Genome size dynamics in Artemisia L. (Asteraceae): following the track of polyploidy

Polyploidy is a key factor in the evolution of higher plants and plays an important role in the variation of plant genomes, leading to speciation in some cases. During polyploidisation, different balancing processes take place at the genomic level that can promote variation in nuclear DNA content. We estimated genome size using flow cytometry in 84 populations of 67 Artemisia species and one population of Crossostephium chinense. A total of 73 sequences of nrDNA ITS and 3′‐ETS were newly generated and analysed, together with previously published sequences, to address the evolution of genome size in a phylogenetic framework. Differences in 2C values were detected among some lineages, as well as an increase of genome size heterogeneity in subgenera whose phylogenetic relationships are still unclear. We confirmed that the increase in 2C values in Artemisia polyploids was not proportional to ploidy level, but 1Cx genome size tended to decrease significantly when high ploidy levels were reached. The results lead us to hypothesise that genome size in polyploids tends to a maximum as it follows saturation behaviour, in agreement with the Michaelis–Menten model. We tested different arithmetic functions with our dataset that corroborated a non‐linear relationship of genome size increase in polyploids, allowing us to suggest a theoretical upper limit for the DNA content of this genus.     

Size matters in Triticeae polyploids: larger genomes have higher remodeling

Polyploidization is one of the major driving forces in plant evolution and is extremely relevant to speciation and diversity creation. Polyploidization leads to a myriad of genetic and epigenetic alterations that ultimately generate plants and species with increased genome plasticity. Polyploids are the result of the fusion of two or more genomes into the same nucleus and can be classified as allopolyploids (different genomes) or autopolyploids (same genome). Triticeae synthetic allopolyploid species are excellent models to study polyploids evolution, particularly the wheat-rye hybrid triticale, which includes various ploidy levels and genome combinations. In this review, we reanalyze data concerning genomic analysis of octoploid and hexaploid triticale and different synthetic wheat hybrids, in comparison with other polyploid species. This analysis reveals high levels of genomic restructuring events in triticale and wheat hybrids, namely major parental band disappearance and the appearance of novel bands. Furthermore, the data shows that restructuring depends on parental genomes, ploidy level, and sequence type (repetitive, low copy, and (or) coding); is markedly different after wide hybridization or genome doubling; and affects preferentially the larger parental genome. The shared role of genetic and epigenetic modifications in parental genome size homogenization, diploidization establishment, and stabilization of polyploid species is discussed.     

FISH mapping of 35S and 5S rRNA genes in Artemisia subgenus Dracunculus (Asteraceae): changes in number of loci during polyploid evolution and their systematic implications

Fluorescence in situ hybridization (FISH) with 35S and 5S rDNA probes was used to characterize cytogenetically representatives of Artemisia subgenus Dracunculus and allied species and to explore their evolution following polyploidization. At the diploid level two rDNA loci were observed in most species belonging to the A. dracunculus complex, a pattern considered to be the ancestral state for diploid Artemisia. In contrast, representative species from the Eurasian grade which belong to the other major lineage of the subgenus had more heterogeneous rDNA profiles, with three to five loci at the diploid level. Divergent patterns of locus evolution were also detected in polyploids, with the number and distribution of rDNA loci broadly fitting the two main diversification lineages in the subgenus. In the polyploid complex of A. dracunculus, the number of rDNA loci was almost proportional to ploidy, although monoploid genome size was shown to decrease with increasing ploidy. However, in polyploids from the Eurasian grade we found a remarkable reduction in the number of rDNA sites, suggesting that these species might have experienced either a complete loss of loci or a significant reduction in the number of repeats following polyploid formation. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013 , 171 , 655–666.     

Microsatellite evidence for low genetic diversity and reproductive isolation in tetraploid Centaurea seridis (Asteraceae) coexisting with diploid Centaurea aspera and triploid hybrids in contact zones

Survival of polyploids in nature depends on several factors, including competition from diploid relatives and increased genetic diversity. Unlike other reported Centaurea polyploid complexes, diploid Centaurea aspera and tetraploid Centaurea seridis coexist in hybrid zones with frequent triploid individuals. The polyploid origin of C. seridis, the genetic diversity and population structure of the three cytotypes, and the degree of genetic differentiation among them were analyzed in seven mixed‐ploidy zones, involving different subspecies and ecological conditions. Ploidy was determined by flow cytometry. Microsatellite data suggested an allopolyploid origin of C. seridis. In the contact zones, diploids and tetraploids were genetically differentiated. When compared with the related C. aspera, a low genetic diversity was observed in C. seridis, which is uncommon in tetraploids. Furthermore, although diploid individuals were grouped in a single widespread genetic cluster, tetraploids were grouped in two highly differentiated clusters and showed significant isolation by distance. This genetic pattern in C. seridis may be related to a minimal gene flow with diploid relatives and/or other genetic factors, such as rare polyploidization events, founder effects or an increased selfing rate. Neither taxonomic assignment at subspecies level, nor ecological conditions could explain the genetic differentiation between tetraploid clusters. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 176 , 82–98.     

A high frequency of allopolyploid speciation in the gymnospermous genus Ephedra and its possible association with some biological and ecological features

The origin and evolution of polyploids have been studied extensively in angiosperms and ferns but very rarely in gymnosperms. With the exception of three species of conifers, all natural polyploid species of gymnosperms belong to Ephedra, in which more than half of the species show polyploid cytotypes. Here, we investigated the origin and evolution of polyploids of Ephedra distributed in the Qinghai–Tibetan Plateau (QTP) and neighbouring areas. Flow cytometry (FCM) was used to measure the ploidy levels of the sampled species that are represented by multiple individuals from different populations, and then, two single‐copy nuclear genes (LFY and DDB2) and two chloroplast DNA fragments were used to unravel the possible origins and maternal donors of the polyploids. The results indicate that the studied polyploid species are allopolyploids, and suggest that allotetraploidy is a dominant mode of speciation in Ephedra. The high percentage of polyploids in the genus could be related to some of its biological attributes such as vegetative propagation, a relatively high rate of unreduced gamete formation, and a small genome size relative to most other gymnosperms. Significant ecological divergences between allotetraploids and their putative progenitors were detected by PCAs and anova and Tukey's tests, with the exception of E. saxatilis. The overlap of geographical distributions and ecological niches of some diploid species could have provided opportunities for interspecific hybridization and allopolyploid speciation.     

Wild and agronomically important Agave species (Asparagaceae) show proportional increases in chromosome number,genome size,and genetic markers with increasing ploidy

Agave (Asparagaceae) includes cultivated and wild varieties of henequen used for hard fibre production. As part of a breeding programme to improve Agave production, species with different ploidy levels were genetically characterized: two diploids [A. tequiliana Weber and the hybrid H11648 ((A. amaniensis Trel. & Nowell × A. angustifolia Haw.) × A. amaniensis)], a triploid (A fourcroydes Lem. var. kitam ki), a tetraploid (A. angustifolia var. letona), three pentaploids (A. fourcroydes var. sac ki, A. fourcroydes var. yaax ki, and A. sisalana Perrine), and two hexaploids (A. angustifolia var. chelem ki from two locations). Chromosome spreading was used to determine the chromosome number, flow cytometry was employed to measure the genome size, and fluorescent in situ hybridization was performed using 45S and 5S ribosomal DNA (rDNA) and the telomeric sequences (TTAGGG)_n and (TTTAGGG)_n as genetic markers. There were proportional increases with ploidy level of the following: (1) chromosome number (from diploid 2n = 2x = 60 to hexaploid 2n = 6x = 180), including the number of large and small chromosomes in the bimodal karyotype of Agave; (2) genome size, with a mean monoploid genome size (1Cx) of 7.5 pg (range, 7.36–7.61 pg); and (3) the number and distribution of 45S and 5S rDNA loci, with one locus of each per basic, monoploid genome. Thus there was complete additivity in genome structure with increasing ploidy, as reported in some angiosperm polyploids. However, as other analyses of polyploids have revealed a decrease in 1Cx values with increased ploidy, possible explanations for the observed genomic stability were considered. With the (TTAGGG)_n probe, the signal was localized at the telomeres, consistent with published data showing that many species in the order Asparagales have this type of telomere sequence. It is speculated that sporadic telomeric signals using the (TTTAGGG)_n probe are probably derived from either errors in telomerase activity or relic ancestral‐type telomeric sequences. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 158 , 215–222.     

Polyploid evolution and biogeography in Chelone (Scrophulariaceae): morphological and isozyme evidence

Chelone is a genus of perennial herbs comprising three diploid species (C. cuthbertii, C. glabra, and C. lyonii) and a fourth species (C. obliqua) that occurs as tetraploid and hexaploid races. To assess patterns of isozyme and morphological variation, and to test hypotheses of hybridization and allopolyploidy, we analyzed variation among 16 isozyme loci from 61 populations and 16 morphological characters from 33 populations representing all taxa and ploidy levels. Based on morphological analyses using clustering (unweighted pair group method using an arithmetic average) and ordination (principal components analysis and canonical variance analysis) methods, we recognize three diploid species without infraspecific taxa. Polyploids in the C. obliqua complex were most similar morphologically to diploid populations of C. glabra and C. lyonii. Patterns of isozyme variation among polyploids, which included fixed heterozygosity and recombinant profiles of alleles present in diploids, suggested polytopic origins of tetraploids and hexaploids. Our data indicate independent origins of polyploids in or near the southern Blue Ridge, Interior Highlands and Plains, and Atlantic Coastal Plain regions from progenitors most similar to C. glabra and C. lyonii. Extant tetraploids were not implicated in evolution of hexaploids, and plants similar to C. cuthbertii appeared unlikely as diploid progenitors for polyploids. We propose multiple differentiation and hybridization/polyploidization cycles in different geographic regions to explain the pattern of allopatry and inferred polytopic origins among polyploids.     

Spatio‐temporal patterns of genome evolution in allotetraploid species of the genus Oryza

Despite knowledge that polyploidy is widespread and a major evolutionary force in flowering plant diversification, detailed comparative molecular studies on polyploidy have been confined to only a few species and families. The genus Oryza is composed of 23 species that are classified into ten distinct ‘genome types’ (six diploid and four polyploid), and is emerging as a powerful new model system to study polyploidy. Here we report the identification, sequence and comprehensive comparative annotation of eight homoeologous genomes from a single orthologous region (Adh1–Adh2) from four allopolyploid species representing each of the known Oryza genome types (BC, CD, HJ and KL). Detailed comparative phylogenomic analyses of these regions within and across species and ploidy levels provided several insights into the spatio‐temporal dynamics of genome organization and evolution of this region in ‘natural’ polyploids of Oryza. The major findings of this study are that: (i) homoeologous genomic regions within the same nucleus experience both independent and parallel evolution, (ii) differential lineage‐specific selection pressures do not occur between polyploids and their diploid progenitors, (iii) there have been no dramatic structural changes relative to the diploid ancestors, (iv) a variation in the molecular evolutionary rate exists between the two genomes in the BC complex species even though the BC and CD polyploid species appear to have arisen <2 million years ago, and (v) there are no clear distinctions in the patterns of genome evolution in the diploid versus polyploid species.     

Photosynthetic characteristics of three ploidy levels of Allium oleraceum L. (Amaryllidaceae) differing in ecological amplitude

To elucidate the mechanisms of the adaptive advantages of polyploidy, there is a need to identify physiological traits that participate in the success of polyploids. We studied selected photosynthetic characteristics, stomatal density, and specific leaf area of three ploidy levels (2n = 4x, 5x, 6x) of the geophyte Allium oleraceum that partially differ in their ecological niches. Although the cytotypes were on average similar with regard to most of the measured photosynthetic traits, the hexaploids showed more rapid initial photosynthetic induction and a tendency for a higher maximum photosynthetic rate per unit area. The stomatal density was not affected by ploidy, though the specific leaf area was reduced for the hexaploids compared to the other cytotypes. A lower intracytotype variation was found for most of the studied photosynthetic and anatomical traits for the hexaploids compared to the large variation found within other cytotypes. A comparison of the photosynthetic traits between the cytotypes showed that the ecological differentiation between cytotypes is only weakly related to the characteristics of their photosynthetic apparatus. However, contrasting ranges of variability in the measured traits between the cytotypes can be related to previously observed differences between cytotypes with regard to the ranges of intracytotype genetic variation, genome size variation, and niche breadth. A higher variability of photosynthetic traits in tetraploids and pentaploids may be related to the existence of a spectrum of types adapted to different environmental conditions. Hexaploids may represent a recently formed cytotype adapted to open environmental conditions.     

The Microgeographical Patterns of Morphological and Molecular Variation of a Mixed Ploidy Population in the Species Complex Actinidia chinensis

Polyploidy and hybridization are thought to have significant impacts on both the evolution and diversification of the genus Actinidia, but the structure and patterns of morphology and molecular diversity relating to ploidy variation of wild Actinidia plants remain much less understood. Here, we examine the distribution of morphological variation and ploidy levels along geographic and environmental variables of a large mixed-ploidy population of the A. chinensis species complex. We then characterize the extent of both genetic and epigenetic diversity and differentiation exhibited between individuals of different ploidy levels. Our results showed that while there are three ploidy levels in this population, hexaploids were constituted the majority (70.3%). Individuals with different ploidy levels were microgeographically structured in relation to elevation and extent of niche disturbance. The morphological characters examined revealed clear difference between diploids and hexaploids, however tetraploids exhibited intermediate forms. Both genetic and epigenetic diversity were high but the differentiation among cytotypes was weak, suggesting extensive gene flow and/or shared ancestral variation occurred in this population even across ploidy levels. Epigenetic variation was clearly correlated with changes in altitudes, a trend of continuous genetic variation and gradual increase of epigenomic heterogeneities of individuals was also observed. Our results show that complex interactions between the locally microgeographical environment, ploidy and gene flow impact A. chinensis genetic and epigenetic variation. We posit that an increase in ploidy does not broaden the species habitat range, but rather permits A. chinensis adaptation to specific niches.     

Cytogeography and genome size variation in the Claytonia perfoliata (Portulacaceae) polyploid complex

Background and Aims

Genome duplication is a central process in plant evolution and contributes to patterns of variation in genome size within and among lineages. Studies that combine cytogeography with genome size measurements contribute to our basic knowledge of cytotype distributions and their associations with variation in genome size.

Methods

Ploidy and genome size were assessed with direct chromosome counts and flow cytometry for 78 populations within the Claytonia perfoliata complex, comprised of three diploid taxa with numerous polyploids that range to the decaploid level. The relationship between genome size and temperature and precipitation was investigated within and across cytotypes to test for associations between environmental factors and nuclear DNA content.

Key Results

A euploid series (n = 6) of diploids to octoploids was documented through chromosome counts, and decaploids were suggested by flow cytometry. Increased variation in genome size among populations was found at higher ploidy levels, potentially associated with differential contributions of diploid parental genomes, variation in rates of genomic loss or gain, or undetected hybridization. Several accessions were detected with atypical genome sizes, including a diploid population of C. parviflora ssp. grandiflora with an 18 % smaller genome than typical, and hexaploids of C. perfoliata and C. parviflora with genomes 30 % larger than typical. There was a slight but significant association of larger genome sizes with colder winter temperature across the C. perfoliata complex as a whole, and a strong association between lower winter temperatures and large genome size for tetraploid C. parviflora.

Conclusions

The C. perfoliata complex is characterized by polyploids ranging from tetraploid to decaploid, with large magnitude variation in genome size at higher ploidy levels, associated in part with environmental variation in temperature.     

Genome size rather than content might affect call properties in toads of three ploidy levels (Anura: Bufonidae: Bufo viridis subgroup)

In vertebrates, genome size has been shown to correlate with nuclear and cell sizes, and influences phenotypic features, such as brain complexity. In three different anuran families, advertisement calls of polyploids exhibit longer notes and intervals than diploids, and difference in cellular dimensions have been hypothesized to cause these modifications. We investigated this phenomenon in green toads (Bufo viridis subgroup) of three ploidy levels, in a different call type (release calls) that may evolve independently from advertisement calls, examining 1205 calls, from ten species, subspecies, and hybrid forms. Significant differences between pulse rates of six diploid and four polyploid (3n, 4n) green toad forms across a range of temperatures from 7 to 27 °C were found. Laboratory data supported differences in pulse rates of triploids vs. tetraploids, but failed to reach significance when including field recordings. This study supports the idea that genome size, irrespective of call type, phylogenetic context, and geographical background, might affect call properties in anurans and suggests a common principle governing this relationship. The nuclear‐cell size ratio, affected by genome size, seems the most plausible explanation. However, we cannot rule out hypotheses under which call‐influencing genes from an unexamined diploid ancestral species might also affect call properties in the hybrid‐origin polyploids. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 105 , 584–590.     

Polyploidy creates higher diversity among Cynodon accessions as assessed by molecular markers

Developing a better understanding of associations among ploidy level, geographic distribution, and genetic diversity of Cynodon accessions could be beneficial to bermudagrass breeding programs, and would enhance our understanding of the evolutionary biology of this warm season grass species. This study was initiated to: (1) determine ploidy analysis of Cynodon accessions collected from Turkey, (2) investigate associations between ploidy level and diversity, (3) determine whether geographic and ploidy distribution are related to nuclear genome variation, and (4) correlate among four nuclear molecular marker systems for Cynodon accessions’ genetic analyses. One hundred and eighty-two Cynodon accessions collected in Turkey from an area south of the Taurus Mountains along the Mediterranean cost and ten known genotypes were genotyped using sequence related amplified polymorphism (SRAP), peroxidase gene polymorphism (POGP), inter-simple sequence repeat (ISSR), and random amplified polymorphic DNA (RAPD). The diploids, triploids, tetraploids, pentaploids, and hexaploids revealed by flow cytometry had a linear present band frequency of 0.36, 0.47, 0.49, 0.52, and 0.54, respectively. Regression analysis explained that quadratic relationship between ploidy level and band frequency was the most explanatory (r = 0.62, P < 0.001). The AMOVA results indicated that 91 and 94% of the total variation resided within ploidy level and provinces, respectively. The UPGMA analysis suggested that commercial bermudagrass cultivars only one-third of the available genetic variation. SRAP, POGP, ISSR, and RAPD markers differed in detecting relationships among the bermudagrass genotypes and rare alleles, suggesting more efficiency of combinatory analysis of molecular marker systems. Elucidating Cynodon accessions’ genetic structure can aid to enhance breeding programs and broaden genetic base of commercial cultivars. O. Gulsen and S. Sever-Mutlu contributed equally to this work.     

Isozymes,Speciation and Evolution in the Polyploid Complex Cochlearia L. (Brassicaceae)

Within the genus Cochlearia L. sect. Cochlearia three isozyme systems (aspartate aminotransferase, leucine aminopeptidase and phosphoglucomutase) have been analysed at the populational level for eleven species with different ploidy levels. Nine loci complexes were resolved with a total of 33 alleles from 89 populations. In general, polyploids displayed higher numbers of alleles per species and per population. A notable exception is diploid C. pyrenaica with as many alleles as some polyploids. Data support: (1) the origin of tetraploid C. officinalis from C. pyrenaica s.l., particularly from C. aestuaria; (2) the allopolyploid origin of octoploid C. anglica from C. officinalis; (3) the allopolyploid origin of hexaploid inland taxa C. polonica, C. tatrae and C. bavarica via hybridization between C. pyrenaica and C. officinalis; (4) a putative allopolyploid origin of hexaploid coastal species C. danica by hybridization between C. pyrenaica s.l. and C. officinalis accompanied by a shift in chromosome complement. A scenario summarizing our present understanding of the evolution within Cochlearia sect. Cochlearia is presented.     

Divergent genome sizes reflect the infrafamilial subdivision of the neotropical woody Marcgraviaceae

Neotropical Marcgraviaceae comprise about seven genera and 130 species of lianas and shrubs. They predominantly occur in lowland or montane rainforests and are characterized by a variety of pollination systems. Early classifications subdivided Marcgraviaceae into subfamilies Marcgravioideae and Noranteoideae, a concept supported by molecular data. Using flow cytometry and chromosome numbers, we investigated the role of genome size and polyploidization in the evolution of Marcgraviaceae and how genome sizes are distributed between the proposed infrafamilial groups. To do this we determined genome sizes and chromosome counts for six genera and 22 species for the first time. Our study supports the subfamilial classification of the family, revealing contrasting genome sizes in Noranteoideae (2C = 5.5–21.5 pg) and Marcgravioideae (2C = 2.3–6.2 pg). Polyploidy is considered to be the main source of genome size variation as in each subfamily the higher nuclear DNA amounts were associated with higher ploidy. In addition, genome size changes independent of polyploidy were also observed in some genera, suggesting an additional role for changes in repetitive DNA abundance in the evolution of Marcgraviaceae. A high chromosome base number (x = 18; 2n = 36 to ～70) points to an undetected lower diploid level or to palaeopolyploidy. Marcgraviaceae show a remarkable (nine‐fold) variation in genome size, and several Noranteoideae have genome sizes among the highest reported for tropical woody angiosperms worldwide. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 177 , 1–14.     

The detection and estimation of linkage in polyploids using single-dose restriction fragments

Summary Restriction fragment length polymorphism (RFLP) linkage maps have been constructed in several major diploid crops. However, construction of RFLP maps directly in polyploids has lagged behind for several reasons: (1) there are a large number of possible genotypes for each DNA probe expected in a segregating population, and these genotypes cannot always be identified readily by their banding phenotypes; and (2) the genome constitutions (allopolyploidy versus autopolyploidy) in many high polyploids are not clearly understood. We present here an analysis of these problems and propose a general method for mapping polyploids based on segregation of single-dose restriction fragments (SDRFS). SDRFs segregate 1:1 (presence: absence) in gametes of heterozygous plants. Hypothetical allopolyploid and autopolyploid species with four ploidy levels of 2n = 4x, 6x, 8x, and 10x, are used to illustrate the procedures for identifying SDRFs, detecting linkages among SDRFs, and distinguishing allopolyploid versus autopolyploids from polyploids of unknown genome constitution. Family size required, probability of linkage, and attributes of different mapping populations are discussed. We estimate that a population size of 75 is required to identify SDRFs with 98% level of confidence for the four ploidy levels. This population size is also adequate for detecting and estimating linkages in the coupling phase for both allopolyploids and autopolyploids, but linkages in the repulsion phase can be estimated only in allopolyploids. For autopolyploids, it is impractical to estimate meaningful linkages in repulsion because very large family sizes (>750) are required. For high-level polyploids of unknown genome constitution, the ratio between the number of detected repulsion versus coupling linkages may provide a crude measurement of preferential chromosome pairing, which can be used to distinguish allopolyploidy from autopolyploidy. To create a mapping population, one parent (P₁) should have high heterozygosity to ensure a high frequency of SDRFs, and the second parent (P₂) should have a low level of heterozygosity to increase the probability of detecting polymorphic fragments. This condition could be satisfied by choosing outcrossed hybrids as one parental type and inbreds, haploids, or doubled haploids as the other parental type.Published as Paper No. 730 in the Journal Series of the Experiment Station, Hawaiian Sugar Planters' Association, and as Paper No. 792 in the Plant Breeding Series of Cornell University, Ithaca, NY 14853, USA     

Physical mapping of 35S rRNA genes and genome size variation in polyploid series of Sisyrinchium micranthum and S. rosulatum (Iridaceae: Iridoideae)

Sisyrinchium micranthum and S. rosulatum are part of a species complex in which S. micranthum displays considerable morphological variation. S. rosulatum is a tetraploid species, whereas S. micranthum plants may present three different ploidy levels (2x, 4x, and 6x), so that polyploidy might have an important role in the diversification of this group. Notwithstanding, most cytogenetic studies on these species are based on chromosome counting. Aiming to understand how polyploidy may have impacted the genomes of these species, the DNA content of 184 specimens was estimated; fluorochrome banding with chromomycin A₃ and fluorescent in situ hybridization using an 18S-5.8S-26S ribosomal DNA (rDNA) probe were also performed. The results showed a reduction in monoploid genome size (1Cx), as well as in the number of heterochromatin bands and rDNA sites per monoploid genome, from diploids to polyploids. Additionally, intraspecific and within-ploidy variations in genome size and number of rDNA sites were observed. The source of varying structure in genome organization of these plants may be the multiple independent formations of polyploids along with an ongoing diploidization process. However, the intraspecific and within-ploidy polymorphisms indicate genetic mechanisms other than genome duplication and diploidization to be important to the genome evolution of these taxa.     

Flow cytometric,chromosomal and morphometric analyses challenge current taxonomic concepts in the Portulaca oleracea complex (Portulacaeae,Caryophyllales)

Portulaca oleracea is a noxious annual weed of worldwide distribution in temperate to tropical climates. Its taxonomy has been treated in contradictory ways in the past. Various microspecies have been described, lumped into a single species by other authors. We re‐examined the importance of seed size and ploidy variation, previously applied as the most important taxonomic characters, for systematic classification based on accessions from Europe, Asia, Africa and South America using flow cytometry, chromosome counting and morphometry. Sixteen microspecies and six transitional forms, covering the ploidy and seed character variation, proposed for the complex, were studied from 178 populations. Portulaca grandiflora was included as a reference species from outside the complex. DNA hyper‐pentaploidy or hexaploidy were inferred for the majority of accessions which exhibited the full range of seed size. It is recommended that the only species of lower ploidy (either diploid based on x = 12 or tetraploid based on x = 12) encountered, P. nicaraguensis, should be separated from the P. oleracea complex as it deviates in base chromosome number and monoploid genome size. The frequency distribution of seed size was continuous and unimodal within the wild taxa of the complex and in pairs of taxa defined by testa sculpture. Seed size of DNA hexaploids was slightly negatively correlated with sample/standard fluorescence intensities. Our results conflict with the current microspecies concept. Possible reasons underlying the discrepancy are discussed and strategies for future systematic research are suggested. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 179 , 144–156.     

Polyploidy in Dalea formosa (Fabaceae) on the Chihuahuan Desert

Dalea formosa consists of diploids (n = 7), tetraploids (n = 14), and hexaploids (n = 21), the polyploids restricted to the Chihuahuan Desert region or its immediate borders. There is very little morphological differentiation between the three chromosome races and, therefore, the polyploids are assumed to be primarily autoploid. Tetraploids discovered were few and were very similar to hexaploids; the two ploidy levels were combined as “polyploids” for analyses of geographically and cytologically correlated morphological variation. Pollen length generally was found to be greater in known polyploids than in known diploids. Through the use of pollen length and geographic origin, chromosomally unknown specimens were estimated as to ploidy level. This produced four groups, known diploids and polyploids, and putative diploids and polyploids, which were then subjected to stepwise discriminant analysis (SDA) to search for other morphological characters that might indicate ploidy level, to evaluate the assignments to putative ploidy level in unknown plants, and to assess correlation of these plants of putative ploidy level to geographic regions. SDA also indicated that pollen length, among ten morphological features, is the primary discriminator between ploidy levels, and that putative polyploids are confined primarily to the Chihuahuan Desert. Chromosomally unknown specimens that were originally assigned to one ploidy level, but were classified by SDA as another, are viewed as indicative of areas where further cytological sampling is particularly needed. These areas are southeastern Arizona, where pollen among known diploids is comparatively large, northeastern New Mexico, where polyploids might occur off the Chihuahuan Desert, east edge of the Chihuahuan Desert in Texas, a cytologically poorly sampled contact zone between diploids and polyploids, and central Coahuila, where no cytological sampling has been done. Canonical variate analysis is used to aid in the visualization of the general morphological relationship between diploids and polyploids.     

Phenotypic differentiation of peripheral populations of Santolina rosmarinifolia (Asteraceae)

Phenotypic differentiation of two tetraploid (2n = 4x = 36, 36+1B, 36+2B) populations of Santolina rosmarinifolia geographically isolated from diploid populations was investigated. The karyotype was relatively homogeneous, meiosis was regular and pollen was fertile in both cytotypes. An autopolyploid or allopolyploid origin for tetraploid cytotypes is discussed. Overall, 80.82% of all variance in achene weight, time t₀, t₅₀ and t₉₀ of germination and accumulated germination rate was due to achene age at each ploidy level. Partition of the total phenotypic variance showed that there was extensive variation between ploidy levels. The mean of morphological characters was generally higher in polyploids. For diploid cytotypes, flower number, achene production and fruiting percentage were significantly higher than for tetraploid cytotypes. Cluster analysis indicated that the patterns of seedling morphology and development were similar in three diploid individuals and several tetraploids; the same analysis showed high similarity between diploid individuals of the natural populations, whereas tetraploid individuals showed high dissimilarity among themselves and with diploid individuals. Multiple correspondence analysis and logistic regression analysis indicated that qualitative characters contribute strongly to cytotype differentiation. The results support recognition of the tetraploid cytotypes at the subspecies level. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 158 , 650–668.