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Frontiers in Plant Science
Plant Development and EvoDevo
Genetic Regulatory Mechanisms Underlying Developmental Shifts in Plant Evolution View all 19 Articles
Annette Becker
University of Giessen, Germany
Masaru Ohme-Takagi
Saitama University, Japan
Charles Gasser
University of California, Davis, United States
The editor and reviewers' affiliations are the latest provided on their Loop research profiles and may not reflect their situation at the time of review.
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Original Research ARTICLE
Front. Plant Sci., 19 September 2018 | https://doi.org/10.3389/fpls.2018.01352
Evidence for the Extensive Conservation of Mechanisms of Ovule Integument Development Since the Most Recent Common Ancestor of Living Angiosperms
Gontran Arnault1, Aurélie C. M. Vialette1, Amélie Andres-Robin1, Bruno Fogliani2, Gildas Gâteblé2 and Charles P. Scutt1*
1Laboratoire Reproduction et Deìveloppement des Plantes, École Normale Supérieure de Lyon, Université Claude Bernard Lyon 1, CNRS, INRA, Université de Lyon, Lyon, France
2Équipe ARBOREAL, "Agriculture Biodiversité et Valorisation", Institut Agronomique Néo-Calédonien (IAC), Païta, New Caledonia
The ovules and seeds of most angiosperm groups are enclosed by two integuments, whose evolutionary origins are considerably separated in time, as the inner integument arose over 300 million years ago (MYA) in an ancestor of all living seed plants, while the outer integument arose, perhaps as recently as 164 MYA, in an ancestor of all living angiosperms. Studies of the model angiosperm Arabidopsis thaliana indicate that the mechanisms of development of the inner and outer integuments depend on largely different sets of molecular players. However, it was not known, in most cases, whether these differences were already present in early flowering plants, or arose later in the Arabidopsis lineage. Here, we analyze the expression patterns of integument regulators in Amborella trichopoda, the likely sister to all other living angiosperms. The data obtained indicate that regulators of the YABBY, KANADI, and homeodomain-leucine zipper class III transcription factor families have largely conserved their integument-specific expression profiles in the Amborella and Arabidopsis lineages since the most recent common ancestor (MRCA) of living angiosperms. We identified only one case, involving the paralogous genes ETTIN and AUXIN RESPONSE FACTOR4, in which integument-specific expression patterns had clearly diverged between Amborella and Arabidopsis. We use the data obtained to partially reconstruct molecular mechanisms of integument development in the MRCA of living angiosperms and discuss our findings in the context of alternative hypotheses for the origin of the angiosperm outer integument.
The ovules and seeds of most seed plants are covered by one or two integuments. These maternal tissues function to (1) protect the internal tissues of the ovule and later the seed, (2) define a route, via the micropyle, for pollen or pollen-tube entry, and (3) contribute in many cases to the regulation of seed hydration and dormancy (Linkies et al., 2010). Gymnosperms possess a single integument, whereas the majority of angiosperm groups, including the most basally diverging of these, possess two integuments, of which the inner integument is considered homologous to the single integument of gymnosperms. The origin of the inner integument therefore dates from before the separation of the living angiosperm and gymnosperm lineages, believed to have occurred over 300 million years ago (MYA), while the outer integument must have arisen somewhere along the angiosperm stem lineage, perhaps as recently as ∼164 MYA: a reasonable estimated date for the most recent common ancestor (MRCA) of living angiosperms (Moore et al., 2010). Within angiosperms, several groups have undergone a secondary reduction to a single integument (Endress, 2011), notably including the asterids, which contain over 80,000 species.
Much of what is known of the molecular mechanisms of integument development comes from the study of the model angiosperm Arabidopsis thaliana (hereafter referred to as Arabidopsis), which possesses anatropous, bitegmic ovules. Ovule initiation in Arabidopsis begins with the expression of the homeobox transcription factor WUSCHEL (WUS) (see Mathews and Kramer, 2012), which is known to promote cell proliferation in meristematic tissues. The ovule thus shares characteristics of meristems, such as the stem apical meristem and floral meristem, both of which produce lateral organs on their flanks. In the case of the bitegmic ovule, the lateral organs produced in this way are the inner and outer integuments, which arise from the chalazal tissue in the central region of the elongating ovule primordium. The inner integument arises first, followed closely by the outer integument (Endress, 2011).
Early expression of WUS at the apex of the Arabidopsis ovule primordium defines the presumptive nucellar tissue. The outgrowth of the integuments from the chalazal region, immediately below the nucellus, depends on the expression of three homeodomain-leucine zipper class III (HD-ZIP III) transcription factors, PHABULOSA (PHB), PHAVOLUTA (PHV), and CORONA (CNA) (Kelley et al., 2009), which act redundantly, and together with other factors, to limit the basal spread of WUS expression (Yamada et al., 2016). Expression of these HD-ZIP III factors defines the adaxial (towards the growth axis) zone of the presumptive inner integument. The inner and outer integument primordia are associated with the formation of auxin maxima, and it has been proposed that ETTIN/AUXIN RESPONSE FACTOR3 (ETT/ARF3) and ABERRANT TESTA SHAPE/KANADI4 (ATS/KAN4), both of which are expressed in the abaxial (away from the growth axis) domain of the inner integument, act to remove auxin from the zone between the two integument primordia (Kelley et al., 2012). Interestingly, Lora et al. (2015) found that in certain species of Prunus whose ovules contain only one integument, expression of the ETT ortholog was absent from the chalaza and inner integument. These authors accordingly proposed the loss of ETT expression to represent a potential evolutionary mechanism for the reduction from two integuments to one in these species.
HD-ZIP III and KANADI family factors are believed to control abaxial–adaxial tissue polarity in lateral organs through the regulation of a common set of direct target genes, many of which are related to auxin signaling or dynamics (Reinhart et al., 2013; Huang et al., 2014). HD-ZIP III factors regulate these targets positively to promote adaxial tissue identity, while KANADI factors regulate the same genes negatively to promote abaxial tissue identity. In Arabidopsis, there is a clear division of labor between the inner and outer integuments, as ATS is the principal KANADI family member expressed in the inner integument (McAbee et al., 2006), while this role is played redundantly by KAN1 and KAN2 in the outer integument (Eshed et al., 2004; McAbee et al., 2006). In the HD-ZIP III family, PHB, PHV, and CNA are expressed specifically in the inner integument, while REV is expressed in both integuments (Sieber et al., 2004b; Kelley et al., 2009).
Tissue outgrowth in angiosperm lateral organs, including for example leaves and carpels, is typically associated with the expression of YABBY transcription factors. These factors are expressed in the abaxial tissue domain, and can act either positively or negatively on the transcription of downstream targets, apparently through protein–protein interactions with distinct sets of co-factors (Bonaccorso et al., 2012; Simon et al., 2017). Despite their abaxial-specific expression profiles, YABBYs are believed not to define abaxial tissue identity per se, but rather to facilitate lateral organ outgrowth by promoting the expression of WUS-related transcription factors in marginal meristems (Nakata et al., 2012). The YABBY gene INNER NO OUTER (INO), is expressed specifically in the abaxial zone of the Arabidopsis outer integument, and an orthologue of this gene shows a largely similar expression pattern in the basally diverging angiosperm Nymphaea (Nymphaeales, Nymphaeaceae) (Yamada et al., 2003, 2011), suggesting INO's tissue-specific role to have been conserved from early stages of angiosperm evolution.
With very few exceptions (such as that of INO, mentioned above), the mechanisms of integument development have been investigated to date uniquely in Arabidopsis, and it is therefore not known, in most cases, which of these were already present in early flowering plants and which evolved later in the Arabidopsis lineage. In the present work, we investigate the conservation of mechanisms of integument development from very early stages of angiosperm evolution by analyzing the expression patterns of the orthologs of Arabidopsis integument regulators in the basally diverging angiosperm Amborella trichopoda (hereafter referred to as Amborella). Most recent molecular phylogenetic studies place Amborella, the only known representative of Amborellales, as sister to all other living angiosperms (Stevens, 2001 onwards). According to these studies, Amborellales is the first of the three earliest diverging "ANA-grade" angiosperm orders, the remaining two being Nymphaeales and Austrobaileyales. Amborella is a dioecious scrambling shrub, endemic to the subtropical rainforests of the southern pacific island of New Caledonia. Its female flowers produce 5–6 unfused carpels, each containing a single, pendant, bitegmic ovule. These ovules are of more-or-less orthotropous symmetry, though show a distinct longitudinal curvature, which may be due to developmental constraints within the ovary, or may represent the residual effect of an ancestral anatropous symmetry (Endress and Igersheim, 2000).
Here, we focus on integument regulators of the YABBY, KANADI, HD-ZIP III, and ARF transcription factor families. We use gene expression patterns to identify molecular mechanisms of integument development that have probably been conserved since the MRCA of living flowering plants, and others that appear to have undergone changes in either the Amborella or Arabidopsis lineages. We discuss the data obtained in the context of alternative hypotheses for the origin of the outer integument in a distant common ancestor of living angiosperms.
Phylogenetic Reconstruction
Phylogenetic reconstructions in the KANADI and HD-ZIP III families were performed using a wide taxonomic sampling of angiosperms. Sequences for inclusion in phylogenetic analyses were selected by TBLASTN searching (Altschul et al., 1997) of the NCBI/NIH non-redundant nucleotide database1 and the Amborella genome database2 using Arabidopsis integument development proteins. Protein sequences were aligned using MUSCLE in the SeaView phylogeny package (Gouy et al., 2010). Sites were selected using G-BLOCKS, using all three options provided to minimize the stringency of selection. Phylogenies were generated from amino acid alignments in PhyML (Guindon et al., 2009) using the following parameters: model: LG; branch support: aLRT; amino acid equilibrium frequencies: model-given; invariable sites: optimized; across-site rate variation: optimized; tree searching operations: best of NNI and SPR; and starting tree: BioNJ.
Gene Expression Studies
Full-length nucleotide sequences corresponding to Amborella orthologs or pro-orthologs of Arabidopsis integument regulators were PCR-amplified from an Amborella female flower cDNA library (Fourquin et al., 2005) using the primers shown in Supplementary Table S1. Digoxigenin-labeled riboprobes were generated from these and used in non-radioisotopic in situ hybridization to tissue sections of Amborella flower buds at approximately Stage 7 (Buzgo et al., 2004), and of female flowers at anthesis, using the protocol given by Vialette-Guiraud et al. (2011). Gene expression patterns were observed and photographed under bright field illumination using a Leica Axio Imager M2 inverted microscope fitted with a Leica AxioCam MRc digital camera.
Anatomical Observations
Amborella ovule anatomy was revealed in sections of fixed female flowers, prepared and photographed as for in situ hybridization. These sections were stained with 0.05% (w/v) toluidine Blue-0 in 0.1 M sodium phosphate buffer (pH 6.8).
Clear Orthologs of Several Arabidopsis Integument Regulators Can Be Identified in Amborella
Several previous studies have focused on the molecular phylogeny of gene families involved in the regulation of integument development. Accordingly, the Amborella ortholog of the Arabidopsis YABBY-family integument regulator INO has previously been reported (Finet et al., 2016), as have those of the paralogous and partially redundant auxin response family regulators ETT and AUXIN RESPONSE FACTOR4 (ARF4) (Finet et al., 2010). However, published phylogenies of the KANADI and HD-ZIP III families have not to our knowledge previously included sequences from Amborella.
Phylogenetic reconstructions of the HD-ZIP III family in the present study (Supplementary Figure S1), generated from the protein alignment shown in Supplementary Figure S2, succeeded in identifying clear Amborella orthologs of the Arabidopsis genes REV and CNA, and a clear pro-ortholog of the Arabidopsis genes PHB and PHV, which accordingly appear to be derived from a duplication within the crown group of living angiosperms. In all these cases, the Amborella genes identified as orthologs or pro-orthologs of Arabidopsis integument regulators occupied basal positions within their respective clades, in agreement with the likely phylogenetic placement of Amborella.
Phylogenetic reconstruction of the KANADI family (Supplementary Figure S3), generated from the protein alignment shown in Supplementary Figure S4, succeeded in identifying a clear Amborella ortholog of the Arabidopsis inner integument regulator ATS (KAN4), which grouped in a small basal clade of ATS-like sequences. Relationships of gene orthology between Amborella sequences and the Arabidopsis outer integument regulators KAN1 and KAN2 were less clear in phylogenetic reconstructions.
The results obtained in these reconstructions, combined with those of previously published work, permitted detailed analyses in the present study of the expression in Amborella female flower tissues of the genes: Atr.INO (XM_006840616), Atr.ATS, (XM_006856161.3), AtrPHB/PHV (XM_006845952.3), Atr.REV (XM_006846714.3), Atr.ARF4 (XM_020671056), and Atr.ETT (XM_011627988.2). The longer alternative splicing variant of Atr.ARF4 (Finet et al., 2010) was used for in situ hybridization studies.
The Amborella Orthologue of the YABBY Gene INO Is Narrowly Expressed in the Abaxial Domain of the Outer Integument, As Is INO in Arabidopsis
Atr.INO proved to be specifically expressed in the most abaxial cell layer of the outer integument. Its expression was first noted in the outer integument, towards the chalazal-end of the ovary (Figure 1A), spreading towards the micropyle-end with the growth of the outer integument (Figure 1B). This expression pattern closely resembles that of INO in Arabidopsis. Atr.INO expression was stronger in early stages of Amborella ovule development, becoming absent in the mature ovule (Figure 1C). These data reinforce observations in Nymphaea (Yamada et al., 2003) that already suggested Arabidopsis INO to have conserved its function in outer integument development from early stages of angiosperm evolution. Expression patterns of INO orthologs in Arabidopsis and Amborella ovules, together with those of the other factors studied (below), are summarized in Figure 2.
FIGURE 1. In situ hybridizations of integument regulators to female Amborella flower sections. (A) Atr.INO expression in the growing outer integument at early Stage 7. (B) Atr.INO expression in the near fully grown outer integument at late Stage 7. (C) Atr.INO expression is no longer apparent in the outer integument (LS) in mature flowers. (D) Atr.ATS expression in the inner integument at floral bud Stage 7. (E) Atr.ATS expression in the inner integument persists in the mature flower. (F) Narrow Atr.PHB/PHV expression in the chalaza and strong expression in the adaxial domain of the ovary wall at early Stage 7. (G) Weak Atr.PHB/PHV expression persists in the abaxial domain of the inner integument at late Stage 7. (H) Atr.REV is extensively expressed in the chalaza, and strongly expressed in the adaxial domain of the ovary wall at early Stage 7. (I) Weak Atr.REV expression persists in the adaxial domains of both integuments at late Stage 7. (J) Atr.ARF4 expression in the abaxial domain of both integuments at early Stage 7. (K) Very weak Atr.ETT expression in the abaxial domain of both integuments at early Stage 7. (L) Atr.ARF4 expression is no longer apparent in the integuments in the mature flower. (M) Atr.ETT expression is no longer apparent in the integuments in the mature flower. (N,O) Toluidine-blue stained sections of Stage 7 and mature ovules, respectively, showing anatomical details. Tissues zones in panels (N,O) have been delimited with dotted white lines for clarity. Developmental stages are as defined by Buzgo et al. (2004). abw, abaxial domain of the ovary wall; adw, adaxial domain of the ovary wall; c, chalaza; cp, chalazal pole; f, funiculus; ii, inner integument; mpp, micropyle pole; n, nucellus; and oi, outer integument. Scale bars = 100 μm.
FIGURE 2. Summary of expression patterns of ovule integument regulators in Arabidopsis thaliana and Amborella trichopoda. Abaxial-promoting genes are shown in red in enlarged diagrams, adaxial-promoting genes are shown in blue. Mature ovules are shown, though the genes indicated may be maximally expressed in the tissues shown at earlier stages. Entire ovules (on the left) are shown in their natural orientations relative to the longitudinal axis of the gynoecium/carpel. c, chalaza; f, funiculus; ii, inner integument; and oi = outer integument.
The Amborella Orthologue of the KANADI Family Gene ATS (KAN4) Is Expressed Abaxially in the Inner Integument, As Is ATS in Arabidopsis
Atr.ATS was found to be expressed specifically in the abaxial domain of the inner integument (Figure 1D), as is ATS in Arabidopsis. This expression persisted into the final stages of ovule development in almost mature flowers (Figure 1E). No expression of Atr.ATS was apparent in any female reproductive tissue other than the inner integument. These data strongly suggest ATS orthologs to have conserved their function in inner integument development in both the Amborella and Arabidopsis lineages since the MRCA of living flowering plants.
Amborella HD-ZIP III Genes Show Integument-Specific Profiles Similar to Those of Their Arabidopsis Orthologs
We found Atr.PHB/PHV, the Amborella pro-ortholog of Arabidopsis PHB and PHV, to be strongly expressed at early stages of development in the chalazal region of the ovule, which gives rise to the integuments, and very strongly expressed in the adaxial tissues of the ovary wall (Figure 1F). At later developmental stages, Atr.PHB/PHV appeared weakly expressed in the adaxial zone of the inner integument (Figure 1G). At early stages of ovule development, Atr.REV showed a similar expression pattern to Atr.PHB/PHV in the chalaza and ovary wall, though arguably with a slightly wider expression in the chalaza (Figure 1H). At later stages of ovule development, reduced levels of Atr.REV expression were visible in the adaxial regions of both integuments (Figure 1I). The association of Atr.PHB/PHV expression with the inner integument, and that of Atr.REV with both integuments (Figures 1G,I), suggests the paralog-specific conservation of function between the Amborella and Arabidopsis lineages since their separation at the base of the living flowering plant clade. Atr.PHB/PHV and Atr.REV expression patterns are nonetheless very similar, and these genes are clearly expressed together in parts of the chalaza, the inner integument and the abaxial region of the ovary wall.
The Amborella Orthologs of the Paralogous Auxin Response Factors ETT and ARF4 Are Expressed Abaxially in Both Integuments, Showing Partial Conservation With Expression Patterns in Arabidopsis
ETT and ARF4 act redundantly in the definition of abaxial tissue identity in the Arabidopsis leaf (Pekker et al., 2005), though ETT appears to play the major role in this process in both the carpel wall and inner integument. An earlier study of the orthologs of these factors in Amborella (Finet et al., 2010) suggested that Atr.ARF4 may show a higher level of expression than Atr.ETT in female reproductive tissues and perhaps therefore play the major developmental role. In the present study, both Atr.ARF4 and Atr.ETT were found to be expressed at early stages in the abaxial domain of both integuments (Figures 1J,K), with Atr.ARF4 giving the stronger hybridization signal. Expression levels of both genes diminished in mature integuments (Figures 1L,M). A switch may therefore have occurred in the major paralog active in the inner integument, with ETT playing the principal role in Arabidopsis, while Atr.ARF4 is more highly expressed in Amborella. In addition, whereas the role of ETT is limited to the inner integument in Arabidopsis, Atr.ARF4 and Atr.ETT are expressed in the abaxial domains of both integuments in Amborella.
It is interesting to note that Arabidopsis ETT has been found to interact physically with ATS, both of which are expressed specifically in the inner integument (Kelley et al., 2012). Atr.ETT and Atr.ARF4 may similarly interact with Atr.ATS in the Amborella inner integument, but this interaction could not occur in the outer integument as Atr.ATS is not apparently expressed in that tissue (Figures 1D,E). Of course, Atr.ETT and Atr.ARF4 might interact in the outer integument with other Amborella KANADI proteins (Supplementary Figure S4) whose expression patterns were not investigated in the present study.
Toluidine-blue stained sections of the carpel wall and ovule in a Stage 7 (Figure 1N) and a mature (Figure 1O) female flower are provided to help interpret anatomical details of in situ hybridization images shown in Figures 1A–M.
Reconstructed Gene Expression Patterns Suggest That Largely Distinct Sets of Regulators Controlled Inner and Outer Integument Development in Early Flowering Plants, As In Present-Day Arabidopsis
The data presented here strongly suggest the extensive conservation of integument-specific developmental roles of genes of the YABBY, KANADI, and HD-ZIP III families in both the Amborella and Arabidopsis lineages since the MRCA of living flowering plants. It was previously known that inner and outer integument development in Arabidopsis depended on substantially distinct sets of molecular players. The current study provides evidence that most of these differences in molecular controls are ancient, and already existed in early flowering plants. The only clear exception noted in the present study, in which molecular mechanisms seem to have diverged in one or both of the Amborella and Arabidopsis lineages, concerns the paralogous ARF family members ETT and ARF4. ETT contributes to abaxial tissue identity uniquely in the inner integument of Arabidopsis, whereas orthologs of both ETT and ARF4 are expressed in the abaxial domain of both integuments in Amborella. The inner integument expression of these factors may, as for that of ETT in Arabidopsis, act to prevent the coalescence of the two integument primordia. However, these factors may play a further role in the establishment and/or maintenance of abaxial/adaxial polarity in the Amborella outer integument, compared to their orthologs in Arabidopsis. Analysis of gene expression patterns in further ANA-grade taxa could be used to determine whether these ARF genes evolved more restricted inner integument-specific roles in the Arabidopsis lineage, or whether, conversely, their expression patterns became more generalized to both integuments, specifically in the Amborella lineage.
It is interesting to note that ETT expression is limited to the abaxial domain of the inner integument in bitegmic species of Prunus (rosid I clade) (Lora et al., 2015), as it is in Arabidopsis. If ETT/ARF4 expression was lost from the outer integument in the Arabidopsis lineage, rather than gained in the Amborella lineage, it would therefore seem likely that this change happened before the divergence of the rosid I and II clades, which is estimated to have occurred some 105 MYA (Moore et al., 2010).
We have used the data presented here, together with information from published studies, to construct a partial molecular model for the control of integument development in the MRCA of living angiosperms (Figure 3). Predicted intermolecular interactions shown in this model are based on functional data from Arabidopsis alone and the conservation of tissue-colocalization of the molecules concerned in Amborella.
FIGURE 3. A partial reconstruction of the molecular mechanisms of integument development in the most recent common ancestor of living flowering plants. Pointed and barred arrows represent cases of positive and negative regulation, respectively. Abaxial-promoting factors are shown in red, adaxial-promoting factors are shown in blue.
Implications of Reconstructed Integument Development Mechanisms for the Origin of the Outer Integument and Flowering Plants
Like the carpel, the outer integument is a pleisiomorphic feature of angiosperms. Two contrasting hypotheses have been proposed to account for the origin of this structure, prior to the radiation of living angiosperms. One of these hypotheses (reviewed by Doyle, 2008), based mainly on paleobotanical data, proposes that the angiosperms evolved from cupulate seed–ferns, possibly resembling the known fossil groups Caytoniales or Glossopteridales. In most known members of these taxa, several ovules occur within laminar cupules that are either borne on the margins of a female rachis (in Caytoniales), or emerge from the axil or midrib of a megasporophyll (in Glossopteridales). According to this "cupule hypothesis", the bitegmic angiosperm ovule would have evolved from a unitegmic gymnosperm ovule by a reduction in the number of ovules-per-cupule to one, the cupule thereby becoming the outer integument. The Caytoniales-type cupule may be a particularly strong candidate as a precursor to the angiosperm outer integument as the orientation of the ovule within this type of cupule could have led directly to the anatropous ovule arrangement (Doyle, 2008), which may have been pleisiomorphic in angiosperms.
The main alternative hypothesis for the origin of the outer integument, as discussed by Mathews and Kramer (2012), is that this structure simply arose through a modular reiteration of the single integument already present in species along the angiosperm stem lineage. Indeed, the development of the outer and inner integuments proceeds in a remarkably similar manner in most angiosperms, and the overexpression of WUS leads, in transgenic Arabidopsis, to the production of supernumerary integuments (Gross-Hardt et al., 2002; Sieber et al., 2004a), demonstrating that extra integuments can be generated from pre-existing developmental mechanisms by a simple molecular change in an upstream regulator. Not all strong candidates for ancestors, or close stem-lineage relatives of the angiosperms, possessed cupules. Notably, Bennettitales, which possessed many angiosperm-like characteristics, including a perianth, a bisexual axis (in some taxa), non-saccate pollen, net-veined leaves, and oleananes (highly resistant terpenoid compounds, also present in angiosperms), possessed no structure resembling an ovule-enclosing cupule. A mechanism based on the modular reiteration of a pre-existing single integument might, therefore, explain the origin of the angiosperm outer integument in a potential bennettialian ancestor.
The data presented here strongly suggest that the precise mechanisms of integument development were already substantially distinct in the inner and outer integuments some 164 MYA in the MRCA of living flowering plants. This conclusion clearly favors the hypothesis of a distant or indirect homology between the inner and outer integuments, such as might be explained by the more direct origin of the outer integument from an ovule-containing seed–fern cupule. It does not provide support for the possible origin of the outer integument by a modular reiteration of the inner integument, as this mechanism would be expected to yield two integuments whose development initially depended, in early flowering plants, on near-identical sets of molecular regulators.
GA performed the experiments and phylogenetic analyses. AV supervised the experimental work and phylogenetic analyses. AA-R supervised the experimental work. BF and GG provided the plant material and contributed to writing the paper. CS planned and supervised the research and wrote the paper.
This research was supported by ANR grant "ORANGe" 2013-18 to CS. GA was supported by a normalien undergraduate scholarship, and AV was employed as an agrégée préparatrice (teacher/researcher) of the ENS-Lyon.
We are grateful to the regional authorities of the province Nord and province Sud of New Caledonia for permission to collect living Amborella material.
The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fpls.2018.01352/full#supplementary-material
FIGURE S1 | Maximum likelihood phylogeny of angiosperm HD-ZIP III family proteins. aLRT branch-support values are shown at nodes. Amborella and Arabidopsis sequences mentioned in the text are highlighted. Key to species names: Ara tha: Arabidopsis thaliana; AmTr: Amborella trichopoda; Bra dis: Brachypodium distachyon; Bra nap: Brassica napus; Bra rapa: Brassica rapa; Cam sat: Camelina sativa; Cit sin: Citrus sinensis; Fra ves: Fragaria vesca; Gly max: Glycine max; Med tru: Medicago truncatula; Mus acu: Musa acuminata; Nel nuc: Nelumbo nucifera; Ory bra: Oryza brachyantha; Ory sat: Oryza sativa; Pop tri: Populus trichocarpa; Pru per: Prunus persica; Raph sat: Raphanus sativus; Ric com: Ricinus communis; Sol lyc: Solanum lycopersicum; Sol tub: Solanum tuberosum; The cac: Theobroma cacao; Vig ang: Vigna angularis; Vig rad: Vigna radiate; Vit vin: Vitis vinifera; and Zea may: Zea mays.
FIGURE S2 | Alignment of angiosperm HD-ZIP III family proteins used to produce the phylogeny shown in Supplementary Figure 1. Sites used in phylogenetic analysis are underlined.
FIGURE S3 | Maximum likelihood phylogeny of angiosperm KANADI family proteins. aLRT branch-support values are shown at nodes. Amborella and Arabidopsis sequences mentioned in the text are highlighted. Key to species names: Ara tha: Arabidopsis thaliana; AmTr: Amborella trichopoda; Bra dis: Brachypodium distachyon; Bra nap: Brassica napus; Bra rapa: Brassica rapa; Cam sat: Camelina sativa; Cit sin: Citrus sinensis; Fra ves: Fragaria vesca; Gly max: Glycine max; Med tru: Medicago truncatula; Mus acu: Musa acuminate; Nel nuc: Nelumbo nucifera; Ory bra: Oryza brachyantha; Ory sat: Oryza sativa; Pop tri: Populus trichocarpa; Pru per: Prunus persica; Raph sat: Raphanus sativus; Ric com: Ricinus communis; Sol lyc: Solanum lycopersicum; Sol tub: Solanum tuberosum; The cac: Theobroma cacao; Vig ang: Vigna angularis; Vig rad: Vigna radiate; Vit vin: Vitis vinifera; and Zea may: Zea mays.
FIGURE S4 | Alignment of angiosperm KANADI family proteins used to produce the phylogeny shown in Supplementary Figure 3. Sites used in phylogenetic analysis are underlined.
TABLE S1 | Oligonucleotide primers used to amplify Amborella integument regulators.
^https://blast.ncbi.nlm.nih.gov/Blast.cgi
^http://genomevolution.org/CoGe/
Altschul, S. F., Madden, T. L., Schaffer, A. A., Zhang, J. H., Zhang, Z., Miller, W., et al. (1997). Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 25, 3389–3402. doi: 10.1093/nar/25.17.3389
PubMed Abstract | CrossRef Full Text | Google Scholar
Bonaccorso, O., Lee, J. E., Puah, L., Scutt, C. P., and Golz, J. F. (2012). FILAMENTOUS FLOWER controls lateral organ development by acting as both an activator and a repressor. BMC Plant Biol. 12:176. doi: 10.1186/1471-2229-12-176
Buzgo, M., Soltis, P. S., and Soltis, D. E. (2004). Floral developmental morphology of Amborella trichopoda (Amborellaceae). Int. J. Plant Sci. 165, 925–947. doi: 10.1086/424024
CrossRef Full Text | Google Scholar
Doyle, J. A. (2008). Integrating molecular phylogenetic and paleobotanical evidence on origin of the flower. Int. J. Plant Sci. 169, 816–843. doi: 10.1086/589887
Endress, P. K. (2011). Angiosperm ovules: diversity, development, evolution. Ann. Bot. 107, 1465–1489. doi: 10.1093/aob/mcr120
Endress, P. K., and Igersheim, A. (2000). Reproductive structures of the basal angiosperm Amborella trichopoda (Amborellaceae). Int. J. Plant Sci. 161, S237–S248. doi: 10.1086/317571
Eshed, Y., Izhaki, A., Baum, S. F., Floyd, S. K., and Bowman, J. L. (2004). Asymmetric leaf development and blade expansion in Arabidopsis are mediated by KANADI and YABBY activities. Development 131, 2997–3006. doi: 10.1242/dev.01186
Finet, C., Floyd, S. K., Conway, S. J., Zhong, B., Scutt, C. P., and Bowman, J. L. (2016). Evolution of the YABBY gene family in seed plants. Evol. Dev. 18, 116–126. doi: 10.1111/ede.12173
Finet, C., Fourquin, C., Vinauger, M., Berne-Dedieu, A., Chambrier, P., Paindavoine, S., et al. (2010). Parallel structural evolution of auxin response factors in the angiosperms. Plant J. 63, 952–959. doi: 10.1111/j.1365-313X.2010.04292.x
Fourquin, C., Vinauger-Douard, M., Fogliani, B., Dumas, C., and Scutt, C. P. (2005). Evidence that CRABS CLAW and TOUSLED have conserved their roles in carpel development since the ancestor of the extant angiosperms. Proc. Natl. Acad. Sci. U.S.A. 102, 4649–4654. doi: 10.1073/pnas.0409577102
Gouy, M., Guindon, S., and Gascuel, O. (2010). SeaView version 4: a multiplatform graphical user interface for sequence alignment and phylogenetic tree building. Mol. Biol. Evol. 27, 221–224. doi: 10.1093/molbev/msp259
Gross-Hardt, R., Lenhard, M., and Laux, T. (2002). WUSCHEL signaling functions in interregional communication during Arabidopsis ovule development. Genes Dev. 16, 1129–1138. doi: 10.1101/gad.225202
Guindon, S., Dufayard, J.-F., Hordijk, W., Lefort, V., and Gascuel, O. (2009). PhyML: fast and accurate phylogeny reconstruction by maximum likelihood. Infect. Genet. Evol. 9, 384–385.
PubMed Abstract | Google Scholar
Huang, T., Harrar, Y., Lin, C., Reinhart, B., Newell, N. R., Talavera-Rauh, F., et al. (2014). Arabidopsis KANADI1 acts as a transcriptional repressor by interacting with a specific cis-element and regulates auxin biosynthesis, transport, and signaling in opposition to HD-ZIPIII factors. Plant Cell 26, 246–262. doi: 10.1105/tpc.113.111526
Kelley, D. R., Arreola, A., Gallagher, T. L., and Gasser, C. S. (2012). ETTIN (ARF3) physically interacts with KANADI proteins to form a functional complex essential for integument development and polarity determination in Arabidopsis. Development 139, 1105–1109. doi: 10.1242/dev.067918
Kelley, D. R., Skinner, D. J., and Gasser, C. S. (2009). Roles of polarity determinants in ovule development. Plant J. 57, 1054–1064. doi: 10.1111/j.1365-313X.2008.03752.x
Linkies, A., Graeber, K., Knight, C., and Leubner-Metzger, G. (2010). The evolution of seeds. New Phytol. 186, 817–831. doi: 10.1111/j.1469-8137.2010.03249.x
Lora, J., Hormaza, J. I., and Herrero, M. (2015). Transition from two to one integument in Prunus species: expression pattern of INNER NO OUTER (INO), ABERRANT TESTA SHAPE (ATS) and ETTIN (ETT). New Phytol. 208, 584–595. doi: 10.1111/nph.13460
Mathews, S., and Kramer, E. M. (2012). The evolution of reproductive structures in seed plants: a re-examination based on insights from developmental genetics. New Phytol. 194, 910–923. doi: 10.1111/j.1469-8137.2012.04091.x
McAbee, J. M., Hill, T. A., Skinner, D. J., Izhaki, A., Hauser, B. A., Meister, R. J., et al. (2006). ABERRANT TESTA SHAPE encodes a KANADI family member, linking polarity determination to separation and growth of Arabidopsis ovule integuments. Plant J. 46, 522–531. doi: 10.1111/j.1365-313X.2006.02717.x
Moore, M. J., Soltis, P. S., Bell, C. D., Burleigh, J. G., and Soltis, D. E. (2010). Phylogenetic analysis of 83 plastid genes further resolves the early diversification of eudicots. Proc. Natl. Acad. Sci. U.S.A. 107, 4623–4628. doi: 10.1073/pnas.0907801107
Nakata, M., Matsumoto, N., Tsugeki, R., Rikirsch, E., Laux, T., and Okada, K. (2012). Roles of the middle domain-specific WUSCHEL-RELATED HOMEOBOX genes in early development of leaves in Arabidopsis. Plant Cell 24, 519–535. doi: 10.1105/tpc.111.092858
Pekker, I., Alvarez, J. P., and Eshed, Y. (2005). Auxin response factors mediate Arabidopsis organ asymmetry via modulation of KANADI activity. Plant Cell 17, 2899–2910. doi: 10.1105/tpc.105.034876
Reinhart, B. J., Liu, T., Newell, N. R., Magnani, E., Huang, T., Kerstetter, R., et al. (2013). Establishing a framework for the Ad/Abaxial regulatory network of Arabidopsis: ascertaining targets of class III HOMEODOMAIN LEUCINE ZIPPER and KANADI regulation. Plant Cell 25, 3228–3249. doi: 10.1105/tpc.113.111518
Sieber, P., Gheyselinck, J., Gross-Hardt, R., Laux, T., Grossniklaus, U., and Schneitz, K. (2004a). Pattern formation during early ovule development in Arabidopsis thaliana. Dev. Biol. 273, 321–334. doi: 10.1016/j.ydbio.2004.05.037
Sieber, P., Petrascheck, M., Barberis, A., and Schneitz, K. (2004b). Organ polarity in Arabidopsis. NOZZLE physically interacts with members of the YABBY family. Plant Physiol. 135, 2172–2185. doi: 10.1104/pp.104.040154
Simon, M. K., Skinner, D. J., Gallagher, T. L., and Gasser, C. S. (2017). Integument development in Arabidopsis depends on interaction of YABBY protein INNER NO OUTER with coactivators and corepressors. Genetics 207, 1489–1500. doi: 10.1534/genetics.117.300140
Stevens, P. F. (2001). Angiosperm Phylogeny Website. Version 14. Available at: http://www.mobot.org/MOBOT/research/APweb/
Vialette-Guiraud, A. C., Alaux, M., Legeai, F., Finet, C., Chambrier, P., Brown, S. C., et al. (2011). Cabomba as a model for studies of early angiosperm evolution. Ann. Bot. 108, 589–598. doi: 10.1093/aob/mcr088
Yamada, T., Ito, M., and Kato, M. (2003). Expression pattern of INNER NOOUTER homologue in Nymphaea (water lily family, Nymphaeaceae). Dev. Genes Evol. 213, 510–513. doi: 10.1007/s00427-003-0350-8
Yamada, T., Sasaki, Y., Hashimoto, K., Nakajima, K., and Gasser, C. S. (2016). CORONA, PHABULOSA and PHAVOLUTA collaborate with BELL1 to confine WUSCHEL expression to the nucellus in Arabidopsis ovules. Development 143, 422–426. doi: 10.1242/dev.129833
Yamada, T., Yokota, S., Hirayama, Y., Imaichi, R., Kato, M., and Gasser, C. S. (2011). Ancestral expression patterns and evolutionary diversification of YABBY genes in angiosperms. Plant J. 67, 26–36. doi: 10.1111/j.1365-313X.2011.04570.x
Keywords: integument, Amborella trichopoda, ovule, angiosperms, HD-ZIP III, YABBY, KANADI, AUXIN RESPONSE FACTOR
Citation: Arnault G, Vialette ACM, Andres-Robin A, Fogliani B, Gâteblé G and Scutt CP (2018) Evidence for the Extensive Conservation of Mechanisms of Ovule Integument Development Since the Most Recent Common Ancestor of Living Angiosperms. Front. Plant Sci. 9:1352. doi: 10.3389/fpls.2018.01352
Received: 23 June 2018; Accepted: 28 August 2018;
Published: 19 September 2018.
Annette Becker, Justus-Liebig-Universität Gießen, Germany
Charles Gasser, University of California, Davis, United States
Masaru Ohme-Takagi, Saitama University, Japan
Copyright © 2018 Arnault, Vialette, Andres-Robin, Fogliani, Gâteblé and Scutt. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: Charles P. Scutt, charlie.scutt@ens-lyon.fr


text:
Q: How can I find kernel of linear map? What is the kernel of linear map $f\colon\mathbb{R}^{2\times2}\to \mathbb{R}^{3\times2}$ if:
$$f
  \begin{pmatrix}
    1 & 1\\
    1 & 1\\
  \end{pmatrix}
=
\begin{pmatrix}
    1 & 2 & 3\\
    1 & 2 &3\\
  \end{pmatrix},
$$
$$f
  \begin{pmatrix}
    0 & 1\\
    1 & 1\\
  \end{pmatrix}
=
\begin{pmatrix}
    3 & 3 & 5\\
    3 & 3 &5\\
  \end{pmatrix},
$$
$$f
  \begin{pmatrix}
    0 & 0\\
    1 & 1\\
  \end{pmatrix}
=
\begin{pmatrix}
    1 & 1 & 1\\
    1 & 1 &1\\
  \end{pmatrix},
$$
$$f
  \begin{pmatrix}
    0 & 0\\
    0 & 1\\
  \end{pmatrix}
=
\begin{pmatrix}
    1 & 0 & 1\\
    1 & 0 &1\\
  \end{pmatrix}.
$$
and how can I find it?

A: To find the kernel of a linear map $f:V\to W$ is the same as solving the equation
$
f(x)=0\;.
$
What you know in your problem are the values of $f(\alpha_k)$ where $\alpha =(\alpha_k:k=1,2,3,4)$ is a basis of $V$.
You can then write out the matrix representation of $f$ with respect to the basis  $\alpha$ of $V$ and the standard basis of $W$. Then you can work with the representation matrix by standard procedures.

A: Hint:
$$f(x)=f\left(\alpha\begin{pmatrix}
    1 & 1\\
    1 & 1\\
  \end{pmatrix}+\beta\begin{pmatrix}
    0 & 1\\
    1 & 1\\
  \end{pmatrix}+\gamma\begin{pmatrix}
    0 & 0\\
    1 & 1\\
  \end{pmatrix}+\delta\begin{pmatrix}
    0 & 0\\
    0 & 1\\
  \end{pmatrix}\right)=$$
$$=\alpha f\begin{pmatrix}
    1 & 1\\
    1 & 1\\
  \end{pmatrix}+\beta f\begin{pmatrix}
    0 & 1\\
    1 & 1\\
  \end{pmatrix}+\gamma f\begin{pmatrix}
    0 & 0\\
    1 & 1\\
  \end{pmatrix}+\delta f\begin{pmatrix}
    0 & 0\\
    0 & 1\\
  \end{pmatrix}$$

A: The input matrices form a basis of $\Bbb R^{2\times2}$.
Write $v\in\Bbb R^{2\times2}$ as $a_1\begin{bmatrix}1&1\\1&1\end{bmatrix}+a_2\begin{bmatrix}0&1\\1&1\end{bmatrix}+a_3\begin{bmatrix}0&0\\1&1\end{bmatrix}+a_4\begin{bmatrix}0&0\\0&1\end{bmatrix}$. Then $v\in\ker f\iff f(v)=0_{\Bbb R^{2\times3}}$ i.e.$$a_1\begin{bmatrix}1&2&3\\1&2&3\end{bmatrix}+a_2\begin{bmatrix}3&3&5\\3&3&5\end{bmatrix}+a_3\begin{bmatrix}1&1&1\\1&1&1\end{bmatrix}+a_4\begin{bmatrix}1&0&1\\1&0&1\end{bmatrix}=\begin{bmatrix}0&0&0\\0&0&0\end{bmatrix}$$from where you get the three independent equations$$a_1+3a_2+a_3+a_4=0\\2a_1+3a_2+a_3=0\\3a_1+5a_2+a_3+a_4=0$$yielding $v=a_1\left(\begin{bmatrix}1&1\\1&1\end{bmatrix}-\begin{bmatrix}0&1\\1&1\end{bmatrix}+\begin{bmatrix}0&0\\1&1\end{bmatrix}+\begin{bmatrix}0&0\\0&1\end{bmatrix}\right)$ and $\ker f=\text{Span}\left\{\begin{bmatrix}1&0\\1&2\end{bmatrix}\right\}$.

A: Start by showing that the four $2\times 2$ matrices $A_1, A_2, A_3, A_4$ you know the images under $f$ of form a basis of $\mathbb R^{2\times 2}$. Then any $A\in\mathbb R^{2\times 2}$ can uniquely be expressed as $A = \lambda_1 A_1 + \dots + \lambda_4 A_4$. Using linearity, $f(A)=0$ is equivalent to
$$
\lambda_1 \begin{pmatrix}
    1 & 2 & 3\\
    1 & 2 & 3
  \end{pmatrix} +
\lambda_2 \begin{pmatrix}
    3 & 3 & 5\\
    3 & 3 & 5
  \end{pmatrix} +
\lambda_3 \begin{pmatrix}
    1 & 1 & 1\\
    1 & 1 & 1
  \end{pmatrix} +
\lambda_4 \begin{pmatrix}
    1 & 0 & 1\\
    1 & 0 & 1
  \end{pmatrix} =
\begin{pmatrix}
    0 & 0 & 0\\
    0 & 0 & 0
  \end{pmatrix}.
$$
This is a linear system of $6$ equations in $4$ variables, but it is rather redundant. It is equivalent to
$$
\lambda_1 \begin{pmatrix}
    1 \\ 2 \\ 3
  \end{pmatrix} +
\lambda_2 \begin{pmatrix}
    3 \\ 3 \\ 5
  \end{pmatrix} +
\lambda_3 \begin{pmatrix}
    1 \\ 1 \\ 1
  \end{pmatrix} +
\lambda_4 \begin{pmatrix}
    1 \\ 0 \\ 1
  \end{pmatrix} =
\begin{pmatrix}
    0 \\ 0 \\ 0
  \end{pmatrix}.
$$
Row reduction yields
$$
\begin{pmatrix}
1 & 3 & 1 & 1 \\
2 & 3 & 1 & 0 \\
3 & 5 & 1 & 1
\end{pmatrix}
\sim
\begin{pmatrix}
1 & 0 & 0 & -1 \\
0 & 1 & 0 &  1 \\
0 & 0 & 1 & -1
\end{pmatrix}
$$
And hence the solution coefficient vectors are
$$
(\lambda_1, \lambda_2, \lambda_3, \lambda_4) \in \operatorname{span}\big((1,-1,1,1)\big).
$$
Thus, the kernel of $f$ is spanned by
$$
\begin{pmatrix}
    1 & 1\\
    1 & 1\\
  \end{pmatrix} -
\begin{pmatrix}
    0 & 1\\
    1 & 1\\
  \end{pmatrix} +
\begin{pmatrix}
    0 & 0\\
    1 & 1\\
  \end{pmatrix} +
\begin{pmatrix}
    0 & 0\\
    0 & 1\\
  \end{pmatrix}
=
\begin{pmatrix}
    1 & 0\\
    1 & 2\\
  \end{pmatrix}.
$$

A: Hint:
Consider the standard bases of$\mathcal M_2(\mathbf R)$
and $\mathcal M_{2\times 3}(\mathbf R)$::
$$E_1=\begin{bmatrix}1&0 \\0&0\end{bmatrix},\enspace E_2=\begin{bmatrix}0&1 \\ 0&0\end{bmatrix},\enspace E_3=\begin{bmatrix}0&0 \\1&0\end{bmatrix},\enspace E_4=\begin{bmatrix}0&0 \\0&1\end{bmatrix} $$
$$e_1=\begin{bmatrix}1&0&0 \\0&0&0\end{bmatrix},\enspace e_2=\begin{bmatrix}0&1 &0 \\ 0&0&0\end{bmatrix},\enspace e_3=\begin{bmatrix}0&0 &0 \\ 0&0&1\end{bmatrix}, \cdots$$
The matrix of the linear map $f$ is
$$T_f=\begin{bmatrix}-2 & 2 & 0 & 1\\-1 & 2 & 1 & 0 \\-2 & 4 & 0 &1 \\ -2 & 2 & 0 & 1 \\ -1 & 2 & 1 & 0 \\ -2 & 1 & 0 & 1\end{bmatrix},$$
and, as it has obviously rank $\le 3$, you can simply consider the matrix
$$A=\begin{bmatrix}-2 & 2 & 0 & 1\\-1 & 2 & 1 & 0 \\-2 & 4 & 0 &1 \end{bmatrix}$$
to determine the kernel, which can easily be done by row reduction.

redpajama_set_name: RedPajamaStackExchange


text:
SupercupNI 2019 a huge success for the Academy
15th August 2019/News
Rhys Williams, Lead Youth Development Phase Coach reports back on the tournament that saw the boys reach an exciting final.
The Charlton Athletic Academy, backed by Valley Gold funding sent a squad of U15 and U16 players (2004 born or younger) to compete in this prestigious tournament formally known as the "Milk" Cup in Northern Ireland. The club had previously won the competition way back in 2000 and until last season had not attended the event for some years. Last year's squad had a tough week so the group travelled well aware of the challenge ahead, particularly as six of the squad had participated last time out.
The group flew to Dublin, but were based in Ballymena from where they travelled to a variety of venues for their fixtures. They took part in the pre-tournament Parade on the Sunday evening attended by thousands of local fans in Coleraine – a wonderful start to the week. 22 teams from all over the world participated in the competition and qualification to the semi-finals would be determined by collating as many points and goals as possible via the three random games, rather than through the traditional 4 team group. BBC Northern Ireland covered the tournament via the Sport website and I-Player and our third fixture against Manchester United was a game selected for coverage.
Our first game saw a comfortable 5-0 win against local side County Fermanagh before overcoming a difficult test against Chilean side Colina. Falling a goal behind during the first half, we had to do it the hard way and showed great character to come back to win 2-1 against an aggressive and cynical opponent who tried everything to disrupt the flow of the game once they were ahead. With two wins on the board, it looked like a point against United could be enough to qualify for the semi-final, dependent on goal difference, but a win would definitely see us through.
Under the spotlight of the BBC TV coverage, we took a little while to settle into our rhythm but scored from a corner just before half time to lead. The second half saw us at our best and we scored a second and could have won more comfortably had we taken our chances.
The semi-final saw us play Pumas Unam from Mexico, again with BBC coverage. This was a hard-fought game where we needed energy and competitive spirit to find a way to win. Having cruised to a 2-0 lead just before the break, the Mexicans reduced the deficit and scored again to equalise. Our team showed great composure not to panic and scored a dramatic late winner to reach the final.
The final saw us meet Glasgow Rangers at Ballymena Utd's Showground stadium again on TV. Our first-half performance was as good as anything we produced during the week and we dominated the ball, pressed excellently to regain and did not allow our opponents to settle. The only thing missing was a goal and this was to cost us dear as against the run of play, Rangers scored right at the end of the first half. In truth, we never really recovered and could have gone further behind in the second half other than a superb goal-keeping performance that included a penalty save to keep us alive. Rangers scored a second in stoppage time to seal the title and break our hearts.
The experience that the boys gained from this trip was incredible. The support of Valley Gold to enable both trips to Northern Ireland in the last twelve months has offered a real development opportunity for our young players. Some fantastic lessons were learnt in adversity last season and the opportunity to go and compete again and evolve the resilience and competitive character required under pressure can only support the development of our players.
A number of the group also attended the Valley Gold funded South American tour last February. In enabling our attendance at these tournaments, the Valley Gold membership is not only offering incredible memories for our players and staff, but also the ability to develop key cultural, life and football experiences that will enhance their career opportunities in the future. Without this support, we would otherwise be unable to offer these experiences. We are incredibly grateful for your support.

redpajama_set_name: RedPajamaCommonCrawl


text:
Q: Cannot implicitly convert type 'System.EventHandler' to 'System.Windows.RoutedEventHandler' in c# In windows phone application I want to add a button dynamically like below:
Button btn = new Button();
btn.Content = tb_groupname.Text;
btn.Width = 200;
btn.Height = 200;
btn.Click += new EventHandler(btn_Click);//Click event

But when I add click event on my button I am getting below error:
Cannot implicitly convert type 'System.EventHandler' to 'System.Windows.RoutedEventHandler'

And below is the click event method of button:
private void btn_Click(object sender, EventArgs e)
{
  textbox1.text = "ABC"; // For Example
}

I don't understand why its getting this error.
Kindly suggest me, waiting for reply.
Thanks.

A: The signature of your event handler is wrong.
It should be:
private void btn_Click(object sender, RoutedEventArgs e)

And the Click event assignment should be changed to:
btn.Click += new RoutedEventHandler(btn_Click);//Click event

A: You need to use a RoutedEventHandler (found on the System.Windows assembly).
In your case, you should be able to change btn.Click += new EventHandler(btn_Click); with btn.Click += new System.Windows.RoutedEventHandler(btn_Click);; and then change the EventArgs ob btn_Click to RoutedEventArgs.
Make sure you add a reference to the System.Windows assembly or it won't compile!
Looking at the MSDN, I got this:

The RoutedEventHandler delegate is used for any routed event that does
  not report event-specific information in the event data. There are
  many such routed events; prominent examples include Click and Loaded.
The most noteworthy difference between writing a handler for a routed
  event as opposed to a general common language runtime (CLR) event is
  that the sender of the event (the element where the handler is
  attached and invoked) cannot be considered to necessarily be the
  source of the event. The source is reported as a property in the event
  data (Source). A difference between sender and Source is the result of
  the event being routed to different elements, during the traversal of
  the routed event through an element tree.

Link to the msdn.

A: Button Click is Routed Event as mentioned by Patrick Hofman.
You can even shorten if don't want new event by  just writing
btn.Click += btn_Click;


text:
static bool plyHasMesh(QString fileName)
{
    std::unique_ptr<t_ply_, int(*)(p_ply)> ply(
            ply_open(fileName.toUtf8().constData(), logRplyError, 0, NULL), ply_close);
    if (!ply || !ply_read_header(ply.get()))
        return false;
    for (p_ply_element elem = ply_get_next_element(ply.get(), NULL);
         elem != NULL; elem = ply_get_next_element(ply.get(), elem))
    {
        const char* name = 0;
        long ninstances = 0;
        if (!ply_get_element_info(elem, &name, &ninstances))
            continue;
        if (strcmp(name, "face") == 0 || strcmp(name, "edge") == 0 ||
            strcmp(name, "polygon") == 0)
        {
            return true;
        }
    }
    return false;
}

//------------------------------------------------------------------------------
Geometry::Geometry()
    : m_fileName(),
    m_offset(0,0,0),
    m_centroid(0,0,0),
    m_bbox(),
    m_VAO(),
    m_VBO(),
    m_Shaders()
{ }

Geometry::~Geometry()
{
    destroyBuffers();
}

std::shared_ptr<Geometry> Geometry::create(QString fileName)
{
    if (fileName.endsWith(".ply") && plyHasMesh(fileName))
        return std::shared_ptr<Geometry>(new TriMesh());
    else if(fileName.endsWith(".hcloud"))
        return std::shared_ptr<Geometry>(new HCloudView());
    else
        return std::shared_ptr<Geometry>(new PointArray());
}

void Geometry::initializeGL()
{
    destroyBuffers();

    initializeBboxGL(shaderId("boundingbox"));
}

void Geometry::destroyBuffers()
{
    // destroy any previously created buffers (in case we are re-initializing this geometry)
    // this should avoid recreating more and more opengl buffers

    for (auto& it: m_VBO)
    {
        GLuint vbo = it.second;
        glDeleteBuffers(1, &vbo);
    }

    m_VBO.clear();

    for (auto& it: m_VAO)
    {
        GLuint vao = it.second;
        glDeleteVertexArrays(1, &vao);
    }

    m_VAO.clear();

}

void Geometry::initializeBboxGL(unsigned int bboxShader)
{

    // tfm::printfln("Geometry :: initializeGL - %i", bboxShader);

    if (!bboxShader)
    {
        tfm::printfln("Bounding box shader was not defined for geometry.");
        return;
    }

    // Transform to box min for stability with large offsets
    Imath::Box3f box2(V3f(0), V3f(m_bbox.max - m_bbox.min));

    GLfloat verts[] = {
            (GLfloat)box2.min.x, (GLfloat)box2.min.y, (GLfloat)box2.min.z,
            (GLfloat)box2.min.x, (GLfloat)box2.max.y, (GLfloat)box2.min.z,
            (GLfloat)box2.max.x, (GLfloat)box2.max.y, (GLfloat)box2.min.z,
            (GLfloat)box2.max.x, (GLfloat)box2.min.y, (GLfloat)box2.min.z,
            (GLfloat)box2.min.x, (GLfloat)box2.min.y, (GLfloat)box2.max.z,
            (GLfloat)box2.min.x, (GLfloat)box2.max.y, (GLfloat)box2.max.z,
            (GLfloat)box2.max.x, (GLfloat)box2.max.y, (GLfloat)box2.max.z,
            (GLfloat)box2.max.x, (GLfloat)box2.min.y, (GLfloat)box2.max.z
    };
    unsigned char inds[] = {
            // rows: bottom, sides, top
            0,1, 1,2, 2,3, 3,0,
            0,4, 1,5, 2,6, 3,7,
            4,5, 5,6, 6,7, 7,4
    };

    // create VBA VBO for rendering ...
    GLuint bboxVertexArray;
    glGenVertexArrays(1, &bboxVertexArray);
    glBindVertexArray(bboxVertexArray);

    setVAO("boundingbox", bboxVertexArray);

    GlBuffer positionBuffer;
    positionBuffer.bind(GL_ARRAY_BUFFER);
    glBufferData(GL_ARRAY_BUFFER, 3*8*sizeof(float), verts, GL_STATIC_DRAW);

    GLint positionAttribute = glGetAttribLocation(bboxShader, "position");
    glVertexAttribPointer(positionAttribute, 3, GL_FLOAT, GL_FALSE, sizeof(float)*(3), (const GLvoid *)0);
    glEnableVertexAttribArray(positionAttribute);

    GlBuffer elementBuffer;
    elementBuffer.bind(GL_ELEMENT_ARRAY_BUFFER);
    glBufferData(GL_ELEMENT_ARRAY_BUFFER, 2*4*3*sizeof(char), inds, GL_STATIC_DRAW);

    glBindVertexArray(0);
}

const unsigned int Geometry::getVAO(const char * vertexArrayName) const
{
    // always call this from an active OpenGL context
    if(m_VAO.find(std::string(vertexArrayName)) != m_VAO.end())
    {
        return m_VAO.at(std::string(vertexArrayName));
    }
    tfm::printfln("Geometry :: vertexArrayObject was not found - %s", vertexArrayName);
    return 0;
}

const unsigned int Geometry::getVBO(const char * vertexBufferName) const
{
    // always call this from an active OpenGL context
    if(m_VBO.find(std::string(vertexBufferName)) != m_VBO.end())
    {
        return m_VBO.at(std::string(vertexBufferName));
    }
    tfm::printfln("Geometry :: vertexBufferObject was not found - %s", vertexBufferName);
    return 0;
}

const unsigned int Geometry::shaderId(const char * shaderName) const
{
    if(m_Shaders.find(shaderName) != m_Shaders.end())
    {
        return m_Shaders.at(shaderName);
    }
    tfm::printfln("Geometry :: shaderId not found - %s", shaderName);
    return 0;
}

redpajama_set_name: RedPajamaGithub


text:
Les Proconodontida sont un ordre éteint d'Euconodonta (« vrais conodontes »).

Description

Histoire géologique

Phylogénie

▲
 └─o Conodonta (éteint)
     ├─? Paraconodontida (éteint)
     └─o Euconodonta (éteint)
         ├─o Proconodontida (éteint)
         └─o Conodonti (éteint)
             ├─o Protopanderodontida (éteint)
             └─o Prioniodontida (éteint)

Les Proconodontida est le taxon frère des Conodonti, parmi les Euconodonta.

Références

Liens externes

Ordre de vertébrés préhistoriques (nom scientifique)
Conodonte (nom scientifique)
Taxon fossile décrit en 1988

redpajama_set_name: RedPajamaWikipedia


Heucheras are known for their spectacular foliage that provide months of of color and texture in the garden. Leaves are semi-evergreen and somewhat rounded with colorful veining. Pink Panther has a low mounding habit reaching 10" tall and 16" wide that quickly provides foliage color up to 3 seasons in the garden. Highly adaptable to containers. Dainty pink flowers on slender stalks rise above the plants are are attractive to hummingbirds and butterflies. The slender stalks add an airy tough to any floral arrangements.
Cooler temperatures (early spring and fall) bring out a deep, rich almost purple color, transforming to a deep rose during the heat and humidity of summer. Such a transition of color is not only enjoyable to watch but also serves as a barometer of an upcoming season.
Foliage color is one of the secrets to long lasting color in the shade garden providing up to 3 seasons of interest while withstanding the rigors of heat & humidity in the summer garden. This shade loving perennial require almost no little maintenance or work on your part, and will live for many years as long as you provide good drainage.
Coral Bells are a low maintenance shade ground cover that thrives in moist, well-drained soil. They are not troubled by pests or diseases.
Heucheras are well suited as an edging to beds and pathways. They grow well under trees. Use as a front of the border plant to frame a bed. Tough and durable in combination planters or planted in mass. Great planting partners include Violas, Hosta, Ferns & Astilbes, or Hydrangeas.


text:
Mislabeled Kiwis Get the Boot in France
By Susanne Kuehne
Twelve percent of expensive "French grown" kiwis are not grown in France after all.
Records involving fraud can be found in the Food Fraud Database.
Kiwis are popular in France; they are one of the top 10 fruit consumed. France has only a fraction of the number of kiwi growers compared to Italy. French kiwis are 40% more expensive than Italian fruit, making it worthwhile to fraudulently change the country of origin. A sudden flood of "French" kiwis on the market at the end of the French kiwi season caught the attention of investigators, and 12% of these kiwis were discovered to originate from Italy. The fraud netted several million Euros in additional profit for the involved Italian kiwi producers. What is especially concerning is that the fruit was treated with products that are banned in France to increase the yield.
Willsher, K. (March 25, 2019). French officials target alleged "kiwigate" fraudsters. The Guardian.
Accessed March 25, 2019. Retrieved from https://www.theguardian.com/world/2019/mar/25/just-desserts-french-officials-target-alleged-kiwigate-fraudsters
Cassel, B., Pelloli, M., Plichon, O.(March 25, 2019): Alerte aux faux kiwis francais. Accessed March 25, 2019. Retrieved http://www.leparisien.fr/economie/consommation/alerte-aux-faux-kiwis-francais-24-03-2019-8038731.php
Mitigating Food Fraud: Top Tips
Get the latest on regulatory and GFSI requirements.
Decernis Acquires USP's Food Fraud Database
The database helps companies make decisions about food fraud vulnerability and economically motivated adulteration.
Rabbit Food?
The latest violations come from a ketchup factory in Pakistan.
Decernis Announces Food Fraud Quick Bites and Resource Center
Food fraud is receiving increased attention and growing market awareness.
Focus Article France Italy kiwi mislabeled
Susanne Kuehne
Sr. Manager, Business Development
Decernis
Susanne Kuehne joined Decernis in 2016 as senior manager, business development. She has 20+ years of experience in the chemicals, plastics, coatings and beverage spaces. Kuehne is located at the Washington, D.C. office, but is originally from the Stuttgart, Germany area. She studied chemistry and business in Germany, then worked for Grace GmbH in Worms, Germany before moving to the United States in 2000. She worked for Grace in the United States before joining the beverage industry for eight years. Kuehne's focus is food contact and chemical industry clients world-wide, across the multiple disciplines Decernis covers.
Kuehne holds a Dipl.-Ing (FH) Farbe/Chemie from Fachhochschule fuer Druck, Stuttgart, and a Dipl.-Betriebswirt (FH) from AKAD Fachhochschule, Lahr.


The Seattle Seahawks struggled on offense against the scrappy St. Louis Rams, so they had to leave Monday night's game in the hands of one of the NFL's most stingy defenses.
ST. LOUIS — The Seattle Seahawks struggled on offense against the scrappy St. Louis Rams, so they had to leave Monday night's game in the hands of one of the NFL's most stingy defenses.
"The defense did a tremendous job, coming up with a huge stop there," said Seahawks star Russell Wilson, who was sacked a career-high seven times.
The outcome capped a lousy night for St. Louis sports fans. The Cardinals lost 3-1 to the Boston Red Sox in Game 5 of the World Series, played just up Broadway at Busch Stadium.
Seattle (7-1) extended the best start in franchise history despite gaining just 135 yards, with 80 coming on Wilson's second TD pass to Golden Tate. It was the third-fewest yards in a victory for the Seahawks, and their seven first downs were the fewest in a win in franchise history.
The World Series no doubt contributed to the stale atmosphere inside the Edward Jones Dome, where the announced crowd of 55,966 was in reality much smaller. Many of the fans who did show up wore Cardinals gear, and Cardinals-Red Sox highlights were shown on the big screen.
Greg Zuerlein staked St. Louis to an early lead with his first of three field goals, but Sherman's pick put the Seahawks in business. Seattle scored six players later, when Wilson hit Tate from the 2-yard line.
NOTES: Rams DE Robert Quinn had three sacks, all in the first half. ... Seattle had minus-1 yard of offense in the first quarter, and 38 yards at the half, its fewest since gaining 37 in the first half against Kansas City in 1998, according to STATS LLC. ... Seahawks WR Sidney Rice left late in the first half with a knee injury and did not return.


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Radhika Gurung death: Two people summonsed to court accused of causing death by driving without due care and attention
Surrey Police said the man and woman are due to appear at Guildford Magistrates' Court on Tuesday (September 26)
Beth Duffell
Two people have been summonsed to court following a fatal collision in which a pedestrian died in Maybury, Woking.
Surrey Police said a 27-year-old man from Woking and a 26-year-old woman from Reading have been summonsed for causing death by driving without due care and attention in relation to the death of pedestrian Radhika Gurung on December 9 2016 in Monument Road, Maybury.
A police spokesman said: "They are due to appear at Guildford Magistrates' Court on Tuesday (September 26)."
Hundreds of people gathered at Woking Crematorium for the funeral of Mrs Gurung on December 22 2016.
The latest court news from the county
Surgery made errors with prescription
Dad escapes driving ban
Man terrified women with calls and texts
Coach Trip star's dad jailed
Relatives and residents travelled from across the UK for a service at the crematorium in Hermitage Road.
Clutching bunches of flowers, the mourners paid their respects to their family member, friend or colleague as the scent of incense wafted through the chilly winter air.
As the community said farewell to Mrs Gurung, a much-loved mother and wife, they took part in a traditional Buddhist ceremony.
Woman dies after car mounts pavement in Woking
Music was played and Lama leaders conducted the service.
Mrs Gurung's daughters threw petals and coins onto the floor as they walked into the crematorium.
Keep up to date with the latest news from around the county via the free Get Surrey app.
You can set up your app to see all the latest news and events from your area, plus receive push notifications for breaking news.
Available to download from the App Store or Google Play for Android .


Securitas Mobile Guarding utilizes expertly trained patrol officers and partnerships with top ATM manufacturers to help mitigate theft caused by ATM Card Skimming, Shimming, and PIN Capturing devices.
Securitas Mobile uses a thorough and systematic inspection process to check each ATM for signs of alteration to include any skimming, shimming, and/or PIN capturing devices. With continuous research on trends and devices, officer training, and information sharing, Securitas Mobile will ensure our attention to detail is second to none.
Securitas Mobile Guarding provides web-based reporting in real time. This enables Securitas Patrol officers to quickly and accurately record location, date, time, pictures and inspection results which can then be instantly accessed via a secure online portal.
Securitas partners with some of the largest ATM manufacturers in the country. This provides in-depth knowledge about the specific machines to Mobile Patrol officers who leverage this knowledge during inspections.
Since 2008, the use of skimming devices has grown approximately 10% annually. It is the most common type of physical fraud and costs over $8 billion in losses per year.


This brand has been more prominent in the fleet of Her Majesty's Secret Service; it is noteworthy that six models of the Aston Martin brand have been exhibited in about twelve of the twenty-four Bond movies till present. For instance, the 1964 Aston Martin DB5 made its debut in GoldFinger, the Academy Award-winning movie. The classic vehicle which has the reputation as the most fantastic car in the franchise has impressive features such as an aluminum 282 hp, 4-litre, straight 6-engine as well as a 5-speed transmission. It can move at a fast pace from zero to sixty mph within eight seconds thereby reaching a premium speed of 145mph.
The vehicle has incredible features such as a revolving license plate, machine guns, oil-slick, smoke-screen apparatuses, passenger-side ejector seat, and bulletproof windshield. It is also important to note that these remarkable attributes are not part of the regular package.
In a bid to mark the beginning of a robust 50-year relationship between Aston Martin and the Eon Productions and Bond, two 1965 DB5s would be utilized during the filming process. However, the vehicle would feature in most of Bond's movies namely: Thunderball, GoldenEye, Tomorrow Never Dies, Casino Royale, Skyfall, and The World Is Not Enough.
This vehicle is the closest automobile that the author of the Casino Royale, Ian Fleming wants Bond to drive in the first version and this is the 1931 Bentley 4.5 Litre. The car was discovered in the 1963 movie, From Russia with Love. Part of the amazing features of the 1935 model is its 110hp, straight 6-engine and its power to run 90mph. It is a perfect choice for those who want to drive without a roof as it is convertible and ladies, especially will jump at the opportunity to cruise it. Bentley was not used on the movies of Bond until Sean Connery reenacted his role in the 1983 classic film, Never Say Never Again.
This vehicle is a unique brand among its equals. The 1967 Toyota 2000GT that was used in the movie, You Only Live Twice in its year of release was just one of the only two variants produced. During the filming phase, it was discovered that Sean Connery's height is too tall for the original hardtop version that was made for the film, but Toyota came to the rescue by providing the unique breed. In fact, only three and fifty-one types of the two-seat coupe were manufactured as it is rated as the first high-performance sports car from Japan.
Bond was seated in the passenger seat as the pretty Japanese SIS agent Aki avoided the attacks from villains. Some of the innovative features of the vehicle are a closed-circuit television, two-way radios, voice-controlled electronics as well as a 150 hp, 2-liter, straight 6-engine and fast speed of 135mph. It has an appealing touch as the present 007 actor; Daniel Craig reportedly said that the 2000GT is his preferred choice when it comes to Bond car.
If there is a vehicle with close ties with the film franchise, it must be the 1962 Rolls-Royce Silver Cloud II in the 1985 movie; A View to a Kill as it is the last ride of Roger Moore as 007. The car also referred to as the CUB 1 as it was named after its original license plate. The co-founder of the Eon Productions who also double as the late Bond film producer, Albert "Cubby" Broccoli owned the classic vehicle. The four-door sedan automobile has a 6.2-liter V-8 engine; it can cover a distance of 60 mph within eleven seconds and a premium speed of 104mph.
In the movie, Bond was driven by Sir Godfrey Tibbet, (Patrick Macnee) in the Rolls Royce to the estate of the ridiculous business mogul; Max Zorin played by Christopher Walken. He remains the only Bond to survive in the crash as the driver and car were found at the base of a lake. Another Silver Cloud II was used for that particular scene, but it is important to note that CUB 1 is a favorite car of the Broccoli family to date.
Out of the entire high-performance vehicles used in the 007 series, no car has the unique depth of the 1976 Lotus S1 Esprit which was featured in the movie, The Spy Who Loved Me. One of the memorable scenes in the film is when Roger Moore and Barbara Bach who acted as the KGB agent, Anya Amasova avoided a helicopter gunship by moving into the Mediterranean. The vehicle developed wings and turned into a submersible means of transportation, and it was one of the best scenes that show the prowess of Bond. The vehicle has incredible attributes like torpedoes, cement sprayer, magnetic mines, surface-to-air missiles, and a black-dye dispenser that looks like an octopus.
Without the fictional characteristics, the car is a wonderful vehicle to possess as it has a 160hp, Lotus 907 2-liter engine and a 5-speed manual transmission. Its publicized acceleration rate was zero to 60mph in less than seven seconds and a top speed of 138 mph.
The real Esprit body used as a disguise in the movie's submarine sequence was seen in a storage container bought for a paltry sum of hundred dollars in 2013. However, the Tesla mogul, Elon Musk offered almost a million dollars at its auction in the same year as he has plans to merge it with his electric powertrain and gears to ensure the submersion is successful.
Bond was reintroduced to Aston Martin by actor, George Lazenby as he had a suave look and seated in the 1968 DBS on Her Majesty's Secret Service. The coupe designed by William Towns (with hood scoop) can travel 150mph as it possesses a robust 282 hp, 4-litre, straight 6-engine mated with a 5-speed manual transmission.
The vehicle was driven to the beach by Bond to prevent Contessa Teresa di Vicenzo from drowning; the car was also utilized as the flower-festooned breakout car for the lovers after their wedding ceremony. On a sad note, the wife of Bond was shot dead in the passenger seat by Blofeld and assassin Irma Bunt while the couple parked on the other side of the road- and the next to the last bachelor was birthed.
Renowned for its guts rather than allure, this amazing 1985 Renault Turbo 11 had an incredible effect despite not having an extended screen life in the 1985 classic movie, A View To a Kill. The Renault which is a common taxi was taken forcefully by Bond to give an assassin, May Day (Grace Jones) who jumped off the Eiffel Tower a hot chase. The murderer was on the run as he just assassinated Bond's acquaintance. While Bond was pursuing May, the hardtop four-door sedan hovers down the flights of stairs in a backward position, and moved over a bus and became a convertible automobile as a traffic blockade removed its roof.
The hatchback hero has a 158 hp, Cleon-Fonte turbocharged engine that can move the car from zero to 62mph within 9.2 seconds and can get to a high speed of 115mph or more. In 1985, the 1985 Renault Turbo 11 with Alain Oreille at the wheels won its class at the Rallye Automobile Monte Carlo race in the same year.
Despite the continuity mistake, it does not take anything from the vehicle's powerful, appealing features which are twin ram-air intakes on the hood which reinforced the hefty 370 hp, 429 Cobra Jet V-8 engine beneath. Numerous popular fastbacks were exploited in filming, and each of the last ones has the value of a few diamonds.
After the Summer of Love, the seductively influential 1969 Mercury Cougar XR-7 convertible accelerated the engines of car lovers with its presence in Her Majesty's Secret Service. The 1969 Mercury Cougar XR-7 is an attractive beast; it possesses a 335 hp, Cobra Jet 428 V-8 engine with a fantastic power to move from zero to sixty mph in less than six seconds.
The vehicle's secret defence is its driver, Contessa Teresa Di Vicenzo, a character played by the inexpressible Diana Rigg. Even though Contessa who has the reputation as the only woman to marry Bond and her vehicle survive by the final credits but the classic Mercury would later become an iconic brand that will command huge demand just like the 77-year-old Dame Diana is featured in the current series of Game of Thrones.
When Pierce Brosnan acted as Bond in the movie, The World Is Not Enough; the 1999 BMW Z8 was the sophisticated roadster. However, the aluminum body designed by Henrik Fisker has a 400 hp V-8 engine in combination with a six-speed manual transmission. Within 4.2 seconds, it can accelerate from zero to sixty mph as its top speed is around 155 mph.
Q had fortified the vehicle with surface-to-air missiles which are coordinated by a guidance system in the steering wheel, and it is always behind Bond. A smart key can be used to operate the car on a remote level. Irrespective of its high-tech cache, the convertible vehicle was cut into two by substantial spinning blades from one of the wicked Elektra's helicopters.


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Home News President of Brazil Jair Bolsonaro: 'I Do Not Know What Bitcoin Is'
President of Brazil Jair Bolsonaro: 'I Do Not Know What Bitcoin Is'
The president of Brazil, Jair Bolsonaro, has stated that he does not know what bitcoin (BTC) is, and endorsed the suspension of a project that would create a crypto for indigenous people to use, Cointelegraph Brazil reported on June 4.
Bolsonaro supported the Minister of Women, Family and Human Rights, Damares Alves in the termination of a project worth $44.9 million Brazilian reals ($11.5 million) between the National Indian Foundation (FUNAI) and the Federal Fluminense University (UFF), which promoted the creation of a cryptocurrency for indigenous people to use. Bolsonaro delivered his comments during an interview with Ratinho's SBT show on June 4.
After saying that the Minister acted properly in blocking the project that "wanted to teach the Indian to use bitcoin," the president was asked by a show participant if he knew what bitcoin was. Bolsonaro replied:
"I do not know what bitcoin is."
Shortly after the speech, Bolsonaro rectified his statement and said that bitcoin was a "virtual currency." The project offered by FUNAI and the UFF will purportedly neither receive financing and support, nor integrate bitcoin.
Over his career, Bolsonaro has made several controversial statements regarding indigenous people in Brazil, and promised to roll back protections on their lands.
As previously reported, the government suspended the project in early January claiming that the contract was issued improperly and lacked technical analysis, such as a detailed description of the project. The contract for the project was allegedly signed directly between FUNAI and UFF, instead of through a legal bidding process. Moreover, the government said that the contract had been approved too quickly and entailed considerable expenditure.
In late May, the President of the Chamber of Deputies of Brazil, Deputy Rodrigo Maia ordered the establishment of a commission to consider cryptocurrency regulation in the country. The commission will be composed of 34 members in accordance with the House Rules of Procedure.
Previous articleUS SEC Adds Fraud to List of Charges Against 'Crypto' Firm Longfin
Next articleGalaxy Digital Fund Leads Funding Round for Blockchain-Based Game Platform


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QualityHealth Home > Diet & Weight Loss Health Center > 10 Most Fattening Ballpark Foods
10 Most Fattening Ballpark Foods
By Valerie Banner
Reviewed by QualityHealth's Medical Advisory Board
There was a time when the national pastime was synonymous with peanuts and cracker jacks. Now, the ballpark menu has expanded-to include one fattening item after another. Stadiums often serve foods made famous by their hometowns (such as cheesesteaks in Philly, crabcakes in Baltimore, or pierogies in Pittsburgh), but many of them should come with an exercise plan on the side. Here, the 10 most fattening ballpark foods in the country.
1. Chili Cheese Dog. To begin with, most hot dogs contain several grams of saturated fat (the kind of fat that causes high cholesterol, according to the American Heart Association). Add to it the greasy, gooey cheese and loads of spicy chili (and if it's Cincinnati's famous Skyline chili, served at the Great American Ball Park, there may be some chocolate added). The final caloric score of these treats? Somewhere in the ballpark of 400 calories.
2. Pretzel Dog. Maybe they're going to for a triple play here: high in fat, high in sodium, and high in carbs. It's not like the hot dog is a health food to begin with, but wrapping a 200-plus-calorie pretzel around it is just adding insult to injury. If a pretzel dog isn't your thing, but you still want all the fatty goodness of thepretzel and sausage, you can order the bratzel (a bratwurst wrapped in a pretzel) the next time you're at a St. Louis Cardinals home game. But beware: You'll be consuming a quarter of your daily calorie intake.
3. Beer. It's technically not a food, but a cold brew is a ubiquitous ballpark staple nonetheless. In fact, the vendors are everywhere, shouting, "Get your beer, here," into the stands. Whether you're having a cold one at Miller Park or Coors Field or any of the other fields not named after beers, each eight-ounce glass weighs in at approximately 100 calories. Light beers could save you about 25 to 30 calories a glass, but they'll still cost an arm and a leg.
4. Garlic fries. These specialty French fries are such a hit, they can be found in multiple stadiums, particularly on the West Coast, including Bank One Ballpark (home of the Arizona Diamondbacks), PETCO Park (home of the San Diego Padres), and AT&T Park (home of the San Francisco Giants). With these fries, you'll be batting 500-calories, that is.
5. Italian sausages. It doesn't matter if you top your sandwich with the hottest peppers or the sweet kind, Italian sausages are a strikeout, for sure. Although the caloric count and fat grams vary widely depending on where you're ordering, they're never low, sometimes swinging as high as 700 calories and more than 40 grams of fat.
6. Anything smothered in barbecue sauce. Pick your poison: the barbecue pork ribs in Atlanta, the barbecue beef sandwich while watching the Royals play at home, or some of Manny's barbecue pit beef in Pittsburgh's PNC Park. To be fair, it's not the sauce that will wreak havoc on your waistline; it's everything underneath it.
7. Cuban sandwich. You can get a Cuban while rooting for the Tampa Bay Rays, the Boston Red Sox, or the Cleveland Indians. Regardless of who they're playing, one pre-game prediction is sure to be true: At about 700 calories a sandwich, this ballpark food will ruin your diet.
8. Cheesesteak. It's no surprise that they serve this famous Philadelphia staple at Citizen's Bank Park where the Phillies play. While Geno's Steaks and Pat's King of Steaks continue to battle across the street from each other for dominance in the Philly cheesesteak market, whether you grab one of those or the kind at the park, one thing is clear: You're going to need to loosen your belt after you finish eating it.
9. Pizza.Two slices of Little Caesar's pepperoni pizza stack up to 780 calories. And if that's not enough to convince you to steer clear, consider this the next time you're at a Detroit Tigers home game: Mike Ilitch owns both the team and the pizza franchise, which has made him enough money to earn a spot on the Forbes Richest Americans list. Maybe you should save your money.
10. Chocolate covered cheesecake. As if the other foods haven't already done enough harm, the final inning brings dessert. And at the Great American Ballpark in Cincinnati, that means chocolate-covered cheesecake, which is sure to issue to the final blow to your waistline.
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text:
Q: htaccess with Angular and Phalcon I'm trying to switch the frontend of my Phalcon app to AngularJS. I'm having some troubles with the .htaccess file though. Below is what I currently have.
RewriteEngine On
RewriteBase /
RewriteCond %{REQUEST_FILENAME} !-d
RewriteCond %{REQUEST_FILENAME} !-f
RewriteRule ^api/.* api/index.php [L,QSA]

RewriteCond $1 !^(api)
RewriteRule ^(.*)$ public/index.php?_url=/$1 [QSA,L]

It routes to Phalcon, but all the .css and .js files in the public folder now throw a 404. What am I doing wrong here?

A: If you want to ignore files and directories you need to add extra rewrite rules to your .htaccess. if you want to ignore extensions as well, there's another rewrite rule for that.
https://stackoverflow.com/a/17029882 will give you the information you need. In your case
RewriteCond %{REQUEST_URI} !(\.css|\.js|\.png|\.jpg|\.gif)$ [NC]
is enough and in most cases also the files and directories are not rewritten with this:
RewriteCond %{REQUEST_FILENAME} !-f
RewriteCond %{REQUEST_FILENAME} !-d


text:
Podium for Alonso and friends at Dubai Kartdrome 24-Hours
While most race drivers are chilling on an exotic beach somewhere relaxing after a tough season, Fernando Alonso relaxes by going racing.
The double F1-World Champion is driving the FA Racing #14 kart in the Kartdrome 24-Hours final round of the Dubai Kartdrome Endurance Championship Powered by Motorsport Wheels.
Alonso and his team contested the one-make Sodi W-Series (SWS), using Sodi karts, in the Pro-Am endurance event with an all Spanish lineup that includes former F1 driver Pedro de le Rosa as well as Alberto Albilares and Ángel Burgueño.
In Qualifying, Alonso did the business by claiming pole, fastest driver of all among the 33 teams in action at the event that has been running for over a decade and is considered the most prestigious of this genre of endurance kart racing.
Kartdrome Manager David Bright said of Alonso's entry, "Since 2013, Fernando has been a regular visitor to Dubai Autodrome as, over the years, he has spent more time in Dubai.
"He entered a team in 2015 which also included Stoffel Vandoorne and Pedro who is here again with Fernando. Each year they take it a little more serious. But it is a very tough contest with a dozen or so serious teams all capable of victory. All with far more experience in this specialised format of karting.
"However Fernando takes it all in his stride, very friendly with his rivals and fans off-track. But when the start-lights go out, with his helmet on it is all about the racing and teamwork in a 24-hour race that is all about strategy, consistency of drivers and of course speed.
"His professionalism and passion for the sport are second to none, affirmed by the fact that he and his team are willing to swap their million-dollar race machinery for karts and simply enjoy racing. This also demonstrates the world-class level of racing at Dubai Kartdrome which has attracted top racing talent from around the globe," added Bright.
At the Kartdrome on Friday, Alonso claimed the top spot start for FA Racing with a late fast lap in a closely fought qualifying session, in which a mere second covered the top 20 teams on the grid of 33.
In the race, perhaps lack of experience in this format of karting within FA Racing took its toll against the more experienced outfits, among them SWS world champions.
Nevertheless, they stuck to the mandate of consistency and pace to claim third-place overall and taking top honours in the Nations Cup for the all-Spanish outfit.
On Saturday, victory in the 15th edition of the race went to series stalwarts Emirates Racing – consisting of Arjun Manjunath, Romaldo Labrooy, Ramez Azzam and Tom Bale – who completed 1049 laps followed by Bin Drai Karting Team in second place three laps adrift of the winners and only 17 seconds ahead of third-placed FA Racing, for whom Alonso also claimed the fastest lap of the race.
go kartgokartkartkartingsodisodi-w-seriesswsw-serieswseries
Camilleri: Why give credibility to a 22-year-old boy?
Binotto: Leclerc has been outstanding in his leadership


Here at Car Loans Georgia, we enable you to secure the auto loan in Dalton GA you need, even if your credit has seen better days. We connect you with loan creditors who frequently give auto loans to consumers in Dalton, GA. The application process will take under 3-4 minutes.
For the most part, car loan providers will want you to have a secure job with $1500 income per month. Besides that, the amount you spend on existing debts, including your auto loan, must not surpass half of your earnings. For folks in Dalton, GA, this equates to $804, on average.
Income or fico scores too low? You should still qualify with a co-signer, more money down, or even through buy here pay here financing in Dalton, GA.
Yes, we can help you to get auto financing in Dalton with no down payment, but down payments are always a good option. The majority of finance companies will be happy with a down payment of 15-20%. An additional aspect to consider is your monthly payments. Never devote more than 18-20% of your income for your Georgia auto loan. Here are average numbers for Georgia car shoppers.
Do you have poor credit? Discover more details about car loans for people with bad credit below.
It is very important to plan the right way for your car loan in Dalton, GA. So please don't spend more than 15% of your income per month toward paying off your bad credit car loan.
$80: 5% for gas, upkeep, etc.
In addition, you'll want to supply a considerable down payment whenever possible. Sure, we could help you obtain Georgia auto loans with no money down, but such loans come with a more serious risk of negative equity.
Make sure that you explore any GA buy here pay here dealer on the web. You'll want to speak with a friend or relative who has bought a car or truck from them before, and find out if they would recommend them.


Forefront Identity Manager 2010 R2 SP1 and SharePoint Server 2013 has introduced the ability to leverage FIM for User Profile Synchronization with Active Directory, versus the built-in version of FIM included with SharePoint Server.
The SharePoint Connector officially supports SharePoint Server 2013, but will unofficially work with SharePoint Server 2010.
You will need a few Domain accounts. An account to run the FIM Service (s-fim), an account to run the FIM Management Agents (s-fimma), the SharePoint farm administrator account (s-sp2013farm), and finally a synchronization account for Active Directory (s-sp2013sync). For the last account, this guide will be using the same account as the one used for the UPA connection. Configure the permissions appropriately for s-sp2013sync.
Provision the UPA and UPSS per the standard instructions. Once both services have been configured, stop the FIM services on the SharePoint server and set them to Disabled. In the UPA under Configure Synchronization Settings, you have Enable External Identity Manager selected.
First, we'll start out with a SQL Server running SQL Server 2012 SP1 with the Database Engine, Integration Services, and Management Studio. All other settings are at their defaults. If you are using a SQL Server that is not running on the same server as the FIM services, make sure to install the SQL Server Native Client on the server running the FIM services.
The FIM server will run SharePoint Foundation 2013, the FIM Synchronization Service as well as FIM Service and Portal, along with the SharePoint User Profile Connector.
Install SharePoint Foundation 2013 and create a Classic Web Application for the FIM Portal. The FIM Portal does not currently work with Claims-based Authentication. Next, install the FIM Synchronization Service. During the installation, specify the FIM Synchronization Service account.
Next, install the FIM Service and Portal. The Portal will leverage our SharePoint Foundation installation and Classic Web Application. The Classic Web Application has been configured with an Alternate Access Mapping of "FIM02" in this example.
Enter the SharePoint site collection URL.
Enter the hostname of the FIM Service server. We're installing the Portal and Service on the same server, so again we'll use "FIM02" here.
Enter the hostname of the Synchronization Service, along with the Management Agent account.
Again, enter the FIM Service service account information.
You can either let FIM generate a self-signed certificate, or use a certificate signed by a Certificate Authority. For purposes of synchronization, a self-signed certificate will work.
Enter the mail server information. Since we're just after synchronization, the remaining options are unchecked (leaving the polling option checked, if not configured properly, will generate Event Log warnings).
Enter the database server name and database name.
Finish the installation of the FIM Service and Portal. Next, install the KB2832389 update for the FIM Synchronization Service and FIM Portal and Service. This update is required prior to installing the SharePoint User Profile Connector. Download the Connector for SharePoint User Profile Store. Install the SharePoint User Profile Connector on the FIM Synchronization Service server.
The next step is to use the FIM client and FIM Portal to set up our Management Agents, Synchronization Rules, Workflows, and Management Policy Rules. This will cover the basics required, but you will want to adjust the attributes used and users targeted based on business requirements. Lastly, this will only cover User objects, but Contact and Group objects are also available for synchronization.
Click Next until you complete the Management Agent.
You may also add other attributes, such as WorkEmail, WorkPhone, and so forth. This example will use some of these other attributes later in the Synchronization Rules. Complete the SharePoint Management Agent.
Configure Run Profiles for each Management Agent. The Active Directory Management Agent requires Full Import, Full Synchronization, Delta Import, and Delta Synchronization. The FIM Management Agent requires Full Import, Full Synchronization, Export, Delta Import, Delta Synchronization. The SharePoint Management Agent requires Full Synchronization, Export, Delta Synchronization.
In the Synchronization Service Manager, go to Tools -> Options and select Enable Synchronization Rule Provisioning.
This completes the Management Agent setup.
Prior to creating the Synchronization Rules, you will want to run a PowerShell script to determine the Custom Expression for the domain attribute. You can get the PowerShell script from Using PowerShell To Generate The Custom Expression For The Domain Attribute Flow. Edit the variables at the beginning of the script to match your domain and forest. Next, save the output value.
accountName => UserName; IIF(Eq(Left(ConvertSidToString(objectSid),40),"S-1-5-21-X-X-X"),"NAUPLIUS","Unknown") => domain; IIF(Eq(Left(ConvertSidToString(objectSid),40),"S-1-5-21-X-X-X"),"NAUPLIUS","Unknown")+"\"+accountName => AccountName; firstName => FirstName*; lastName => LastName*; IIF(Eq(Left(ConvertSidToString(objectSid),40),"S-1-5-21-X-X-X"),"NAUPLIUS","Unknown")+"\"+accountName => ProfileIdentifier; firstName+" "+lastName => PreferredName*; email => WorkEmail*; officePhone => OfficePhone*; ConvertSidToString(objectSid)** => dn; ConvertSidToString(objectSid)** => Anchor.
Values marked with an * may have the option "Allow null attributes" checked and values marked with ** are set to Initial Flow Only.
Note: If you set the Synchronization Rule to Apply Rule "to all metaverse resources of this type…", you can skip creating a Workflow, MPR, and Set. However, you do lose further flexibility by doing so. For standard import purposes, this flexibility is likely not required.
Next, create a Workflow. On the left, under Management Policy Rules, select Workflows. This workflow will be used to export objects into SharePoint. Name the workflow appropriately. Leave Run on Policy Update unchecked. For the Activity, add a new Synchronization Rule Activity type, then select the SharePoint Management Agent, and select Add as the Action Selection. Save and exit the workflow.
Create the Management Policy Rule. Name the rule appropriately. Select Transition Set, then select Transition In with an appropriate Transition Set. I will be using "All People" in this example. Under the Policy Workflows, select the workflow you just created.
Check the User Profile Service Application. There should be a number of User Profiles.
If there is a change in the Management Agent rules, re-run the initial synchronization. On the Configure Run Profiles window for each Management Agent, you can Script each run profile to VBScript. This will allow you to create a scheduled task to execute the run profile automatically.
That is all there is to it. Note that when upgrading SharePoint (Cumulative Update or Service Pack), you must re-provision the User Profile Synchronization Service, then stop it again. This is required in order to update the User Profile databases.
Leveraging FIM as your Identity Manager will allow you to create more complex business rules, as well as import from a variety of sources not supported directly via SharePoint, or write your own custom code for import purposes.