Structural Study of Nano-Sized Gahnite (ZnAl2O4): From the Average to the Local Scale

Spinel gahnite (ZnAl2O4) has been obtained through a hydrothermal synthesis method with a grain size of about 2 nm. The sample was calcined for a few hours at two different temperatures (800 and 900 °C) in order to obtain larger grain sizes to be analyzed by means of powder diffraction with the Rietveld method, and by means of total scattering with the Pair Distribution Function (PDF) method. The idea is to compare the average to the local structure, as a function of increasing grain size. The total scattering data were collected at the European Synchrotron Radiation Facility (ESRF), Grenoble. The samples have been also characterised by means of high resolution Transmission Electron Microscopy (TEM), showing an increasing grain size up to about 9 nm. The average structure presented variations in the inversion degree and an increase in grain size. TEM observations demonstrated that the small crystals are well crystallised: the high resolution images neatly showed the atomic planes, even in the smallest particles. However, the average structure did not properly fit the PDF data in the local region, owing to a slightly different coordination among the octahedra. A new structural model is proposed for the local region of the PDF, that helped our understanding of the differences between a real nanostructured sample and that of a microcrystalline one. The oxygen disorder, due to the inversion grade of the spinel, is demonstrates to be at the basis of the local deviation. No signals of interstitial Zn atoms were detected

Unraveling the diversification history of grasshoppers belonging to the <i>"Trimerotropis pallidipennis"</i> (Oedipodinae: Acrididae) species group : A hotspot of biodiversity in the Central Andes

The Andean Mountain range has been recognized as one of the biodiversity hotspotsof the world. The proposed mechanisms for such species diversification, among others, are due to the elevation processes occurring during the Miocene and the intensiveglacial action during the Pleistocene. In this study we investigated the diversificationhistory of the grasshopper Trimerotropis pallidipennis species complex which showsa particularly wide latitudinal and altitudinal distribution range across the northern, central and southern Andes in South America. Many genetic lineages of this complexhave been so far discovered, making it an excellent model to investigate the role of thecentral Andes Mountains together with climatic fluctuations as drivers of speciation. Phylogenetics, biogeographic and molecular clock analyses using a multi-locus datasetrevealed that in Peru there are at least two, and possibly four genetic lineages. Twodifferent stocks originated from a common ancestor from North/Central Americawould have dispersed toward southern latitudes favored by the closure of the PanamaIsthmus giving rise to two lineages, the coastal and mountain lineages, which still coexistin Peru (i.e., T. pallidipennis and T. andeana). Subsequent vicariant and dispersalevents continued the differentiation process, giving rise to three to six genetic lineages(i.e., clades) detected in this study, which were geographically restricted to locationsdispersed over the central Andes Mountains in South America. Our results provideanother interesting example of ``island diversification" motored by the topographyplus unstable climatic conditions during the Pleistocene, pointing out the presence ofa hotspot of diversification in the Andean region of Peru.Centro de Estudios Parasitológicos y de Vectore

The Effect of Reproductive System on Invasiveness: Lessons from South American Weevils

Successful invasion of a species into novel, marginal areas often requires the ability to face different ecological characteristics than those prevailing in its native environment. In insects, one of the factors that affect invasiveness is the reproductive system. Unisexuality provides advantages because a single specimen can initiate a new population. Unisexual reproduction precludes breakup of genetic combinations that promote ecological specialization, although it may limit evolutionary potential for colonization. In order to assess the importance of the reproductive mode in the colonization ability of the weevils that are native to South America, we compared 1 bisexual and 2 parthenogenetic species that expanded their ranges in the last 2 centuries. First, for parthenogenetic species we tested clonality of the sample. Second, we proposed central and marginal areas through phylogeographic and habitat modeling analyses, and identified the pathways of dispersal for each species. Bisexual Naupactus xanthographus (Germar) (Coleoptera: Curculionidae) expanded its range westward to areas with similar environmental constraints than in its native ecosystem. Conversely, parthenogenetic Naupactus leucoloma Boheman and Naupactus cervinus Boheman (both Coleoptera: Curculionidae) invaded other continents where they had low to null predicted habitat suitability. While a single clone of N. cervinus successfully established around the world in areas with apparently adverse conditions, clones of N. leucoloma expanded their range to areas only moderately suitable. We conclude that parthenogenesis is a driver in these particular species for colonization of marginal habitats. However, N. cervinus also would have pre-existing adaptations that allowed it to establish in areas with apparently low potential to survive.Fil: Rodriguero, Marcela Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; ArgentinaFil: Guzman, Noelia Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; ArgentinaFil: Lanteri, Analia Alicia. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; ArgentinaFil: Confalonieri, Viviana Andrea. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Ecología, Genética y Evolución. Grupo de Investigación en Filogeografía y Filogenias Moleculares; Argentin

CRISPR/Cas9-Mediated Targeted Mutagenesis of CYP93E2 Modulates the Triterpene Saponin Biosynthesis in Medicago truncatula

In the Medicago genus, triterpene saponins are a group of bioactive compounds extensively studied for their different biological and pharmaceutical properties. In this work, the CRISPR/Cas9-based approach with two single-site guide RNAs was used in Medicago truncatula (barrel medic) to knock-out the CYP93E2 and CYP72A61 genes, which are responsible for the biosynthesis of soyasapogenol B, the most abundant soyasapogenol in Medicago spp. No transgenic plants carrying mutations in the target CYP72A61 gene were recovered while fifty-two putative CYP93E2 mutant plant lines were obtained following Agrobacterium tumefaciens-mediated transformation. Among these, the fifty-one sequenced plant lines give an editing efficiency of 84%. Sequencing revealed that these lines had various mutation patterns at the target sites. Four T0 mutant plant lines were further selected and examined for their sapogenin content and plant growth performance under greenhouse conditions. The results showed that all tested CYP93E2 knock-out mutants did not produce soyasapogenols in the leaves, stems and roots, and diverted the metabolic flux toward the production of valuable hemolytic sapogenins. No adverse influence was observed on the plant morphological features of CYP93E2 mutants under greenhouse conditions. In addition, differential expression of saponin pathway genes was observed in CYP93E2 mutants in comparison to the control. Our results provide new and interesting insights into the application of CRISPR/Cas9 for metabolic engineering of high-value compounds of plant origin and will be useful to investigate the physiological functions of saponins in planta

Are flightless Galapaganus weevils older than the Galapagos islands they inhabit?

The 15 species in the weevil genus Galapaganus Lanteri 1992 (Entiminae: Curculionidae: Coleoptera) are distributed on coastal Peril and Ecuador and include 10 flightless species endemic to the Galapagos islands. These beetles thus provide a promising system through which to investigate the patterns and processes of evolution on Darwin's archipelago. Sequences of the mtDNA locus encoding cytochrome oxidase subunit I (COI) were obtained from samples of seven species occurring in different ecological zones of the oldest south-eastern islands: San Cristobal, Espanola and Floreana, and the central island Santa Cruz. The single most parsimonious tree obtained shows two well-supported clades that correspond to the species groups previously defined by morphological characters. Based on a mtDNA clock calibrated for arthropods, the initial speciation separating the oldest species, G. galapagoensis (Linell) on the oldest island, San Cristobal, from the remaining species in the Galapagos occurred about 7.2 Ma. This estimate exceeds geological ages of the extant emerged islands, although it agrees well with molecular dating of endemic Galapagos iguanas, geckos and lizards. An apparent explanation for the disagreement between geological and molecular time-frames is that about 7 Ma there were emerged islands which subsequently disappeared under ocean waters. This hypothesis has gained support from the recent findings of 11 -Myr-old submarine seamounts (sunken islands), south-east of the present location of the archipelago. Some species within the darwini group may have differentiated on the extant islands, 1-5 Ma.Facultad de Ciencias Naturales y Muse

Are flightless Galapaganus weevils older than the Galapagos islands they inhabit?

The 15 species in the weevil genus Galapaganus Lanteri 1992 (Entiminae: Curculionidae: Coleoptera) are distributed on coastal Peril and Ecuador and include 10 flightless species endemic to the Galapagos islands. These beetles thus provide a promising system through which to investigate the patterns and processes of evolution on Darwin's archipelago. Sequences of the mtDNA locus encoding cytochrome oxidase subunit I (COI) were obtained from samples of seven species occurring in different ecological zones of the oldest south-eastern islands: San Cristobal, Espanola and Floreana, and the central island Santa Cruz. The single most parsimonious tree obtained shows two well-supported clades that correspond to the species groups previously defined by morphological characters. Based on a mtDNA clock calibrated for arthropods, the initial speciation separating the oldest species, G. galapagoensis (Linell) on the oldest island, San Cristobal, from the remaining species in the Galapagos occurred about 7.2 Ma. This estimate exceeds geological ages of the extant emerged islands, although it agrees well with molecular dating of endemic Galapagos iguanas, geckos and lizards. An apparent explanation for the disagreement between geological and molecular time-frames is that about 7 Ma there were emerged islands which subsequently disappeared under ocean waters. This hypothesis has gained support from the recent findings of 11 -Myr-old submarine seamounts (sunken islands), south-east of the present location of the archipelago. Some species within the darwini group may have differentiated on the extant islands, 1-5 Ma.Facultad de Ciencias Naturales y Muse

A tale of swinger insects: Signatures of past sexuality between divergent lineages of a parthenogenetic weevil revealed by ribosomal intraindividual variation

Naupactus cervinus (Boheman) (Curculionidae, Naupactini) is a parthenogenetic weevil native to the Paranaense Forest which displays high levels of genetic variation. Two divergent clades were identified, one ranging in forest areas (Forest clade), and the other in open vegetation areas (Grassland clade). Both of them have individuals with high levels of heterozygosity in ribosomal sequences. Investigation of intraindividual variation in ITS1 sequences through cloning and posterior sequencing suggested that mating between both groups most likely occurred in the Paranaense Forest after a secondary contact, which led to fixed heterozygotes as a consequence of parthenogenesis. Otherwise, sexual segregation would have disrupted multilocus genotypes. Only a small number of heterozygous genotypes of all the possible combinations are found in nature. We propose the occurrence of a hybrid zone in the Paranaense Forest. The fact that it is one of the most important biodiversity hotspots of the world, together with its key role for investigating evolutionary processes, makes it worthy of conservation. This is the first genetic evidence of bisexuality in N. cervinus.Fil: Rodriguero, Marcela Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; ArgentinaFil: Wirth, Sonia Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Alberghina, Josefina Silvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; ArgentinaFil: Lanteri, Analía Alicia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. División Entomología; ArgentinaFil: Confalonieri, Viviana Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; Argentin

Host-specific gene expression as a tool for introduction success in Naupactus parthenogenetic weevils

Food resource access can mediate establishment success in invasive species, and generalist herbivorous insects are thought to rely on mechanisms of transcriptional plasticity to respond to dietary variation. While asexually reproducing invasives typically have low genetic variation, the twofold reproductive capacity of asexual organisms is a marked advantage for colonization. We studied host-related transcriptional acclimation in parthenogenetic, invasive, and polyphagous weevils: Naupactus cervinus and N. leucoloma. We analyzed patterns of gene expression in three gene categories that can mediate weevil-host plant interactions through identification of suitable host plants, short-term acclimation to host plant defenses, and long-term adaptation to host plant defenses and their pathogens. This approach employed comparative transcriptomic methods to investigate differentially expressed host detection, detoxification, immune defense genes, and pathway-level gene set enrichment. Our results show that weevil gene expression responses can be host plant-specific, and that elements of that response can be maintained in the offspring. Some host plant groups, such as legumes, appear to be more taxing as they elicit a complex gene expression response which is both strong in intensity and specific in identity. However, the weevil response to taxing host plants shares many differentially expressed genes with other stressful situations, such as host plant cultivation conditions and transition to novel host, suggesting that there is an evolutionarily favorable shared gene expression regime for responding to different types of stressful situations. Modulating gene expression in the absence of other avenues for phenotypic adaptation may be an important mechanism of successful colonization for these introduced insects.Fil: Mackay Smith, Ava. Wellesley College; Estados UnidosFil: Dornon, Mary Kate. Wellesley College; Estados UnidosFil: Lucier, Rosalind. Wellesley College; Estados UnidosFil: Okimoto, Anna. Wellesley College; Estados UnidosFil: Sousa, Flavia Mendonca de. Wellesley College; Estados UnidosFil: Rodriguero, Marcela Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Confalonieri, Viviana Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Lanteri, Analia Alicia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. División Entomología; ArgentinaFil: Sequeira, Andrea. Wellesley College; Estados Unido

Dependence of egg hatching on Wolbachia density in a parthenogenetic weevil revealed by antibiotic treatment

Naupactini is a tribe of Neotropical broad-nosed weevils highly diverse in South America. This group includes several parthenogenetic species, some of them harmful for agriculture and invasive around the world. Although some hypotheses based on polyploidy and hybridization have been proposed to explain the origin of parthenogenesis in weevils, the infection with the bacterium Wolbachia pipientis may be involved in the origin of parthenogenetic reproduction of some species. In this contribution, we studied the role of Wolbachia in the reproductive biology of Pantomorus postfasciatus Hustache (Coleoptera: Curculionidae) through a curing experiment using tetracycline. This weevil has a mixed mode of reproduction including sexual and parthenogenetic populations. Exposure to an antibiotic did not affect fecundity, but did reduce egg hatching in comparison with untreated individuals. Consequently, we inferred that Wolbachia most probably takes part in the reproduction of P. postfasciatus, either by exerting nutritive functions in oogenesis necessary for egg hatching, or by induction of thelytokous parthenogenesis. Although infection was not totally cured, Wolbachia load was significantly lower in treated than in control females. Thereby, we hypothesize that a minimum threshold density of Wolbachia is required for weevil reproduction. We conclude that all analyses support a role of Wolbachia in P. postfasciatus reproduction.Fil: Rodriguero, Marcela Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; ArgentinaFil: Scannapieco, Alejandra Carla. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Genética; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Monti, Daniela Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; ArgentinaFil: Chifflet, Lucila. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; ArgentinaFil: Elías Costa, Agustín Jorge. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; ArgentinaFil: Lanteri, Analia Alicia. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; ArgentinaFil: Confalonieri, Viviana Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; Argentin

Unraveling the diversification history of grasshoppers belonging to the <i>"Trimerotropis pallidipennis"</i> (Oedipodinae: Acrididae) species group : A hotspot of biodiversity in the Central Andes

The Andean Mountain range has been recognized as one of the biodiversity hotspotsof the world. The proposed mechanisms for such species diversification, among others, are due to the elevation processes occurring during the Miocene and the intensiveglacial action during the Pleistocene. In this study we investigated the diversificationhistory of the grasshopper Trimerotropis pallidipennis species complex which showsa particularly wide latitudinal and altitudinal distribution range across the northern, central and southern Andes in South America. Many genetic lineages of this complexhave been so far discovered, making it an excellent model to investigate the role of thecentral Andes Mountains together with climatic fluctuations as drivers of speciation. Phylogenetics, biogeographic and molecular clock analyses using a multi-locus datasetrevealed that in Peru there are at least two, and possibly four genetic lineages. Twodifferent stocks originated from a common ancestor from North/Central Americawould have dispersed toward southern latitudes favored by the closure of the PanamaIsthmus giving rise to two lineages, the coastal and mountain lineages, which still coexistin Peru (i.e., T. pallidipennis and T. andeana). Subsequent vicariant and dispersalevents continued the differentiation process, giving rise to three to six genetic lineages(i.e., clades) detected in this study, which were geographically restricted to locationsdispersed over the central Andes Mountains in South America. Our results provideanother interesting example of ``island diversification" motored by the topographyplus unstable climatic conditions during the Pleistocene, pointing out the presence ofa hotspot of diversification in the Andean region of Peru.Centro de Estudios Parasitológicos y de Vectore