Cytogeographic Distribution and Genome Size Variation in Prairie Cordgrass (Spartina pectinata Bosc ex Link)

Prairie cordgrass plants (Spartina pectinata Bosc ex Link) were examined from 61 locations representing the geographic distribution of prairie cordgrass in the U.S. Using flow cytometry, the genome size of 183 individual plants of prairie cordgrass was determined, and the chromosome counts were obtained. Three distinct ploidy levels were observed: tetraploid ( $ \overline x = {1}.{56} {\text{pg}},{2}n = {4} \times = {4}0 $ ), hexaploid ( $ \overline x { = 2}.{33} {\text{pg}},{2}n = {6} \times = {6}0 $ ), and octoploid ( $ \overline {\text{x}} { = 3}.0{6} {\text{pg}},{2}n = {8} \times { = 8}0 $ ). In the sampled areas, the tetraploid populations extended from the East North Central to the New England regions of the U.S., while the octoploid cytotypes were mostly distributed in the West North Central regions. Populations of the tetraploids and octoploids were found in close proximity in the West North Central (Iowa and Kansas) and the West South Central (Oklahoma) regions. The hexaploid cytotype was found in one mixed population (4x?+?6x) occurring in Illinois. No statistically significant intraploidy genome size variation was found in the tetraploid populations, while significant intraploidy genome size variation was found in the octoploid populations. This study precisely defined the geographic distribution of cytotypes in prairie cordgrass throughout the different regions of the U.S. These results provide critical genome size and ploidy distribution information needed to design efficient breeding schemes for high yielding cultivars of prairie cordgrass with local adaption.     

Chloroplast DNA variation within prairie cordgrass (Spartina pectinata Link) populations in the U.S.

Chloroplast DNA (cpDNA) is most often maternally inherited and highly conserved leading to previous observation of little to no sequence variation. Comparing cpDNA haplotypes have provided valuable insight into the establishment and migration of polyploid populations. However, to use chloroplast haplotypes to their full potential intrapopulational variation needs to be addressed. In this study, cpDNA haplotype variation was surveyed within 16 natural populations of prairie cordgrass (Spartina pectinata Link) located east of the 100th west meridian and north of the 35th north parallel in the U.S.A. using two non-coding, polymorphic chloroplast regions. Two main clades were defined with subclades as follows: haplotype 1 and haplotype 2A and 2B. It was discovered that seven populations showed intrapopulational chloroplast genome variation. Of the total amount of variation, 95.5% occurred within the octoploid populations and 4.5% occurred within the tetraploid populations. Both variant haplotypes, 2A and 2B, were found in a larger sampling of one of the natural populations, but no variation was found in a mixed ploidy population. The intrapopulational cpDNA variation we found in this study cannot directly be related to mechanisms of introduction of the non-native populations into native populations. Therefore, this cpDNA variation could be novel natural variation that has been fixed as the octoploid populations were established and moved northwest. This analysis provides insight into determining the usefulness of indels and single nucleotide polymorphisms for population identification and may provide information in regards to the origin of chloroplast variation and its subsequent fixation and establishment in natural prairie cordgrass populations.     

Above- and Belowground Prairie Cordgrass Response to Applied Nitrogen on Marginal Land

Prairie cordgrass (Spartina pectinata, Link.) has been evaluated for its biomass potential because of its high yield, relatively low nutrient demand, and diverse geographical adaptation. Our objectives were to determine (1) biomass production potential of prairie cordgrass in South Dakota and Kansas under varying nitrogen levels, (2) the effect of N on prairie cordgrass yield components (tillers m^?2 and tiller mass), and (3) the effect of N on yield and N concentration of belowground biomass. Older stands of Red River prairie cordgrass (RR-PCG) in South Dakota and Atkins prairie cordgrass (AT-PCG) in Kansas were fertilized with 0, 56, 112 and 168 kg N ha^?1 from 2008 to 2011 in South Dakota and in 2009 and 2010 in Kansas. Experimental design at all locations was a 4?×?4 Latin square. Prairie cordgrass was harvested around a killing frost in October and early November. Biomass production ranged from 5.50 to 13.69 Mg ha^?1 in South Dakota and 5.33 to 12.51 Mg ha^?1 in Kansas. Prairie cordgrass yield did not increase significantly with N application at any location or year. Across years, tiller density ranged from 536 to 934 tillers m^?2 for RR-PCG in South Dakota and from 234 to 315 tillers m^?2 for AT-PCG in Kansas. Neither tiller density or tiller mass was affected by N rate at any location in any year. Belowground biomass production to a depth of 25 cm was equal to or greater than aboveground biomass. However, it was not affected by N rate in all locations by any year. Understanding prairie cordgrass nitrogen-use dynamics to improve biomass and nutrient management will be essential for future investigations. Findings of this study are important to support the notion that prairie cordgrass biomass production in two different environments can be achieved with minimal N inputs.     

Chloroplast DNA phylogeography and cytotype geography in autopolyploid Plantago media

In order to gain insight into the causes of parapatric diploid and tetraploid distributions in Plantago media chloroplast DNA (cpDNA) restriction site polymorphism was studied in 36 European populations. Parapatric distributions are often explained by adaptive differences between cytotypes to an underlying heterogeneity in environmental factors. Alternatively, such distribution patterns may be explained nonadaptively, through frequency-dependant production of hybrids with low fitness. However, nonadaptive explanations have been neglected in polyploid literature. In this study nine chloroplast haplotypes were found. Their phylogeny suggests that tetraploids arose at least three times from diploids. In general, related haplotypes were also geographically clustered, although there were some marked geographical discontinuities. In the Pyrenees, diploids and tetraploids carried diverged haplotypes throughout their parapatric ranges. At the contact zone the level of cpDNA introgression in a mixed diploid-tetraploid population was low. It is discussed that the cpDNA phylogeography supports the nonadaptive hypothesis that parapatric cytotype distributions may be explained by postPleistocene range expansions followed by mutual minority cytotype exclusion, due to hybrid unfitness.     

Neopolyploidy in Spartina pectinata Link: 1. Morphological analysis of tetraploid and hexaploid plants in a mixed natural population

Prairie cordgrass has been reported as a multi-polyploidy species having three cytotypes: tetra- (2n?=?4x?=?40), hexa- (2n?=?6x?=?60), and octoploid (2n?=?8x?=?80). A mixed-ploidy population comprising tetraploids and hexaploids was recently found at a single location in Illinois. However, adaptation and morphological differences between tetra- and hexaploids occurring in natural conditions as well as the contact zones of these cytotypes have yet to be determined. In this study, the cytotypes of 147 individuals of prairie cordgrass collected across the contact zone (4x?+?6x) were determined by flow cytometry using somatic G1 nuclei, and the results were confirmed by chromosome counts. Nineteen morphological characteristics were compared between the cytotypes. Tetra- and hexaploid plants have 2C genome sizes of 1.57 and 2.36?pg with chromosome counts of 40 and 60, respectively. This increase in polyploidy resulted in a greater variability of morphological expression in Illinois prairie cordgrass. Substantial differences in the flowering time, stomatal size, and plant morphological characteristics were observed between tetra- and hexaploids. The results indicate that the increasing of ploidy level in prairie cordgrass resulted in increased plant size in ploidy mixtures. The recent event of ploidy mixtures in prairie cordgrass natural populations offers unique opportunities for studying the formation and establishment of neopolyploidy.     

Molecular evidence for multiple polyploidization and lineage recombination in the Chrysanthemum indicum polyploid complex (Asteraceae)

The Chrysanthemum indicum polyploid complex comprises morphologically differentiated diploids, tetraploids and hybrids between C. indicum and C. lavandulifolium. The relationships between species and cytotypes within this complex remain poorly understood. Random amplified polymorphic DNAs (RAPDs), intersimple sequence repeats (ISSRs) and chloroplast SSR markers were used to elucidate the genetic diversity and relationships of the C. indicum polyploid complex. Molecular analysis of three diploid and nine tetraploid populations provided strong evidence for recurrent origins and lineage recombination in the C. indicum polyploid complex. The high similarity in molecular marker profiles and cpDNA haplotypes between the diploids and tetraploids distributed in the Shen-Nong-Jia Mountain area of China suggested an autopolyploid origin of the tetraploids, while the tetraploids from other populations may have originated via allopolyploidization. Lineage recombination was revealed by the extensive sharing of chloroplast haplotypes and genetic markers among the tetraploid populations with different origins. Multiple differentiation and hybridization/polyploidization cycles have led to an evolutionary reticulation in the C. indicum polyploid complex, and resulted in the difficulties in systematic classification.     

Linkage mapping in prairie cordgrass (<Emphasis Type="Italic">Spartina pectinata</Emphasis> Link) using genotyping-by-sequencing

Prairie cordgrass (Spartina pectinata Link) is a polyploid Chloridoid grass with tetraploid (2n = 40), hexaploid (2n = 60), and octoploid (2n = 80) cytotypes and is a potential dedicated energy crop with promising yields in marginal environments. Efforts to breed prairie cordgrass are currently hampered by the lack of a linkage map, the lack of a Chloridoid reference genome, and the lack of information on inheritance patterns (disomic versus polysomic). Genotyping-by-sequencing (GBS) was applied to a population of 85 progenies from a reciprocal cross of heterozygous tetraploid parents. A total of 26,418 SNPs were discovered, with a distribution of allele frequencies suggesting disomic inheritance. A filtered set of 3034 single-dose, high-coverage SNPs was used for pseudo-testcross mapping with 63 progenies, resulting in two parental maps of 20 linkage groups containing 1522 and 1016 SNPs and a nearly 1:1 ratio of coupling to repulsion phase linkages, again suggesting disomic inheritance. Genomic contigs from tef, another Chloridoid grass, were used as a bridge to associate genetic markers in prairie cordgrass with unique positions in the sorghum genome, providing a glimpse into synteny between Chloridoids and other grasses. GBS enabled rapid generation of a linkage map that will aid in future breeding and genomics efforts in prairie cordgrass.     

Multiple origins of polyploidy and the geographic structure of Heuchera grossulariifolia

Multiple origins of polyploidy from an ancestral diploid plant species were investigated using restriction site polymorphism and sequence variation in the chloroplast DNA (cpDNA) of Heuchera grossulariifolia (Saxifragaceae). Phylogenetic analysis indicated that autopolyploidy has arisen at least twice in the evolutionary history of this species and potentially up to as many as seven times. These results suggest a greater range of independent polyploid origins as compared to a previous study of H. grossulariifolia using cpDNA restriction sites that indicated a minimum of three independent origins. Moreover, most polyploid populations did not contain cpDNA haplotypes from a single origin, but rather combined haplotypes from at least two polyploid origins. Past migration among polyploid populations of independent origin or localized polyploid formation may explain the distribution of polyploid haplotypes within and among populations. The analysis also revealed a discrepancy between relatedness and geographical location. In nearly all sympatric populations of diploids and polyploids, polyploids had the same cpDNA haplotypes as diploids from a geographically remote population. This geographical discordance has several possible explanations, including small sample sizes, extinction of parental diploid haplotypes, chloroplast introgression, and homoplasy in the cpDNA sequence data. We conclude that the recurrent formation of polyploids is an important evolutionary mechanism in the diversification of H. grossulariifolia .     

Morphology and biomass production of prairie cordgrass on marginal lands

Prairie cordgrass (Spartina pectinata Link.) is indigenous throughout most of the continental United States and Canada to 60°N latitude and is well suited to marginal land too wet for maize (Zea mays L.) and switchgrass (Panicum virgatum L.). Evaluations of prairie cordgrass in Europe and North America indicated it has high potential for biomass production, relative to switchgrass, in short‐season areas. Our objective was to describe morphology and biomass production and partitioning in mature stands of ‘Red River’ prairie cordgrass and determine biomass production of natural populations on marginal land. This study was conducted from 2000 to 2008 in eastern South Dakota. Mean biomass production of mature stands of Red River was 12.7 Mg ha^?1. Leaves composed >88% of the biomass, and 60% of the tillers had no internodes. Belowground biomass to a depth of approximately 25 cm, not including roots, was 21 Mg ha^?1. Tiller density ranged from 683 tillers m^?2 for a 10‐year‐old stand to 1140 tillers m^?2 for a 4‐year‐old stand. The proaxis was composed of about eight phytomers, with rhizomes originating at proximal nodes and erect tillers at distal nodes. Vegetative propagation was achieved by both phalanx and guerilla growth. Differences among natural populations for biomass were expressed on gravelly marginal land. However, production, averaged across populations, was low (1.37 Mg ha^?1) and comparable to ‘Cave‐In‐Rock’ switchgrass (1.67 Mg ha^?1) over a 4‐year period. The large carbon storage capacity of prairie cordgrass in proaxes and rhizomes makes it useful for carbon sequestration purposes. Prairie cordgrass should be compared with switchgrass and other C₄ perennial grasses along environmental gradients to determine optimum landscape positions for each and to maximize bioenergy production and minimize inputs.     

LOCAL POLYPLOID VARIATION IN THE NATIVE PRAIRIE GRASS ANDROPOGON GERARDII

The microscale distribution of polyploid variants of the dominant grass big bluestem (Andropogon gerardii) in virgin tallgrass prairie was mapped using flow cytometry. The correlation between DNA content and polyploidy allows the use of flow cytometry for nondestructive determination of polyploidy in intact plants. At Konza Prairie, local plots contained from 0 to 100% hexaploid cytotypes but most showed fine-scale mixing of the polyploid variants. The relationship of cytotype frequency to moisture availability or burning history was nonsignificant.     

Mitochondrial DNA variation and reticulate evolution of the genus Abies

The sequences of three regions of mitochondrial DNA (mtDNA) of a total length of 5226 bp were used to study the phylogeography of the genus Abies. The mtDNA haplotype network, comprising 36 studied Abies taxa, consisted of two branches; the first represented all American species plus two Asian, and the second included the remaining Eurasian species. Within these clusters, the haplotypes formed nine major groups, generally corresponding to the clades of the previously obtained phylogeny based on chloroplast DNA (cpDNA), but the relationships of these groups were significantly different; species assignment to the particular mtDNA haplotype group was more in line with its geographical distribution. In addition, the mtDNA haplotype network contains cycles indicating the recombination. It is assumed that the incongruence of cpDNA and mtDNA phylogenies is caused by the introgression capture of alien mtDNA during species hybridization and thus contains information about past migrations. The cases of incongruence of mitochondrial and chloroplast DNA suggesting a migration of Abies between Asia and North America are discussed.     

Polyploidy,phylogeography and Pleistocene refugia of the rockfern Asplenium ceterach: evidence from chloroplast DNA

Chloroplast DNA sequences were obtained from 331 Asplenium ceterach plants representing 143 populations from throughout the range of the complex in Europe, plus outlying sites in North Africa and the near East. We identified nine distinct haplotypes from a 900 bp fragment of trnL-trnF gene. Tetraploid populations were encountered throughout Europe and further afield, whereas diploid populations were scarcer and predominated in the Pannonian-Balkan region. Hexaploids were encountered only in southern Mediterranean populations. Four haplotypes were found among diploid populations of the Pannonian-Balkans indicating that this region formed a northern Pleistocene refugium. A separate polyploid complex centred on Greece, comprises diploid, tetraploid and hexaploid populations with two endemic haplotypes and suggests long-term persistence of populations in the southern Mediterranean. Three chloroplast DNA (cpDNA) haplotypes were common among tetraploids in Spain and Italy, with diversity reducing northwards suggesting expansion from the south after the Pleistocene. Our cpDNA and ploidy data indicate at least six independent origins of polyploids.     

A species-level phylogenetic study of the Verbena complex (Verbenaceae) indicates two independent intergeneric chloroplast transfers

Two major impediments to infer plant phylogenies at inter- or intra- species level include the lack of appropriate molecular markers and the gene tree/species tree discordance. Both of these problems require more extensive investigations. One of the foci of this study is examining the phylogenetic utility of a combined chloroplast DNA dataset (>5.0kb) of seven non-coding regions, in comparison with that of a large fragment (ca. 3.0kb) of a low-copy nuclear gene (waxy), in a recent, rapidly diversifying group, the Verbena complex. The complex includes three very closely related genera, Verbena (base chromosome number x=7), Glandularia (x=5), and Junellia (x=10), comprising some 150 species distributed predominantly in South and North America. Our results confirm the inadequacy of non-coding cpDNA in resolving relationships among closely related species due to lack of variation, and the great potential of low-copy nuclear gene as source of variation. However, this study suggests that when both cpDNA and nuclear DNA are employed in low-level phylogenetic studies, cpDNA might be very useful to infer organelle evolutionary history (e.g., chloroplast transfer) and more comprehensively understand the evolutionary history of organisms. The phylogenetic framework of the Verbena complex resulted from this study suggests that Junellia is paraphyletic and most ancestral among the three genera; both Glandularia and Verbena are monophyletic and have been derived from within Junellia. Implications of this phylogenetic framework to understand chromosome number evolution and biogeography are discussed. Most interestingly, the comparison of the cpDNA and nuclear DNA phylogenies indicates two independent intergeneric chloroplast transfers, both from Verbena to Glandularia. One is from a diploid North American Verbena species to a polyploid North American Glandularia species. The other is more ancient, from the South American Verbena group to the common ancestor of a major Glandularia lineage, which has radiated subsequently in both South and North America. The commonly assumed introgressive hybridization may not explain the chloroplast transfers reported here. The underlying mechanism remains uncertain.     

Chloroplast sharing in the Tasmanian eucalypts

The biogeographic pattern of chloroplast DNA (cpDNA) haplotypes in Eucalyptus on the island of Tasmania is consistent with reticulate evolution, involving at least 12 Tasmanian species from the subgenus Symphyomyrtus. Intraspecific cpDNA polymorphism in 14 of 17 species is coupled with extensive sharing of identical haplotypes across populations of different species in the same geographic area. Haplotype diversity is lowest in central regions of Tasmania formerly occupied by alpine vegetation during glacial intervals and in northern regions that were periodically linked to continental Australia by land bridges. The observed distribution of several cpDNA haplotypes unique to Tasmania coincides with modeled locations of glacial refugia in coastal areas of Tasmania and shows the power of cpDNA in unraveling the complex history of past distributions of Eucalyptus. The results suggest that the model of evolution of the eucalypts should be reassessed to allow for the anastomosing effects of interspecific hybridization and introgression.     

The hierarchical spatial distribution of chloroplast DNA polymorphism across the introduced range of Silene vulgaris

Silene vulgaris was introduced into North America sometime prior to 1800. In order to document the population structure that has developed since that time, collections were made from 56 local populations distributed among 9 geographical regions in eastern North America. Individual plants were characterized for chloroplast DNA (cpDNA) haplotype by restriction fragment size analysis of four noncoding regions of cpDNA amplified by polymerase chain reaction. A total of 19 cpDNA haplotypes were detected using this method. The overall gene diversity of 0.85 is quite similar to the diversity detected in these same regions of cpDNA in a previously published sample of S. vulgaris taken from across much of Europe. The spatial distribution of the North American cpDNA diversity was quantified by hierarchical F-statistics that partitioned the genetic variance into variation among local populations within regions, and variation among regions. The average FST among populations within regions was 0.66 and the FST among regions was 0.09. The among-region variation was due to both differences among regions in the frequency of two most common haplotypes, and to the presence of a number of region-specific haplotypes. In order to test for isolation by distance at the regional level, FST values were calculated for all possible pairs of regions, and regressed against the geographical distance between those regions. There was no evidence for isolation by distance. It is suggested that the local population structure is generated by recent extinction/colonization dynamics, and that the among-region structure reflects demographic events associated with range expansion following introduction to North America.     

Comparative phylogeography of red maple (Acer rubrum L.) and silver maple (Acer saccharinum L.): impacts of habitat specialization,hybridization and glacial history

Aim We analysed variation in chloroplast DNA (cpDNA) in red maple (Acer rubrum L.) and silver maple (Acer saccharinum L.) across a large part of their geographic ranges. Acer rubrum is one of the most common and morphologically variable deciduous trees of eastern North America, while its sister species A. saccharinum has a more restricted habitat distribution and displays markedly less morphological variation. Our objective was to infer the impact of biogeographic history on cpDNA diversity and phylogeographic structure in both species. Location Deciduous forests of eastern North America. Methods We sequenced 1289 to 1645 bp of non‐coding cpDNA from A. rubrum (n = 258) and A. saccharinum (n = 83). Maximum parsimony networks and spatial analysis of molecular variance (SAMOVA) were used to analyse phylogeographic structure. Rarefaction analyses were used to compare genetic diversity. Results A total of 40 cpDNA haplotypes were recovered from A. rubrum (38 haplotypes) and A. saccharinum (7 haplotypes). Five of the seven A. saccharinum haplotypes were shared with nearby samples of A. rubrum. SAMOVA recovered four phylogeographic groups for A. rubrum in: (1) south‐eastern USA, (2) the Gulf and south‐eastern Coastal Plain, (3) the lower Mississippi River Valley, and (4) the central and northern regions of eastern North America. Acer saccharinum had significantly lower haplotype diversity than A. rubrum, and novel haplotypes in post‐glaciated northern limits of its range were shared with A. rubrum. Main conclusions This is the first study of A. rubrum to report a distinct phylogeographic group centred on the lower Mississippi River, and the first to examine data comparatively with A. saccharinum. We hypothesized that A. rubrum would display stronger phylogeographic structure and greater haplotype diversity than A. saccharinum because of its greater geographic range, and ecological and morphological variation. This hypothesis was supported by the cpDNA analysis. The sharing of cpDNA and chloroplast simple sequence repeat (cpSSR) haplotypes in areas of geographic overlap provides evidence of introgression, which led to an increase in haplotype diversity in both species, and to novel phylogeographic structure in A. rubrum. We recommend that introgression be considered, along with other potential causes, as an explanation for the phylogeographic structure of cpDNA in plants.     

Reciprocal hybrid formation of Spartina in San Francisco Bay

Diversity in the tRNALEU1 intron of the chloroplast genome of Spartina was used to study hybridization of native California cordgrass, Spartina foliosa, with S. alterniflora, introduced to San Francisco Bay approximately 25 years ago. We sequenced 544 bases of the tRNALEU1 intron and found three polymorphic sites, a pyrimidine transition at site 126 and transversions at sites 382 and 430. Spartina from outside of San Francisco Bay, where hybridization between these species is impossible, gave cpDNA genotypes of the parental species. S. foliosa had a single chloroplast haplotype, CCT, and this was unique to California cordgrass. S. alterniflora from the native range along the Atlantic coast of North America had three chloroplast haplotypes, CAT, TAA, and TAT. Hybrids were discriminated by random amplified polymorphic DNA (RAPD) phenotypes developed in a previous study. We found one hybrid that contained a cpDNA haplotype unknown in either parental species (TCT). The most significant finding was that hybridization proceeds in both directions, assuming maternal inheritance of cpDNA; 26 of the 36 hybrid Spartina plants from San Francisco Bay contained the S. foliosa haplotype, nine contained haplotypes of the invading S. alterniflora, and one had the cpDNA of unknown origin. Furthermore, cpDNA of both parental species was distributed throughout the broad range of RAPD phenotypes, suggesting ongoing contributions to the hybrid swarm from both. The preponderance of S. foliosa cpDNA has entered the hybrid swarm indirectly, we propose, from F1s that backcross to S. foliosa. Flowering of the native precedes by several weeks that of the invading species, with little overlap between the two. Thus, F1 hybrids would be rare and sired by the last S. foliosa pollen upon the first S. alterniflora stigmas. The native species produces little pollen and this has low viability. An intermediate flowering time of hybrids as well as pollen that is more vigourous and abundant than that of the native species would predispose F1s to high fitness in a vast sea of native ovules. Thus, spread of hybrids to other S. foliosa marshes could be an even greater threat to the native species than introductions of alien S. alterniflora.     

Intraspecific diversification in North American Boechera stricta (= Arabis drummondii), Boechera xdivaricarpa, and Boechera holboellii (Brassicaceae) inferred from nuclear and chloroplast molecular markers--an integrative approach

We performed a combined evolutionary analysis of North American Boechera stricta, Boechera holboellii, and their hybrid Boechera ×divaricarpa using information on ploidy level estimators, allelic microsatellite variation, noncoding regions of the plastidic genome (cpDNA), and sequences of the internal transcribed spacers 1 and 2 of the nuclear ribosomal DNA (ITS). Somatic ploidy levels of herbarium specimens were estimated based on comparison of pollen size and the number of alleles per locus at seven microsatellites. Results indicate that B. stricta and B. holboellii are genetically distinct from each other, although we also find evidence for occasional introgression between both parental species. Microsatellite patterns for B. stricta from northeastern North America are genetically distinct from western populations, suggesting isolation in glacial refugia along the southeastern margin of the continuous ice shield. Microsatellites supported recent origin of B. ×divaricarpa. Correspondence of nrDNA with cpDNA genetic variation for the majority of diploid B. holboellii accessions suggests a basal, sexual evolutionary unit within a polymorphic B. holboellii group. Hybridization of genetically distinct lineage(s) evidently played an important role in the establishment of polyploid B. holboellii. Frequency of polyploid B. holboellii is substantially higher in the southern United States. This trend corresponds to a southerly distribution of derived chloroplast haplotypes, suggesting an evolutionary advantage of polyploidy and associated apomixis in the colonization of the Sierra Nevada and the Southern Rocky Mountains.     

Phylogeography of Eriotheca species complex: insights into the origin and range expansion of apomictic and polyploid trees in Neotropical Savannas

• Polyploidy and whole genome duplication are major evolutionary drivers in plants. Climate variations during the Pleistocene have influenced distribution and range expansion worldwide. Similar trends have been reported for Cerrado plants, but no attempt has been made to link phylogeography with ploidy and breeding changes. Thus, we aimed to (i) assess ploidy and genome size of Eriotheca estevesiae Carv.-Sobr., and compare it with E. pubescens (Mart.) Schott & Endl. (Both included into the Eriotheca Stellate Trichome Species Complex – ESTSC). (ii) Subsequently, we investigated their phylogeography to see whether genetic structure and range expansion trends were similar to those previously described for the Cerrado biome. Finally (iii), we discuss whether ESTSC phylogeographic patterns could be associated with geographic parthenogenesis processes.
• Common cytogenetic techniques and flow cytometry were used to confirm chromosome number and genome size of E. estevesiae. We used three cpDNA regions to analyse 14 ESTSC Cerrado populations, for which we also obtained ploidy level and breeding information. We investigated haplotype diversity, population structure and tested neutrality, aiming to reconstruct phylogeographic scenarios.
• We found three ploidy levels and eight cpDNA haplotypes in ESTSC, one shared by most populations. Haplotype and ploidy distribution corroborated that E. pubescens, the widely distributed polyploid and apomictic species, may have originated from northern diploid and probably sexual E. estevesiae.
• Matrilinear cpDNA links support the idea that apomixis and polyploidy in ESTSC may have allowed range expansion during the Pleistocene, in a process analogous to the geographic parthenogenesis described elsewhere.