy shallow coast, nearly as good as for the planktonic developer Littorina littorea (L.) which is found on generally the same sites in Belgium. The breakwater populations of L. saxatilis, however, tend to be somewhat less variable (level of heterozygosity about 10% less) than non-Belgian L. saxatilis populations of natural sites. This suggests that the breakwater populations have passed through bottlenecks when founded, but probably restored population sizes fairly rapidly afterwards. No relationship is found between geographic and genetic distances between populations of L. saxatilis.     

Demographic bottlenecks and low gene flow in remnant populations of the critically endangered <Emphasis Type="Italic">Berchemiella wilsonii</Emphasis> var<Emphasis Type="Italic">. pubipetiolata</Emphasis> (Rhamnaceae) inferred from microsatellite markers

Berchemiella wilsonii var. pubipetiolata (Rhamnaceae) is an endangered plant with only four remnant populations in eastern China. Population genetic information is essential for understanding population history and formulating conservation strategies for this species. Thirteen microsatellite loci were used to investigate genetic variation and population structure of the four remnant populations. Moderate levels of expected heterozygosity (H _E = 0.466–0.543) and low allelic diversity (A = 3.1–3.6 and A _R = 2.2–2.4, respectively) were observed within populations. Bottleneck tests found three out of four populations to deviate from mutation-drift equilibrium under the two-phase model (TPM), suggesting a recent population decline, which is congruent with known demographic history. The evolutionary history of the species seems dominated by genetic drift rather than gene flow. Low historical gene flow was inferred from several different approaches and N _m ranged from 0.582 by the private allele method to 0.783 by the coalescent method. Contemporary gene flow was also found to be even lower for only one first generation migrant was detected with individual-based assignment analysis. Restricted pollen and seed dispersal as well as a recent decline in population size associated with habitat fragmentation may have contributed to low levels of historical and contemporary gene flow, and resulted in a high genetic differentiation. Under this scenario, Berchemiella wilsonii var. pubipetiolata populations are expected to display more pronounced population genetic structure in the future as a result of increased inbreeding and genetic drift.     

Understanding biological conservation strategies: a molecular-genetic approach to the case of myrtle (<Emphasis Type="Italic">Myrtus communis</Emphasis> L.) in two Italian regions: Sardinia and Calabria

Myrtle (Myrtus communis L.), a shrub widespread in the Mediterranean area, is the only species belonging to the Myrtaceae family growing in Europe. The pharmacological and aromatic properties of myrtle have caused a growing interest in this plant. The use of myrtle as an aromatic plant is traditionally established in the Italian regions of Sardinia and Calabria, where it is subjected to an ecological stress due to the large biomass removal for liquor production and environmental damage. The reduction in size and number of wild populations is often related to a loss of genetic variation and reproductive potential. In this study fluorescent amplified fragment length polymorphisms (fAFLPs) were employed to assess the genetic variation within and among natural populations of myrtle from Sardinia and Calabria to gain new insights into their fitness and survival potential. AMOVA analysis indicated that genetic variation was greater within populations (51.86%) than among populations (16.99%), as previously reported for outcrossing species. A significant amount of variation (31.15%) was attributed to variation between Sardinia and Calabria population groups, suggesting a genotypic differentiation between the myrtle populations of these two regions. Intra-population genetic variation, assessed by estimating expected unbiased heterozygosity (H _E), ranged from 0.0595 to 0.2595. These values resulted correlated with population extension (r = 0.918; P < 0.01) and with two reproductive parameters: seed germinability (r = 0.793; P < 0.01) and number of seeds per fruit (r = 0.631; P < 0.05). A moderate gene flow among Sardinia myrtle populations and Calabria populations (1.2719 and 1.0478, respectively) counteracts the low level of genetic variation observed in some␣populations and avoids their differentiation and isolation.     

Population variation and phylogeny in the endangered <Emphasis Type="BoldItalic">Chamaesyce skottsbergii</Emphasis> (Euphorbiaceae) based on RAPD and ITS analyses

Chamaesyce skottsbergii var. skottsbergii is federally listed as an endangered taxon, and is found in small and isolated populations restricted to calcareous soils in dry shrubland habitats on the Hawaiian islands of Oahu and Molokai. Concern over the genetic relationship among these disjunct populations arose as a result of threats to the habitat of the Oahu population. The populations were examined using random amplified polymorphic DNA (RAPD) markers and sequence analysis of the internal transcribed spacer (ITS) region of the rDNA cistron. Chamaesyce skottsbergii var. vaccinioides, a closely related variety found in several small populations on Molokai, was used for baseline comparison of the genetic divergence among populations. RAPD analysis demonstrated that variation within and among populations is the highest for any Hawaiian species examined. Polymorphism was greater than 95% within populations and was 99.4% at the species level. Similarly, measures of genetic similarity indicate that differentiation among these populations is higher than is known for some species. Both RAPD and ITS sequence analysis indicate that populations of C. skottsbergii var. skottsbergii on Oahu and Molokai are genetically distinct, and the extent of this genetic differentiation supports the recognition of these populations as distinct varieties. The Molokai population is in fact much more closely related to var. vaccinioides than to var. skottsbergii on Oahu, and thus should be recognized by the previously used variety name, C. skottsbergii var. audens. Further conservation measures for each of the varieties are addressed.     

Polymorphic microsatellite DNA markers from the marine gastropod Littorina saxatilis

Seven polymorphic microsatellite loci are described in the marine intertidal gastropod Littorina saxatilis. Preliminary data on allelic variation of these loci in a White Sea population of L. saxatilis are presented in order to assess their potential utility as high‐resolution genetic markers for this model species.     

Phenetic diversity in the <Emphasis Type="Italic">Fritillaria camschatcensis</Emphasis> population grown on the Sapporo campus of Hokkaido University

Triploid Fritillaria camschatcensis (L.) Ker-Gawler (2n = 3x = 36) is a wild species growing in the low-lying areas of Hokkaido Island, Japan, including the Sapporo campus of Hokkaido University. Many F. camschatcensis plants grew on the campus about a century ago, but we seldom find the plants nowadays and so a project to restore this species is being planned. Because preservation of genetic diversity and composition in populations has become a major target of conservation, this study compared variation in the F. camschatcensis population on the Sapporo campus with that in two other populations in Hokkaido. Phenetic variation assessed by 57 randomly amplified polymorphic DNA markers showed that the three populations were significantly distinct from each other; analysis of molecular variance showed 64.3% of variation (P < 0.001) existed among the three populations. Comparison of phenetic diversity on the Sapporo campus population with that in the two other populations showed that the Sapporo campus population contained large genetic variation despite reduced plant numbers. These results indicate that multiplying F. camschatcensis individuals on the Sapporo campus is adequate to restore the Sapporo campus population because this population contains enough genetic diversity, and that transplanting from other populations should be avoided so as not to introduce different genotypes into the campus. These results will be used to design the restoration strategy.     

细叶云南松天然种源林遗传多样性的SSR分析

利用12对SSR引物对三个不同种源的细叶云南松群体遗传多样性进行研究。结果表明:共检测到13个位点37个等位基因,每个位点平均观察等位基因数(A)为2.85,多态率为100%;平均有效等位基因数(Ne)1.45。各群体内的有效等位基因数平均为1.447,观察杂合度平均为0.341,期望杂合度平均为0.281。三个群体的Nei’s基因多样度指标的变化范围为0.256~0.297,Shannon多样性指数变化范围为0.448~0.484,各群体间的多态性水平差异不大。细叶云南松群体间的基因分化系数(Gst)为0.089,群体间的遗传分化水平较低,大部分变异均存在群体内。细叶云南松群体间的基因流(Nm)在不同位点的变化范围从4.693~122.189,平均为11.17。说明细叶云南松群体间存在比较充分的基因交流。     

Molecular phylogeny of relict-endemic <Emphasis Type="Italic">Liquidambar orientalis</Emphasis> Mill based on sequence diversity of the chloroplast-encoded <Emphasis Type="Italic">mat</Emphasis>K gene

The genetic diversity and evolutionary divergence in Liquidambar species and Liquidambar orientalis varieties were compared with respect to the matK gene. A total of 66 genotypes from 18 different populations were sampled in southwestern Turkey. The matK region, which is about 1,512 bp in length, was sequenced and studied. L. orientalis, L. styraciflua, and L. formosana had similar magnitude of nucleotide diversity, while L. styraciflua and L. acalycina possessed higher evolutionary divergence. The highest evolutionary divergence was found between L. styraciflua and eastern Asian Liquidambar species (0.0102). However, the evolutionary divergence between L. orientalis and other species was of a similar magnitude. The maximum-parsimony phylogenetic tree showed that L. styraciflua and L. orientalis formed a closer clade while East Asian species were in a separate clade. This suggests that the North Atlantic Land Bridge through southern Greenland may have facilitated continuous distribution of Liquidambar species from southeastern Europe to eastern North America in early Tertiary period. The maximum-parsimony tree with only 18 Oriental sweetgum populations indicated that there were two main clusters: one with mainly L. orientalis var. integriloba and the other with var. orientalis and undetermined populations. High nucleotide diversity (0.0028) and divergence (0.00072) were found in L. orientalis var. integriloba populations and Muğla-1 geographical region. This region could be considered as the major refugium and genetic diversity center for the species. The low genetic diversity and divergence at intraspecies level suggest that L. orientalis populations in Turkey share an ancestral polymorphism from which two varieties may have evolved.     

Microsatellite cross-species amplification in the genus Littorina and detection of null alleles in Littorina saxatilis

Microsatellite DNA is widely used as population genetic marker,but the cost of using microsatellites is high, as they usuallyneed to be developed and optimized for each species separately.Cross-species amplification of microsatellites is thereforecommonly applied to bring down the cost, but it can also involvegenotyping errors. We studied cross-species amplification ofmicrosatellites in four species of the Atlantic group of Littorina(Neritrema): L. saxatilis (Olivi, 1792), L. obtusata (Linnaeus,1758), L. fabalis (Turton, 1825) and L. arcana Hannaford Ellis,1978 to investigate whether markers originally developed fora more distantly related Pacific species [L. subrotundata (Carpenter,1864)] suffered from more amplification problems than markersdeveloped for one of the species in the Atlantic group (L. saxatilis).We also compared variation in amplification success among thespecies and among different regions in the NE Atlantic. Approximatelyhalf of the 12 primers developed for L. subrotundata and theseven primers developed for L. saxatilis were successfully amplifiedin other species of the subgenus. The success was dependenton phylogenetic distance among species within the subgenus.On the other hand, the variation in performance of the locibetween geographically remote populations of the same specieswas as high as variation among the species. In earlier studiesstatistical analyses indicated that several loci showed a heterozygotedeficiency due to null alleles. The presence of null alleleswas confirmed by a segregation analysis of the microsatelliteloci in eight half-sib families of L. saxatilis. (Received 2 April 2007; accepted 19 November 2007)     

A population genetic evaluation of ecological restoration with the case study on <Emphasis Type="Italic">Cyclobalanopsis myrsinaefolia</Emphasis> (Fagaceae)

The ultimate goal of ecological restoration is to create a self-sustaining ecosystem that is resilient to perturbation without further assistance. Genetic variation is a prerequisite for evolutionary response to environmental changes. However, few studies have evaluated the genetic structure of restored populations of dominant plants. In this study, we compared genetic variation of the restored populations with the natural ones in Cyclobalanopsis myrsinaefolia, a dominant species of evergreen broadleaved forest. Using eight polymorphic microsatellite loci, we analyzed samples collected from restored populations and the donor population as well as two other natural populations. We compared the genetic diversity of restored and natural populations. Differences in genetic composition were evaluated using measurements of genetic differentiation and assignment tests. The mean number of alleles per locus was 4.65. Three parameters (A, A _R, and expected heterozygosity) of genetic variation were found to be lower, but not significantly, in the restored populations than they were in the natural populations, indicating a founder effect during the restoration. Significant but low F _ST (0.061) was observed over all loci, indicating high gene flow among populations, as expected from its wind-pollination. Differentiation between the two restored populations was smallest. However, differences between the donor population and the restored populations were higher than those between other natural populations and the restored populations. Only 13.5% and 25.7% individuals in the two restored populations were assigned to the donor population, but 54.1 and 40% were assigned to another natural population. The genetic variation of the donor population was lowest, and geographic distances from the restoration sites to the donor site were much higher than the other natural populations, indicating that the present donor likely was not the best donor for these ecological restoration efforts. However, no deleterious consequences might be observed in restored populations due to high observed heterozygosity and high gene flow. This study demonstrates that during the restoration process, genetic structures of the restored populations may be biased from the donor population. The results also highlight population genetic knowledge, especially of gene flow-limited species, in ecological restoration.     

A geographically-based study of shell shape in small rough periwinkles

A study using principal component analysis and discriminant analysis was carried out on shell shape variation in 3093 specimens of rough periwinkles, 2500 of which were below 5.5 mm in columella length, from around the North Atlantic. Using a combination of colour plus sculpture, and life history trait, the snails were classified by inspection and examination into Littorina nigrolineata, L. arcana, L. saxatilis and L. neglecta. Principal component analyses indicated that similar aspects of variation were important in the different taxa, but these were sometimes of differing levels of importance between L. saxatilis and L. neglecta. Crossvalidation in a discriminant analysis showed classification of shells larger than 5.5 mm to have at least an 88% accuracy. That of shells below 5.5 mm showed an accuracy of 49% in L. arcana, increasing to 54% in L. saxatilis and 63% in L. neglecta, with 76% accuracy for small L. nigrolineata. This last was a special case as only one site was sampled, therefore comparative data are not available. This geographically-based study reveals that L. neglecta is more homogeneous over its range than recently reported by other workers and shows greater differences from L. saxatilis than the latter does from either L. nigrolineata or L. arcana. Size effects do not account for these differences because L. neglecta is morphometrically distinct from both large and small L. saxatilis. Furthermore, small, mid-shore L. saxatilis classify with large high-shore L. saxatilis in discriminant analysis, not with L. neglecta. These results provide evidence that the taxon L. neglecta is more distinct than has sometimes been suggested.     

Population Genetics of Indigenous Quercus robur L. Populations and of Derived Half‐Sib Families has Implications for the Reproductive Management of the Species

Abstract: In the Netherlands indigenous Quercus robur L. populations are rare and have been maintained as patches in ancient woodland. For adequate conservation of these populations, information about genetic variation and population structure is necessary. In order to assess the genetic variation and structure of these populations, microsatellite polymorphisms were studied in two autochthonous populations. These two populations differed slightly for their gene diversity, which was as high as was observed for Q. robur populations in France and Germany. For reforestation purposes there is an interest in the genetic variation of a half‐sib family harvested from one tree. The gene diversity of the two studied half‐sib families ‐ obtained from a forest and an urban area ‐ was similar, but relatively low. This indicates that, for reforestation purposes, seeds should be harvested from many different trees in order to obtain a population with a genetic variation as high as was observed for an autochthonous population.     

Conservation Genetics of Mexican Beech, Fagus grandifoliavar. mexicana

Fagus grandifolia var. mexicana(Fagaceae) is a Mexican endemic tree, currently threatened with extinction. In order to assess the level and structure of genetic variation in four remaining populations, leaf samples were analysed using random amplified polymorphic DNA (RAPD) and cpDNA PCR-RFLP markers. A sample of the more widespread congener, F. grandifoliavar. grandifolia from the USA was also analysed for comparison. Thirty-three polymorphic RAPD bands were produced using 18 10-mer primers. AMOVA of RAPD data indicated significant (P < 0.002) population differentiation, with 15.6% of variation recorded between Mexican populations. PCR-RFLP analysis enabled three cpDNA haplotypes to be identified, denoted types A, B, and C. Types A and B were each restricted to an individual Mexican population, whereas Type C was fixed for two Mexican populations, and the population from the USA. Within-population genetic variation, quantified as percentage polymorphic bands, Shannon's Diversity Index and Nei's gene diversity measure, was found to be lower in Mexican populations than in that from the USA, and was positively related to population size. These results suggest that an unexpectedly high degree of genetic variation exists within Mexican beech, and this variation should be considered in developing the conservation strategy that is urgently required if extinction of this taxon is to be prevented.     

Genetic structure of Japanese Chattonella marina (Raphidophyceae) populations revealed using microsatellite markers

Chattonella marina var. antiqua and C. marina var. marina (Raphidophyceae) are red tide‐forming, harmful phytoplankton species. We investigated the genetic diversity and genetic relationship among the populations using microsatellite markers to identify putative sources of C. marina var. antiqua and C. marina var. marina in Japanese coastal populations. A positive correlation between genetic divergence and geographical distance (isolation by distance) was recognized for C. marina var. antiqua. The C. marina var. antiqua populations were established throughout a geological time scale, and genetic divergence had progressed in each population with gene flow depending on geographic distances. In contrast, isolation by distance was not observed for C. marina var. marina populations, and the genetic divergence among populations was extremely high. The Tokyo Bay population of C. marina var. marina, which was first recognized in 2008, had many private alleles but was related to the Kagoshima Bay population. The Tokyo Bay population may have been established by several invasions from the Kagoshima Bay population and other regions.