Past, present and future in the geographical distribution of Mexican Tepezmaite cycads: Genus Ceratozamia.

Ceratozamia morettii, C. brevifrons, and C. tenuis are cycads considered endangered in montane forests in the center of Veracruz state. However, the amount of theoretical and empirical information available on the historical distribution of these species and how they could be affected in the future by the effects of climate change still needs to be increased. Our objective was to generate information on the spatial distribution of the species since the last glacial maximum, present, and future. To map the spatial distribution of species, we created a potential distribution model for each species. The spatial data used for the models included 19 bioclimatic data variables in the present, at the last glacial maximum using two models (CCSM4 and MIROC), and in the future (2080) using two models of the RCP 8.5 scenario of climate change (HadGEM2-CC and MIROC5). We found that each species occupies a unique ecoregion and climatic niche. Ceratozamia morettii and C. tenuis have a similar pattern with an expansion of their distribution area since the last glacial maximum with a larger distribution area in the present and a projected reduction in their distribution under future climatic conditions. For C. brevifrons, we also showed an increase in their distributional area since the last glacial maximum. We also showed that this expansion will continue under future climatic conditions when the species reaches its maximum distributional area. Projections about the future of these endemic cycad species show changes in their habitat, highlighting that temperate zone species (C. morettii and C. tenuis) will face imminent extinction if no effort is made to protect them. On the other hand, the tropical climate species (C. brevifrons) will be favored.

Discriminatory of some morphometry and biological aspects of the Cycad blue butterfly, Chilades pandava reared on Cycas and Zamia ornamental palms / Discriminação de alguns aspectos morfométricos e biológicos da borboleta azul Cycad, Chilades pandava, criada em palmeiras ornamentais Cycas e Zamia

The serious blue butterfly, Chilades pandava -Horsfield, 1829- (Lepidoptera: Lycaenidae) is consider one of the main destructive insect pests for ornamental palms Cycas and Zamia. Biological and morphological measurements were carried out of C. pandava stages reared on Cycas revoluta (Cycadaceae) and Zamia encephalartoides (Zamiaceae). In description details, non-significant variations were recorded between the two gender of cycad blue butterfly in the obtained data, but the male adult was more densely blue or violet than female adult. By the aid of SEM, C. pandava all stages were distinct by long and thick hairs covered all the body. The morphometric characters namely, length, width and venation of wings, body length, forewing, hindwing could be as a guide for taxonomic discrimination. The data showed that the life cycle duration of C. pandava was ranged between 20.64 to 21.7 days. The developmental periods of different C. pandava stages are slightly higher on zamia than cycas palms. This investigation detected that a high survival rate was found on Cycas palms (86%) than the survived rate recording on Zamia palms (82%). In the present study, the described morphometric characters could be used as a guide for taxonomic discrimination of this pest. Consequently, this study added a valuable knowledge about C. pandava to have sound decisions for proposal of its management and conservation in Egypt.

A borboleta azul séria, Chilades pandava - Horsfield, 1829 (Lepidoptera: Lycaenidae), é considerada uma das principais pragas de insetos destrutivas para as palmeiras ornamentais Cycas e Zamia. Medidas biológicas e morfológicas foram realizadas em estádios de C. pandava criados em Cycas revoluta (Cycadaceae) e Zamia encephalartoides (Zamiaceae). Nos detalhes da descrição, variações não significativas foram registradas entre os dois gêneros de borboleta azul cicadácea nos dados obtidos, mas o adulto masculino era mais densamente azul ou violeta do que o adulto feminino. Com o auxílio do MEV, C. pandava, todos os estágios foram diferenciados por pelos longos e grossos que cobriam todo o corpo. Os caracteres morfométricos a saber, comprimento, largura e venação das asas, comprimento do corpo, asa anterior e posterior, podem servir de guia para a discriminação taxonômica.Os dados mostraram que a duração do ciclo de vida de C. pandava variou entre 20,64 e 21,7 dias. Os períodos de desenvolvimento dos diferentes estágios de C. pandava são ligeiramente maiores em zamia do que em cicas. Essa investigação detectou que uma alta taxa de sobrevivência foi encontrada em palmeiras Cycas (86%) do que a taxa de sobrevivência registrada em palmeiras Zamia (82%). No presente estudo, os caracteres morfométricos descritos podem servir de guia para a discriminação taxonômica dessa praga. Consequentemente, este estudo agregou um conhecimento valioso sobre C. pandava para ter decisões acertadas para a proposta de seu manejo e conservação no Egito.

Two new species of Pharaxonothinae beetles (Coleoptera: Erotylidae) inhabiting cones of the cycad Ceratozamia santillanii Prez-Farr. amp; Vovides (Cycadales: Zamiaceae) in Mexico.

Two beetles, Ceratophila (Vovidesa) chipjonesi, new species, and Pharaxonotha perezi, new species, (Coleoptera: Erotylidae: Pharaxonothinae) are described from the male cones of the New World cycad Ceratozamia santillanii Prez-Farr. Vovides (Cycadales: Zamiaceae). Morphological analysis and keys are provided for distinguishing these from related beetles inhabiting Ceratozamia in Mexico.

Discriminatory of some morphometry and biological aspects of the Cycad blue butterfly, Chilades pandava reared on Cycas and Zamia ornamental palms.

The serious blue butterfly, Chilades pandava -Horsfield, 1829- (Lepidoptera: Lycaenidae) is consider one of the main destructive insect pests for ornamental palms Cycas and Zamia. Biological and morphological measurements were carried out of C. pandava stages reared on Cycas revoluta (Cycadaceae) and Zamia encephalartoides (Zamiaceae). In description details, non-significant variations were recorded between the two gender of cycad blue butterfly in the obtained data, but the male adult was more densely blue or violet than female adult. By the aid of SEM, C. pandava all stages were distinct by long and thick hairs covered all the body. The morphometric characters namely, length, width and venation of wings, body length, forewing, hindwing could be as a guide for taxonomic discrimination. The data showed that the life cycle duration of C. pandava was ranged between 20.64 to 21.7 days. The developmental periods of different C. pandava stages are slightly higher on zamia than cycas palms. This investigation detected that a high survival rate was found on Cycas palms (86%) than the survived rate recording on Zamia palms (82%). In the present study, the described morphometric characters could be used as a guide for taxonomic discrimination of this pest. Consequently, this study added a valuable knowledge about C. pandava to have sound decisions for proposal of its management and conservation in Egypt.

Niche conservatism promotes speciation in cycads: the case of Dioon merolae (Zamiaceae) in Mexico.

Niche conservatism is the tendency of lineages to retain the same niche as their ancestors. It constrains biological groups and prevents ecological divergence. However, theory predicts that niche conservatism can hinder gene flow, strengthen drift and increase local adaptation: does it mean that it also can facilitate speciation? Why does this happen? We aim to answer these questions. We examined the variation of chloroplast DNA, genome-wide single nucleotide polymorphisms, morphological traits and environmental variables across the Dioon merolae cycad populations. We tested geographical structure, scenarios of demographic history, and niche conservatism between population groups. Lineage divergence is associated with the presence of a geographical barrier consisting of unsuitable habitats for cycads. There is a clear genetic and morphological distinction between the geographical groups, suggesting allopatric divergence. However, even in contrasting available environmental conditions, groups retain their ancestral niche, supporting niche conservatism. Niche conservatism is a process that can promote speciation. In D. merolae, lineage divergence occurred because unsuitable habitats represented a barrier against gene flow, incurring populations to experience isolated demographic histories and disparate environmental conditions. This study explains why cycads, despite their ancient lineage origin and biological stasis, have been able to diversify into modern ecosystems worldwide.

Evolution of Ceratozamia cycads: A proximate-ultimate approach.

Evidence suggests that past climatic fluctuations affected speciation of extant cycads. However, empirical genetic and morphological evidence explaining patterns and processes of species diversification are scarce. There are some explanations for the origin and evolution of the genus Ceratozamia, but with inconclusive results. To elucidate the evolution of Ceratozamia, we used genetic and phenotypic sources as empirical data, which were applied in a 'proximate-ultimate' framework (ecological and evolutionary scale, respectively). Our results suggested that the evolutionary mechanisms of speciation were shaped by deterministic (natural selection-adaptation) driven by climatic conditions associated to water stress, and probably enhanced by stochastic processes (gene drift and inbreeding). In general terms, punctuated evolution models were those that best explained the patterns of speciation throughout the phylogenetic history of the lineages encompassed in the genus Ceratozamia. Finally, we provide empirical evidence on the tempo and mode of the evolution of a 'living plant fossil'.

Unlocking a high bacterial diversity in the coralloid root microbiome from the cycad genus Dioon.

Cycads are among the few plants that have developed specialized roots to host nitrogen-fixing bacteria. We describe the bacterial diversity of the coralloid roots from seven Dioon species and their surrounding rhizosphere and soil. Using 16S rRNA gene amplicon sequencing, we found that all coralloid roots are inhabited by a broad diversity of bacterial groups, including cyanobacteria and Rhizobiales among the most abundant groups. The diversity and composition of the endophytes are similar in the six Mexican species of Dioon that we evaluated, suggesting a recent divergence of Dioon populations and/or similar plant-driven restrictions in maintaining the coralloid root microbiome. Botanical garden samples and natural populations have a similar taxonomic composition, although the beta diversity differed between these populations. The rhizosphere surrounding the coralloid root serves as a reservoir and source of mostly diazotroph and plant growth-promoting groups that colonize the coralloid endosphere. In the case of cyanobacteria, the endosphere is enriched with Nostoc spp and Calothrix spp that are closely related to previously reported symbiont genera in cycads and other early divergent plants. The data reported here provide an in-depth taxonomic characterization of the bacterial community associated with coralloid root microbiome. The functional aspects of the endophytes, their biological interactions, and their evolutionary history are the next research step in this recently discovered diversity within the cycad coralloid root microbiome.

Ethnobotany of Mexican and northern Central American cycads (Zamiaceae).

BACKGROUND: This study documents cycad-human relationships in Mexico, Belize, Guatemala, El Salvador, and Honduras over the last 6000 years. The impetus was acute need for a better understanding of previously undocumented uses of cycads in this region, and the need to improve cycad conservation strategies using ethnobotanical data. We hypothesized that cycads are significant dietary items with no long-term neurological effects, are important to religious practice, and contribute to cultural identity and sense of place, but that traditional knowledge and uses are rapidly eroding. Guiding questions focused on nomenclature, food and toxicity, relationships to palms and maize, land management issues, roles in religious ceremony, and medicinal uses, among others, and contributions of these to preservation of cycads. METHODS: From 2000 to 2017, the authors conducted 411 semi-structured ethnographic interviews, engaged in participant-observation in Mexican and Honduran communities, and carried out archival research and literature surveys. RESULTS: We documented 235 terms and associated uses that 28 ethnic groups have for 57 species in 19 languages across 21 Mexican states and 4 Central American nations. Carbohydrate-rich cycads have been both famine foods and staples for at least six millennia across the region and are still consumed in Mexico and Honduras. Certain parts are eaten without removing toxins, while seed and stem starches are detoxified via several complex processes. Leaves are incorporated into syncretic Roman Catholic-Mesoamerican religious ceremonies such as pilgrimages, Easter Week, and Day of the Dead. Cycads are often perceived as ancestors and protectors of maize, revealing a close relationship between both groups. Certain beliefs and practices give cycads prominent roles in conceptions of sense of place and cultural heritage. CONCLUSIONS: Cycads are still used as foods in many places. Though they do not appear to cause long-term neurological damage, their health effects are not fully understood. They are often important to religion and contribute to cultural identity and sense of place. However, because most traditional knowledge and uses are rapidly eroding, new community-based biocultural conservation efforts are needed. These should incorporate tradition where possible and seek inspiration from existing successful cases in Honduras and Mexico.

The phylogeography of the cycad genus Dioon (Zamiaceae) clarifies its Cenozoic expansion and diversification in the Mexican transition zone.

Background and Aims: Biogeographic transition zones are promising areas to study processes of biogeographic evolution and its influence on biological groups. The Mexican transition zone originated due to the overlap of Nearctic and Neotropical biota, which promoted great biological diversification. However, since most previous studies in this area were focused on revealing the phylogeography of Nearctic plants, how historical biogeographic configuration influenced the expansion and diversification of the Neotropical flora remains almost unknown. Using the cycad genus Dioon (Zamiaceae), this study aimed to test whether the biogeographic provinciality of the Mexican transition zone reflects the history of diversification of Neotropical plants. Methods: Two chloroplast DNA (cpDNA) regions were analysed from 101 specimens of 15 Dioon species to reveal the distribution of haplogroups. In addition, genome-wide single nucleotide polymorphisms (SNPs) from 84 specimens were used to test the concordance between phylogenetic clusters and the biogeographic provinces. An ultrametric tree was constructed from the sequences containing SNPs to reconstruct the biogeographic events of vicariance and dispersal of Dioon across the Neotropical biogeographic provinces. Key Results: Four Dioon lineages with strong phylogeographic structures were recognized using both cpDNA and SNP data. The lineages correspond to two clades that originated from a common ancestor in Eastern Mexico. One clade expanded and diversified in South-east Mexico and Central America. Another clade diversified into three lineages that dispersed to North-east, South and North-west Mexico. Each lineage was biogeographically delimitated. Biogeographic provinces might have provided disparate ecological conditions that facilitated speciation in Dioon since the Miocene. Conclusions: The current genetic structure and species diversity of Dioon depict the history of expansion and diversification of the northernmost Neotropical provinces. Past biogeographic connectivities were favoured by elevated topographies, since mountain systems served as corridors for the migration of Dioon and as refugia of tropical communities that diversified during the formation of modern Neotropical forests.

Conservation Genetics of Two Highly Endangered and Poorly Known Species of Zamia (Zamiaceae: Cycadales) in Colombia.

Genetic diversity is key in providing the variation needed to face stochastic change. Increased habitat loss alters population size and dynamics posing serious threats to the conservation of wild species. Colombia has undergone massive deforestation over the last century, but harbors extraordinary high species diversity of genus Zamia (Cycadales); however, most of the species are under threat. In this study, we targeted the largest accessible remaining populations of 2 closely related species growing as endemics in the Magdalena Valley region of Colombia. We successfully transferred the SSR loci used in previous Zamia studies to these species. In total, we amplified 13 microsatellite loci in 3 wild populations, aiming at: 1) assessing genetic diversity and 2) understanding if the structure found between the 3 populations reflected species and population boundaries due to ecological and historical genetic isolation. We found that the actual population size does not reflect population genetic diversity with a small population (Perales) harboring the highest genetic diversity. In addition, all populations are highly structured regardless of species containment, all showing signs of genetic isolation. Given the high degree of ecological threat, and the inherent biological traits of Cycads, we provide information regarding the prioritization of populations for ex situ management.

Shifting Quaternary migration patterns in the Bahamian archipelago: Evidence from the Zamia pumila complex at the northern limits of the Caribbean island biodiversity hotspot.

PREMISE OF THE STUDY: The Bahamas archipelago is formed by young, tectonically stable carbonate banks that harbor direct geological evidence of global ice-volume changes. We sought to detect signatures of major changes on gene flow patterns and reconstruct the phylogeographic history of the monophyletic Zamia pumila complex across the Bahamas. METHODS: Nuclear molecular markers with both high and low mutation rates were used to capture two different time scale signatures and test several gene flow and demographic hypotheses. KEY RESULTS: Single-copy nuclear genes unveiled apparent ancestral admixture on Andros, suggesting a significant role of this island as main hub of diversity of the archipelago. We detected demographic and spatial expansion of the Zamia pumila complex on both paleo-provinces around the Piacenzian (Pliocene)/Gelasian (Pleistocene). Populations evidenced signatures of different migration models that have occurred at two different times. Populations on Long Island (Z. lucayana) may either represent a secondary colonization of the Bahamas by Zamia or a rapid and early-divergence event of at least one population on the Bahamas. CONCLUSIONS: Despite changes in migration patterns with global climate, expected heterozygosity with both marker systems remains within the range reported for cycads, but with significant levels of increased inbreeding detected by the microsatellites. This finding is likely associated with reduced gene flow between and within paleo-provinces, accompanied by genetic drift, as rising seas enforced isolation. Our study highlights the importance of the maintenance of the predominant direction of genetic exchange and the role of overseas dispersion among the islands during climate oscillations.

Estimation of genetic variation in closely related cycad species in Ceratozamia (Zamiaceae: Cycadales) using rapds markers / Variación genética en especies de cícadas estrechamente relacionadas en Ceratozamia (Zamiaceae: Cycadales) mediante el uso de RAPDs

AbstractThe Ceratozamianorstogii complex from Southern Mexico is made up of four closely related taxa and occurs in similar habitats (Quercus forest). All have linear-lanceolate leaflets with great similarity between them, especially in juvenile stages, but differentiate with age. There has been debate regarding delimitation of species due to character loss in herbarium specimens. The aim of this study was to determine the genetic variation, and to measure genetic similarity between the four taxa. We studied populations in Cintalapa (Chiapas) for C. alvarezii and C. norstogii; the Sierra Atravesada (Oaxaca) for C. chimalapensis, and Villa Flores (Chiapas) for C. mirandae. One population for each taxon was sampled (only one population is known for C. alvarezii) 11-15 randomly chosen adult individuals were sampled. Twenty-eight primers were tested of which five were polymorphic using the RAPD'S technique. The data were analyzed using Bayesian methods. Results revealed low genetic diversity, and a differentiation was found between species, suggesting a recent divergence. A previous morphological and anatomical study on the complex has found the taxa to be distinct. However, the results of this study have shown that the C. norstogii species complex is in a divergence process, probably through genetic drift and founder effects. Rev. Biol. Trop. 65 (1): 305-319. Epub 2017 March 01.

ResumenLos cuatro taxa que componen el complejo Ceratozamia norstogii de especies en el sur de México están estrechamente relacionados y se dan en hábitats similares (bosque de Quercus). Todos tienen folíolos linear-lanceolados con gran similitud entre ellos, sobre todo en las etapas juveniles, pero se diferencian con la edad. Ha habido un debate en relación con la delimitación de especies debido a la pérdida de caracteres en especímenes de herbario. Los objetivos de este estudio son determinar la variación genética y medir la similitud genética entre los cuatro taxones en el complejo. Las poblaciones estudiadas están en; Cintalapa, Chiapas para C. alvarezii y C. norstogii, la Sierra Atravesada, Oaxaca para C. chimalapensis y Villa Flores, Chiapas para C. mirandae. Se tomaron muestras de una población de cada taxón (sólo una población es conocida para C. alvarezii) 11-15 individuos adultos elegidos al azar fueron muestreados. Veintiocho primers fueron probados, de los cuales cinco fueron polimórficos mediante la técnica RAPD's. Los datos fueron analizados utilizando métodos bayesianos. Los resultados revelaron baja diversidad genética y la diferenciación encontrada entre las especies sugiere una divergencia reciente. Un estudio morfológico y anatómico anterior en el complejo encontró que los taxa son distintos. Sin embargo, los resultados del presente estudio han demostrado que el complejo C. norstogii aun se encuentra en un proceso de divergencia, probablemente a través de deriva genética y efectos de fundador.

Estimation of genetic variation in closely related cycad species in Ceratozamia (Zamiaceae: Cycadales) using RAPDs markers.

The Ceratozamia norstogii complex from Southern Mexico is made up of four closely related taxa and occurs in similar habitats (Quercus forest). All have linear-lanceolate leaflets with great similarity between them, especially in juvenile stages, but differentiate with age. There has been debate regarding delimitation of species due to character loss in herbarium specimens. The aim of this study was to determine the genetic variation, and to measure genetic similarity between the four taxa. We studied populations in Cintalapa (Chiapas) for C. alvarezii and C. norstogii; the Sierra Atravesada (Oaxaca) for C. chimalapensis, and Villa Flores (Chiapas) for C. mirandae. One population for each taxon was sampled (only one population is known for C. alvarezii) 11-15 randomly chosen adult individuals were sampled. Twenty-eight primers were tested of which five were polymorphic using the RAPD'S technique. The data were analyzed using Bayesian methods. Results revealed low genetic diversity, and a differentiation was found between species, suggesting a recent divergence. A previous morphological and anatomical study on the complex has found the taxa to be distinct. However, the results of this study have shown that the C. norstogii species complex is in a divergence process, probably through genetic drift and founder effects.

Comparative anatomy of leaflets of Zamia acuminata and Z. pseudomonticola (Zamiaceae) in Costa Rica.

The genus Zamia is morphologically and ecologically the most diverse of the order Cycadales. Throughout its history this genus has been restricted to the New World and is presently almost entirely restricted to the Neotropics. Unusual anatomical traits of the leaflets, such as the sunken stomata and thick cuticle, are common in this and related genera. The objective of this research was to study and compare the leaflet anatomy of Zamia acuminata and Z pseudomonticola and establish possible phylogenetic relationships between the anatomical traits and the near relatives of these species. The leaf material was obtained from living plants and then processed for electron microscopy study. We found that both species are very similar to each other and to Z fairchildiana, and that they share several unusual traits with other species of the genus, such as the parenchyma morphology, the spatial distribution of tissues between the veins and the stomata morphology. The main differences between these species were seen in their fiber clusters and in the abundance oftrichome basal cells on the epidermis. The anatomical similarities between the three species could be the result of their close phylogenetic relationship and the divergences between them could be the result of recent speciation during the Pleistocene, resulting from geological changes in Southern Costa Rica.

Comparative anatomy of leaflets of Zamia acuminata and Z. pseudomonticola (Zamiaceae) in Costa Rica

The genus Zamia is morphologically and ecologically the most diverse of the order Cycadales. Throughout its history this genus has been restricted to the New World and is presently almost entirely restricted to the Neotropics. Unusual anatomical traits of the leaflets, such as the sunken stomata and thick cuticle, are common in this and related genera. The objective of this research was to study and compare the leaflet anatomy of Zamia acuminata and Z. pseudomonticola and establish possible phylogenetic relationships between the anatomical traits and the near relatives of these species. The leaf material was obtained from living plants and then processed for electron microscopy study. We found that both species are very similar to each other and to Z. fairchildiana, and that they share several unusual traits with other species of the genus, such as the parenchyma morphology, the spatial distribution of tissues between the veins and the stomata morphology. The main differences between these species were seen in their fiber clusters and in the abundance of trichome basal cells on the epidermis. The anatomical similarities between the three species could be the result of their close phylogenetic relationship and the divergences between them could be the result of recent speciation during the Pleistocene, resulting from geological changes in Southern Costa Rica.

Zamia es morfológica y ecológicamente el género más diverso del orden Cycadales. Este género siempre ha estado restringido a América, pero en la actualidad habita principalmente en la región neotropical. Características anatómicas inusuales en los foliolos como los estomas hundidos y las cutículas gruesas son comunes en Zamia y géneros afines. El objetivo de este trabajo consiste en comparar la anatomía de los foliolos de Zamia acuminata y Z. pseudomonticola y establecer posibles relaciones filogenéticas entre las características anatómicas y los parientes cercanos de esta especie. Las hojas de las especies seleccionadas fueron obtenidas de plantas vivas y luego procesadas para el estudio por microscopía electrónica. Ambas especies son muy similares entre sí y respecto a Z. fairchildiana y comparten varias características en común con otras especies del género como son la morfología del parénquima, de los estomas y la distribución espacial de tejidos alrededor de las haces vasculares. Las diferencias más notables entre especies se vieron a nivel de sus paquetes de fibras y en la abundancia de células basales de los tricomas en la epidermis. Las similitudes anatómicas entre estas tres especies pueden ser el resultado de su cercanía filogenética y las diferencias podrían ser el resultado de especiación durante el Pleistoceno, producto de los eventos geológicos y cambios sucedidos en el sur de Costa Rica en esa época.

Respuesta de Zamia incognita L. al cultivo in vitro, una alternativa para su conservación / In vitro culture response of Zamia incognita, an alternative for preservation

Las Zamiaceas son plantas relictuales consideradas fósiles vivientes. En Colombia, el 65% de esta familia se encuentra en alguna categoría de amenaza, por la destrucción del hábitat e intensa recolección. Teniendo en cuenta que entre las ventajas de la propagación in vitro está la conservación ex situ de germoplasma, el presente trabajo tuvo como objetivo evaluar el potencial de regeneración de plantas de Z. incognita a partir de explantes foliares y embriones cigoticos. Se evaluó el efecto de diferentes combinaciones de Auxinas (2,4-D y ANA) y citoquininas (KIN, BAP y TDZ) sobre la formación de callo y la regeneración de brotes (directa o indirecta), utilizando como medio basal MS (MB1) y medio basal B5 modificado (MB2). La formación de callo se presentó sobre un amplio rango de concentraciones de 2,4-D con KIN y 2,4-D con BAP, independientemente del medio basal, pero no en los explantes tratados con ANA más KIN o TDZ. Para los explantes foliares no hubo respuesta a la formación de embriones somáticos y/o brotes con las combinaciones y concentraciones hormonales evaluadas, no obstante los callos inducidos en MB2 con 2,4-D (0,22 mg/l) y BAP (0, 1, 2, 3 mg/l) fueron diferentes, su aspecto nodular, color crema y apariencia proembriogénica coincidió con una gran cantidad de células meristemáticas potenciales para el proceso de regeneración. A partir de embriones cigoticos inmaduros se logró la formación de embriones somáticos en el medio MB2 exento de reguladores o conteniendo 2,4-D solo (0,22 mg/l) y en combinación con BAP (1 mg/l), sin lograr el proceso de conversión a plántulas.

Zamiaceas are relict plants considered living fossils. In Colombia, 65% of this family is under some threat category due to their habitat destruction and their intense collection. Given that the advantages of in vitro propagation is ex situ conservation of germoplasm, this study aimed to evaluate the regeneration potential of Z incognita plants from leaf explants and zygotic embryos. The effect of different combinations of auxin (2.4-D and NAA) and cytokinins (KIN, BAP and TDZ) was evaluated on the formation of callus and shoot regeneration (direct or indirect), using MS (MB1) basal medium and B5 (MB2) basal modified medium. The callus formation was presented over a wide concentration range of 2.4-D with KIN and 2.4-D with BAP, regardless of the basal medium, but not in explants treated with ANA more TDZ or KIN. For leaf explants there was no response to the formation of somatic embryos or shoots with hormonal combinations and concentrations evaluated; however, MB2 calluses induced with 2.4-D (0.22 mg / l) and BAP (0. 1 , 2. 3 mg / l) were different, their nodular aspect, cream color and pro-embryogenic appearance coincided with a lot of potential meristematic cells for the regeneration process. From immature zygotic embryos, somatic embryo formation in the MB2 medium was achieved without growth regulators or containing 2.4-D alone (0.22 mg /l) or 2.4-D in combination with BAP (1 mg/l) without achieving the conversion process to seedlings.

Zamia (Cycadales: Zamiaceae) on Puerto Rico: asymmetric genetic differentiation and the hypothesis of multiple introductions.

PREMISE OF THE STUDY: This study of Zamia in Puerto Rico is the most intensive population genetics investigation of a cycad to date in terms of number of markers, and one of few microsatellite DNA studies of plants from the highly critical Caribbean biodiversity hotspot. Three distinctive Zamia taxa occur on the island: Z. erosa on the north coast, and Z. portoricensis and Z. pumila, both in the south. Their relationships are largely unknown. We tested three hypotheses about their genetic diversity, including the possibility of multiple introductions. METHODS: We used 31 microsatellite loci across 10 populations and analyzed the data with AMOVA, Bayesian clustering, and ABC coalescent modeling. KEY RESULTS: Puerto Rican zamias exhibit an amalgam of patterns of genetic differentiation that have been reported for cycads. Overall, the taxa are slightly inbred, with high infra-populational variation and little evidence of recent bottlenecks. Zamia erosa exhibits a more than threefold greater degree of population differentiation than the other two taxa. Admixture is evident only between Z. portoricensis and Z. pumila. Zamia portoricensis is inferred to be the youngest taxon on the island, on the basis of estimates of coalescence time and effective population size. A selective sweep may be underway in a small population of Z. erosa in a saline environment. CONCLUSIONS: Zamia erosa may represent an independent introduction into Puerto Rico; Z. portoricensis and Z. pumila fit a scenario of allopatric speciation. This will be explored further in the context of genetic analysis across the entire Caribbean region.

Two genera of Aulacoscelinae beetles reflexively bleed azoxyglycosides found in their host cycads.

Aulacoscelinae beetles have an ancient relationship with cycads (Cycadophyta: Zamiaceae), which contain highly toxic azoxyglycoside (AZG) compounds. How these "primitive" leaf beetles deal with such host-derived compounds remains largely unknown. Collections were made of adult Aulacoscelis appendiculata from Zamia cf. elegantissima in Panama, A. vogti from Dioon edule in Mexico, and Janbechynea paradoxa from Zamia boliviana in Bolivia. Total AZG levels were quantified in both cycad leaves and adult beetles by high performance liquid chromatography (HPLC). On average, cycad leaves contained between 0.5-0.8% AZG (frozen weight, FW), while adult beetles feeding on the same leaves contained even higher levels of the compounds (average 0.9-1.5% FW). High AZG levels were isolated from reflex bleeding secreted at the leg joints when beetles were disturbed. Nuclear magnetic resonance and mass spectroscopy identified two AZGs, cycasin and macrozamin, in the reflex bleeding; this is the first account of potentially plant-derived compounds in secretions of the Aulacoscelinae. These data as well as the basal phylogenetic position of the Aulacoscelinae suggest that sequestration of plant secondary metabolites appeared early in leaf beetle evolution.

Character-based, population-level DNA barcoding in Mexican species of Zamia L. (Zamiaceae: Cycadales).

BACKGROUND AND AIMS: With the recent proposal of matK and rbcL as core plant DNA barcoding regions by the Consortium for the Barcoding of Life Plant Working Group, the construction of reference libraries in the botanical DNA barcoding initiative has entered a new phase. However, in a recent DNA barcoding study in the three Mexican genera of the gymnosperm order Cycadales, we found that neither matK nor rbcL allow high levels of molecular identification of previously established species. MATERIALS AND METHODS: Our data analysis in that study rested on the "Characteristic Attributes Organization System" (CAOS), a character-based algorithm for the definition of "DNA diagnostics." Here, we use CAOS to analyze a population-level molecular data set in Zamia, one of the three cycad genera occurring in Mexico, whose populations display contrasting biogeographic patterns. Our population-level study, which includes all species in the region formally known as Megamexico, is restricted to the genome region, which showed the best single-locus molecular identification performance in our previous study-namely, the noncoding intergenic chloroplast spacer psbK-I. RESULTS: Our comparison of single-individual vs. population-level psbK-I datasets in Zamia indicates that CAOS analyses are sensitive to slight alignment changes, which in turn derive from the different amounts of molecular variation present in each matrix type. CONCLUSION: We, therefore, suggest that character-based studies that involve population-level data should contemplate this type of comparison between data matrices, before a set of DNA diagnostics in a given DNA barcoding reference library is considered definitive.

La familia Zamiaceae en Guatemala / The family Zamiaceae in Guatemala

La actualización del conocimiento de las Coniferas de Guatemala comprende 57 especies de plantas, de las cuales 28 especies son nativas y 29 son exóticas, agrupadas en diferentes familias, una de ellas, la familia Zamiaceae, que es un grupo de plantas parecidas a las palmas y que se estudiaron a través del Proyecto Universitario de Investigación en Ciencias Básicas PUICB.057 Actualización de la flora de Guatemala, capítulo I Pinophyta. Se colectaron y georeferenciaron ejemplares de las distintas especies que conforman las Zamiáceas, ello permitió preparar mapas de distribución de las especies nativas, incluidas en los géneros Ceratozamia (1 especie) y Zamia (4 especies) además se documentaron especies exóticas como Dioon mejiae y Zamia furfuraceae, empleadas como ornamentales...