Morphological and geographic variation of the cycad Dioon edule Lindl. (Zamiaceae): ecological and evolutionary implications

The relationship in geographical distribution and morphological variation of leaflet width and length (diagnostic trait), between and within populations of Dioon edule Lindl., has been investigated throughout its known range in eastern Mexico (from the states of Nuevo León to Veracruz, north to south, respectively). A total of 1832 leaflets were measured for width and length from 154 plants distributed amongst five populations using four leaflet replicas from each of three leaves per plant. For leaflet width and length the variation among populations indicated significant stat-istical differences ( F _4,147 = 125.83; P  < 0.0001; R ² = 92.17% and F _4,147 = 9.04; P  < 0.001; R ² = 26.8%), respectively. With respect to leaflet width, the multiple range test showed three groups with a north to south distributional relationship along the range of the species. The correlation coefficient among paired populations, respect to geographical distance and the absolute value of the mean difference of leaflet width in each population, was positive, and different from zero ( r  = 0.82; P  = 0.013). A great variation of important ecological and evolutionary parameters was shown.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society , 2003, 141 , 465–470.     

Surviving background extinction: Inferences from historic and current dynamics in the contrasting population structures of two endemic Mexican cycads

Ecological and microevolutionary perspectives were used to investigate signs of background extinction in two endemic species. We studied relict populations of the cycad Dioon caputoi, contrasting its population structure and neighborhood size with those of Dioon planifolium as a demographically healthier reference population. Population dynamics analysis was performed on two populations of D. caputoi through Integral Projection Models and genetic neighborhoods compared between species through 172 Inter Simple Sequence Repeat loci performed in one population of each of the two study species. The D. caputoi populations mostly comprised adult plants, while D. planifolium presented mainly juvenile individuals. Dioon caputoi showed equilibrium in population growth (i.e., λ ≈ 1), with low recruitment, and its genetic neighborhood revealed highly related individuals in a unique distance class (r_ij = 0.407, 33 m). A contrasting pattern was found in D. planifolium, which showed higher relatedness in the first distance class (r_ij = 0.543, 5 m), gradually decreasing to 20 m. The discrepancies between the two species reflect different strategies of persistence. Dioon caputoi conserves a relict dynamics with signs of a multigenerational, attritional loss of reproductive fitness, while D. planifolium does not. This study furthers our understanding of the background extinction process and the information will thus contribute to the management and conservation of this endangered species.     

Diversity and genetic structure of three species of Dioon Lindl. (Zamiaceae, Cycadales) from the Pacific seaboard of Mexico

We have estimated levels of genetic diversity and partitioning in the Mexican endemic cycad species Dioon sonorense, Dioon tomasellii, and Dioon holmgrenii, whose populations are exclusively distributed along the Pacific seaboard. For the three species, the patterns of variation at 19 allozyme loci in a total of 11 populations were evaluated. The average number of alleles per locus was in the range 2.05–1.68, corresponding to the northernmost population of D. sonorense (Mazatán), and the southernmost population of Dioon holmgrenii (Loxicha), respectively. In turn, the percentage of polymorphic loci peaked (94.73) in the El Higueral and Altamirano populations of Dioon tomasellii, and was estimated to be lowest (57.89) in the Loxicha population of D. holmgrenii. The mean expected heterozygosis varied markedly between taxa, with relatively high indices for D. sonorense and D. tomasellii (H_E = 0.314 and 0.295, respectively) and substantially lower values for D. holmgrenii (H_E = 0.170). Comparison of the inferred genetic structure based on F‐statistics for the three species also indicated differences along the north‐south Pacific seaboard axis. For D. sonorense and D. tomasellii, local inbreeding (F_IS) was zero but global inbreeding (F_IT) values were positive and significantly different from zero (0.130 and 0.116, respectively). By contrast, values of both F_IT and F_IS were negative and significantly different from zero (?0.116 and ?0.201, respectively) for D. holmgrenii. The genetic differentiation between populations (F_ST) had positive values in all taxa and corresponded with their geographic location along the north‐south axis: according to this statistic, D. sonorense was the most differentiated species (F_ST = 0.151), D. tomasellii had intermediate values (F_ST = 0.145), and D. holmgrenii was the less differentiated taxon (F_ST = 0.069). Finally, a phenogram representing Nei's genetic distances among populations displayed three major groups, each one corresponding to each of the studied species. Within D. tomasellii (of intermediate geographic distribution), a further division into two clusters corresponded precisely to the pair of populations that are geographically divided by the Trans Mexican Neovolcanic Mountains. © 2008 The Linnean Society of London, Biological Journal of the Linnean Society, 2008, 94 , 765–776.     

VARIATION IN LEAFLET MORPHOLOGY AMONG POPULATIONS OF CARIBBEAN CYCADS (ZAMIA)

Caribbean cycads (Zamia L.) are well known for variation in leaflet morphology. Here, I examine the variation in leaflet morphology among five populations of Zamia in Florida. Variables measured were leaflet length and width, leaflet length: width ratio, leaflet surface area, rachis and petiole lengths, number of leaflets per leaf, and total leaf area. In addition to comparisons among the Florida populations, these populations are compared with three previously studied populations in Puerto Rico (Newell, 1986). For the most part, the populations exhibited significant differences in all variables. In spite of the extensive morphological variation in this group of plants, leaflet morphology has long been important in Zamia taxonomy. The Florida populations would be considered by some to be either one, two, or three different species. The Florida and Puerto Rico populations together might be considered as few as one species or as many as five species. Based upon the data presented here, the five Florida populations appear to represent a single species; and the three Puerto Rico populations appear to represent two additional species. Further clarification of the taxonomy of Caribbean Zamia will require additional characters such as cone morphology and protein or DNA patterns.     

SPECIES DELIMITATION,TAXONOMY, AND BIOGEOGRAPHY OF DICTYOTA IN EUROPE (DICTYOTALES,PHAEOPHYCEAE)1

Taxonomy of the brown algal genus Dictyota has a long and troubled history. Our inability to distinguish morphological plasticity from fixed diagnostic traits that separate the various species has severely confounded species delineation. From continental Europe, more than 60 species and intraspecific taxa have been described over the last two centuries. Using a molecular approach, we addressed the diversity of the genus in European waters and made necessary taxonomic changes. A densely sampled DNA data set demonstrated the presence of six evolutionarily significant units (ESUs): Dictyota dichotoma (Huds.) J. V. Lamour., D. fasciola (Roth) J. V. Lamour., D. implexa J. V. Lamour., D. mediterranea (Schiffn.) G. Furnari, D. spiralis Mont., and the newly described D. cyanoloma sp. nov., which was previously reported as D. ciliolata from the Mediterranean Sea. Species distributions, based on DNA‐confirmed occurrence records, indicate that all species are geographically confined to the NE Atlantic Ocean with the exception of D. dichotoma and D. implexa, which also occur in South Africa and Bermuda, respectively. To investigate potential hybridization between D. dichotoma and D. implexa, which were previously shown to be sexually compatible in culture, we compiled and analyzed sets of mitochondrial, plastid, and nuclear markers to detect putative hybrids or introgression in natural populations. Failure to detect natural hybrids indicates that effective pre‐ and postzygotic isolation mechanisms are at play in natural populations and supports the by‐product hypothesis of reproductive isolation.     

Elliptic Fourier Analysis of leaf outline shape in forest fragment populations of <Emphasis Type="Italic">Anthurium sinuatum</Emphasis> and <Emphasis Type="Italic">A. pentaphyllum</Emphasis> (Araceae) from Northeast Brazil

A field-based comparative study of leaflet outline shape in five populations of Anthurium sinuatum Benth. ex Schott from Amazonia and humid brejo forests of Ceará, Northeast Brazil, and ten populations of A. pentaphyllum (Aubl.) G. Don var. pentaphyllum from Amazonia and the Brazilian Atlantic forest is reported. 1,120 leaflets from 561 individuals in 15 populations were sampled. Using images, leaflet outlines were captured by digitisation and subjected to Elliptic Fourier Analysis (EFA) to produce matrices of Fourier coefficients based on 40 harmonics; lateral and central leaflets were analysed separately. Twelve shape variables, extracted from the Fourier coefficient matrices, described the leaflet outlines of 531 individuals. Between-population relationships were estimated using cluster analysis, Canonical Variates Analysis and non-parametric MANOVA. Amazonian populations showed the greatest within-population variability. In A. sinuatum the Pacatuba population was the most distinct in Ceará and the Pará population (Amazonia) was not significantly different from those at Ibiapaba and Maranguape (both Ceará). In A. pentaphyllum 80% of population pairs were significantly different and three significantly different groups of populations were observed. Although the populations of A. pentaphyllum displayed considerable variation, the morphological patterns did not correlate with previously reported molecular ones. The two species were found to be slightly but significantly different on leaf outline data.     

Comparison of DNA restriction fragment length polymorphisms of Nostoc strains in and from cycads

DNA was prepared from cyanobacteria freshly isolated from coralloid roots of natural populations of five cycad species: Ceratozamia mexicana mexicana (Mexico), C. mexicana robusta (Mexico), Dioon spinulosum (Mexico), Zamia furfuraceae (Mexico) and Z. skinneri (Costa Rica). Using the Southern blot technique and cloned Anabaena PCC 7120 nifK and glnA genes as probes, restriction fragment length polymorphisms of these cyanobacterial symbionts were compared. The five cyanobacterial preparations showed differences in the sizes of their DNA fragments hybridizing with both probes, indicating that different cyanobacterial species and/or strains were in the symbiotic associations. On the other hand, a similar comparison of cyanobacteria freshly collected from a single Encephalartos altensteinii coralloid root and from three independently subcultured isolates from the same coralloid root revealed that these were likely to be one and the same organism. Moreover, the complexity of restriction patterns shows that a mixture of Nostoc strains can associate with a single cycad species although a single cyanobacterial strain can predominate in the root of a single cycad plant. Thus, a wide range of Nostoc strains appear to associate with the coralloid roots of cycads.Non-standard abbreviations bp base pairs - kbp kilobase pairs - RFLP's restriction fragment length polymorphisms     

Interbreeding versus morphological and ecological differentiation in Mediterranean Diamysis (Crustacea,Mysidacea), with description of four new taxa

Hybridization experiments in the laboratory, performed until the F1 free-living juvenile stage, showed failure of embryonic development in crossbreedings between certain populations of mysids from meso- to metahaline waters of the Mediterranean that were previously lumped together under the name Diamysis bahirensis (G. O. Sars). Western Mediterranean populations did not successfully crossbreed with those from the eastern basin and vice versa. This is consistent with the W-E morphological differentiation in this genus and with different salinity affinities of the western as compared to the eastern forms. From the study of type material, the diagnosis of D. bahirensis is redefined to populations in the SW-Mediterranean showing large spiniform projections of the sixth pleonite, flanking the telson, and numerous small fringes on the posterior margin of the male carapace. Crossbreeding was unsuccessful with populations of D. mesohalobia n. sp. in the E-Mediterranean, which show smooth posterior margin of the carapace in both sexes and distally pectinate paradactylary setae on certain pereiopods, at least in females. Within this species, three morphotypes with different salinity affinities in nature are distinguished; as they showed successful laboratory interbreeding under mesohaline conditions, the taxa, therefore, are defined at subspecific level (D. mesohalobia mesohalobia, D. mesohalobia gracilipes n. ssp., D. mesohalobia heterandra n. ssp.). Populations of these three subspecies failed to crossbreed with a W-Mediterranean (NE-Tyrrhenian) population of D. lagunaris n. sp., which is characterized by a smooth carapace in both sexes, cuticular `windows' on the eyestalks, and rounded projections of the sixth pleonite. In D. lagunaris, D. mesohalobia mesohalobia and D. mesohalobia gracilipes, survival of brood pouch larvae in the laboratory was higher under mesohaline compared with euhaline conditions. This along with biomineralogical and morphological similarities of the statoliths with fossil representatives from Miocene deposits of the brackish Paratethys, suggest a brackish water origin, even in euhalobiontic forms that may have immigrated into the sea from low salinity environments. A key to the 10 species and two subspecies in this genus is given.     

Evaluating hypotheses on the origin and diversification of the ringneck snake Diadophis punctatus (Colubridae: Dipsadinae)

Close affinities recognized between taxa in Mexico and the contiguous USA have led to a variety of biogeographical scenarios. One such hypothesis suggests that species that occur in both countries have an origin in central Mexico followed by dispersal into the USA. This study expands upon previous phylogeographical work of the ringneck snake Diadophis punctatus by incorporating new data from previously unsampled areas appropriate to critically assess hypotheses regarding a Mexican origin for this species. Maximum likelihood and maximum parsimony analyses inferred a derived position for the lineage from southern Mexico with constraint tests for alternate evolutionary hypotheses resulting in significantly worse likelihood values. Ancestral area reconstructions inferred an origin for D. punctatus in the south‐eastern USA followed by a south‐east to north‐east then westward directionality of historical migration. The position within the phylogeny and date estimate for the south‐western + Mexico clade suggests a recent invasion into central Mexico with expansion into the Nearctic/Neotropic transition zone. The extensive lineage diversity inferred from the mtDNA suggests that the genus is a complex of cryptic species whose conservational status should be re‐evaluated on both the national and regional levels. © 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 158 , 629–640.     

Population genetics of Dioon edule Lindl. (Zamiaceae, Cycadales): biogeographical and evolutionary implications

Dioon edule Lindl. (Zamiaceae) is a cycad endemic to Mexico, that occurs as one species D. edule and the geographical variety D. edule var. angustifolium (Miq.) Miq. Dioon edule has a north to south distribution in eastern Mexico. In this study, we analysed 14 allozymic loci in eight populations of D. edule from its total distribution range by sampling all known populations. Patterns of diversity and genetic variability, within and among populations, were obtained. The mean number of alleles per locus ( A ) was 1.44 and the percentage of polymorphic loci was relatively high ( P  = 54.78). The mean observed ( H _O ) and expected heterozygosity ( H _E ) were 0.27 and 0.24, respectively. F -statistics revealed an excess of heterozygous genotypes, locally and globally ( F  = −0.17 and f  = −0.27, respectively). The genetic variation explained by differences among populations was only 7.5%. We also detected a negative relationship between genetic diversity and latitude. On average, the gene flow between population pairs was relatively high ( Nm  = 2.98); furthermore, gene flow between population pairs was significantly correlated with geographical distances ( r  = −0.38, P  = 0.025). Therefore, patterns of genetic diversity in D. edule appear to be associated with the post-Pleistocene spread of the species, from its southerly (origin) to its northerly range (derived populations, including its central distribution). The biogeographical and evolutionary aspects of the results of this study are discussed. We recognize Dioon angustifolium Miq. for the northernmost disjunct populations.  © 2003 The Linnean Society of London, Biological Journal of the Linnean Society , 2003, 80 , 457–467     

Heterozygote excess in ancient populations of the critically endangered Dioon caputoi (Zamiaceae,Cycadales) from central Mexico

Dioon caputoi is a long‐lived cycad known from only four populations that range in size from 50 to 120, mostly adult individuals. Dioon caputoi has the most narrow geographical range of all Dioon spp. (less than 10 km), existing completely within the boundaries of the Tehuacán–Cuicatlán Biosphere Reserve, Mexico. Negative inbreeding values were found in all four populations (F_IT = ?0.242) and within subpopulations (F_IS = ?0.379). Only c. 10% of the total genetic variation was partitioned among populations (F_ST = 0.099). We also found that most mean values of genetic variation (A = 1.91 ± 0.12; P = 78.9 ± 10.2; H_E = 0.35 ± 0.01) are within the range reported for other Dioon species with larger populations and with wider geographical ranges. These results support recent findings that rare plant species maintain high levels of genetic diversity. The heterozygote excess found at all loci is discussed in detail from a neutral evolutionary perspective, leaving arguments as working hypotheses for further research. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 158 , 436–447.     

Morphometric and genetic differentiation in isolated populations of the endangered Mesoamerican stingless bee <Emphasis Type="Italic">Melipona yucatanica</Emphasis> (Hymenoptera: Apoidea) suggest the existence of a two species complex

The stingless bee Melipona yucatanica is a rare species only found in preserved forests across Mesoamerica. Morphometric and molecular analyses (DNA barcoding of the cytochrome oxidase (cox1) and microsatellites) were combined to characterize and compare populations from Mexico and Guatemala. We aim to test the hypothesis predicting that populations from these two geographic regions could be considered as distinct taxonomic units. Morphometric analyses revealed geographic differences, Guatemalan bees being larger than Mexican specimens. Bayesian analyses of the mitochondrial cox1 region and the microsatellite loci demonstrated that M. yucatanica form two clades corresponding to the Mexican and Guatemalan populations. These results suggest that M. yucatanica from Mexico and Guatemala could represent two distinct species. However, more studies are needed on their ecology and behavior to determine the possibility of gene flow between them.     

Morphological and molecular data support recognition of a new rupicolous species of Pinguicula (Lentibulariaceae) from the Iberian Peninsula

A new rupicolous species, Pinguicula saetabensis, belonging to P. sect. Pinguicula is described from calcareous cliffs of central-southern Valencia province, in the eastern Iberian Peninsula. It has previously been confused with P. mundi and P. vallisneriifolia, two close allies endemic to southern Spain which share some morphological traits and a similar habitat. However, some peculiarities allow recognition of those Valencian plants at the specific rank. Data on morphology, ecology, biogeography and conservation are reported for the new species, and its taxonomic affinities are discussed on the basis of phylogenetic analyses of the internal transcribed spacer region of nuclear ribosomal DNA. Furthermore, the presence of one population of P. vallisneriifolia is confirmed in that province, also based on morphological and molecular data.     

A PHYLOGENETIC ANALYSIS OF DIOON (ZAMIACEAE)

A phylogenetic analysis of all the intrageneric taxa of the genus Dioon Lindley (Zamiaceae) was undertaken by using chloroplast DNA restriction fragment length polymorphism analysis. Wagner parsimony analysis on a 187 character matrix yielded two equally parsimonious trees, differing only for the position of D. caputoi. The consensus tree has two well-defined major clades. The first is composed of D. mejiae, D. rzedowskii, and D. spinulosum; the second is composed of D. califanoi, D. caputoi, D. edule var. angustifolium, D. edule var. edule, D. holmgrenii, D. merolae, D. purpusii, D. tomasellii var. sonorense, and D. tomasellii var. tomasellii. A phenetic analysis of the same data showed results broadly congruent with the cladistic analysis. This resulting phylogeny is partially congruent with morphological data and is also compatible with the biogeography of the genus. Modem species of Dioon may have evolved as a consequence of a very fast succession of vicariance events that mainly occurred during the early Cenozoic. The short time between each of these events may not have allowed the accumulation of a large number of morphological synapomorphies for the groups of species.     

Seri Indian traditional knowledge and molecular biology agree: no express train for island‐hopping spiny‐tailed iguanas in the Sea of Cortés

Aim The role of human activities in species biogeography can be difficult to identify, but in some cases molecular techniques can be used to test hypotheses of human‐mediated dispersal. A currently accepted hypothesis states that humans mediated the divergence of two species of spiny‐tailed iguanas in the Ctenosaura hemilopha species complex, namely C. conspicuosa and C. nolascensis, which occupy islands in the Sea of Cortés between the peninsula of Baja California and mainland Mexico. We test an alternative hypothesis that follows the traditional knowledge of the Seri Indians and states that the divergence of these species was not mediated by humans. Location Mexico, including Baja California, Sonoran and Sinaloan coastal regions, and Isla San Esteban and Isla San Pedro Nolasco in the Sea of Cortés. Methods We analysed mitochondrial (cytochrome b and cytochrome c oxidase subunit III) DNA sequences from four species in the C. hemilopha species complex. Maximum parsimony and Bayesian inference were used to infer matriarchal genealogical relationships between the species and several outgroup taxa. Bayesian methods were used to estimate divergence times for the major nodes on the trees based on previously published, fossil‐calibrated priors. Results Our analysis indicated that lineages within the C. hemilopha species complex diverged long before human colonization of the Americas. The divergence of C. nolascensis and C. conspicuosa could not be attributed to Seri translocations. The matriarchal genealogy of the species complex currently defies a simple biogeographical interpretation. Main conclusions We conclude that humans did not mediate the divergence of C. nolascensis and C. conspicuosa. This conclusion is consistent with the traditional knowledge of the Seri people. These results demonstrate the utility of molecular techniques in investigating potential cases of human‐mediated dispersal of plants and animals, and reinforce the importance of considering traditional knowledge in the formation of scientific hypotheses and the interpretation of results.