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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.

Premise of research. The genus Zamia L. (Zamiaceae), consisting of 79 species, is the most species-rich and widely distributed cycad genus in the New World and arguably the most morphologically and ecologically diverse genus in the Cycadales. However, a strong phylogenetic framework for this genus is still lacking. Methodology. We used a multilocus sequence data set of 10 independent loci (nine single-copy nuclear genes [SCNGs] and one plastid) and extensive taxon sampling (ca. 90% of species) to infer phylogenetic relationships within Zamia. We implemented a concatenated matrix analysis with maximum parsimony, a partitioned maximum likelihood (ML) analysis, and a time-calibrated Bayesian species tree-estimation approach. Diversification, historical biogeography, and ancestral character state reconstruction analyses were conducted using the species tree topology that was the most morphologically and geographically congruent. Pivotal results. We inferred a robust phylogenetic tree for the genus with a strong geographic delimitation of clades and found that four morphological characters typically used for diagnostic purposes in the genus exhibit a high degree of homoplasy. The stem group of Zamia was estimated to have originated at 68.28 Ma (95% highest posterior density [HPD] 51.0–84.5) and the crown group at 9.54 Ma (95% HPD 9.0–10.62). The majority of species richness in the genus appeared during the Pliocene and Pleistocene, with the highest diversification rates occurring in clades comprising Caribbean and South American species. Biogeographic analysis suggests a Caribbean or Mesoamerican origin for Zamia with subsequent dispersal to the Central American Isthmus and South America, where the genus reaches its maximum species and morphological diversity. Conclusions. The high degree of convergence found in multiple morphological characters has historically confounded efforts to elucidate species relationships using nonphylogenetic methods. This study presents the most species-comprehensive, well-resolved hypothesis of phylogenetic relationships within Zamia and provides a strong phylogenetic framework for further studies.

Background Bayesian relaxed-clock dating has significantly influenced our understanding of the timeline of biotic evolution. This approach requires the use of priors on the branching process, yet little is known about their impact on divergence time estimates. We investigated the effect of branching priors using the iconic cycads. We conducted phylogenetic estimations for 237 cycad species using three genes and two calibration strategies incorporating up to six fossil constraints to (i) test the impact of two different branching process priors on age estimates, (ii) assess which branching prior better fits the data, (iii) investigate branching prior impacts on diversification analyses, and (iv) provide insights into the diversification history of cycads. Results Using Bayes factors, we compared divergence time estimates and the inferred dynamics of diversification when using Yule versus birth-death priors. Bayes factors were calculated with marginal likelihood estimated with stepping-stone sampling. We found striking differences in age estimates and diversification dynamics depending on prior choice. Dating with the Yule prior suggested that extant cycad genera diversified in the Paleogene and with two diversification rate shifts. In contrast, dating with the birth-death prior yielded Neogene diversifications, and four rate shifts, one for each of the four richest genera. Nonetheless, dating with the two priors provided similar age estimates for the divergence of cycads from Ginkgo (Carboniferous) and their crown age (Permian). Of these, Bayes factors clearly supported the birth-death prior. Conclusions These results suggest the choice of the branching process prior can have a drastic influence on our understanding of evolutionary radiations. Therefore, all dating analyses must involve a model selection process using Bayes factors to select between a Yule or birth-death prior, in particular on ancient clades with a potential pattern of high extinction. We also provide new insights into the history of cycad diversification because we found (i) periods of extinction along the long branches of the genera consistent with fossil data, and (ii) high diversification rates within the Miocene genus radiations.

As a gymnosperm group, cycads are known for their ancient origin and specialized coralloid root, which can be used as an ideal system to explore the interaction between host and associated microorganisms. Previous studies have revealed that some nitrogen-fixing cyanobacteria contribute greatly to the composition of the endophytic microorganisms in cycad coralloid roots. However, the roles of host and environment in shaping the composition of endophytic bacteria during the recruitment process remain unclear. Here, we determined the diversity, composition, and function prediction of endophytic bacteria from the coralloid roots of a widely cultivated cycad, Cycas revoluta Thunb. Using next-generation sequencing techniques, we comprehensively investigated the diversity and community structure of the bacteria in coralloid roots and bulk soils sampled from 11 sites in China, aiming to explore the variations in core endophytic bacteria and to predict their potential functions. We found a higher microbe diversity in bulk soils than in coralloid roots. Meanwhile, there was no significant difference in the diversity and composition of endophytic bacteria across different localities, and the same result was found after removing cyanobacteria. Desmonostoc was the most dominant in coralloid roots, followed by Nostoc, yet these two cyanobacteria were not shared by all samples. Rhodococcus, Edaphobacter, Niastella, Nordella, SH-PL14, and Virgisporangium were defined as the core microorganisms in coralloid roots. A function prediction analysis revealed that endophytic bacteria majorly participated in the plant uptake of phosphorus and metal ions and in disease resistance. These results indicate that the community composition of the bacteria in coralloid roots is affected by both the host and environment, in which the host is more decisive. Despite the very small proportion of core microbes, their interactions are significant and likely contribute to functions related to host survival. Our study contributes to an understanding of microbial diversity and composition in cycads, and it expands the knowledge on the association between hosts and symbiotic microbes.

The current research aimed to evaluate the potential effects of Solanum mammosum, Dioon mejiae, and Amanita caesarea on Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus survival and performance after exposure to different harsh conditions such as bile, acid, gastric juice, and lysozyme to mimic the digestive system from mouth to the intestine. Probiotic protease activity was observed to evaluate the proteolytic system. Probiotics were cultured in a broth mixed with plant material, and after incubation, the results were compared to the control sample. Therefore, plant material’s total phenolic compound, total carotenoid compound, antioxidant activity, sugar profile, and acid profile were obtained to discuss their impact on the survival of probiotics. The results indicate that Amanita caesarea negatively affected probiotic survival in the bile tolerance test and positively affected Lactobacillus bulgaricus in the protease activity test. Otherwise, the other plant material did not change the results significantly (p > 0.05) compared to the control in different tests. Consequently, Solanum mammosum and Dioon mejiae had no significant effects (p > 0.05) in increasing probiotic survival.

Heterocyte-forming cyanobacteria form symbiotic relationships with several lineages of plants. Here, twenty (20) strains of endosymbiotic cyanobacteria (cyanobionts) with Nostoc-like morphologies were isolated from the highly specialized coralloid roots of five host species in Cycadales—Cycas debaoensis, C. fairylakea, C. elongata, Ceratozamia robusta, and Macrozamia moorei. Molecular phylogeny based on the 16S rRNA gene placed these strains into seven different taxa within the Nostocaceae, specifically under the genera Desmonostoc and Dendronalium. The percent dissimilarity and unique patterns in the secondary structures of the D1-D1′, Box-B, V2, and V3 helices, which were based on the 16S–23S rRNA internal transcribed spacer (ITS) regions, supported three distinct species in Desmonostoc. These three morphologically distinct novel species are described in this report: Desmonostoc debaoense sp. nov., Desmonostoc meilinense sp. nov., and Desmonostoc xianhuense sp. nov. Other investigated strains were phylogenetically identified as members of the recently discovered genus Dendronalium and represent the first report of association of that genus with cycads. Our findings suggest that the order Cycadales hosts diverse species of cyanobionts in their coralloid roots and that many potential unreported or novel taxa are present in cycads occurring in their natural habitat and await discovery.

 Ceratozamia (Zamiaceae, Cycadales), is a member one of the most endangered seed plant groups. Species of Ceratozamia grow from near sea level up to 2,100 meters in Mexico and Mesoamerica. We present a modern taxonomic treatment of Ceratozamia , based on fieldwork combined with detailed study of herbarium specimens in and from Mexico and Central America. This new revision is based on incorporation of morphological, molecular and biogeographic data that have been previously published in circumscriptions of species complexes by our group. Detailed morphological descriptions of the 36 species of Ceratozamia are provided and relevant characters for the genus are discussed and described. A majority are endemic to Mexico and are concentrated at high elevations in mountainous areas. Synonymies, lectotypifications, etymologies, taxonomic notes, distribution maps, illustrations and detailed species-level comparisons are included, as well as a dichotomous key for identification of all species. Data on distributional ranges and habitats of all species are summarized. Ceratozamia osbornei D.W.Stev., Mart.-Domínguez & Nic.-Mor., sp. nov. is described from evergreen tropical forests of Belize and we highlight new populations and distributional ranges for C. subroseophylla Mart.-Domínguez & Nic.-Mor. and C. vovidesii Pérez-Farr. & Iglesias in the Mesoamerican region. Keywords: Circumscription, cryptic species, cycads, lectotypification, Mesoamerica, Mexico

A useful DNA barcode requires sufficient sequence variation to distinguish between species and ease of application across a broad range of taxa. Discovery of a DNA barcode for land plants has been limited by intrinsically lower rates of sequence evolution in plant genomes than that observed in animals. This low rate has complicated the trade-off in finding a locus that is universal and readily sequenced and has sufficiently high sequence divergence at the species-level. Here, a global plant DNA barcode system is evaluated by comparing universal application and degree of sequence divergence for nine putative barcode loci, including coding and non-coding regions, singly and in pairs across a phylogenetically diverse set of 48 genera (two species per genus). No single locus could discriminate among species in a pair in more than 79% of genera, whereas discrimination increased to nearly 88% when the non-coding trnH-psbA spacer was paired with one of three coding loci, including rbcL. In silico trials were conducted in which DNA sequences from GenBank were used to further evaluate the discriminatory power of a subset of these loci. These trials supported the earlier observation that trnH-psbA coupled with rbcL can correctly identify and discriminate among related species. A combination of the non-coding trnH-psbA spacer region and a portion of the coding rbcL gene is recommended as a two-locus global land plant barcode that provides the necessary universality and species discrimination.

It has been observed that the leaves of some Zamia species undergo a kind of “reverse ripening”; that is, they change from their original brown color to green during development. We assumed that this strange color change was due to the change in carotenoid composition, so we followed the changes for several weeks. The detailed carotenoid composition and content at different stages of development of the leaves was determined with HPLC-DAD focusing on the changes in red and yellow carotenoids. The total and relative amounts of red and yellow carotenoids were determined simultaneously from one measurement from a saponified and/or unsaponified extract. At the beginning of development, the concentration of red carotenoids was higher than that of the yellow ones; it decreased drastically until 22 days and continued to decrease slowly until they completely disappeared. The concentration of yellow carotenoids decreased at the beginning as well, but after 22 days it started to increase. The amount of red carotenoids started to decrease when the leaflet stopped growing. Lutein is the main component in old leaflets, which is not a red carotenoid precursor. Red carotenoids can always be found in their esterified form in the leaves. These findings support the hypothesis that red and yellow carotenoid accumulation are independent and probably have different functions in the leaflet. The strange color change was explained based on the compartmentalization of red and yellow carotenoids and on the changing activity of the enzyme capsanthin–capsorubin synthase responsible for the synthesis of red carotenoids capsorubin and capsanthin.