Genotypic variation in agamospermous Erigeron compositus (Asteraceae)

Few integrative analyses of the structure of agamospermous plant populations have been conducted. Erigeron compositus occurs in montane western North America and comprises both sexual and agamospermous populations. Sexual E. compositus has previously been characterized as outcrossing and predominantly diploid (2n = 18). Agamic E. compositus is usually hexaploid (2n = 54), though counts herein range from 2n = 36 to 2n = 80. Starch-gel electrophoresis, cytology, and analysis of pollen production were used to evaluate variation within and among agamospermous populations. Fifteen enzyme loci were used to identify 24 unique multilocus genotypes in seven populations, an average of 3.4 genotypes per population. Proportion of distinct genotypes per population sample size (GIN) and measures of genetic diversity (D) and evenness (E) are 0.10, 0.48, and 0.61, respectively, which indicate that E. compositus maintains levels of diversity similar to other agamospermous taxa. Most agamospermous populations are mosaics comprising groups of genetically distinct individuals that are frequently distinguished by cytotype and capacity for pollen production. The geographical and ecological separation of sexual and agamospermous populations make it unlikely that gene flow from sexual populations is a direct source of genetic variation in agamospermous populations. Instead, crossing between genetically distinct facultative agamosperms probably accounts for most variation. Genetic and morphological evidence document one such putative crossing event. Agamospermous E. compositus is very similar genetically to sexual E. compositus. Allozyme analysis further shows that genetic variation in agamospermous populations is partitioned among a few highly heterozygous genotypes, whereas sexual populations maintain numerous genotypes of relatively low heterozygosity.     

Evolution of Agamospermous Races in Boehmeria and Eupatorium

Abstract Recent progress in research on the evolution of agamospermy in Boehmeria (Urticaceae) and Eupatorium (Asteraceae) is reviewed. Specific questions addressed are: (1) Is there any trend in distributions and habitat preferences of sexual and agamospermous races? (2) Is hybridity or a high level of heterozygosity a prerequisite for the evolution of agamospermous races? (3) Do agamospermous plants produce genetically variable offsprings through hybridization with sexual relatives? and (4) Do agamospermous plants have the theoretically predicted reproductive advantage over sexuals? From the results presented, the following conclusions can be drawn. (1) Sexual and agamospermous relatives are usually ecologically differentiated; sexuals tend to require extreme habitats, while agamosperms prefer disturbed places. (2) Hybridity or a high level of heterozygosity is not a prerequisite to the evolution of agamospermous races. (3) Agamospermous races produce genetically variable offspring through hybridization with sexual relatives and subsequent segregation. (4) Agamospermous plants have a reproductive advantage often greater than twofold; part of this advantage is attributed to polyploidy. The question of why agamospermous races do not replace sexuals remains open. Male recovery in agamosperms through hybridization and/or the disadvantage of polyploidy associated with agamospermy might work as mechanism(s) enabling sexuals to resist colonization of reproductively superior agamosperms under some extreme ecological conditions.     

Evidence for multiple, autoploid origins of agamospermous populations in Eupatorium sessilifolium (Asteraceae)

Comparative analyses were made of agamospermous populations of Eupatorium sessilifolium, which have previously been documented to be polyploid, to determine whether they are alloploid or autoploid in origin and to assess the possibility that they have arisen more than once. There was no variability in ITS sequences among seven agamospermous and eight sexual diploid populations of E. sessilifolium, which is consistent with morphological observations in suggesting that the agamospermous populations were autoploids. The ITS sequence characteristic of E. sessilifolium differs from all other North American species by a minimum of 15 changes, and heterogeneity or polymorphism would be expected if the agamospermous populations were alloploids. Analysis of the chloroplast-based trnC-psbM spacer region showed variability among both sexual diploid and agamospermous populations of E. sessilifolium, which suggested that the agamospermous populations stem from multiple origins. Analysis of ISSR data revealed considerable intraspecific variability within E. sessilifolium, and the distribution of variability, with agamospermous populations showing variability from one another, added further evidence for multiple origins of agamospermous populations. The results in conjunction with distributional evidence that the sexual diploid populations of E. sessilifolium are geographically restricted and uncommon suggest that monitoring of populations might be warranted to evaluate whether measures are needed to enhance their continued survival.     

Distribution of Sexual and Agamospermous Populations of Eupatorium (Compositae) in Asia

Abstract Examinations of chromosome number voucher specimens show that sexual diploid and agamospermous polyploid plants of Eupatorium in Asia can be distinguished by morphology, fertility, and stainability of pollen grains. Using these criteria, reproductive systems (sexual vs. agamospermous) were estimated for 558 herbarium specimens of Eupatorium in East Asia. Of 22 taxa examined, six included both sexual and agamospermous specimens; those of one taxon were all agamospermous; and those of 15 taxa were all sexual. This result shows that 21 taxa are not agamospermous microspecies but are differentiated at the diploid level. Sexual populations of most taxa had restricted geographical distributions. Eupatorium chinense var. chinense and var. oppositifolium consisted of both sexual and agamospermous populations. Sexual populations of these two varieties were allopatric and distinct in external morphology, suggesting differentiation at the species level. The specimens of E. heterophyllum s. str. were all agamospermous, while those of E. mairel , often treated as synonymous with E. heterophyllum , were sexual.
In comparing East Asian and North American Eupatorium for the distributional patterns of sexual and agamospermous populations, three differences can be pointed out: (1) Agamospermous plants of autopolyploid origin have evolved in eight species and are widespread in North America; while most agamospermous plants in East Asia may be of allopolyploid origin, have relatively restricted ranges, and are less frequent than the diploid plants with the exception of in E. chinense var. oppositifolium , (2) The number of sexual species with wide range is greater in North America than in East Asia. (3) The number of sexual species with restricted ranges is greater in East Asia than in North America. The results obtained suggest that speciation among the plants of Eupatorium has occurred more recently in East Asia than in North America.     

A taxometric analysis ofLimonium (Plumbaginaceae) in Western Europe

A taxometric analysis ofLimonium species in Western Europe has provided evidence for the subgeneric groupings proposed by other authors. A cluster analysis suggests that the origin of sexual and agamospermous species has taken place separately. Sexual and agamospermous species are usually clustered separately. The lack of a sexual process in many agamospermous species, as indicated by the distribution of incompatibility morphs and by very low pollen fertility, suggests that the evolution of sexual and agamospermous taxa has been by different processes. It is suggested that following the origin of agamospermous species or species groups by hybridisation, subsequent evolution in the agamospermous taxa has occurred by asexual means. Evidence from the distribution of pollen/stigma incompatibility morphs, especially the existence of self-compatible agamospermous species supports this hypothesis.     

A comparison of within-plant karyological heterogeneity between agamospermous and sexualTaraxacum (Compositae) as assessed by the nucleolar organiser chromosome

Morphological variation for the NOR chromosome was studied for four half-siblings of a sexual outbreedingTaraxacum, for three siblings of the obligate agamospermT. pseudohamatum, and for two individuals of the agamospermT. brachyglossum. No rearrangement was detected for the 113 chromosomes of sexuals, or for 41 chromosomes of two agamospermous individuals. In the other three agamospermous individuals, 3/16, 5/50, and 5/20 chromosomes showed evidence of chromosomal rearrangement. The majority of rearrangement events (10/13) occurred to the satellite rather than to the body of the NOR-chromosome. It is considered that such high levels of somatic chromosomal rearrangement in agamospermousTaraxacum may be the result of activity by transposable genetic elements. This recombination may be of selective advantage to asexual plants which cannot generate genetic variability through the sexual process.     

Polymorphic microsatellite loci for the marine angiosperm Cymodocea nodosa

The seagrass Cymodocea nodosa (UCRIA) Ascherson represents a good model to assess the relative contribution of clonal and sexual reproduction to genetic structure in marine plant populations. Seven microsatellite loci with repeat units consisting of one trinucleotide, four simple dinucleotides and two complex dinucleotides are described here. The seven loci are characterized by high number of alleles (from three to 13) and high heterozygosity (H_O ranging from 0.240 to 0.860) in the tested populations. Conditions for multiplex polymerase chain reactions are also described.     

Distribution of reproductive systems and taxonomical concepts in the genus Taraxacum F.H. Wigg. (Asteraceae,Lactuceae) in Germany

Reproductive studies of the genus Taraxacum F.H. Wigg. (Asteraceae) were carried out in the Eastern part of Germany (approx. 1,300 individuals). Using the method of variability of pollen diameter we observed almost in all cases only obligately agamospermous plants in the area studied. With regard to Germany, regions with 1. obligately sexual and obligately agamospermous 2. facultatively and obligately agamospermous 3. exclusively obligately agamospermous individuals, could be distinguished. From these facts, it is concluded that the microspecies concept is preferable to the traditional macrospecies concept, especially in regions with exclusively obligately agamospecies. The microspecies concept is also applicable in a differentiated form in the remaining regions.     

Clonal diversity in the agamospermous polyploids ofTaraxacum hondoense in northern Honshu,Japan

Chromosome numbers and allozyme variations were surveyed in 74 polyploid populations ofTaraxacum hondoense, in northern Honshu, Japan. Most of the populations (94.4%) consisted of triploid (2n=24), indicating the predominance of this ploidy level. Approximately 42.6% were found to contain tetraploid (2n=32), and a few plants were pentaploid (2n=40). Electrophoretic analysis at6 Pgdh-1 revealed twelve phenotypes with four alleles (including one putative null allele). The triploids showed excessive heterozygosity (82.4%) and all of the tetraploids and pentaploids were heterozygote. Phenotype IV was the most frequent and widely distributed in northern Honshu. Forty five percent of the populations were found to contain multiple phenotypes at 6Pgdh-1. A total of 21 clones were distinguished using three polymorphic loci (6Pgdh-1, Got andMdh), and a considerable amount of clonal diversity was detected both within and among polyploid populations ofT. hondoense. Factors causing multiclonality in agamospermous polyploids are discussed.     

Race structure within the Mesoamerican gene pool of common bean (Phaseolus vulgaris L.) as determined by microsatellite markers

Common bean (Phaseolus vulgaris L.) cultivars are distinguished morphologically, agronomically and ecologically into specific races within each of the two gene pools found for the species (Andean and Mesoamerican). The objective of this study was to describe the race structure of the Mesoamerican gene pool using microsatellite markers. A total of 60 genotypes previously described as pertaining to specific Mesoamerican races as well as two Andean control genotypes were analyzed with 52 markers. A total of 267 bands were generated with an average of 5.1 alleles per marker and 0.297 heterozygosity across all microsatellites. Correspondence analysis identified two major groups equivalent to the Mesoamerica race and a group containing both Durango and Jalisco race genotypes. Two outlying individuals were classified as potentially of the Guatemala race although this race does not have a defined structure and previously classified members of this race were classified with other races. Population structure analysis with K = 1–4 agreed with this classification. The genetic diversity based on Nei’s index for the entire set of genotypes was 0.468 while this was highest for the Durango–Jalisco group (0.414), intermediate for race Mesoamerica (0.340) and low for race Guatemala (0.262). Genetic differentiation (G _ST) between the Mesoamerican races was 0.27 while genetic distance and identity showed race Durango and Jalisco individuals to be closely related with high gene flow (N _m) both between these two races (1.67) and between races Durango and Mesoamerica (1.58). Observed heterozygosity was low in all the races as would be expected for an inbreeding species. The analysis with microsatellite markers identified subgroups, which agreed well with commercial class divisions, and seed size was the main distinguishing factor between the two major groups identified.Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Genetic interpretation of enzyme variation in sexual and agamospermous taxa of Taraxacum sections Vulgaria and Mongolica

The inheritance of 15 enzymes, comprising at least 22 genetic loci, was investigated in crosses between sexual diploid individuals of Taraxacum sections Vulgaria and Mongolica. Patterns were consistent with simple Mendelian segregation. From the inheritance information isozyme phenotypes in agamospermous plants from natural populations were inferred. In some crosses part or all of the progeny originated from self-fertilization, sofar a very rare phenomenon in the sections Vulgaria and Mongolica. It is possible that the probability of self-fertilization increases after pollination by triploid pollen, affecting the cohabitation dynamics of the various ploidy components in mixed natural stands.     

THE LEVEL OF AGAMOSPERMY IN A NEBRASKA POPULATION OF SPIRANTHES CERNUA (ORCHIDACEAE)

This study investigated reproduction in a prairie population of Spiranthes cernua in eastern Nebraska. Spiranthes cernua reproduces both sexually and asexually, through adventitious embryony. Agamospermous seeds are monoembryonic, polyembryonic, have extruded or free embryos, or lack embryos. These kinds of seeds are extremely rare in Spiranthes vernalis, a close relative that relies exclusively on sexual reproduction. Sexually derived seeds are monoembryonic or lack embryos. One objective was to quantify the amount of sexual reproduction in a natural population. In 1985 and 1986, maximum estimates of sexual seed production per capsule were 19.2% and 33.6%, respectively. Thus, every capsule examined contained a high proportion of agamospermous seeds. In addition, attempts were made to manipulate the reproductive system with experimental crosses performed in the field. The treatments examined agamospermy, autogamy, and outcrossing. In all treatments the majority of seeds in a capsule were agamospermous.     

SEX CHROMOSOME LINKED GENETIC VARIANCE AND THE EVOLUTION OF SEXUAL DIMORPHISM OF QUANTITATIVE TRAITS

Theory predicts that sex chromsome linkage should reduce intersexual genetic correlations thereby allowing the evolution of sexual dimorphism. Empirical evidence for sex linkage has come largely from crosses and few studies have examined how sexual dimorphism and sex linkage are related within outbred populations. Here, we use data on an array of different traits measured on over 10,000 individuals from two pedigreed populations of birds (collared flycatcher and zebra finch) to estimate the amount of sex‐linked genetic variance (h²_z). Of 17 traits examined, eight showed a nonzero h²_Z estimate but only four were significantly different from zero (wing patch size and tarsus length in collared flycatchers, wing length and beak color in zebra finches). We further tested how sexual dimorphism and the mode of selection operating on the trait relate to the proportion of sex‐linked genetic variance. Sexually selected traits did not show higher h²_Z than morphological traits and there was only a weak positive relationship between h²_Z and sexual dimorphism. However, given the relative scarcity of empirical studies, it is premature to make conclusions about the role of sex chromosome linkage in the evolution of sexual dimorphism.     

Allozyme Variation and Phylogenetic Relationships in Picea jezoensis (Pinaceae) Populations of the Russian Far East

Genetic variation and differentiation of 12 populations of Picea jezoensis from the Russian Far East were studied using 20 allozyme loci. The mean number of alleles per locus was 2.63, the percent of polymorphic loci was 88.1%, the observed heterozygosity was 0.181, and the mean value of expected heterozygosity amounted to 0.189. The values of expected heterozygosity of the northern and central mainland populations were higher than in the southern part of the natural range. A significant bias of Hardy–Weinberg heterozygosity to equilibrium heterozygosity (H_eq) suggests that most of the mainland populations have recently experienced a severe expansion in population size while populations from Kamchatka Peninsula have undergone a reduction in population size. Unbiased Nei’s genetic distance values were low within and between the mainland and Sakhalin Island populations (D_N=0.008). The largest values (D_N=0.063) were found between the mainland/Sakhalin and Kamchatka Peninsula populations. Based on genetic distance, P. jezoensis and P. kamtschatkensis could be considered as distinct taxa, but P. ajanensis, P. microsperma, and P. komarovii do not warrant taxonomic recognition.     

GROWTH AND MATING OF GONZUM PECTORALE (VOLVOCALES) IN DEFINED MEDIA1

Growth in defined media of 32 populations of Gonium pectorale was studied to learn more about the sexual isolation reported for this species. The 23 populations containing both mating types (+ & -) were also studied for the ability to form zygotes in defined media. A preliminary study showed that some populations grew and reproduced sexually in a defined mineral medium, whereas others appeared to require exogenous organic materials for growth and/or zygote formation. The diverse reactions exhibited by the populations indicate physiological races which may explain, in part, the occurrence of sexual isolation.     

Isozymes, and the status of Taraxacum (Asteraceae) agamospecies

HUGHES, L. & RICHARDS, A. J., 1989. Isozymes and the status of Taraxacum (Asteraceae) agamospecies. Genetic identities I and Similarity Indices SI are calculated between 12 samples of Taraxacum, on the basis of 40 isozymes at 15 loci for 10 enzyme systems. Samples included three polyploid agamospermous populations from northern England (group 1), three sexual diploid populations from south-central France (group 2), and six accessions of ‘primitive’ diploid self-compatible sexual taxa from southern Europe and Australia. Samples could be assigned to eight species, classified in seven sections of the genus. Two clusters of high relationship were evident. All the group 1 and group 2 species were very closely related, with pairwise comparisons for I in excess of 0.93. The three group 3 accessions identified as T. bessarabicum showed pairwise comparisons for SI in excess of 0.71. Comparisons for SI between the other group 3 species, and between all the group 3 species and the group 1 and 2 species were all very low, not exceeding 0.45. It is concluded that dissimilarity between samples as assessed by isozymes is probably related to the time of evolutionary divergence of those samples. Although allopolyploid, and morphologically very diverse, the group 1 agamospecies may have very recently diverged asexually from a common stock. The group 2 diploids may have resulted from rediploidization and regained sexuality from the same originally agamospermous stock. In areas of Europe in which such ‘modern’ sexuality is common, it is probable that all ‘modern’ Taraxaca, including at least five sections of the genus, should be included within a single taxon. In contrast, ‘primitive’ self-compatible sexual species in group 3 appear to have diverged from each other several million years ago, and with the exception of the disjunct accessions of T. bessarabicum, are genetically highly distinct. Such species should be maintained in the taxonomies of all areas. It is probable that an agamospecies classification of ‘modern’Taraxacum will continue to convey much useful information in areas, such as northern Europe, in which sexuality is absent.     

Frequency of hybridization between Ostrinia nubilalis E‐and Z‐pheromone races in regions of sympatry within the United States

Female European corn borer, Ostrinia nubilalis, produce and males respond to sex pheromone blends with either E‐ or Z‐Δ11‐tetradecenyl acetate as the major component. E‐ and Z‐race populations are sympatric in the Eastern United States, Southeastern Canada, and the Mediterranean region of Europe. The E‐ and Z‐pheromone races of O. nubilalis are models for incipient species formation, but hybridization frequencies within natural populations remain obscure due to lack of a high‐throughput phenotyping method. Lassance et al. previously identified a pheromone gland‐expressed fatty‐acyl reductase gene (pgfar) that controls the ratio of Δ11‐tetradecenyl acetate stereoisomers. We identified three single nucleotide polymorphism (SNP) markers within pgfar that are differentially fixed between E‐ and Z‐race females, and that are ≥98.2% correlated with female pheromone ratios measured by gas chromatography. Genotypic data from locations in the United States demonstrated that pgfar‐z alleles were fixed within historically allopatric Z‐pheromone race populations in the Midwest, and that hybrid frequency ranged from 0.00 to 0.42 within 11 sympatric sites where the two races co‐occur in the Eastern United States (mean hybridization frequency or heterozygosity (H_O) = 0.226 ± 0.279). Estimates of hybridization between the E‐ and Z‐races are important for understanding the dynamics involved in maintaining race integrity, and are consistent with previous estimates of low levels of genetic divergence between E‐ and Z‐races and the presence of weak prezygotic mating barriers.     

The cytology of the genus Briza L. (Gramineae)

Meiosis has been studied in five European and four South American species of Briza. The various species have different chiasma frequencies and different patterns of chiasma localisation. Population studies show that there are significant differences in mean plant chiasma frequency between populations of B. maxima whereas B. media and B. spicata do not show interpopulation differences. B. media is shown to have diploid and autotetraploid races and the distribution of these chromosome races in Europe has been plotted. The South American species studied are all bivalent forming tetraploids and would therefore appear to have an allopolyploid origin. Interchange heterozygosity has been found in B. media and B. elatior, different populations of B. media have been shown to be heterozygous for different interchanges. These topics are discussed in relation to the regulation of recombination.     

GROWTH AND MATING OF GONIUM PECTORALE (VOLVOCALES) IN DEFINED MEDIA1

Growth in defined media of 32 populations of Gonium pectorale was studied to learn more about the sexual isolation reported for this species. The 23 populations containing both mating types (+ & -) icere also studied for the ability to form zygotes in defined media. A preliminary study showed that some populations grew and reproduced sexually in a defined mineral medium, whereas others appeared to require exogenous organic materials for growth and/or zygote formation. The diverse reactions exhibited by the populations indicate physiological races which may explain, in part, the occurrence of sexual isolation.     

A comparative analysis of population structure of a forest tree, Eucalyptus globulus (Myrtaceae), using microsatellite markers and quantitative traits

Eucalyptus globulus (Myrtaceae) is a forest tree native to southeastern Australia, but is grown globally for pulpwood and timber. Eight microsatellite loci were used to determine the degree of selectively neutral differentiation between native populations of the geographic races of E. globulus that are used in a national breeding programme. Spatial differentiation was detected among 340 samples from across the species range (F _ST=0.09±0.02). Analysis of molecular variance showed that there was significant variation between the races, and an unweighted pair group method with arithmetic mean analysis of Nei’s genetic distance between races showed that geographically proximal races tended to be more closely related than geographically distant races. This contrasted markedly with analyses based on quantitative genetic data, where some races appeared to be highly divergent from their geographically closest neighbours. Comparison of racial differentiation based on quantitative (Q _ST) and molecular (F _ST) data suggested that at least five of the quantitative traits used for defining races of E. globulus have been influenced by natural selection, resulting in cases of both phenotypic divergence of parapatric races and phenotypic convergence of allopatric races. We conclude that selectively neutral molecular markers are more useful than quantitative genetic data for identifying the evolutionary affinities and lineages within E. globulus. However, both sources of information should be used in defining evolutionarily important units for conservation. The population structure observed in E. globulus has important consequences for future association studies and may also affect breeding strategies if significant genome co-adaptation has occurred.