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اهمیت تکاملی پلیپوئیدی پس از یک قرن مطالعه

Spreading Winge and flying high: The evolutionary importance of polyploidy after a century of study

Since the discovery of polyploidy in plants just more than a century ago, research on polyploidy is as vigorous as ever. At the Botany 2015 meeting in Edmonton, Alberta, six researchers participated in the symposium “The Evolutionary Importance of Polyploidy” to address recent debates on polyploid speciation. These authors are joined by others in this special issue of the American Journal of Botany to highlight recent advances in polyploid research. The 19 articles collected here represent a diverse range of research from established and nascent scientists. Here we provide a guide to the articles in this issue and highlight their major results.

Perhaps no topic of study has received a greater boost from the genomics era than polyploidy. Speciation by genome duplication was recognized early in the study of evolutionary genetics and “represented the first major triumph in the genetics of speciation” (Coyne and Orr, 2004, p. 322). Plants with sets of doubled chromosomes were encountered in early cytological studies (Lutz, 1907), and Winge (1917) provided one of the most influential hypotheses for the origins of these numbers. He proposed the now familiar concept that these polyploid plants were fertile because genome doubling restored chromosome pairing in otherwise sterile hybrids. The restoration of fertility to sterile hybrids by doubling their genomes was a potent and elegant demonstration of how postzygotic reproductive isolation could be solved (Clausen and Goodspeed, 1925). Further support for Winge’s hypothesis came from Müntzing (1930) who recreated a naturally occurring polyploid species. These studies demonstrated one of the first mechanisms of how new species may arise from genomic changes of existing species. The pioneering polyploidy research of these botanists played a leading role in shaping research on speciation genetics. Their experiments even inspired a young Dobzhansky (1933) to test whether polyploidy restored fertility to sterile fruit flies (it did not). By the time of the modern synthesis, polyploidy was already a relatively well-studied topic and recognized as the biggest difference between plant and animal speciation (Dobzhansky, 1937)—and was the most shocking and important correction to Darwin’s theory of the origin of species (Haldane, 1959).

By the latter half of the 20th century, however, polyploidy had been relegated as largely unimportant by many researchers. Many plant species were certainly recognized to have polyploid origins (Stebbins, 1950; Grant, 1981), but the prevalence and persistence of diploids raised questions about the evolutionary contribution of polyploidy (Stebbins, 1971). Although ancient polyploidy was hypothesized and implicitly recognized in many “basic” chromosome number estimates (Stebbins, 1950, 1971; Klekowski and Baker, 1966; Ohno, 1970; Grant, 1981), many biologists came to regard polyploidy as “evolutionary noise” (Wagner, 1970). During his talk at the Botany 2015 symposium on polyploidy, Doug Soltis recounted that as a young researcher he was told to avoid polyploidy because it was a black hole of research! Today such advice would probably not be given. Research beginning with the molecular era revealed that even plants with high chromosome numbers have diploid patterns of gene expression (Gastony and Gottlieb, 1982; Gastony and Darrow, 1983; Barker and Wolf, 2010; Haufler, 2014). One hypothesis proposed to explain these patterns of chromosome number and gene expression was diploidization following ancient polyploidy (Stebbins, 1985; Haufler and Soltis, 1986; Haufler, 1987; Gastony, 1991). Analyses of numerous plant (Vision et al., 2000; Bowers et al., 2003; Blanc and Wolfe, 2004; Cui et al., 2006; Jaillon et al., 2007; Barker et al., 2008, 2009, 2012, 2016b; Rensing et al., 2008; Soltis et al., 2009; Schmutz et al., 2010; Shi et al., 2010; Tang et al., 2010; Velasco et al., 2010; Wang et al., 2011; Rodgers-Melnick et al., 2012; Tomato Genome Consortium, 2012; Amborella Genome Project, 2013; Barker, 2013; Chalhoub et al., 2014; Kagale et al., 2014; Liu et al., 2014; Vanneste et al., 2014; Edger et al., 2015; Marques et al., 2016), animal (Gregory and Mable, 2005; Santini et al., 2009; Hallinan and Lindberg, 2011; Cañestro et al., 2013; Berthelot et al., 2014; McLysaght et al., 2014; Nossa et al., 2014; Clarke et al., 2015; Pasquier et al., 2016), and other eukaryotic genomes (Aury et al., 2006; Scannell et al., 2006; Fares et al., 2013; Conant, 2014; McGrath et al., 2014; Scienski et al., 2015) revealed that many eukaryotes do indeed have a polyploid ancestry. Although there have been recent debates on the average fate of polyploid species (Mayrose et al., 2011, 2015; Soltis et al., 2014a), there is consensus that polyploidy has contributed to the evolution of nearly all lineages of vascular plants (Jiao et al., 2011; Arrigo and Barker, 2012; Soltis et al., 2014b; Li et al., 2015; Wendel, 2015) and that polyploids comprise a large fraction of contemporary plant diversity (Wood et al., 2009; Soltis et al., 2010; Mayrose et al., 2011; Scarpino et al., 2014; Barker et al., 2016a).

The resurgence of interest in polyploidy over the past 15 yr has been dramatic. Although polyploidy has long been a favored topic of many botanists, the Botany 2000 meeting of the Botanical Society of America in Portland, Oregon, featured only six abstracts, or 1% of the total, with “polyploidy” as a key word (Fig. 1). At Botany 2015, 36 talks and posters listed “polyploidy” as a key word, for 3% of the total. Although 3% may seem like a relatively small fraction of the submissions, polyploidy was the third-ranked key word at Botany 2015. Only submissions on “phylogeny” with 59 and “fungi” with 37 ranked higher than those on “polyploidy”. Polyploidy will likely remain an important topic in the near future as botanists continue to explore the consequences of polyploidy at multiple scales of ecology and time.

Figure 1.

Percentage of abstracts from the annual meetings of the Botanical Society of America featuring “polyploidy” as a key word. Data were collected from a search of key words for the abstracts of each meeting from 2000–۲۰۱۶ (data are not available for 2002) at website http://www.botany.org/conferences/.

Nearly a century after Winge’s (1917) landmark publication, research on polyploidy remains vibrant and diverse. Although frequently discussed as a topic of genome research, polyploidy impacts the biology of plants in many dimensions. Many of these different avenues of polyploid research are featured in this special issue of the American Journal of Botany. The 19 papers published in this special issue provide a snapshot of the breadth of research on polyploidy in plants. We have organized the manuscripts into three thematic areas to aid navigation of the research discussed in this special issue. These include papers on the evolutionary consequences of polyploidy, the origins of polyploid species, and the impacts of polyploidy on plant ecology. Next, we highlight the major findings of these papers and place them in context.

EVOLUTIONARY CONSEQUENCES OF POLYPLOIDY
Analyses of plant genomes have revealed the prevalence and importance of polyploidy in the evolutionary history of plants, but many questions remain about the consequences of genome duplication. Soltis et al. (2016, in this issue) review the recent history of research on polyploidy and highlight many of the obstacles to establishing a paradigm of polyploid biology. Many barriers to a paradigm reflect gaps in our knowledge about particular study systems. Echoing previous reviews of polyploidy (Wendel, 2000; Osborn et al., 2003; Doyle et al., 2008; Edger and Pires, 2009; Soltis et al., 2010; Barker et al., 2012; Conant et al., 2014), Soltis et al. (2016) also recognize that inconsistent patterns of evolution following polyploidy have made it challenging to develop a “unified theory” of polyploidy. Recent progress has been made toward better predicting the outcomes of polyploidy (Hollister and Gaut, 2009; Xiong and Pires, 2011; Coate et al., 2014; Conant et al., 2014; Garsmeur et al., 2014; Woodhouse et al., 2014; Renny-Byfield et al., 2015; Wendel, 2015; Steige and Slotte, 2016) and resolving the biology behind these patterns of genome evolution. The review by Soltis et al. (2016) provides a comprehensive roadmap for polyploid research. Many of the questions that they raise are addressed in other papers in this issue.A quartet of papers explore different aspects of how genomes evolve following polyploidization at different scales. Numerous changes in genome organization follow polyploidy, and these changes often obscure syntenic relationships among plant genomes (Sankoff et al., 2010). Salse (2016, in this issue) reviews the current state of the field in plant genomics and the problems that polyploidy introduces in reconstructing the ancestral genome organizations of angiosperms. The reconstruction and use of ancestral genomes will be of great value in understanding plant evolution, and Salse (2016) highlights the many promises and challenges of developing these resources. A second paper by Zenil-Ferguson et al. (2016, in this issue) employs a phylogenetic approach to test hypotheses of genome size evolution across ploidal levels. Their results provide new perspectives on the interaction between ploidal level, chromosome numbers, and genome size across angiosperms. Going to the laboratory bench, Mandáková et al. (2016, in this issue) use chromosome painting to demonstrate that Cardamine cordifolia, a species with a putative triploid chromosome count, is in fact a diploidized tetraploid. Analyses of chromosomal rearrangements indicate that four terminal translocations reduced the number of chromosomes in this species. Although an analysis of only one species, their results remind us that chromosome counts alone do not tell the entire story of polyploidy. Taking a population genomic approach to genome evolution, Ågren et al. (2016, in this issue) evaluated the dynamics of transposable element (TE) evolution in the allotetraploid Capsella bursa-pastoris. In particular, they tested whether changes in TE composition are more consistent with hybrid breakdown of TE silencing or relaxation of natural selection. They find evidence that the amount and nature of TE proliferation in C. bursa-pastoris is most consistent with relaxed selection. Their results suggest that relaxed selection following ploidal increases likely have multiple impacts on plant genome evolution that may persist for millions of years following polyploidy. A major aim of future research will be reconciling the results of these different research programs to improve our models of plant genome evolution.Genomic analyses suggest that plants have experienced many cycles of polyploidy during their evolutionary history (Cui et al., 2006; Soltis et al., 2009; Arrigo and Barker, 2012; Wendel, 2015). Although many analyses have documented paleopolyploidies throughout plant evolution, our knowledge about most of these events is limited. Barker et al. (2016b, in this issue) combine new data and methods to update our understanding of the paleopolyploidy in the ancestry of the Compositae (Asteraceae) (Barker et al., 2008). Using a phylogenomic approach, MAPS (Li et al., 2015), they found evidence that the Compositae and Calyceraceae shared an ancient whole-genome duplication, followed closely by a second round of genome duplication (paleohexaploidization) in the ancestry of most Compositae. Although previous studies have documented paleohexaploidies in plant genomes (Jiao et al., 2012; Tang et al., 2012; Tomato Genome Consortium, 2012; Cheng et al., 2013), the study by Barker et al. (2016b) is notable because it provides the first evidence for extant paleotetraploid and paleohexaploid descendants from the same ancient polyploid complex. The consequences of ancient polyploidy are further explored by van den Bergh et al. (2016, in this issue). They evaluate the contribution of paleopolyploidy to the evolution and diversification of genes associated with the glucosinolate pathway in the Brassicaceae and Cleomaceae. Recent analyses have demonstrated that neofunctionalization of paralogs from ancient polyploidy have driven a chemical arms race between plants of the Brassicales and their pierid butterfly herbivores (Edger et al., 2015). Here, van den Bergh et al. (2016) demonstrate that most genes in the glucosinolate pathway are derived from paleopolyploidies in the history of Cleomaceae and Brassicaceae (Barker et al., 2009; Cheng et al., 2013). These papers highlight that while we have documented many paleopolyploidies throughout plant evolution, we have only just begun to understand the nature and consequences of these events.Another question addressed with a fresh perspective is the association between polyploidy and sexual systems. Flowering plants have an array of sexual systems (Barrett, 2002; Bachtrog et al., 2014). Most flowering plants are hermaphroditic with flowers that contain both female and male sexual organs. Monoecious species have both sexes in separate flowers on one individual. At the other end of this spectrum are dioecious species with an individual plant producing either male or female flowers. Polyploidy has long been hypothesized to be associated with transitions in sexual systems in flowering plants (Ashman et al., 2013), and alternatively hypothesized to be associated with both monoecy and dioecy. Using a large data set of ploidal levels and sexual systems for >1000 species, Glick et al. (2016, in this issue) analyzed the association of polyploidy and transitions in sexual systems with new coevolutionary phylogenetic models. Overall, their analyses suggest a correlation between polyploidy and sexual dimorphism. The complexity of the interactions between polyploidy and sexual systems is further revealed by the analyses of Glick et al. (2016). No doubt this phylogenetically broad empirical update will stimulate many future studies exploring the numerous evolutionary consequences of polyploid associated transitions in sexual systems.
THE ORIGINS OF POLYPLOID SPECIES
A second collection of papers in this special issue address how polyploid species originate. Fowler and Levin (2016, in this issue) provide a thoughtful introduction to this section in their analyses modeling the establishment of allopolyploid species. Most of the past models of polyploid establishment have focused exclusively on autopolyploids (Felber and Bever, 1997; Husband, 2004; Baack, 2005; Rausch and Morgan, 2005; Oswald and Nuismer, 2011). Most named polyploid species are actually allopolyploids (Barker et al., 2016a), and the biology of their establishment is quite different from autopolyploids given their hybrid origins (Levin, 2012; Barker et al., 2016a). In their population dynamic models, Fowler and Levin (2016) found that population sizes of all species involved is important to overcome the demographic stochasticity that faces these incipient species. Although they only analyzed allopolyploid species, these results likely also apply to autopolyploids. Despite more than two decades of modeling the biology of polyploid establishment, Fowler and Levin (2016) demonstrate that there is still much that these approaches can teach us about polyploid speciation.Polyploidy has long been proposed as a mechanism of “instant speciation” (Lutz, 1907; Winge, 1917), but empirical analyses of this contention are relatively rare. Postzygotic reproductive isolation between ploidal levels is expected to be strong and rapid because of chromosomal pairing problems (Grant, 1981), and there are a number of examples of reduced fertility in heteroploid hybrids (Ramsey and Schemske, 1998). However, there are many open questions about how genome duplication contributes to reproductive isolation and speciation. Three papers use varied methods to further explore the connection between polyploidy and reproductive isolation. Zhan et al. (2016, in this issue) develop a phylogenetic approach to explore the common assumption that polyploidization causes instant speciation. Their models find that polyploidy is frequently cladogenic rather than anagenic and largely supports the assumption that polyploidization equals speciation. The latter two studies use synthetic polyploids and controlled crosses to evaluate the impact of polyploidy on pre- and postzygotic reproductive isolation in different study systems. Husband et al. (2016, in this issue) examine the degree of reproductive isolation that arises immediately upon genome duplication and finds this to be significantly less than in natural tetraploids of Chamerion angustifolium. Unique among many analyses of polyploidy, they collect data on numerous aspects of prezygotic isolation to provide a more complete picture of total reproductive isolation caused by genome duplication. Their comparison of synthetic and natural polyploids provides a powerful test of the immediate contribution of polyploidy to prezyogtic reproductive isolation. Vallejo-Marín et al. (2016, in this issue) use reciprocal crosses and sequenced cytoplasmic genomes, including the first Mimulus chloroplast genome assembly, to explore how the strength of asymmetric hybridization barriers contribute to the formation of a new allohexaploid species, Mimulus peregrinus. Hybrid speciation frequently displays differences in fertility depending on the direction of the cross. In the case of allohexaploid speciation, the maternal parent is often disproportionately the tetraploid (Stebbins, 1957; Ramsey and Schemske, 1998; Köhler et al., 2010). Surprisingly, Vallejo-Marín et al. (2016) found the opposite pattern in Mimulus peregrinus with an excess of paternal tetraploids and maternal diploids. Together, these three studies demonstrate that although polyploid speciation is often assumed to be rapid with complete reproductive isolation, the reality is more nuanced with many unexplored questions about how polyploidy contributes to reproductive isolation and mediates subsequent evolutionary divergence.A unique aspect of polyploidy is that many polyploid species may trace their ancestry to numerous, independent origins (Werth et al., 1985; Soltis et al., 2010, 2016; Beck et al., 2012). Three papers in this special issue address the origins and taxonomy of polyploid species. Revisiting the classic Babcock and Stebbins (1938) study on the agamic Crepis species complex, Sears and Whitton (2016, in this issue) use flow cytometry and plastid phylogenetics to test the hypothesized relationships of species. They find that the inferences of ploidal variation by Babcock and Stebbins (1938) were largely correct. Significantly, Sears and Whitton (2016) demonstrate that the North American Crepis agamic complex is monophyletic and provide a reasonably well-resolved phylogeny. Rešetnik et al. (2016, in this issue) explore similar questions in the Knautia drymeria species complex, one of the most taxonomically challenging plant genera of Europe. Knautia drymeria includes diploid and tetraploid cytotypes, and tetraploid populations from the Apennines have been treated as an independent species, K. gussonei. Genetic and morphological analyses could not distinguish K. drymeria, the subspecies of K. drymeria, and K. gussonei. Knautia is an excellent example of the taxonomic knots created by the presence of allo- and autopolyploid cytotypes within species. Finally, McAllister and Miller (2016, in this issue) provide a population genomic analysis of the origins of autopolyploids within the iconic plant of North American tallgrass prairies, Andropogon gerardii, or big bluestem grass. Using a genotyping-by-sequencing (GBS) approach, they analyzed thousands of single nucleotide polymorphisms and found that 9x plants likely formed many times from 6x progenitors in distinct geographic regions. This study is one of the first to employ a GBS approach to explore multiple origins of polyploids in natural populations and will serve as a great example for future research. Overall, these results reinforce that recurrent polyploid origins are probably the rule, rather than the exception in nature.
IMPACTS OF POLYPLOIDY ON PLANT ECOLOGY
Although many papers in this issue have focused on the evolutionary aspects of whole-genome duplication, polyploidy also has significant consequences for plant ecology. Four papers in this special issue explore the relationships between polyploidy and ecology. Given the potentially numerous changes in plant phenotype caused by polyploidy (Stebbins, 1940; Levin, 1983; Otto and Whitton, 2000), there exists the possibility that genome duplications have cascading impacts on biotic interactions throughout ecosystems (Thompson et al., 2004). In an excellent review of the literature, Segraves and Anneberg (2016, in this issue) find that, although rare, polyploidy can uniquely change plant interactions with pollinators, herbivores, and pathogens. Echoing other areas of polyploid research, more detailed studies are needed to disentangle whether genome duplications per se or natural selection in new reproductively isolated lineages cause these novel interactions.The final three papers in this issue focus on polyploidy and ecological niches. In an analysis of the South African Cape Flora, Oberlander et al. (2016, in this issue) found that polyploidy is under-represented in the climatically and geologically stable yet hyperdiverse region. In comparisons of the proportions of polyploids from 21 clades, they found significantly fewer polyploids in the Cape region than from other parts of the world. These results support hypotheses that polyploidy may be relatively rare in stable environments and more frequent in more ecologically dynamic regions or times (Stebbins, 1971; Vanneste et al., 2014). At a smaller scale, two studies by Arrigo et al. (2016, in this issue) and Laport et al. (2016, in this issue) found that polyploid cytotypes in Alyssum montanum and Larrea tridentata are often associated with novel ecological niches compared with their parental taxa. In the case of Alyssum montanum, Arrigo et al. (2016) report that allopolyploids had higher rates of niche evolution than autopolyploids. The cytotypes of L. tridentata are all autopolyploids and demonstrated significant ecological differentiation, even though Laport et al. (2016) found evidence that the hexaploids are likely formed continuously. Both of these studies set the stage for future investigations to unravel whether polyploidy per se or subsequent natural selection drive changes in the ecological niche of plants and the relationship with speciation.
CONCLUSION
Research on polyploidy is vibrant and going strong nearly a century after Winge’s (1917) influential paper. Although polyploidy may have experienced a nadir of interest during the latter half of the 20th century, the sequencing and analyses of plant genomes have revealed numerous cycles of polyploidy in the history of nearly all land plants (Wendel, 2015). The manuscripts published in this special issue provide a sampling of the ongoing research on whole-genome duplications and plants. A challenge for the community over the coming years will be to better integrate these disparate threads of research. At the phylogenetic scale, synthesis appears well on the way as large and diverse data sets are being integrated and analyzed in joint frameworks. A larger challenge ahead will likely be the expansion of study of the classic polyploid model systems. Historically, research in these systems has reflected the efforts and interests of particular laboratories and individuals. Some of the most interesting questions in research on plant evolution and polyploidy will be answered by comprehensively analyzing multiple dimensions of species and population biology within single systems. Adding genetics and genomics to classically ecological study systems, or ecological and phenotypic analyses to well-studied genetic model systems will be a recipe for future success in polyploidy.
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