Cytological and hybridization studies inLeucanthemum (Compositae—Anthemideae) from North Africa

A cytological survey ofLeucanthemum species in North Africa has shown that they are all diploid with a somatic chromosome number of 2n = 18. New counts are reported for 8 species and 1 subspecies. 11 taxa were used in a crossing programme and the results show a good agreement between phenetic distance based on morphological characters and genetic differentiation as expressed in the ability of the species to hybridize. A strong barrier to gene exchange exists between European and North African species. Bagging experiments showed that the North African species comprise a self-incompatible group of cytogenetically related species.     

Phenotypic plasticity and gene diversity in Pistacia lentiscus L. along environmental gradients in Israel

The Mediterranean common shrub Pistacia lentiscus is distributed in a wide range of habitats along the climatic gradient in Israel. We studied the factors that may shape its morphological, physiological, and genetic differentiation. We examined the phenotypic and molecular genetic variability among and within the six Israeli populations as correlated with the local environmental conditions. The genetic structure of the shrub on the island of Cyprus was also examined. Plant morphological parameters correlated significantly with the local environmental conditions, especially with the annual precipitation and temperature. Gene diversity did not differ significantly among locations, and, hence, no differentiation among Israeli populations or between populations in Israel and Cyprus was found. The major part of the molecular variance (69%) was found within the populations, 22% of the variance was found between Israel and Cyprus and 9% among the populations within the region. Gene flow estimates among all the tested populations were high with no indication for the isolation by distance. We did not find any pattern of ecologically related genetic differentiation; hence, the morphological and physiological differences are probably due to phenotypic plasticity. It seems that the ability of P. lentiscus to express the different phenotypes in response to the varying conditions in the Mediterranean region is an adaptive trait in a species that is characterized by intensive gene flow.     

Relationships among species inCarex sect.Phyllostachys (Cyperaceae) based on allozyme divergence

Allozyme variation was examined inCarex sect.Phyllostachys (Cyperaceae) to provide insight into phylogenetic relationships hypothesized in an earlier study and to determine the degree of genetic differentiation within and between taxa. Genetic identity values are concordant with the morphological differences found between species. The lowest values are found between species with the greatest morphological dissimilarity. Conversely, the highest values are associated with species pairs distinguished by relatively few morphological differences. Conspecific populations possess high genetic identities, although interpopulation differentiation has characterized the evolutionary history of some species. Geographic patterning is also evident within species, with geographically proximate populations often having the highest identity values. Phylogenetic trees produced using different cladistic methods were poorly supported and varied in their depiction of relationships among species. One cladogram produced using presence/absence allelic data is more or less congruent with a topology recovered from an earlier analysis utilizing molecular and morphological data. The wide- and narrow-scaled clades are maintained as are the sister species pairsC. backii/C. saximontana, C. basiantha/C. superata, andC. jamesii/C. juniperorum. Contrary to the finding of our previous study, however,C. willdenowii is aligned withC. jamesii/C. juniperorum.     

Phylogeography of island canary (Serinus canaria) populations

Island canaries (Serinus canaria) are characterised as a species living exclusively on North Atlantic islands, mainly on the Azores, Madeira and Canary Islands. Although they are very common in their habitats, their behaviour and breeding system has only recently been studied systematically. To advance the understanding of their ecology and to see if the rather isolated archipelagos are already promoting a genetic differentiation, we investigated their phylogeographic relationship as revealed by mtDNA sequences of the cytochrome b gene and investigated whether this measure corresponds to morphological characteristics within the islands. Genetic distances were very low throughout the distribution range of the species. Although the variation of genetic distances within the population of Pico (Azores) was larger than that on Madeira and Canary Islands, the genetic distances between island populations were very low throughout which prevented a clear phylogeographic differentiation. Moreover, morphological measurements did not reveal a consistent pattern to reliably separate the populations, although the measures of beak length and body weight revealed a clear island-specific differentiation. These data lead to the assumption that the colonisation of the Atlantic islands by the canaries occurred very recently, while there is no persisting gene flow between the populations.     

Allozymic Differentiation Among Geographically Distant Populations of Patella vulgata (Mollusca, Patellogastropoda)

Patella vulgata is a boreal cold temperate species and is the dominant limpet in northern Europe. Few works have focussed on the population genetics of this species. Therefore, the aim of this work was to assess the degree of genetic and morphological differentiation of P. vulgata on a macroscale by using 20 allozyme loci and 6 morphological variables. Samples were taken from the following locations: Dingle Peninsula (Southwest Ireland), Port Erin (Southwest Isle of Man), St. Bees Head (north Cumbria, England), St. Agnes Head (north Cornwall, England), Cellar Beach (south Devon, England), Whitley Bay (north Newcastle-Upon-Tyne, England), Sines (Portugal), and Pointe de Chanchardon, La Rochelle (Bay of Biscay, France). Morphological variables were analysed by the multivariate Canonical discriminant analysis. Genetic variation was assessed by diversity measures such as polymorphism and heterozygosity; genetic subdivision of P. vulgata population was determined by the estimator θ of F _ST, and the genetic similarity between populations was measured by Nei’s genetic identity. No significant morphological differentiation was observed among samples. Moderate genetic population subdivision was observed (θ = 0.137±0.074) despite great geographic distances. The minimum genetic identity observed was between Ireland and France (I = 0.942) and maximum was observed between Portugal and north-east England (I=0.998). Two main groups were shown by UPGMA cluster analysis (I = 0.965). One formed by Irish, Manx, north Cumbria, and curiously, south Devon samples, while the second includes Portuguese, French, north-Newcastle-upon-thyne, and north Cornwall samples. No association (g = 0.956; p>0.050) was found between pair-wise genetic divergence and geographic distance separating subpopulations, mainly due to an unexpected pattern of genetic heterogeneity found in Southwest England.     

Maintenance of genetic and morphological identity in two sibling Syrrhopodon species (Calymperaceae,Bryopsida) despite extensive introgression

Bryophytes are a group of land plants in which the role of hybridization has long been challenged. Using genotyping by sequencing to circumvent the lack of molecular variation at selected loci previously used for phylogeny and morphology, we determine the level of genetic and morphological divergence and reproductive isolation between the sibling Syrrhopodon annotinus and S. simmondsii (Calymperaceae, Bryopsida) that occur in sympatry but in different habitats in lowland Amazonian rainforests. A clear morphological differentiation and a low (0.06), but significant Fst derived from the analysis of 183 single nucleotide polymorphisms were observed between the two species. Conspecific pairs of individuals consistently exhibited higher average kinship coefficients along a gradient of geographic isolation than interspecific pairs. The weak, but significant genetic divergence observed is consistent with growing evidence that ecological specialization can lead to genetic differentiation among bryophyte species. Nevertheless, the spatial genetic structures of the two species were significantly correlated, as evidenced by the significant slope of the Mantel test based on kinship coefficients between pairs of interspecific individuals and the geographic distance separating them. Interspecific pairs of individuals are thus more closely related when they are geographically closer, suggesting that isolation‐by‐distance is stronger than the interspecific reproductive barrier and pointing to interspecific gene flow. We conclude that interspecific introgression, whose role has long been questioned in bryophytes, may take place even in species wherein sporophyte production is scarce due to dioicy, raising the question as to what mechanisms maintain differentiation despite weak reproductive isolation.     

Allozyme diversity in relation to geographic distribution and population size inLathyrus vernus (L.) Bernh. (Fabaceae)

Lathyrus vernus (L.) Bernh. is a diploid, long-lived perennial and insect-pollinated herb with no special adaptation to long-distance dispersal. It occurs on neutral soil in deciduous forests throughout western Eurasia. Due to specific habitat preferences,L. vernus has a fragmented distribution with isolated populations. We investigated allozyme variation at eleven loci in 20 populations ofL. vernus from one geographically central region (the Czech Republic and the Slovak Republic) and two geographically marginal regions (southern and central Sweden) in the species present-day distribution. There was a clear differentiation between the three regions and the genetic distance between the populations was highly correlated with geographic distance. The total genetic diversity (H_T) was 0.354. The proportion of genetic diversity due to differentiation between regions, and to differentiation between populations within regions, accounted for 10% each. There was no difference in level of genetic diversity between the three regions. No significant difference in level of genetic diversity was found between small and large populations. The genetic diversity inL. vernus may either be a result of the long generation-time of the species or peculiarities in the post-glacial migration species, e.g. survival only in refugia far east of the sampled populations and/or migration as a continuous process not involving founder-events.     

Patterns of genetic differentiation in two annual bromegrasses,Bromus lanceolatus andB. hordeaceus (Poaceae)

Infraspecific genetic differentiation was analysed in two tetraploid annual bromegrasses,Bromus lanceolatus (from N Africa) andB. hordeaceus (from N Africa and France). Genetic analysis of populations was based on allozyme polymorphisms at 17 loci. Different fixed heterozygous phenotypes were scored in both species, according to their allopolyploid origin. In N Africa, more variation occurred among populations ofB. lanceolatus than ofB. hordeaceus. The variation was not randomly distributed among populations of both species. InB. lanceolatus, differentiation was correlated with climatic variables rather than with geographic distance between populations. Higher correlation of genetic differentiation with geographic distance occurred inB. hordeaceus, particularly at large geographic scale, between French and N African populations. Within each region, the populations appeared weakly genetically differentiated, even when belonging to different subspecies.     

Genetic Variation in Hippophae rhamnoides ssp. sinensis (Elaeagnaceae) Revealed by RAPD Markers

Hippophae rhamnoides ssp. sinensis is endemic to China, and it is a dioecious, outcrossing plant. Although many studies have been undertaken mainly on its agricultural, nutritional, medical, and ornamental value, little is known about its population genetics. This study uses random amplified polymorphic DNA to investigate the genetic diversity and population genetic structure of 13 natural populations of the subspecies sinensis. Fifteen primers amplified 107 reproducible bands, with 95 (88.79%) being polymorphic. The gene diversity within population was 0.168, considerably lower than that of tree species and most perennial, outcrossing species, but higher than that of annual or short-lived, selfing species. The Gst value showed that 18.3% of the total genetic variation resided among populations, a little lower than that of outcrossing species. The present results are quite similar to those previously reported in another subspecies, H ssp. . rhamnoides rhamnoides. The low genetic differentiation among populations in ssp. sinensis may be attributed to the long-distance dispersal of seeds facilitated by birds, in addition to its characteristics of outcrossing, wind pollination, and widespread distribution. No association between genetic distance and geographical distribution was found. The population relationships revealed by the UPGMA dendrogram parallel this result, in that genetic distance did not increase with geographic separation. This pattern of population differentiation may imply the adaptation of ssp. s populations to the local environment, given that its habitats vary greatly across its distribution.     

喀斯特耐阴灌木淡黄金花茶的精细空间遗传结构与基因流

淡黄金花茶(Camelia flavida)是一种具有淡黄色花瓣的金花茶植物,为喀斯特典型的耐阴灌木。基于广西弄岗喀斯特季节性雨林15 hm2(300 m×500 m)监测样地,利用13对微卫星(SSR)标记,探讨喀斯特地貌对淡黄金花茶的精细空间遗传结构(SGS)、基因流和小尺度范围遗传分化的影响。结果表明:(1)淡黄金花茶在50 m距离内产生显著的SGS,在灌木类型和依赖种子传播的物种中具有中等程度的SGS强度(Sp=0.0248);(2)淡黄金花茶种子和花粉传播平均距离较短,分别为12.47 m和29.03 m,72.2%花粉和81.0%种子的传播距离均主要是在20 m内;(3)淡黄金花茶在小尺度范围(种群间距离1 km)的4个种群,甚至相距100 m的两个斑块产生了显著的遗传分化。喀斯特生境地貌对淡黄金花茶的基因流产生阻碍作用,从而导致淡黄金花茶在小尺度范围产生遗传分化。     

Genetic analyses of the federally endangered Echinacea laevigata using amplified fragment length polymorphisms (AFLP)—Inferences in population genetic structure and mating system

Echinacea laevigata (Boynton and Beadle) Blake is a federally endangered flowering plant species restricted to four states in the southeastern United States. To determine the population structure and outcrossing rate across the range of the species, we conducted AFLP analysis using four primer combinations for 22 populations. The genetic diversity of this species was high based on the level of polymorphic loci (200 of 210 loci; 95.24%) and Nei’s gene diversity (ranging from 0.1398 to 0.2606; overall 0.2611). There was significant population genetic differentiation (G_ST = 0.294; Ө^II = 0.218 from the Bayesian f = 0 model). Results from the AMOVA analysis suggest that a majority of the genetic variance is attributed to variation within populations (70.26%), which is also evident from the PCoA. However, 82% of individuals were assigned back to the original population based on the results of the assignment test. An isolation by distance analysis indicated that genetic differentiation among populations was a function of geographic distance, although long-distance gene dispersal between some populations was suggested from an analysis of relatedness between populations using the neighbor-joining method. An estimate of the outcrossing rate based on genotypes of progenies from six of the 22 populations using the multilocus method from the program MLTR ranged from 0.780 to 0.912, suggesting that the species is predominantly outcrossing. These results are encouraging for conservation, signifying that populations may persist due to continued genetic exchange sustained by the outcrossing mating system of the species.     

High levels of genetic structuring as a result of population fragmentation in the tropical tree species <Emphasis Type="Italic">Caesalpinia echinata</Emphasis> Lam.

We have investigated levels of genetic diversity within and among seven remnant populations of Caesalpinia echinata Lam., an endangered species found as fragmented populations in three major areas around the coastal regions of Brazil. Using amplified fragment length polymorphism (AFLP) genetic markers, we detected levels of within-population genetic diversity ranging from 0.092 to 0.163, with the lowest values generally being found in the smallest populations. Estimates of between-population genetic differentiation were strongly correlated with geographical distance (r = 0.884, p < 0.001), which,along with a neighbour-joining phylogenetic analysis, strongly suggested high levels of genetic isolation by distance. Over half (62%) of the total genetic diversity was partitioned between populations, further highlighting the genetic distinctness of individual populations. Taken together, these results suggest that fragmentation has led to an increase in population differentiation between fragments of C. echinata. These formations will be of great value in the development of conservation plans for species exhibiting high levels of genetic differentiation due to fragmentation, such as indication of conservation unit size, which populations should be chosen as priority in conservation plans and which samples should be introduced in areas with a low number of individuals of brazilwood.     

GENETIC CONSEQUENCES OF SEED DISPERSAL IN THREE SYMPATRIC FOREST HERBS. I. HIERARCHICAL POPULATION-GENETIC STRUCTURE

To examine the effects of seed dispersal on spatial genetic structure, we compare three sympatric species of forest herbs in the family Apiaceae whose fruits differ widely in morphological adaptations for animal-attached dispersal. Cryptotaenia canadensis has smooth fruits that are gravity dispersed, whereas Osmorhiza claytonii and Sanicula odorata fruits have appendages that facilitate their attachment to animals. The relative seed-dispersal ability among species, measured as their ability to remain attached to mammal fur, is ranked Sanicula > Osmorhiza > Cryptotaenia. We use a nested hierarchical sampling design to analyze genetic structure at spatial scales ranging from a few meters to hundreds of kilometers. Genetic differentiation among population subdivisions, estimated by average genetic distance and hierarchical F-statistics, has an inverse relationship with dispersal ability such that Cryptotaenia > Osmorhiza > Sanicula. In each species, genetic differentiation increases with distance among population subdivisions. Stochastic variation in gene flow, arising from seed dispersal by attachment to animals, may partly explain the weak relationship between pairwise spatial and genetic distance among populations and heterogeneity in estimates of single locus F-statistics. A hierarchical island model of gene flow is invoked to describe the effects of seed dispersal on population genetic structure. Seed dispersal is the predominant factor affecting variation in gene flow among these ecologically similar, taxonomically related species.     

Differences in allozyme patterns between Diaphanosoma brachyurum and Diaphanosoma mongolianum,as revealed in Central European populations

Recent taxonomic revisions in the genus Diaphanosoma have made the uncertainty in species discrimination with morphological characters obvious. Therefore species characterization on a genetic basis seems to be required.Here we examined the genetic structure of two Diaphanosoma mongolianum populations and three Diaphanosoma brachyurum populations in Central Europe by allozyme electrophoresis. A genetic differentiation between both species was evident. 5 out of 9 tested loci carried diagnostic alleles. Both species differed in their habitat choice: D. mongolianum was adapted to higher trophic levels than D. brachyurum. Co-occurrence was observed in a eutrophic lake. Populations sampled from their preferred habitats showed high genotype diversities.Most genotype frequency distributions were in agreement with Hardy-Weinberg expectations and temporal (between years) changes in genotypic compositions were not common. This indicated annually occurring bouts of sexual reproduction, similar to intermittent Daphnia populations. Genetic differentiation among lakes suggested restricted gene flow.     

Allozyme diversity and systematics inAnnonaceae—a pilot project

Five species ofAnnona and one species fromArtabotrys, Cananga, Polyalthia, andRollinia were investigated in regard to 11 different allozyme loci. Preliminary studies on small population samples ofAnnona suggest genetic uniformity in three species and variability within and between populations in two other species. The allotetraploid origin ofA. glabra is clearly shown by its hybrid enzyme bands. The genetic distance between fiveAnnona species partly corresponds with their morphological relationships; onlyA. muricata appears more separated than is suggested by morphology. A comparison of the five genera demonstrates close relationship betweenAnnona andRollinia. Two enzyme loci are identical within all taxa investigated and possibly may serve as a genetic marker for the family.     

Integrated analyses of chromosome, molecular and morphological variability in the Andean mice Eligmodontia puerulus and E. moreni (Rodentia, Cricetidae, Sigmodontinae)

Patterns of evolution and systematics of sigmodontine rodents are matters of continuous revision and debate. The silky mouse, Eligmodontia, is a phyllotine rodent adapted to arid environments. Chromosomal and molecular data have identified six species in this genus. Among these E. puerulus and E. moreni are sister taxa from the high Andean and lowland deserts, respectively, with large chromosomal differences and intermediate levels of molecular divergence. The purpose of our study was to quantify the degree of variability (morphological, cytogenetic, and molecular) and to analyze its evolutionary implications within, and between, these sister species in the Monte and Puna biomes of Argentina. Our results show a high variability at the chromosomal and molecular level, but low morphological differentiation among populations of E. puerulus. Diploid numbers vary from 31 to 37 due to a complex Robertsonian system, whereas cytochrome-b distances range from 0.15% to 5.75%. On the other hand, E. moreni shows high morphological differentiation between populations, but low intraspecific differentiation at the molecular (from 0.73% to 1.4%) and chromosomal level (2n = 52). Comparison of E. puerulus with E. moreni reveals high morphological and chromosomal distinction between them, but absence of molecular differentiation. Our results suggest that: (1) the high genetic variability of E. puerulus could be associated to its geographic distribution in the complex topography of the high Andean Puna; (2) the high morphological differentiation between E. moreni and E. puerulus could be the result of natural selection; and (3) molecular polyphyly between E. puerulus and E. moreni could be due to introgression or a recent split of these taxa. Finally, our results emphasize the need to integrate different datasets in the analysis of species variability and diversification, as tools for understanding their evolutionary histories.     

不同种源马尾松ISSR遗传结构及影响因素分析

应用ISSR分子标记技术,对来自广西、贵州3个种源的马尾松开展遗传多样性、遗传结构及遗传距离等研究。结果表明：从100条引物中筛选出12条引物,共扩增出92个条带,86条具有多态性。 POPGENE分析显示：马尾松群体水平上的Nei’ s基因多样性指数的变化范围为0.1824~0.2065,Shannon遗传多样性指数的变化范围为0.2818~0.3178,3个群体的多态性水平差异不大;物种水平上的多态性百分率为93.48％, Nei’ s基因多样性指数为0.2842,Shannon信息指数为0.4381;表明马尾松在物种水平上具有较高水平的遗传多样性。遗传结构分析显示：马尾松的基因分化系数( Gst)为0.3153,表明遗传变异主要来源于群体内;基因流Nm为1.0853,表明不同群体间存在一定的基因流动。 AMOVA分析显示：马尾松的遗传分化指数( Fst)为0.246( P＝0.001),表明群体间已出现明显的遗传分化。 UPGMA聚类和Mantel检测结果显示：每个群体内的个体均能很好地首先聚集为一个分支,群体间的遗传距离与地理距离之间存在显著相关性( r＝0.972, P＝0.001)。这说明马尾松在裸子植物界中具有较高水平的遗传多样性,遗传变异主要分布于群体内,群体间已出现了明显的遗传分化,这种分化并非由遗传漂变引起,可能与地理生境的差异有关。     

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.     

Changes in morphological traits of the cabbage aphid (<Emphasis Type="Italic">Brevicoryne brassicae</Emphasis>) associated with the use of different host plants

The genetic and morphological differentiation of insect populations in relation to the use of different host plants is an important phenomenon that predates ecological specialisation and speciation in sympatric conditions. In this study, we describe the morphological variation of populations of Brevicoryne brassicae (Homoptera: Aphididae) associated with two host species, Brassica oleracea and Brassica campestris, which occur sympatrically in the highlands of Chiapas, Mexico. The study is aimed at obtaining evidence regarding phenotypic differentiation induced by, or associated with, the use of distinct but closely related host species. Seven morphological characters were measured in 696 wingless aphids collected from plants of the two host species at four localities. Morphological variation was summarised through principal components analysis (PCA). Sixty-two percent of morphological variation was explained by the first two PCs. The first component (PC1) was related to the general size of appendages, and PC2 was interpreted as the relationship between body size (body and leg size) and antenna length. Aphids growing on B. campestris were bigger than those collected from B. oleraceae. Significant differences between hosts were detected for PC1, whereas a significant effect of locality, host, and the interaction locality × host was detected for PC2. These results indicate that the average phenotype of B. brassicae individuals inhabiting different host-plant species differs as a consequence of the contrasting feeding environments the host species provide.     

Wheat phylogeny determined by RFLP analysis of nuclear DNA. 2. Wild tetraploid wheats

Intra- and inter-specific variations in the nuclear DNA of Triticum dicoccoides Körn. (2n = 28, genome constitution AABB) and T. araraticum Jakubz. (2n = 28, AAGG), wild species, respectively, of the Emmer and Timopheevi group, were studied by restriction fragment length polymorphism (RFLP) analysis. Total DNAs of 32 T. dicoccoides and 24 T. araraticum accessions, collected from throughout the distribution areas of these species, were treated with two 6-bp cutters and hybridized with 30 nuclear DNA clones as probes to detect RFLPs. A total of 167 hybrid bands were observed per accession. All the enzyme-probe combinations showed RFLPs between accessions. The average genetic distance between the T. dicoccoides accessions was 0.0135 ± 0.0031 and that between the T. araraticum accessions 0.0036 ± 0.0015, indicative of about a four-fold intraspecific variation in T. dicoccoides as compared to T. araraticum in terms of genetic distance. No significant genetic differentiation was found for the geographical populations of these species, the genetic distance between the two species being 0.0482 ± 0.0022. The interspecific divergence corrected for intraspecific divergence was 0.0395, about three times that for T. dicoccoides and 11 times that for T. araraticum. The results show that in the wild state the Emmer and Timopheevi groups are clearly differentiated and that T. dicoccoides has much greater variation than T. araraticum, suggesting a relatively recent origin for the latter and therefore a diphyletic origin for these species.Contribution from the Laboratory of Genetics, Faculty of Agriculture, Kyoto University, Japan, No. 535