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Premise of research. Whole-genome duplication is often associated with the evolution of dimorphic sexual systems; however, the association is not universal, and the evolutionary pathway(s) underlyi...

Lycium is the only member of tribe Lycieae (Solanoideae, Solanaceae), and it has a cosmopolitan distribution with its greatest diversity in southern South America, southern Africa, and southwestern North America. To date, there has been no attempt to synthesize and evaluate the significance of the available cytogenetical data from a phylogenetic perspective, which is the objective of this study. Firstly, new data on 27 taxa from all its range of distribution (FISH in all of them, banding in 23, Feulgen technique in 14) were provided to fill gaps in the information. The chromosome numbers of L. australe, L. humile, and L. repens were recorded for the first time. Species showed x = 12 with different ploidy levels (mostly diploid or tetraploid), small chromosomes, and symmetrical karyotypes. Lycium fremontii and L. repens were outstanding for having the highest numbers reported for the genus: 10x and 11x, respectively. North American species showed comparatively longer chromosomes. Secondly, cytogenetical traits were mapped on a phylogenetic tree, using character mapping and ancestral states reconstruction, to understand the dynamics of the evolutionary changes. The main cytotaxonomical features were included: chromosome number, presence of polyploidy, total length of the haploid chromosome set, mean chromosome length, karyotype formula, A1 and A2 asymmetry indices, percentage of heterochromatin, number of CMA+ /DAPI− NORs bands, and number and position of 5S and 18S-5.8S-26S sites. The mean chromosome length, total haploid chromosome length, number of 18S-5.8S-26S loci, number of CMA+/DAPI− NORs bands underwent comparatively few transitions compared with the ploidy level and number of 5S loci. The mapping of the characters on the phylogenetic tree showed that the most probable ancestral chromosome number was 2n = 24 with several independent polyploidization events and one pair of eachrDNA locus. The most likely ancestral condition for the genus would be: diploid, with small chromosomes, scarce heterochromatin, asynteny of rDNA loci, and one pair of both 18S-5.8S-26S and 5S loci. Fil: Stiefkens, Laura Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; Argentina Fil: Las Peñas, Maria Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; Argentina Fil: Levin Rachel A.. Amherst College; Estados Unidos Fil: Miller Jill S.. Amherst College; Estados Unidos Fil: Bernardello, Gabriel Luis Mario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; Argentina

Premise of research. Whereas most Lycium species are diploid and hermaphroditic, an association across Lycium species has been demonstrated between higher ploidy levels and gender dimorphism. Previous monographic work, in combination with a single tetraploid chromosome count, suggested that the Galapagos endemic Lycium minimum may be dimorphic. If substantiated, this would be the only documented Lycium species from South America to be dimorphic in gender expression, which is especially notable given that species diversity is highest in South America. The goals of this study are to (1) test the monophyly, characterize population-level genetic diversity, and determine the closest mainland relatives of L. minimum; (2) infer the sexual system of L. minimum; and (3) assess/verify ploidy using DNA content measurements.Methodology. Phylogenetic, morphological, and cytometric approaches were used to document evolutionary affinities, gynodioecy, and polyploidy. Phylogenetic analyses were conducted with DNA sequenc...

Dinoflagellates within the genus Symbiodinium are photosymbionts of many tropical reef invertebrates, including corals, making them central to the health of coral reefs. Symbiodinium have therefore gained significant research attention, though studies have been constrained by technical limitations. In particular, the generation of viable cells with their cell walls removed (termed protoplasts) has enabled a wide range of experimental techniques for bacteria, fungi, plants, and algae such as ultrastructure studies, virus infection studies, patch clamping, genetic transformation, and protoplast fusion. However, previous studies have struggled to remove the cell walls from armored dinoflagellates, potentially due to the internal placement of their cell walls. Here, we produce the first Symbiodinium protoplasts from three genetically and physiologically distinct strains via incubation with cellulase and osmotic agents. Digestion of the cell walls was verified by a lack of Calcofluor White fluorescence signal and by cell swelling in hypotonic culture medium. Fused protoplasts were also observed, motivating future investigation into intra- and inter-specific somatic hybridization of Symbiodinium. Following digestion and transfer to regeneration medium, protoplasts remained photosynthetically active, regrew cell walls, regained motility, and entered exponential growth. Generation of Symbiodinium protoplasts opens exciting, new avenues for researching these crucial symbiotic dinoflagellates, including genetic modification.

The identification of genomic regions with sufficient variation to elucidate fine-scale relationships among closely related species is a major goal of phylogenetic systematics. However, the accumulation of such multi-locus data sets brings its own challenges, given that gene trees do not necessarily represent the true species tree. Using genomic tools developed for Solanum (Solanaceae), we have evaluated the utility of nuclear conserved ortholog set II (COSII) regions for phylogenetic inference in tribe Lycieae (Solanaceae). Five COSII regions, with intronic contents ranging from 68% to 91%, were sequenced in 10 species. Their phylogenetic utility was assessed and compared with data from more commonly used nuclear (GBSSI, nrITS) and cpDNA spacer data. We compared the effectiveness of a traditional total evidence concatenation approach versus the recently developed Bayesian estimation of species trees (BEST) method to infer species trees given multiple independent gene trees. All of the sampled COSII regions had high numbers of parsimony-informative (PI) characters, and two of the COSII regions had more PI characters than the GBSSI, ITS, and cpDNA spacer data sets combined. COSII markers are a promising new tool for phylogenetic inference in Solanaceae, and should be explored in related groups. Both the concatenation and BEST approaches yielded similar topologies; however, when multiple individuals with polyphyletic alleles were included, BEST was clearly the more robust approach for inferring species trees in the presence of gene tree incongruence.

We examine evolutionary relationships among the three genera in tribe Lycieae using DNA sequence data from the nuclear granule-bound starch synthase gene (GBSSI, waxy). Tribe Lycieae is comprised of the large cosmopolitan genus Lycium and the predominately South American genera Grabowskia and Phrodus. Phylogenetic results strongly suggest that Lycium contains Grabowskia and may also include Phrodus. Further, we examine relationships among several clades of American Lycium and within the monophyletic Old World lineage. This study has the largest taxon sampling to date for tribe Lycieae, with 85% of the named species. Increased sampling within eastern Asia and South America, with the world’s highest species-richness of Lycium, as well as the addition of more rapidly evolving genetic markers, are the areas on which to focus future work.

Species of Solanum subgenus Leptostemonum comprise almost one third of the genus and are distributed worldwide. Members of this group are defined by their sharp epidermal prickles; thus, they are commonly referred to as the ‘‘spiny solanums.’’ This subgenus includes a number of economically important species such as the Old World eggplants, as well as locally cultivated New World species such as the naranjilla and cocona. Given the size and importance of this group we have examined phylogenetic relationships across subgenus Leptostemonum, including a large sampling of species from previously defined species groups within the subgenus. Evolutionary relationships were inferred using DNA sequence data from two nuclear regions (ITS and the granule-bound starch synthase gene [GBSSI or waxy]) and one chloroplast spacer region (trnS-trnG). Results suggest that Solanum subgenus Leptostemonum is monophyletic when the S. wendlandii and S. nemorense species groups are excluded. We have defined 10 clades within subgenus Leptostemonum, some of which correspond to previously circumscribed species groups or sections. Most of the Old World species of subgenus Leptostemonumbelong to a single species-rich clade. Sharp prickles and/or stellate hairs evolved more than once in Solanum, and floral heterandry originated multiple times within the ‘‘spiny solanums.’’ waxy.

The "spiny solanums," Solanum subgenus Leptostemonum (Solanaceae), comprise a large lineage with over 350 species and include the cultivated eggplant, Solanum melongena. Despite the importance of this subgenus, phylogenetic relationships among these taxa are currently unclear. The present research contributes to this understanding while focusing on Solanum section Acanthophora, a group of ca. 19 species defined by the presence of simple hairs, rather than the stellate hairs common across the rest of subgenus Leptostemonum. In this study we inferred phylogenetic relationships among 29 Solanum taxa, including 14 species of section Acanthophora, using DNA sequence data from two nuclear regions (ITS and the granule-bound starch synthase gene [GBSSI or waxy]) and two chloroplast regions (trnT-trnF and trnS-trnG). This combination of gene regions resulted in a well resolved phylogenetic hypothesis, with results strongly suggesting that Solanum sect. Acanthophora is not monophyletic, although the majority of taxa comprise a monophyletic lineage that is sister to Solanum section Lasiocarpa. Of the four gene regions, waxy was especially useful for phylogenetic inference, with both a high percentage of parsimony-informative sites as well as a low level of homoplasy. Further studies in progress will help elucidate relationships of sect. Acanthophora with respect to other members of subgenus Leptostemonum.

We examined relationships between fragrance and phylogeny using a number of approaches to coding fragrance data and comparing the hierarchical information in fragrance data with the phylogenetic signal in a DNA sequence data set. We first used distance analyses to determine which coding method(s) best distinguishes species while grouping conspecifics. Results suggest that interspecific differences in fragrance composition were maximized by coding as presence/absence of fragrance compounds and biosynthetic pathways rather than when quantitative information was also included. Useful systematic information came from both compounds and pathways and from fragrance emitted by both floral and vegetative tissues. The coding methods that emerged from the distance analyses as best distinguishing species were then adapted for use in phylogenetic analysis. Although hierarchical signal among fragrance data sets was congruent, this signal was highly incongruent with the phylogenetic signal in the DNA sequence data. Notably, topologies inferred from fragrance data sets were congruent with the DNA topology only in the most distal portions (e.g., sister group pairs or closely related species that had similar fragrance profiles were often recovered by analyses of fragrance). Examination of consistency and retention indices for individual fragrance compounds and pathways as optimized onto one of the most-parsimonious trees inferred from DNA data revealed that although most compounds were homoplastic, some compounds were perfectly congruent with the DNA phylogeny. In particular, compounds and pathways found in a few taxa were less homoplastic than those found in many taxa. Pathways that synthesize few volatiles also seem to have lower homoplasy than those that produce many. Although fragrance data as a whole may not be useful in phylogeny reconstruction, these data can provide additional support for clades reconstructed with other types of characters. Factors other than phylogeny, including pollinator interactions, also likely influence fragrance composition. (Acleisanthes; character coding; four o'clocks; fragrance; Mirabilis; phylogenetics; scent; volatiles.)

GC-MS analyses of nocturnal and diurnal floral volatiles from nine tobacco species (Nicotiana; Solanaceae) resulted in the identification of 125 volatiles, including mono- and sesquiterpenoids, benzenoid and aliphatic alcohols, aldehydes and esters. Fragrance chemistry was species-specific during nocturnal emissions, whereas odors emitted diurnally were less distinct. All species emitted greater amounts of fragrance at night, regardless of pollinator affinity. However, these species differed markedly in odor complexity and emission rates, even among close relatives. Species-specific differences in emission rates per flower and per unit fresh or dry flower mass were significantly correlated; fragrance differences between species were not greatly affected by different forms of standardization. Flowers of hawkmoth-pollinated species emitted nitrogenous aldoximes and benzenoid esters on nocturnal rhythms. Four Nicotiana species in section Alatae sensu strictu have flowers that emit large amounts of 1,8 cineole, with smaller amounts of monoterpene hydrocarbons and alpha-terpineol on a nocturnal rhythm. This pattern suggests the activity of a single biosynthetic enzyme (1,8 cineole synthase) with major and minor products; however, several terpene synthase enzymes could contribute to total monoterpene emissions. Our analyses, combined with other studies of tobacco volatiles, suggest that phenotypic fragrance variation in Nicotiana is shaped by pollinator- and herbivore-mediated selection, biosynthetic pathway dynamics and shared evolutionary history.

Despite intensive morphological and molecular studies of Onagraceae, relationships within the family are not fully understood. One drawback of previous analyses is limited sampling within the large tribe Onagreae. In addition, the monophyly of two species-rich genera in Onagreae, Camissonia and Oenothera, has never been adequately tested. To understand relationships within Onagraceae, test the monophyly of these two genera, and ascertain the affinities of the newly discovered genus Megacorax, we conducted parsimony and maximum likelihood analyses with rbcL and ndhF sequence data for 24 taxa representing all 17 Onagraceae genera and two outgroup Lythraceae. Results strongly support a monophyletic Onagraceae, with Ludwigia as the basal lineage and a sister-taxon relationship between Megacorax and Lopezia. Gongylocarpus is supported as sister to Epilobieae plus the rest of Onagreae, although relationships within the latter clade have limited resolution. Thus, we advocate placement of Gongylocarpus in a monogeneric tribe, Gongylocarpeae. Most relationships within Onagreae are weakly resolved, suggesting a rapid diversification of this group in western North America. Neither Camissonia nor Oenothera appears to be monophyletic; however, increased taxon sampling is needed to clarify those relationships. Morphological characters generally agree with the molecular data, providing further support for relationships.

The plant genus Lycium (Solanaceae) originated in the Americas and includes approximately 85 species that are distributed worldwide. The vast majority of Old World species occur in southern Africa and eastern Asia. In this study, we examine biogeographic relationships among Old World species using a phylogenetic approach coupled with molecular evolutionary analyses of the S-RNase self-incompatibility gene. The phylogeny inferred from nuclear granule-bound starch synthase I (GBSSI), nuclear conserved ortholog set II (COSII) marker C2_At1g24360, and plastid spacer data (trnH-pbsA, trnD GUC – trnT GGU , rpl32–trnL UAG , and ndhF–rpl32) includes a clade of eastern Asian Lycium nested within the African species, suggesting initial dispersal from the Americas to Africa, with subsequent dispersal to eastern Asia. Molecular dating estimates suggest that these dispersal events occurred relatively recently, with dispersal from the Americas to Africa approximately 3.64 Ma (95% highest posterior density [HPD]: 1.58–6.27), followed by subsequent dispersal to eastern Asia approximately 1.21 Ma (95% HPD: 0.32–2.42). In accordance, the S-RNase genealogy shows that S-RNases isolated from Old World species are restricted to four lineages, a subset of the 14 lineages including S-RNases isolated from New World Lycium species, supporting a bottleneck of S-RNase alleles concomitant with a single dispersal event from the Americas to the Old World. Furthermore, the S-RNase genealogy is also consistent with dispersal of Lycium from Africa to Asia, as eastern Asian alleles are restricted to a subset of the lineages that also include African alleles. Such a multilocus approach, including complementary data from GBSSI, COSII, plastid spacer regions, and S-RNase, is powerful for understanding dispersal histories of closely related species.

We present results of dynamic head-space collections and GC–MS analyses of floral and vegetative fragrances for 20 species in three genera of Nyctaginaceae: Acleisanthes, Mirabilis and Selinocarpus. Most of the species included in this study are either hawkmoth or noctuid moth-pollinated. A wide variety of compounds were observed, including mono- and sesquiterpenoids, aromatics (both benzenoids and phenylpropanoids), aliphatic compounds, lactones, and nitrogen-bearing compounds. Intraspecific variation in fragrance profiles was significantly lower than interspecific variation. Each species had a unique blend of volatiles, and the fragrance of many species contained species-specific compounds. The fragrance profiles presented here are generally consistent with previous studies of fragrance in a variety of moth-pollinated angiosperms.

Recent molecular and morphological studies suggest that neither Acleisanthes nor Selinocarpus (Nyctaginaceae) are monophyletic as currently circumscribed. Further, these data provide no basis for continued recognition of the monotypic genus Ammocodon as separate from these other two genera. Therefore, all 8 Selinocarpus species and the monospecific Ammocodon are transferred to Acleisanthes; 3 additional Selinocarpus species are reduced to synonymy. The genus Acleisanthes is now composed of 16 species, distributed primarily in the Chihuahuan Desert of North America. Diagnostic features of Acleisanthes include an herbaceous or woody perennial habit in arid environments, and the presence of both cleistogamous and chasmogamous flowers often simultaneously on an individual plant. Further, flowers are usually axillary and subtended by 1 to 4 subulate bracts, with chasmogamous flowers frequently white or yellow and funnelform to salverform.

Nyctaginaceae are a small family of mainly New World tropical and subtropical trees, shrubs, and herbs. To date phylogenetic relationships within the family have not been examined. This study provides the first phylogenetic hypothesis of relationships within Nyctaginaceae tribe Nyctagineae based on sequence data from both nuclear (ITS) and chloroplast (accD 5‘coding region and intergenic region between the rbcL and accD genes). Morphological characters are also discussed as they relate to the phylogeny inferred using molecular data. Results suggest that neither Acleisanthes nor Selinocarpus is monophyletic but that together they comprise a monophyletic lineage. The genus Mirabilis is strongly supported as monophyletic, but the monophyly of two of its sections is suspect. Morphology generally agrees with the molecular data and in some instances reinforces clades weakly supported by nuclear and chloroplast data. Further sampling will help clarify relationships of these genera within Nyctaginaceae. ...