THE DISTRIBUTION OF ANTHOCHLOR FLORAL PIGMENTS IN NORTH AMERICAN COREOPSIS (COMPOSITAE): TAXONOMIC AND PHYLETIC INTERPRETATIONS

Anthochlors (chalcones and aurones) occur in floral tissues of all members of North American Coreopsis. The genus in North America consists of 46 species distributed in 11 sections, and a total of five chalcone aglycones are found in it. The chalcone butein occurs in all sections, and is the only anthochlor constituent in the closely related woody Mexican sections Electra and Anathysana. These two sections are viewed as primitive. The morphologically distinct but putatively unspecialized woody Mexican section Pseudo-Agarista sequesters both okanin and butein; the same two anthochlors also characterize two additional sections of herbaceous perennials, Silphidium and Tuckermannia. Sections Euleptosyne and Pugiopappus, comprised of annuals restricted primarily to California, exhibit the rare chalcone stillopsidin in addition to butein and okanin. The remaining four sections, Palmatae, Coreopsis, Calliopsis and Eublepharis are among the more specialized in North America, and are sometimes considered to represent a distinct phyletic line or two related lines. All or some of the members of these sections sequester the very rare chalcone lanceolatin in floral tissues in addition to okanin. Butein is also present in some or all species in each section, and isoliquiritigenin occurs rarely in two sections. Those sections considered more specialized have additional anthochlors relative to primitive sections, and any compound found in a more primitive section also occurs in a more advanced one. Thus, the trend in floral anthochlors in North American Coreopsis is toward increased number of compounds rather than reduction.     

A cladistic study of North AmericanCoreopsis (Asteraceae: Heliantheae)

A cladistic study of all 44 species of North AmericanCoreopsis was performed using 35 characters. The resulting cladogram indicated that all 11 sections are monophyletic. At the intersectional level, two lineages were revealed, one consisting of six sections occurring almost exclusively in Mexico and California, and another comprising five sections restricted largely to the eastern and southeastern United States. The cladogram is similar to phylogenies produced by less explicit methods but it differs in two major respects: the monotypic sect.Silphidium is placed with other sections from the southeastern United States rather than with Mexican sections, and sect.Anathysana from Mexico is more closely allied with the three California sections than with sect.Electra from Mexico.     

The phytomelanin layer in traditional members of Bidens and Coreopsis and phylogeny of the Coreopsideae (Compositae)

The phytomelanin layer on the pericarp of cypselae (achenes) of many members of traditional Bidens and Coreopsis, both considered polyphyletic, was studied with the help of scanning electron (SEM) and light (LM) microscopes. It is found to be more prominent in taxa kept within Bidens than in Coreopsis. The black ‘peg‐like’ phytomelanin found in traditional members of Bidens is also found in some members of Coreopsis. Some traditional members of Coreopsis display distinctive pericarp morphology but lack phytomelanin. The pericarp in Bidens is striated, i.e. it is interrupted by longitudinal bands of parenchyma through which the embryo emerges during seed germination. No striation was found in cypselae of traditional Coreopsis. Emergence of the seed in taxa with this type of pericarp morphology is observed to be by rupturing the carpel wall along the sutures. Characteristic morphology of the phytomelanin layer and other cellular secretions on the pericarp in representative species of these genera and segregates as well as the probable adaptive value of this layer and that of the parenchyma is discussed. Coreopsis sect. Tuckermannia (Nutt.) Blake, C. sect. Pugiopappus (A. Gray) Blake, and C. sect. Euleptosyne (A. Gray) Blake, are elevated to the genus Leptosyne DC., while Coreopsis sect. Electra (DC.) Blake is returned to Electra DC. A key to the segregate genera and the remaining sections of Coreopsis as well as new combinations and synonyms are provided.     

Phylogeny of Eastern North American Coreopsis (Asteraceae-Coreopsideae): insights from nuclear and plastid sequences, and comments on character evolution

A molecular phylogenetic study of eastern North American Coreopsis and representatives of other genera of tribe Coreopsideae was conducted using combined sequences from nuclear ITS and two plastid regions (matK, rpl16). A total of 25-30 species has been recognized in five sections of Coreopsis in eastern North America. Based on morphological characters, these taxa have generally been considered a monophyletic group. Our well-resolved phylogeny supports the monophyly of sections that have been recognized in Coreopsis, but the sections collectively do not comprise a monophyletic group because species of north temperate Bidens occur within one of the two major Coreopsis clades. The most notable departure of present results from prior views of relationships among sections is the lack of a sister group relationship between sections Calliopsis and Eublepharis; the shared presence of four-lobed disk floret corollas had been used to support a close relationship between these two sections. Relationships within sections show both similarities and differences with the results of previous studies based primarily on morphological characters. Mapping of morphological characters used taxonomically in Coreopsis and related genera onto the phylogeny indicates that the evolution of these characters has been complex, and this compromises their value for defining monophyletic groups. Examples include the annual habit, alternate leaves, winged fruits, red or brown basal spots on the yellow ligules, and four-lobed disk floret corollas.     

Evidence for duplication and divergence of the structural gene for phosphoglucoisomerase in diploid species of clarkia

Formal genetic analysis of the mode of inheritance of the electrophoretic phenotypes for phosphoglucoisomerase (PGI) in the annual plants Clarkia rubicunda and C. xantiana showed that these diploid species have two and three genes, respectively, that specify PGI subunits. Electrophoretic examination of seven other diploid species of Clarkia revealed that species assigned to ancestral sections in the current taxonomy have two PGI genes, whereas more specialized species have three PGI genes. Together with evidence that diploid species in two closely related genera have two PGI genes, this suggests the third PGI gene arose within Clarkia. Intergenic heterodimers are formed between polypeptides specified by the third gene and one of the other PGI genes, indicating they have a high degree of structural similarity. The combined genetic, biochemical, and phylogenetic evidence suggests that the third PGI gene resulted from a process of gene duplication. The apparent Michaelis constants (F6P to G6P) of the most common electrophoretic variants of the ancestral gene in C. xantiana and in C. rubicunda are closely similar, but that of the duplicate enzyme is much higher. The intergenic heteromer has an intermediate value. Four alleles have been identified for the duplicate PGI gene in C. xantiana, including a null allele which eliminates the activity of its product. This allele is one of the few examples of a "silenced" duplicate gene. The ancestral and duplicate genes assort independently in C. xantiana. In conjunction with the substantial chromosomal rearrangements that characterize species of Clarkia, this may mean that the duplicate PGI marks a duplicated chromosomal segment that originated from a cross between partially overlapping reciprocal translocations rather than from unequal crossing over.     

Comparative capitular morphology and anatomy of Coreopsis L. and bidens L. (compositae), including a review of generic boundaries

The capitular and floral morphology and anatomy ofBidens L. andCoreopsis L. were studied. All the North American species ofCoreopsis were studied. Selected species ofBidens from North and South America andCoreopsis from South America were included. The results were compared with previous observations on African species ofBidens (incl.Coreopsis). Emphasis was given to character states of the ray florets, paleae, stylearm apices, outer phyllaries, achenes, and pollen grains. Some of the character states are unique features ofCoreopsis, e.g., globular and elongately conical receptacles, deltoid outer phyllaries, truncate and indistinctly 3–5-dentate, 3–4-lobed ray florets, narrowly spathulate paleae, subulate paleae with linear-filiform upper half, hairy and apically 3-cleft paleae, truncate, convex or shallowly conical stylearm apices with the sweeping hairs limited to the area above the stigmatic surfaces and the orbicular to circular achenes. The cylindric setaceous pappus bristles so commonly encountered inBidens are unknown inCoreopsis. The pappus bristles inCoreopsis are paleaceous but similar, though thicker ones are also found in African species ofBidens (incl.Coreopsis) with winged achenes. Twin-celled hairs (setulae) with differing degrees of wall thickness are found on the achenes ofCoreopsis sect.Pseudoagarista (Mexico and South America),Coreopsis sect.Pugiopappus (California), AfricanBidens with winged achenes (e.g.,B. prestinaria, B. macroptera) and some North AmericanBidens (e.g.,B. aristosa). Similar sclerotic parenchyma make up the achenial wings of species in both genera. These may be interpreted as homologous structures, indicating the underlying similarity of these taxa and their derivation from a common ancestral stock.     

PHYLETIC TRENDS IN SECTIONS EUBLEPHARIS AND CALLIOPSIS OF THE GENUS COREOPSIS (COMPOSITAE)

Artificial hybridization, fertility of interspecific and intersectional hybrids, chromosome numbers, and trends in habit and morphology in taxa of sections Eublepharis and Calliopsis of the genus Coreopsis are used to consider the sectional relationships and to derive a presumed phylogeny for the two sections. The two sections are closely related and show a low level of interfertility, but this level is as high as in some interspecific crosses within sections. The sections differ in base chromosome number and achene winging. The problematic C. rosea evidently should remain in section Eublepharis. Both sections probably arose from x = 13 stock resembling C. integrifolia or C. pubescens. Descending aneuploidy was involved in the derivation of section Calliopsis, while chromosome evolution in section Eublepharis has involved polyploidy.     

Leaf flavonoid chemistry and taxonomy of Coreopsis sect. Coreopsis

Seventeen flavonoid glycosides were isolated from the leaves of the nine species of Coreopsis sect. Coreopsis. The compounds include two flavones, four flavonols, a 6-hydroxyflavone, a 6-hydroxyflavonol, two 6-methoxyflavones, a 6-methoxyflavonol and three chalcone-aurone pairs. Chemistry distinguishes all species except C. auriculata and C. intermedia. The mean flavonoid similarity for pair-wise comparison of the four species of annuals (C. basalis, C. nuecensis, C. nuecensoides and C. wrightii) is 0.63 whereas it is 0.66 for the five perennial species (C. auriculata, C. grandiflora, C. intermedia, C. lanceolata, C. pubescens) and 0.54 for comparison of annuals with perennials. This indicates that on the average there is slightly higher flavonoid similarity among species of annuals and among species of perennials than there is between annuals and perennials. Lack of the chalcone-aurone pair coreopsin-sulfurein in perennials is the only consistent chemical difference between annuals and perennials. Leaf flavonoid chemistry supports the hypothesis that C. nuecensis and C. nuecensoides are closely related species but flavonoid data are not concordant with the hypothesis of close relationship between C. basalis and C. wrightii, intermedia, Coreopsis pubescens and C. lanceolata are considered closely related and flavonoid chemistry is concordant with this hypothesis. Coreopsis lanceolata is also viewed as closely allied to C. auriculata, but flavonoids suggest the latter species is more similar to C. intermedia. Coreopsis grandiflora is the only perennial species lacking flavonols and has the fewest number of compounds of any species in the section; chemical data support the view that C. grandiflora is quite distinct from other perennial species of sect. Coreopsis.     

CHROMOSOME NUMBERS AND EVOLUTION IN NORTH AMERICAN SPECIES OF LINUM

Among the North American species of Linum there are three basic chromosome numbers representing invasions from the Old World of three distinct evolutionary lines. N = 9 is found only in the blue-flowered group represented in North America by two species, L. lewisii and L. pratense which are closely related to and may be conspecific with the Old World L. perenne. The basic number for the yellow-flowered species is n = 18 which is characteristic of the Scabrella and Virginiana subgroups. The loss of chromosomes in the Neo-mexicana (n = 13) and Sulcata-Rigida (n = 15) subgroups suggests that the basic haploid number of 18 might be a polyploid derivative of an Old World ancestor with n = 9. The incidence of n = 9 among Old World species of Linum may indicate that this represents an ancestral condition. Linum catharticum has n = 8; this number and features of morphology and distribution suggest that it is not directly related to either the blue-flowered or yellow-flowered groups in North America but represents a separate introduction on this continent.     

Chromosome numbers and taxonomic notes for Mexican coreopsis,sections electra and pseudoagarista (compositae: heliantheae)

Notes on morphology and chromosome numbers are given for several species of MexicanCoreopsis, most of which were poorly known prior to recent collections.Coreopsis parvifolia (sectionElectra) is a large, shrubby, octoploid (2n=112) species apparently restricted to a small area in Puebla. It andC. cuneifolia (a diploid with 2n=28) appear to be closely related but differ by a number of morphological features in addition to ploidy level. A first chromosome report forC. pringlei (sectionPseudoagarista) shows this rare species to be diploid (2n=26), a fact which is in agreement with the base number of the section, i.e.,x=13. Additional collections of the very rareC. rudis andC. mcvaughii show them to be similar yet distinct species. Chromosome determinations forC. petrophila from Nayarit and Durango agree with previous counts for Jalisco populations of the species, i.e., 2n=26. Considerable morphological variation exists within this species but no subspecific entities are recognized.     

Systematics ofLavatera andMalva (Malvaceae,Malveae)—a new perspective

Lavatera andMalva are circumscribed by epicalyx characters, but the distinction is untenable on this basis. Morphological characters and phylogenetic trees from rDNA ITS sequence data indicate the presence of two well-supported species groups different from the traditional genera. One group includes a mixture of closely related species in both genera and forms a more broadly circumscribedMalva, including most disjunctLavatera species—notably four of western North American islands, one Australian and one Canarian species. The other group forms a more narrowly definedLavatera. Another CanarianLavatera species may be a common ancestor to the two groups.     

A genetic contribution to the taxonomy ofOenothera sect.Oenothera (including subsectionsEuoenothera,Emersonia, Raimannia andMunzia)

Based on an analysis of results from experimental hybridization, the plants assigned byMunz toOenothera subg.Oenothera and subg.Raimannia, now divided into approximately 76 species, are referred to a single section,Oenothera. This section is in turn divided into five subsections:Euoenothera, Munzia, Raimannia, Emersonia, and an undescribed group of three species related toOenothera pubescens. Euoenothera is maintained in the traditional sense, and includes about 14 species of North America, widely naturalized elsewhere.Munzia consists of 45 species, comprising three series, and native to South America.Raimannia is restricted to a group of approximately 11 North American species.Emersonia comprises four rather heterogenous species of northern Mexico and southern New Mexico, of whichOenothera macrosceles, O. maysillesii, andO. organensis have been described. Within these four subsections, interspecific hybrids can be made in general, although plastid differentiation often leads to incompatibilities. With varying degrees of difficulty, hybrids were produced in all intersectional combinations involvingEuoenothera, Emersonia, Munzia, andRaimannia, the most difficult being those betweenEmersonia andRaimannia. Based on their habit and distribution,Emersonia species, and especiallyOenothera maysillesii, appear to resemble most closely the common ancestor of the section,Euoenothera andMunzia to have been derived from it or its common ancestor, andRaimannia perhaps to be more closely related to the phylogenetic branch that leads toEuoenothera.     

MOLECULAR DELINEATION OF SPECIES AND SPECIES RELATIONSHIPS IN THE RED ALGAL AGAROPHYTES GRACILARIOPSIS AND GRACILARIA (GRACILARIALES)1

Delineation of species in the economically important agarophyte genera Gracilaria and Gracilariopsis has proven extremely difficult using available morphological characteristics. In this study, we examine the usefulness of two transcribed spacers for molecular systematic studies of these genera. The polymerase chain reaction was used to amplify the internal transcribed spacers (ITSs) and the intervening 5.8S ribosomal DNA of the nuclear ribosomal repeat region. In addition, a plastid spacer region and flanking regions of coding genes were amplified from the RUBISCO operon. Both regions were sequenced for individuals and populations of Gracilariopsis lemaneiformis (Bory) Dawson, Acleto, et Foldvik to determine the usefulness of these spacers in delimiting populations. These studies reveal that there is as much variation among individuals of a population as there is between individuals of geographically separate populations. In addition, the ITS spacer regions were compared between different species of Gracilariopsis and Gracilaria. The nuclear ITS spacer region is conserved at a species level in both genera and provides phylogenetically informative characters that can be used to examine species interrelationships among relatively closely related taxa. However, because of the difficulties of aligning this entire region among species from the two genera, the ITS region is not useful for examining intergenera relationships. ITS interspecies sequence comparisons indicate that Gracilariopsis lemaneiformis from California is significantly different from G. lemaneiformis from China and that a species of Gracilariopsis from Peru is more closely related to G. lemaneiformis from North Carolina than it is to the other Gracilariopsis species examined. In addition, these studies indicate that Gracilaria chilensis Bird, McLachlan, et Oliveira from New Zealand and Gracilaria tenuistipitata Chang et Xia from southeast Asia are as closely related as are Gracilaria verrucosa (Hudson) Papenfuss, G. pacifica Abbott, and Gracilaria robusta Kylin. Phylogenetic analysis of aligned plastid spacer sequences from Gracilaria and Gracilariopsis taxa provide similar conclusions about species relationships.     

An updated classification for the hyper-diverse genus Corydalis (Papaveraceae: Fumarioideae) based on phylogenomic and morphological evidence

The genus Corydalis, with ca. 530 species, has long been considered taxonomically challenging because of its great variability. Previous molecular analyses, based on a few molecular markers and incomplete taxonomic sampling, were clearly inadequate to delimit sections and subgenera. We have performed phylogenetic analyses of Corydalis and related taxa, using 65 shared protein-coding plastid genes from 313 accessions (including 280 samples of ca. 226 species of Corydalis) and 152 universal low-copy nuclear genes from 296 accessions (including 271 samples of Corydalis) covering all 42 previously recognized sections and five independent “series”. Phylogenetic trees were inferred using Bayesian Inference and Maximum Likelihood. Eight selected morphological characters were estimated using ancestral state reconstructions. Results include: (i) of the three subgenera of Corydalis, two are fully supported by both the plastid and nuclear data; the third, subg. Cremnocapnos, is weakly supported by plastid DNA only, whereas in the nuclear data the two included sections form successive outgroups to the rest of the genus; (ii) among all 42 sections and five “series”, 25 sections and one “series” are resolved as monophyletic in both data sets; (iii) the common ancestor of Corydalis is likely to be a perennial plant with a taproot, yellow flowers with a short saccate spur, linear fruits with recurved fruiting pedicels, and seeds with elaiosomes; (iv) we provide a new classification of Corydalis with four subgenera (of which subg. Bipapillatae is here newly described), 39 sections, 16 of which are consistent with the previous classification, 16 sections have been recircumscribed, one section has been reinstated and six new sections are established. Characters associated with lifespan, underground structures, floral spur, fruit and elaiosomes are important for the recognition of subgenera and sections. These new phylogenetic analyses combined with ancestral character reconstructions uncovered previously unrecognized relationships, and greatly improved our understanding of the evolution of the genus.     

Okanin 4′-O-diglucoside from Coreopsis petrophiloides and comments on anthochlors and evolution in Coreopsis

A new chalcone glycoside, okanin 4′-O-diglucoside, is identified from the ray florets of Coreopsis petrophiloides. The known distribution of anthochlors in Coreopsis indicates that two of the most primitive sections produce in their floral parts complex glucosides of butein (and sulfuretin). The more advanced sections synthesize the monoglucosides of okanin (marein), butein (coreospin) and lanceoletin (lanceolin) and their corresponding aurones. The co-occurrence of marein and lanceolin is thus far restricted to members of sect. Coreopsis.     

CHLOROPLAST DNA RESTRICTION SITE VARIATION AND THE PHYLOGENY OF COREOPSIS SECTION COREOPSIS (ASTERACEAE)

Restriction site variation in chloroplast DNAs (cpDNAs) of Coreopsis section Coreopsis was employed to assess divergence and phylogenetic relationships among the nine species of the section. A total of fourteen restriction site mutations and one length mutation was detected. Cladistic analysis of the cpDNA data produced a phylogeny that is different in several respects from previous hypotheses. CpDNA mutations divide the section into two groups, with the two perennial species C. auriculata and C. pubescens lacking any derived restriction site changes. The other seven species are united by five synapomorphic restriction site mutations and the one length mutation. These seven species fall into three unresolved clades consisting of 1) the remaining three perennial species, C. grandiflora, C. intermedia, and C. lanceolata; 2) three annual species, C. basalis, C. nuecensoides, and C. nuecensis; and 3) the remaining annual, C. wrightii. The cpDNA data suggest that, although the perennial habit is primitive within the section, the annual species of section Coreopsis have likely not originated from an extant perennial species. The estimated proportion of nucleotide differences per site (given as 100p) for the cpDNAs of species in the section ranges from 0.00 to 0.20, which is comparable to or lower than values reported for other congeneric species. The low level of cpDNA divergence is concordant with other data, including cross compatibility, interfertility and allozymes, in suggesting that species of the section are not highly divergent genetically.     

CHROMOSOME NUMBERS OF HAWAIIAN FLOWERING PLANTS AND THE SIGNIFICANCE OF CYTOLOGY IN SELECTED TAXA

Chromosome numbers are reported for 128 species of flowering plants indigenous or endemic to Hawaii, including first reports for 13 genera and 82 species. The special significance of reports for Ilex, Tribulus, Keysseria, Pisonia, Boerhavia, Jacquemontia, Claoxylon, Lipochaeta, Railliardia, and Dubautia are discussed. The cytological and morphological variation in Railliardia and Dubautia is considered and their treatment as congeners is advocated. The cytogeogruphic pattern in Dubautia and Railliardia and other factors suggest that the ancestral chromosome number of the Hawaiian tarweeds is n = 14. Their derivation from a western North American progenitor similar to Adenothamnus is considered plausible if not indeed likely.     

EVOLUTION OF RIBOSOMAL DNA: FIFTY MILLION YEARS OF RECORDED HISTORY IN THE FROG GENUS RANA

Evolution of nuclear ribosomal DNA (rDNA) arrays of frogs of the genus Rana was examined among 32 species that last shared a common ancestor approximately 50 million years ago. Extensive variation in restriction sites exists within the transcribed and nontranscribed rDNA spacer regions among the species, whereas rDNA coding regions exhibit comparatively little interspecific variation in restriction sites. The most parsimonious phylogenetic hypothesis for the evolution of the group was constructed based on variation in restriction sites and internal spacer lengths among the 32 species of Rana and one species of Pyxicephalus (examined for outgroup comparison). This analysis suggests that R. sylvatica of North America is more closely related to the R. temporaria group of Eurasia than to other North American Rana. The hypothesized phylogeny also supports the monophyly of the R. boylii group, the R. catesbeiana group, the R. palmipes group, the R. tarahumarae group, and the R. pipiens complex. Furthermore, the restriction site data provide information about the evolution within and among these species groups. This demonstrates that restriction site mapping of rDNA arrays provides a useful molecular technique for the examination of historical evolutionary questions across considerable periods of time.     

Duplicated phosphoglucose isomerase genes in avocado

Summary Avocado (Persea americana) cultivars were assayed for phosphoglucose isomerase (PGI) isozymes using starch gel electrophoresis. Three PGI genes were identified: one monomorphic locus, Pgi-I, coding for the plastid isozyme and two independently assorting loci, Pgi-2 and Pgi-3, coding for the cytosolic isozymes. The genetic analysis was based on comparisons of PGI zymograms from somatic and pollen tissue and on Mendelian analysis of progeny from selfed trees. The isozymic variability for PGI can be used for cultivar identification and for differentiating between hybrid and selfed progeny in avocado breeding.