Mitochondrial genomes of flowering plants (angiosperms) are highly dynamic in genome structure. 133 sequences of a conserved domain in the reverse transcriptase gene of Ty1-copia elements were isolated from 13 varieties of waterlilies and found to be homologous to either Ale, Angela, or TAR lineages. How an organism develops determines its phenotype, and therefore differences among ontogenies are what generate diversity. The Gnetales can be supported as a monophyletic group, with Gnetum and Welwitschia more closely related to each other than either is to Ephedra. Forest trees are long-lived organisms subject to repeated environmental constraints throughout their long lifetimes. These results suggest that several monotypic relict gymnosperm lineages like P. chienii in south-east China did not remain in situ and undiversified for millions of years. Approximately 12% and 0.5% of the total SNPs and InDels identified in three lines were located in genic regions, respectively. Of general evolutionary interest is the protracted "rebound" of the larger vertebrate fauna and the nature of its controlling factors. The differentiation of ACPs into distinct types occurred 85–98 and 45–57 million years ago. Cenomanian-Turonian sediments from the Sarbay locality in north-western Kazakhstan have yielded a rich assemblage of plant fossils including well preserved flowers, fruits, and seeds of angiosperms. through time, extinction and recovery, past climate The results obtained from the study of chloroplast tRNA will greatly help to increase our understanding of tRNA biology at a new level. The results suggest that increased understanding of Triassic Bennettitales, Gnetales, and corystosperms will be of maximum interest in further elucidating the phylogenetic relationships of flowering plants. Whole genome duplication, a prevalent force of evolution in plants, results in massive genome restructuring in different organisms. Besides, we found that polyploidy contributed to the expansion of key functional genes, e.g., vitamin C biosynthesis genes. To investigate relationships and character evolution in the angiosperm family Chloranthaceae, and to integrate fossil specimens into the phylogeny of extant taxa, a parsimony analysis was carried out using morphological data from 38 extant and six fossil species. Asymmetric gene flow and similar population divergence times (~ 3.74 Ma) were characterized using the isolation with migration model. In this study, we investigated whether there is a potential relationship between separated neighboring gene pairs and WGDs, and the mechanism by which neighboring genes are separated. Morphological- and molecular-based outgroup arrangements gave essentially identical ingroup relationships and support values. tenuitesta, are established and new information on the previously described species,L. Cycad-like plants Bennettitales? Hybridization and heterogeneous habitats in the HDM were suggested to play an important role in the polyploidization in subg. Conse- quently, as an entomologist, because of the close asso- ciations between plants and insects, I have been prompted to contribute to the discussion on this "abom- inable mystery" as Charles Darwin called it. They successfully dispersed and became established across the South-East Asian region, initially carried by wind or birds, beginning at about 49 Ma, and with a terrestrial connection after about 41 Ma. Symmetry of flowers is either structurally correlated with the basic merosity, the result of extreme reduction, or developmentally correlated with shifts in organ initiation. 38 or 44 events of inversions and translocations are needed to reconcile the mitogenome of Nymphaea with Amborella or Liriodendron, respectively. We also detected significant molecular footprints of a recent (about 9.04 MYA), medium (about 43.44 MYA) and an ancient (about 116.39 MYA) WGD events that occurred in the evolutionary history of three blueberry species. The following points highlight the top nine theories about ancestors of angiosperms. Tepal aestivation is spiral imbricate in Amborella and Austrobaileyales, with broadly overlapping flanks; aestivation is mainly contort within each of the two tepal whorls in Cabomba (but open within each whorl and imbricate only between whorls in Brasenia) and is whorled imbricate in Nymphaeaceae. Flowers represent a key innovation during plant evolution. 85% are taxa at the magnoliid grade - perhaps also with some basal monocots. In more than 30 analyses of up to 10 million generations each (with an average run time of 45 d), log-likelihood plots showed that runs can stabilize for several million generations before making jumps in likelihood scores. Whole genome duplication events (WGDs) have been recognized as an important evolutionary force. Visitation frequency and pollen load of pollen and oil-collecting bees were also investigated. Using dispersal-vicariance analysis, we infer that the ancestor of Nymphaeales occupied the American and Eurasian continents during the Eocene and that the present distributional patterns require several subsequent dispersal and extinction events. We calibrated the trees by using the minimum age of the angiosperm crown group constrained to 131.8 mya. Results We summarize the primary physiological and structural traits involved in response to cold- and drought stress, outline the phylogenetic distribution of these adaptations, and describe the recurring association of these changes with rapid diversification events that occurred in multiple lineages over the past 15 million years. The current status of studies is briefly summarized here. Parsimony jackknifing was performed to assess the support of clades. In particular, we found strong evidence of two sequential tetraploidization events, with one (Dc-α) approximately 46-52 million years ago (Mya) and the other (Dc-β) approximately 77-87 Mya, both likely allotetraploidization in nature. Here, we review the developmental role of A, B, C, D, and E gene classes and explore the recent advances in understanding the floral development and subsequent organ specification in major cereals with reference to model plants. Species co‐occurrence at fine spatial scales is expected to be nonrandom with respect to phylogeny because of the joint effects of evolutionary (trait convergence and conservatism) and ecological (competitive exclusion and habitat filtering) processes. The gene order in a eukaryotic genome is not random. Polyploidization has long been recognized as one of the most important driving forces of plant evolution. The developmental mechanisms responsible for changes in stamen number have rarely been evaluated in basal angiosperms, in contrast to the eudicots. Theories of geological change. We identified 30,143, 29,922 and 28,891 putative protein coding sequences from 45,535, 42,914 and 43,630 unigenes assembled from the leaves' transcriptome assembly of 19 rabbiteye (T1), 13 southern highbush (T2) and 22 northern highbush (T3) blueberry cultivars. Arbitrary parameters used to implement minimum-bound calibrations were found to have a strong impact upon the prior and posterior of the divergence times. (2016). This chapter also highlights new gaps in our understanding of early evolution of angiosperms and in the implementation of fossil calibrations in Bayesian molecular clock dating, and discusses several areas for future research. Some dispersals from Australasia occurred following its collision with Sunda and following the uplift of New Guinea and the islands of Wallacea, Gondwanan montane taxa also found their way into the region. Moreover, though mostly gene losses were nearly random, largely but not fully described by geometric distribution, we showed that polyploidization contributed divergently to copy number variation of important gene families. The polyphyletic origin of angiosperms is further supported by the fact that primitive orders of both the monocots and dicots do … It followed that primitive woody dicots were seen as the living descendants of early angiosperms. Three genetic clusters were identified. Overexpression of GhERF1‐7A in transgenic Arabidopsis resulted in a substantial increase in the number of siliques and total seed yield. Furthermore, the results indicate that those homologous to the Angela or TAR lineages are more conserved to their counterpart in other plants than those homologous to the Ale lineage. Evolutionary aspects continue to be insufficiently investigated, and although most comparative studies treat growth rings recognized by different markers as though they were a single character, it remains questionable whether they can be regarded as homologous or the result of homoplasy. The origins of angiosperms and their biological consequences . Tepals are absent in Hydatellaceae, Chloranthaceae (other than Hedyosmum), and Ceratophyllaceae. (With Methods)| Industrial Microbiology, How is Cheese Made Step by Step: Principles, Production and Process, Enzyme Production and Purification: Extraction & Separation Methods | Industrial Microbiology, Fermentation of Olives: Process, Control, Problems, Abnormalities and Developments, The best answers are voted up and rise to the top. Current evidence is most congruent with the concept that a process of double fertilization first evolved in a common ancestor of the Gnetales and angiosperms. the phylogeny using two different genomes, this research will investigate the historical construction of East Asian EBLFs an, Krassilov wrote in the beginning of his book (1989), that nearly all eminent botanists considered it their duty to speak on the problem of angiosperm origin. Furthermore, we found that there is a double linear relationship between separated neighboring genes, total genes, and WGDs. Roles of the resultant duplicated genes are poorly understood, both functionally and evolutionarily. Guidelines for submitting commentsPolicy: Comments that contribute to the discussion of the article will be posted within approximately three business days. Only in the latter groups are there genera with basally united tepals (Amborella, Cabomba, and Hedyosmum). Important fossils are depicted and put into the evolutionary context in this review. To assess whether homologues of floral organ identity genes in Persea may play a role in shaping this dimorphic perianth, we compared their expression patterns in the two species. Results of the analysis suggest that the seed ferns as currently circumscribed are not a meaningful group for phylogenetic purposes and permit an evaluation of the possible phylogenetic position of Eremopteris, Nystroemia, Spermopteris, Phasmatocycas, Vojnovskiales, Leptostrobus, and several other enigmatic groups of plant fossils. Many clades receiving moderate to low jackknife support in parsimony analyses received pp values of 1.0. across traditionally separate research areas is critical to continued success. Evidence from the history of other major clades of land plants, such as seed ferns, suggests that the characteristic features of angiosperms were acquired sequentially through time [3]. Angiosperm, any of about 300,000 species of flowering plants, the largest and most diverse group in the plant kingdom. Transcription factors (TFs) play a key role in regulating plant development and response to environmental stimuli. This result refutes the anthophyte hypothesis. Regarding expression patterns, most ACPs were expressed constitutively and tissue-specifically. There are two main models for the origins of eukaryotes on Earth, "genetic merger" which postulates that eukaryotes evolved between 3 bya to 2 bya following the merger of two species of prokaryote, and "deep roots" which posits that eukaryotes and prokaryotes appeared on Earth at the same time. Ceratophyllaceae are sister to the eudicots. Here, we review the most recent comprehensive studies of plant MCFs, incorporating the torrent of genomic data emanating from next-generation sequencing techniques. Tetraploid black locust possessed more tolerance of, and ability to acclimate to, salinity stress than diploids, which may be attributable to the ability to maintain mitochondrial structure and to trigger different expression patterns of mitochondrial proteins during salinity stress. We have extended our studies of angiosperm phylogeny based on a pair of duplicated phytochrome (PHY) genes, PHYA and PHYC. It discusses different approaches for estimating divergence times, with emphasis on Bayesian molecular clock dating methods. Repair of an AAD3 missense mutation at the catalytically essential Tyr73 residue did not result in a functional enzyme. All of them demonstrate the existence of enclosed ovule in the organ, satisfying the criterion for angiosperms. The phylogenetic definitions proposed here should help focus future discussions of the PhyloCode on real definitions rather than simplified hypothetical ones. The resulting cladograms support the monophyly of the Chloranthaceae as well as the four extant genera Chloranthus, Sarcandra, Ascarina, and Hedyosmum. d the relationship between its assembly with the dynamics of the Asian monsoons. The Nymphaea mitogenome resembles the other available early angiosperm mitogenomes by a similarly rich 64-coding gene set, and many conserved gene clusters, whereas stands out by its highly repetitive nature and resultant remarkable intron expansions. Polyploidization is well tolerated in many species and can lead to specific biological functions. We conclude that a population genomics approach within a clearly defined phylogenomic framework is essential not only for understanding the evolution of the pathogenicity mechanism but also for identifying informative candidates associated with pathogenicity that can be developed as targets in disease management programs. Illicium at 25.9 pg, providing further support for recognizing these taxonomic sections. As a model plant to study perennial trees in the Salicaceae family, the poplar (Populus trichocarpa) genome was sequenced, revealing recurrent paleo-polyploidizations during its evolution. These TF duplicates with asymmetrically partitioned ancestral states are likely maintained because one copy retains ancestral functions while the other, at least in some cases, acquires novel cis-regulatory sites that may be important for novel, adaptive traits. given at the numerous symposia are included. Here, we performed a reanalysis of the genome and found clear evidence of 2 tetraploidization events, with one occurring ∼50-57 million years ago (Mya) and the other ∼18-20 Mya. Cytogenetics and Genome Size Evolution in Illicium L. Two reported cytotypes of the emergent orchid model species Erycina pusilla are two different species, Deciphering the Origin, Evolution, and Physiological Function of the Subtelomeric Aryl-Alcohol Dehydrogenase Gene Family in the Yeast Saccharomyces cerevisiae, Functional diversifications of GhERF1 duplicate genes after the formation of allotetraploid cotton: Functional diversifications of GhERF1 duplicate genes, Evidence for an ancient whole genome duplication in the cycad lineage, Comparative mitochondrial proteomic, physiological, biochemical and ultrastructural profiling reveal factors underpinning salt tolerance in tetraploid black locust (Robinia pseudoacacia L.), Bordered pits in xylem of vesselless angiosperms and their possible misinterpretation as perforation plates, Two Highly Similar Poplar Paleo-subgenomes Suggest an Autotetraploid Ancestor of Salicaceae Plants, Multilocus phylogenetic reconstruction informing polyploid relationships of Aconitum subgenus Lycoctonum (Ranunculaceae) in China, Angiosperms at the edge: Extremity, diversity, and phylogeny, Epigenetics in Forest Trees: State of the Art and Potential Implications for Breeding and Management in a Context of Climate Change, Genomic and evolutionary aspects of chloroplast tRNA in monocot plants, Towards understanding the incidence and evolutionary history of seed recalcitrance: An analytical review, Metabolic Roles of Plant Mitochondrial Carriers, Re-sequencing and transcriptome analysis reveal rich DNA variations and differential expressions of fertility-related genes in neo-tetraploid rice, Genetic and Molecular Control of Floral Organ Identity in Cereals, The evolution of angiosperm lianescence: a perspective from xylem structure-function, The growth ring concept: seeking a broader and unambiguous approach covering tropical species, Evolutionary genomics model of chromosome number reduction and B chromosome production, Evolutionary history of a relict conifer, Pseudotaxus chienii (Taxaceae), in south-east China during the late Neogene: old lineage, young populations, Polyploidy in liver development, homeostasis and disease, Fungi in the rear mirror A brief history of the fungi during the last two billion years, The specialization-generalization continuum in oil-bee pollination systems: a case study of six Brazilian species of Tigridieae (Iridaceae), Evolvability of flower geometry: Convergence in pollinator-driven morphological evolution of flowers, Pilze im Rückspiegel – eine kurze Geschichte der Pilze während der letzten zwei Milliarden Jahre, Significance of whole-genome duplications on the emergence of evolutionary novelties. Thus, it is likely that the earliest flowering plants displayed an ab initio cellular pattern of endosperm development, whose expression was inherited from that of the supernumerary embryo of the ancestors of flowering plants. We applied four different methods, a strict molecular clock, nonparametric rate smoothing (NPRS), penalized likelihood (PL), and a Bayesian method, to estimate divergence times. Lastly, isozyme data support the hypothesis that basic chromosome numbers near x = 11 or 12 may mark a convenient point for demarcating polyploids. Rooted species trees inferred from duplicate gene networks by minimizing gene Ceratophyllum 1 water lilies duplications and losses are highly congruent with the gene subtrees and with the results of recent analyses of other genes, even when Ceratophyllum is included. ADVERTISEMENTS: There are two theories regarding Evolution of Bryophytes: (i) The first theory is known as up-grade or progressive evolution theory, (ii) and the second one is called down-grade or regressive evolution theory. Approximate Bayesian computation analyses showed that the three clusters diverged in the late Pliocene (~3.68 Ma) and two admixture events were detected. Crown clade names can be defined by three kinds of definitions, two of which are used here: standard node-based and branch-modified node-based. Lernen Sie in Abschn. They have developed various mechanisms enabling them to cope with fluctuating environmental conditions during their life span, and to survive to current climate change. At present, most investiga- tors consider that these localities belong to the late Jurassic, rather than to the middle Cretaceous, while the general curve remains mainly the same. Furthermore, we discuss the evolutionary relationships among known floral organ identity genes. This was followed by transcriptome assembly, after which these data were used to build paralog age distributions. Conversely, an approximate date for the origin of the first gene can be estimated based on the time frame in which life appeared on Earth and then evolved. Dramatic changes in the flora and fauna were exemplified by the development of angiosperm-dominated plant communities and the rapid diversification of mammals, birds and insects. Simultaneous independent runs reached apparent stationarity as early as 2 million generations and as late as 9.7 million generations, suggesting that (a) 10 million generations are insufficient for data sets of this size and (b) periods of stationarity even as long as 6 million generations should not be taken as an indication that the tree is fully optimized. With new fossil records from the Jurassic, research in this direction becomes more promising. to the same clade within the highly paraphyletic Campanula, the Rapunculus clade, members of the Isophylla group do not form a monophyletic group but fall into three separate clades: (i) C. elatines and C. elatinoides in the Alps; (ii) C. fragilis At each of the five localities between 7 and 26 types of angiosperm pollen have been recognized based on pollen grains in situ within anthers, or on stigmatic or fruit surfaces. We use data from 11 existing vegetation surveys to test whether co‐occurrence in schoenoid sedge assemblages in the Cape Floristic Region shows significant phylogenetic structuring and to examine whether this changes with the phylogenetic scale of the analysis. Trimery and whorled perianth phyllotaxis have played a major role in basal angiosperm perianth evolution and represent the ancestral states for the large clade comprising all angiosperms other than Amborella, Nymphaeaceae, and Austrobaileyales. The main factors affecting the estimates in this study are also discussed. Painting image courtesy Geoffrey Kibby (British Mycological Society). While radiation has led to an enormous variability in flower forms and sizes, recurrent evolutionary patterns can be observed. Information on cytogenetics, coupled with phylogenetic analyses, identifies some limitations, but also considerable potential for the development of plant breeding and improvement programs with this genus. The process of separation of neighboring genes caused by WGDs is also not random but abides by the double linear model. Evidence from the fossil record, comparative morphology, and molecular phylogenetic analyses indicates that Chloranthaceae are among the oldest lineages of flowering plants alive today. In view of the enormous diversity of angiosperms, many authors have found it hard to envisage a mono-phyletic origin of this group. The loss of the non-avian dinosaurs meant a loss of larger herbivorous browsers not replenished for some millions of years into the Paleocene. The evolution of the flower has long been a matter of intense debate. Chloroplasts are one of the most indispensable organelles that make life forms on the earth possible by their capacity to photosynthesize. The molecular basis for these differences is of fundamental importance to plant systematics, yet the topic remains poorly understood. Results suggest that seed plants are a monophyletic group, and Lyginopteris is resolved as the sister taxon to all other seed plants considered. Early Cretaceous fruits with three tepals and Asteropollis pollen belong near the base of Hedyosmum, but Couperites fruits with "Clavatipollenites" pollen, compared with Ascarina, may be either nested within the family or sister to it. a group consisting of all descendants derived from a single ancestor) or polyphyletic (i.e. Conclusions s.l. i. Epigenetics has recently emerged as a powerful set of mechanisms regulating various developmental processes, plant growth and responses to environmental variations. The diversity and abundance of angiosperms in the Portuguese mesofloras contrast strongly with the apparent paucity of angiosperm pollen in dispersed palynofloras, as well as the scarcity of angiosperm wood and leaves in Barremian-Aptian fossil plant assemblages. The genome encoded 41 protein coding genes, 20 tRNA and three rRNA genes with 25 group II introns disrupting 10 protein coding genes. i. Subsequently, in chapter four different strategies for summarizing fossil information to construct calibration priors were assessed employing an a priori procedure for deriving accurate calibration densities in Bayesian divergence dating. The alternative rates place the onset of divergence among crown group (extant) species of Hedyosmum at 60 or 29 Ma, between the Paleocene and the Oligocene; that among extant Chloranthus at 22 or 11 Ma; and that among extant Ascarina at 18 or 9 Ma, implying long-distance dispersal between Madagascar and Australasia-Polynesia. Be careful if copying and pasting from a Word document. An exception to that rule has recently been found in the orchid Erycina pusilla, which has been reported to have two cytotypes with chromosome numbers of 2n = 10 and 2n = 12. Subsequent to the divergence of the angiosperm lineage from its closest relatives (which include the Gnetales), modification of the development of the supernumerary embryo (derived from the second fertilization event) led to the establishment of an embryo-nourishing endosperm. Conclusions: The Stachyospory-Phyllospermae Theory 7. The analysis of mutations affecting flower structure has led to the identification of some of the genes that direct flower development. Background 140 mya (early Cretaceous) but already major radiation! Fungal evolutionary history goes back for at least two billion years. Inside an anther (male part of a flower) each cell experience meiosis (reduction of chromosomes), producing four haploid spore cells. Bayesian analyses of a three-gene, 567-taxon (560 angiosperms, seven outgroups) data set revealed the analytical challenges posed by such large data sets. These emerging areas offer many novel opportunities to link paleoecology and neoecology. FLOWERING plants or angiosperms have dominated the Earth's flora since at least the late Cretaceous1 and were already highly diversified by Barremian times, about 120 million years (Myr) ago. (C) 2003 The Linnean Society of London. The liver is one of the few mammalian organs that display changes in polyploidy during homeostasis, regeneration and in response to damage. Kenrick, Paul. For over a century, theories of Angiosperm origin have been framed within the assumption of seed plant monophyly. The study of cytogenetics of Illicium can clarify various discrepancies and further elucidate chromosome numbers, ploidy, and chromosome and genome size evolution in this basal angiosperm lineage and provide basic information to guide plant breeding and improvement programs. Subsequent climate and habitat changes coincide with the radiation of large herbivorous mammals such as perissodactyls and artiodactyls. One part of the enigma lies in the difficulty of identifying the earliest angiosperms; the other involves the uncertainty regarding the sister group of angiosperms among extant and fossil gymnosperms. Benites-Palomino, Aldo M.1,*; Tejada-Medina, Luz2 The objectives of this study were to use flow cytometry and traditional cytology to determine chromosome numbers, ploidy levels, and relative genome sizes of cultivated Illicium. Results Current phylogenetic hypotheses broadly support previous views that the most basal angiosperm taxa fall within a grade of organization corresponding to the subclass Magnoliidae; however, there are divergent views on the resolution of relationships within the magnoliid grade. However, when Ceratophyllum is included, and then Chloranthaceae diverge from the remaining angiosperms before Austrobaileya 1 Illicium . Homology searches indicate that Saccharomyces cerevisiae strain BY4741 contains seven redundant genes that encode putative aryl-alcohol dehydrogenases (AAD). The results strongly suggest that the studied species are distributed along a specialization-generalization continuum. The wide distribution of angiosperm plants caused the Middle Cretaceous biocoenosis crisis, according to Zherikhin (1978). The separation of neighboring gene pairs is closely related to whole genome duplication events. Background: Different theories for the origin of the angiosperm hermaphrodite flower make different predictions concerning the overlap between the genes expressed in the male and female cones of gymnosperms and the genes expressed in the hermaphrodite flower of angiosperms. There have been many proposals of candidates for the ancestors or closest relatives of angiosperms. Results: To evaluate the ploidy-dependent differences in leaf mitochondria between diploid and tetraploid black locust under salinity stress, we conducted comparative proteomic, physiological, biochemical and ultrastructural profiling of mitochondria from leaves. The common ancestor of angiosperms and Gnetales may be as old as Permian, and some of their shared advances, such as double fertilization (without endosperm formation), may have arisen as adaptations to seasonal temperate climates in Gondwana. Privacy Policy3. What are the general characters of bryophytes? Schmeissneria, Xingxueanthus, Solaranthus, Euanthus, Juraherba and Yuhania are three female or bisexual organs of plants found in the Middle Jurassic in China and the Early Jurassic in Germany. Taken together, these data provide an important resource to guide direct genetic studies aimed at addressing the relevance of these vital carrier proteins. Notably, the average expression levels of ACP1, mtACP3, and mtACP1 were positively correlated with those of ACP3, ACP4, and mtACP2, respectively. Ancestral perianth phyllotaxis and merosity are equivocal for each node of the Amborellaceae, Nymphaeaceae, Austrobaileyales grade; however, an undifferentiated perianth is reconstructed as the ancestral state for the angiosperms. A hallmark of flowering plants is their ability to invade some of the most extreme and dynamic habitats, including cold and dry biomes, to a far greater extent than other land plants. We conclude that the elaboration of a robust but broad definition which can include all the variation observed in growth rings of tropical species is a complex task, which is only possible by overcoming the restrictions adopted by dendrochronologists and wood anatomists of temperate regions for whom growth rings are essentially annual, strongly demarcated, tangentially continuous and restricted to the most pronounced markers. Here we report a phylogenetic analysis of DNA sequences of five mitochondrial, plastid and nuclear genes (total aligned length 8,733 base pairs), from all basal angiosperm and gymnosperm lineages (105 species, 103 genera and 63 families). This view is based at deeper nodes than proposed in earlier studies numerous. Is a process of separation of neighboring genes caused by WGDs is also helpful make... Bases present in the posted comment new AG-like genes from 15 diverse angiosperm species examined either alone or in with... Has resulted in the phylogenetic definitions are provided for the comment and Your affiliation differences! Aad genes are poorly understood genetic program across the angiosperms have typically been as. Chloranthaceae ( other than Hedyosmum ), and the nature of the angiosperms using two prominent fossils! 25 group II introns disrupting 10 protein coding genes, including Podostemaceae, Aphloiaceae, and possible! 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