Floral structure and development in Nartheciaceae (Dioscoreales), with special reference to ovary position and septal nectaries.

We present a comparative study of the floral structure and development of Nartheciaceae, a small dioscorealean family consisting of five genera (Aletris, Lophiola, Metanarthecium, Narthecium, and Nietneria). A noticeable diversity existed in nine floral characters. Analyses of their respective character states in the light of a phylogenetic context revealed that the flowers of Nartheciaceae, whose plesiomorphies occur in Aletris and Metanarthecium, have evolved toward in all or part of Lophiola, Narthecium, and Nietneria: (1) loss of a perianth tube; (2) stamen insertion at the perianth base; (3) congenital carpel fusion; (4) loss of the septal nectaries; (5) unilocular style; (6) unfused lateral carpellary margins in the style; (7) flower with the median outer tepal on the abaxial side; (8) flower with moniliform hairs; and (9) flower with weak monosymmetry. We further found that, as the flowers developed, the ovary shifted its position from inferior to superior. As a whole, their structure changes suggest that the Nartheciaceae flowers have evolved in close association with pollination and seed dispersal. By considering inferior ovaries and the presence of septal nectaries as plesiomorphies of Nartheciaceae, we discussed evolution of the ovary position and septal nectaries in all the monocots.

Embryology of Cardiopteris (Cardiopteridaceae, Aquifoliales), with emphasis on unusual ovule and seed development.

Cardiopteris (Cardiopteridaceae), a twining herb of two or three species distributed from Southeast Asia to Northern Australia, requires an embryological study for better understanding of its reproductive features. The present study of C. quinqueloba showed that the ovule and seed development involves a number of unusual structures, most of which are unknown elsewhere in angiosperms. The ovule pendant from the apical placenta is straight (not orthotropous), ategmic, and tenuinucellate, developing a monosporic seven-celled/eight-nucleate female gametophyte with an egg apparatus on the funicular side. Fertilization occurs by a pollen tube entering from the funicular side, resulting in a zygote on the funicular side. The endosperm is formed by the cell on the funicular side in the two endosperm cell stage. While retaining a (pro)embryo/endosperm as it is, the raphe (differentiating late in pre-fertilization stages) elongates toward the antiraphal side during post-fertilization stages, resulting in an anatropous seed. The two-cell-layered nucellar epidermis (belatedly forming by periclinal divisions), along with the raphe, envelops the embryo/endosperm entirely as the seed coat. The possibility was discussed that the arrested integument development triggers a series of the subsequent unusual structures of ovule and seed development. The fertilization mode in Cardiopteris underpins the hypothesis that the Polygonum‒type female gametophyte comprises two four-celled archegonia.

Embryology of Phyllonoma (Phyllonomaceae, Aquifoliales): characteristics and character evolution.

Phyllonoma, the sole genus of Phyllonomaceae (Aquifoliales) consisting of four Central American species, has not been well-characterized morphologically. Following a previous study of flower and inflorescence morphology, I here report the embryology of the genus based on P. tenuidens and compare its characteristics with those of other aquifolialean families, namely, Aquifoliaceae, Cardiopteridaceae, Helwingiaceae, and Stemonuraceae. Comparisons indicate that although Phyllonoma resembles all the other families embryologically, it more closely resembles Aquifoliaceae and Helwingiaceae in lacking a vascular bundle in its integument and bearing ab initio Cellular endosperm. The genus especially resembles Helwingiaceae by possessing a tenuinucellate ovule. This result corroborates molecular and floral morphological evidence, supporting the distinctness of Phyllonoma as a family and its sister-group relationship to East‒Asian Helwingiaceae. However, Phyllonoma is clearly distinguished from Helwingiaceae by seed coat structure. In Phyllonoma, the seeds (dispersed in berries) have a thick seed coat composed of irregularly enlarged, thick-walled exotestal cells, whereas the seeds (dispersed in drupes) have a thin membranous seed coat in Helwingiaceae. Taken together with earlier information on pollination (entomophily in Phyllonoma versus ambophily in Helwingiaceae), embryological evidence shows that distinct evolution has occurred in reproductive characters relating to pollination and seed dispersal in Phyllonoma.

Floral morphology and structure of Emblingia calceoliflora (Emblingiaceae, Brassicales): questions and answers.

Emblingia calceoliflora, the sole species of the family Emblingiaceae (Brassicales), is a creeping shrub endemic to South Western Australia. The flowers have a characteristic slipper-like corolla (calceolus). Earlier studies using dry specimens have left some questions regarding the flower unresolved. Here I present an anatomical study of fresh flowers to resolve these questions. The flowers are pedicellate, strongly monosymmetric, and pentamerous with the median sepal in the abaxial position. During flower development, a pedicel turns clockwise or anticlockwise, placing the adaxial calceolus (comprising both petals) downward and a transversely dilated androgynophore upward with a large tunnel-like space between them. Two short longitudinal walls develop from the basal part of the petals, enclosing a nectary gland deep in the flower. The vascular anatomy of the androgynophore shows that lateral dédoublement occurs in five stamens, resulting in two pairs of fertile stamens on the adaxial side and (three to) six staminodes as the "hood" on the opposite side. Androecial configuration is obhaplostemony, and the gynoecium is tricarpellate/trilocular. Comparisons with flowers of other Brassicales show that an extrastaminal nectary is a synapomorphy of the core Brassicales including Emblingiaceae. The flower of Emblingia is highly specialized for adaptation to insect-pollination.

Floral morphology and embryology of Helwingia (Helwingiaceae, Aquifoliales): systematic and evolutionary implications.

Helwingia, a shrub or small tree of four species distributed in East Asia, has been assigned to various families, mainly Cornaceae. However, molecular analyses show that the genus belongs to its own family Helwingiaceae which is sister to Phyllonomaceae (Phyllonoma only) in the order Aquifoliales. On the basis of H. japonica, we investigated the poorly understood floral and embryological characters of Helwingia, and compared their features with those of other Aquifoliales, particularly those of Phyllonomaceae. Results showed that perianth leaves of Helwingia represent sepals, because in plesiomorphic pentamerous flowers, they agreed in position with sepals (not with petals) in pentamerous flowers of Phyllonoma. Overall comparisons based on available information show that, while sharing with Phyllonoma the epiphyllous inflorescence, the inferior ovary, and an epigynous disc nectary as syapomorphies, Helwingia is characterized by loss of petals, obhaplostemony, large recurved stigmas, poorly developed disc nectary, tenuinucellate ovules with a mature female gametophyte filled with densely stained cytoplasm, and a thin mature seed coat. Morphological evidence, like molecular evidence, confirms that Helwingia is sufficiently distinct to be placed in its own family. Morphological and field observations suggest wind and insect pollination in H. japonica, which is the first example of ambophily in Aquifoliales.

Embryology of Biebersteinia (Biebersteiniaceae, Sapindales): characteristics and comparisons with related families.

Biebersteinia, a perennial herb of five species distributed from Greece to Central Asia, was long considered to be placed in, or near Geraniaceae. Recent molecular analyses, however, have shown that the genus is the sole member of the family Biebersteiniaceae in Sapindales (not including Geraniaceae). Here, we report the embryological features of Biebersteinia and provide embryological corroboration for the molecular sapindalean affinities of the genus. We compared its embryology with those of eight other families of Sapindales, as well as with those of the related orders Huerteales, Malvales, and Brassicales. Overall comparisons showed that Biebersteinia fits in Sapindales because of the presence of anther tapetal cells with polyploid nuclear mass and non-fibrous exotegmen. Further, the genus is characterized by three-celled pollen grains, tetrasporic 16-nucleate Penaea-type female gametophyte, unitegmic ovules, pseudoporogamy, and the chalaza shifting its position near the concave side in the post-fertilization stage. A considerable number of autapomorphies, combined with the lack of synapomorphies with other sapindalean families, supports placing Biebersteinia in its own family. Biebersteiniaceae appear to represent an early divergent lineage of Sapindales. Previous descriptions of seed coats, which were considered to have developed from "bitegmic" ovules, were revised.

Floral morphology and structure of Phyllonoma (Phyllonomaceae): systematic and evolutionary implications.

Phyllonoma, a small tree genus of four species distributed from Mexico to Peru, has been placed in various families (mainly in Saxifragaceae), but now, based on molecular evidence, is placed in a distinct family Phyllonomaceae in Aquifoliales. To better understand the morphological relationships of the genus and family, I studied its floral morphology, anatomy, and vasculature using P. tenuidens. Most of the external and internal floral characteristics were described more than 120 years ago. Although some of them were confirmed, some were substantially revised, mainly those concerning the gynoecial structure. Flowers are small and basically pentamerous, consisting of five sepals, five petals, five stamens, and a gynoecium composed of two carpels usually in transversal position. Comparisons with other Aquifoliales show that Phyllonomaceae share the inferior ovary, epiphyllous inflorescence and epigynous disc nectary with East-Asian Helwingiaceae (Helwingia only), but clearly differ from Helwingiaceae in having glandular trichomes on the sepal margins and a bicarpellate, unilocular gynoecium bearing many ovules on the parietal placentae. Evidence from floral morphology and structure supports the distinctness of Phyllonomaceae and its sister-group relationship with Helwingiaceae. Its floral characteristics suggest that Phyllonomaceae have evolved by adapting to distinct biological habitats in relation to pollination and seed dispersal.

Floral structure of Cardiopteris (Cardiopteridaceae) with special emphasis on the gynoecium: systematic and evolutionary implications.

Cardiopteris, a small herbaceous genus, had long been placed in its own family, Cardiopteridaceae. However, the family was recently broadly circumscribed to include more genera in Aquifoliales. To better understand the morphological relationships of the genus and the family, I studied the floral anatomy and development of Cardiopteris using C. quinqueloba. As has been previously described, flowers are 5-merous with a unilocular gynoecium. I confirmed that the gynoecium is bicarpellate, possessing two dissimilar styles. An analysis of the development, structure, and vasculature of the gynoecium showed that it is pseudomonomerous, consisting of one fertile adaxial carpel and one solid sterile abaxial carpel. The adaxial carpel forms a thin style with a capitate stigma, whereas the abaxial carpel has a thick style, which develops into a freshy fruit appendage. Comparisons with flowers of other genera (Citronella, Gonocaryum, and Leptaulus) of Cardiopteridaceae as well as the other families (Aquifoliaceae, Helwingiaceae, Phyllonomaceae, and Stemonuraceae) of Aquifoliales showed that the pseudomonomerous gynoecium is very likely a synapomorphy to support the sister-group relationship between Cardiopteridaceae and Stemonuraceae. Moreover, contrary to all previous descriptions, Cardiopteris flowers were found to have an annular nectariferous disk at the base of the gynoecium.

Ecological barriers to gene flow between riparian and forest species of Ainsliaea (Asteraceae).

Understanding the role of habitat-associated adaptation in reducing gene flow resulting in population differentiation and speciation is a major issue in evolutionary biology. We demonstrate a significant role for habitat divergence in species isolation between two naturally hybridizing riparian and nonriparian plants, Ainsliaea faurieana and A. apiculata (Asteraceae), on Yakushima Island, Japan. By analyzing the fine-scale population structure at six sympatric sites, we found that variations in leaf shape, geography, light conditions, and genotype were strongly correlated across riverbank-forest transitions. No evidence of effective gene flow was found between the two species across the majority of the transition zones, although the NewHybrid clustering analysis confirmed interspecific hybridization. However, a relatively high level of gene flow was observed across one zone with a more diffuse ecotone and intermediate flooding and light conditions, possibly generated by human disturbances. These results suggest that the barriers to gene flow between the riparian and forest species are primarily ecological. Additional common garden experiments indicated that the two species are adaptively differentiated to contrasting flooding and light environments. Overall, our study suggests that adaptations to different habitats can lead to the formation of reproductive isolating barriers and the maintenance of distinct species boundaries.

Molecular phylogenetic analyses of Tofieldiaceae (Alismatales): family circumscription and intergeneric relationships.

Tofieldiaceae are a small monocot family comprising about 20 species, mostly distributed in the Northern Hemisphere, with some in northern South America. To clarify the family circumscription, the number of distinguishable genera in the family, and relationships among the genera, we conducted molecular analyses of cpDNA (matK and non-coding trnL-trnL-trnF region) sequences of 17 associated species of Tofieldiaceae, along with 14 species of Acorales, Alismatales, Dioscoreales, Pandanales, and Liliales. Results showed that Tofieldiaceae are monophyletic, comprising all the species assignable to Harperocallis, Isidrogalvia, Pleea, Tofieldia, and Triantha, thus supporting the original family circumscription. Within the family, Pleea is sister to the rest of the family, in which Isidrogalvia is sister to Harperocallis, and Tofieldia to Triantha. Morphological characters supporting the relationships among the genera were briefly discussed.

Embryology of the Irvingiaceae, a family with uncertain relationships among the Malpighiales.

The Irvingiaceae, one of 40 families of the Malpighiales, comprise a small woody family of 10 species in three genera distributed in Old World tropics. Its relationships with other families are unclear, although recent molecular analyses suggest affinities with Linaceae, Caryocaraceae, Erythroxylaceae, and Rhizophoraceae. To gain insight into family relationships, we investigated 63 embryological characters of two previously unstudied African species, Irvingia gabonensis and I. smithii, and compared them with other Malpighiales and the sister group Oxalidales. Embryologically, Irvingia is characterized by the absence of an integumentary tapetum and by having a non-multiplicative inner integument, a multiplicative testa, many discrete fascicles of vascular bundles running in the testa from the raphe to antiraphe (each fascicle comprised several strands arranged in a concentric manner), and a fibrous exotegmen. Comparisons showed that Irvingia did not resemble any of the Linaceae, Caryocaraceae, Erythroxylaceae, Rhizophoraceae, or any of the other malpighialean families for which embryological data are available. The genus rather resembled Huaceae and Connaraceae (Oxalidales) in seed coat structure. However, 18 families (45%) of the Malpighiales are still poorly understood embryologically, and therefore additional studies are required for further critical comparisons.

Endosperm development in the Araceae (Alismatales) and evolution of developmental modes in monocots.

The Araceae, a basal-most family of Alismatales that basally diverged subsequent to Acorales in monocot phylogeny, are known to have diverse modes of endosperm development: nuclear, helobial, and cellular. However, the occurrence of nuclear and helobial endosperm development has long been debated. Here, we report a (re-)investigation of endosperm development in Lysichiton, Orontium, and Symplocarpus of the Orontioideae (a basal Araceae), in which nuclear endosperm development was recorded more than 100 years ago. The results show that all three genera exhibit a cellular, rather than nuclear, endosperm development and suggest that the helobial endosperm development reported as an "unmistakable record" from Ariopsis is likely cellular. Thus the Araceae are very likely characterized by cellular endosperm development alone. An extensive comparison with other monocots in light of phylogenetic relationships demonstrates that a plesiomorphic cellular endosperm development is restricted to the three basal monocot orders Acorales, Alismatales, and Petrosaviales, in which evolutionary changes from cellular to nuclear endosperm development occurred twice as major events, once within Alismatales and once as a synapomorphy of the eight remaining monocot orders, including Dioscoreales, Liliales, Asparagales, and Poales, and that helobial endosperm development, which is known for many monocot families, evolved as homoplasy throughout the monocots.

Embryology of Petrosavia (Petrosaviaceae, Petrosaviales): evidence for the distinctness of the family from other monocots.

The affinities of Petrosavia, a rare, leafless, mycoheterotrophic genus composed of two species indigenous to East to Southeast Asia, have long been uncertain. However, recent molecular analyses show that the genus is sister to Japonolirion osense. Japonolirion and Petrosavia comprise the Petrosaviaceae, which are now placed in its own order, Petrosaviales, distinct from other monocots based on molecular analyses. We conducted an embryological study of Petrosavia, comparing it to Japonolirion, as well as to basal monocots (Acorus and Araceae) and more derived monocots (Nartheciaceae, Velloziaceae, and Triuridaceae). Our results showed that Petrosavia is very similar in embryology to Japonolirion, with both genera sharing a glandular anther tapetum, simultaneous cytokinesis in microspore mother cells, anatropous and crassinucellate ovules, T-shaped tetrads of megaspores, ab initio Cellular-type endosperm, and a mature seed coat composed of the exotesta, endotesta, and endotegmen. The two genera of Petrosaviaceae are clearly distinct from Acorus, and all Araceae, Nartheciaceae, Velloziaceae, and Triuridaceae genera in various combinations of characters. Thus, both molecular and embryological evidence support the distinctness of the Petrosaviaceae from other monocots and its placement in its own order, Petrosaviales.

Syncyte formation in the microsporangium of Chrysanthemum (Asteraceae): a pathway to infraspecific polyploidy.

Polyploidy, which is thought to have played an important role in plant evolution and speciation, is prevalent in Chrysanthemum (x = 9). In fact, polyploid series are known in C. zawadskii (2x, 4x, 6x, 8x, and 10x) and C. indicum (2x, 4x, and 6x), but the mechanism by which polyploidization occurs is unknown. Here we show that in diploid individuals of both C. zawadskii and C. indicum, the fusion between two adjacent pollen mother cells (PMCs) occurs at a frequency of 1.1-1.3% early in the first meiotic division. While possessing the chromosomes of both PMCs, the fused cell or syncyte undertakes subsequent meiotic division processes as a single large PMC, producing four 2n pollen grains that are able to germinate. Despite their low frequency, syncyte formation may have played a major role in the production of infraspecific polyploids in Chrysanthemum.

Embryology of Japonolirion (Petrosaviaceae, Petrosaviales): a comparison with other monocots.

Japonolirion osense, the sole species of the genus, endemic to Japan, which is placed together with Petrosavia in the Petrosaviaceae and the order Petrosaviales, is still poorly known with respect to systematic characters. Here I present an embryological study of the anther, ovule, and seed of J. osense. Japonolirion is characterized by a glandular anther tapetum, simultaneous cytokinesis in the microspore mother cell, two-celled mature pollen grains, anatropous and crassinucellate ovules, a two-cell-layered nucellar cap formed early in ovule development, antipodal cells hypertrophied in post-fertilization stages, the ab initio cellular mode of endosperm formation, and exotegmic seeds. Comparisons with the basal monocots Acorus (Acorales) and Araceae (Alismatales), and with the more derived monocots Nartheciaceae (Dioscoreales) and Velloziaceae/Triuridaceae (Pandanales), showed that Japonolirion is clearly distinct from those basal and more derived monocots, supporting a distinct position for Petrosaviaceae or Petrosaviales within the monocots. Extensive comparisons further suggest that the two-cell-layered nucellar cap, whose cells are rich in cytoplasm at the time of fertilization in Japonolirion and thus obviously function as the obturator, is likely to be a common characteristic of the basal monocots and may even be a link with the magnoliids.

Embryology of Koeberlinia (Koeberliniaceae): Evidence for core-Brassicalean affinities.

Koeberlinia, comprising a single xerophytic species K. spinosa, had previously been placed in various families, mainly Capparaceae. Current molecular evidence now places it in its own family Koeberliniaceae, thought to be related to the Bataceae/Salvadoraceae among the 17 other families of the Brassicales. We investigated 55 embryological characters of the genus, most of which are not understood yet, and thereby assessed its systematic relationships. Koeberlinia has many embryological features in common with the Capparaceae and seven other core-Brassicalean families (i.e., Brassicaceae, Cleomaceae, Emblingiaceae, Gyrostemonaceae, Pentadiplandraceae, Resedaceae, and Tovariaceae), specifically by possessing a campylotropous ovule with a nonmultiplicative (two-cell-layered) outer integument, reniform seeds with a curved embryo, and a fibrous exotegmen in the mature seed coat. However, Koeberlinia is clearly distinguished from them by a tenuinucellate rather than crassinucellate ovule as previously reported, markedly enlarged apical nucellar epidermal cells, and an "exotestal" seed coat. Embryologically, Koeberlinia resembles neither the Bataceae nor the Salvadoraceae, although only limited embryological data are available for these two families. Embryological evidence thus favors its joining the core Brassicales, but additional molecular analyses and embryological studies on the missing data of the Bataceae and Salvadoraceae are needed for final confirmation of its phylogenetic position.

Development and structure of the female gametophyte in Austrobaileya scandens (Austrobaileyaceae).

Austrobaileyales, comprising the four families Austrobaileyaceae, Trimeniaceae, Schisandraceae, and Illiciaceae, are included in the basal angiosperms along with Amborellaceae and Nymphaeaceae. Here, we present the first developmental study of the female gametophyte in Austrobaileya scandens, the only species of Austrobaileyaceae, which are sister to the rest of the Austrobaileyales. Austrobaileya scandens has a four-celled/four-nucleate embryo sac as in the derived families of the order, e.g., Illiciaceae and Schisandraceae. It is monosporic, with the chalazal megaspore of a tetrad developing into the embryo sac composed of an egg cell, two synergids, and one polar nucleus. This mode of embryo sac formation was first reported in Schisandra over 40 years ago and should now be established as the Schisandra type. Its occurrence in A. scandens shows that the Schisandra-type embryo sac is likely common to the whole Austrobaileyales as well as to Nymphaeaceae. Amborellaceae were recently reported to have an eight-celled/nine-nucleate embryo sac, clarifying that none of the basal angiosperms has the seven-celled/eight-nucleate Polygonum-type embryo sac found in the majority of angiosperms, and that the Polygonum-type embryo sac represents a derived character state in angiosperms.

Is Cycas revoluta (Cycadaceae) wind- or insect-pollinated?

Among the Cycadales (Cycadaceae and Zamiaceae), the Zamiaceae are known to be insect-pollinated. In contrast, the Cycadaceae are still considered wind-pollinated, although some doubt has been cast on several species, including Cycas revoluta. Using a large population of C. revoluta on Yonaguni Island (Okinawa, Japan), we performed exclusion experiments, documented insects from male and female cones, and analyzed the morphology of the apical part of the ovule to determine the pollination method of this species. Insect exclusion resulted in a notable reduction in seed set, except in a few individuals growing near male cones. The amount of airborne pollen was abundant within a 2-m radius of male cones but decreased markedly beyond this distance. Pollen grains of C. revoluta were found on the body of Carpophilus chalybeus (Nitidulidae, Coleoptera), one of a few species of insects collected from both male cones and female cones far from males. We conclude that C. revoluta relies on both wind (anemophily) and insect pollination (entomophily), although such anemophily is restricted to female trees growing within a 2-m radius of male trees. The nitidulids are not host specific to this cycad and primarily feed on plant tissue but serve as pollinators during pollen release. Cycas revoluta appears to be in an initial mode of animal pollination, as opposed to the host-specific insect pollination observed in most Zamiaceae.

Phylogenetic analyses of Malpighiales using plastid and nuclear DNA sequences, with particular reference to the embryology of Euphorbiaceae sens. str.

We present phylogenetic analyses of Malpighiales, which are poorly understood with respect to relationships within the order, using sequences from rbcL, atpB, matK and 18SrDNA from 103 genera in 23 families. From several independent and variously combined analyses, a four-gene analysis using all sequence data provided the best resolution, resulting in the single most parsimonious tree. In the Malpighiales [bootstrap support (BS) 100%], more than eight major clades comprising a family or group of families successively diverged, but no clade containing more than six families received over 50% BS. Instead, ten terminal clades that supported close relationships between and among families (>50% BS) were obtained, between, for example, Balanopaceae and Chrysobalanaceae; Lacistemataceae and Salicaceae; and Phyllanthaceae and Picrodendraceae. The monophyly of Euphorbiaceae sens. str. were strongly supported (BS 100%), but its sister group was unclear. Euphorbiaceae sens. str. comprised two basally diverging clades (BS 100%): one leading to the Clutia group (Chaetocarpus, Clutia, Pera and Trigonopleura), and the other leading to the rest of the family. The latter shared a palisadal, instead of a tracheoidal exotegmen as a morphological synapomorphy. While both Acalyphoideae (excluding Dicoelia and the Clutia group) and Euphorbioideae are monophyletic, Crotonoideae were paraphyletic, requiring more comprehensive analyses.

Chromosome evolution in the Laurales based on analyses of original and published data.

We present a summary of currently available chromosome information for all seven families in the order Laurales on the basis of original and previously published data and discuss the evolution of chromosomes in this order. Based on a total of 53 genera for which chromosome data were available, basic chromosome numbers appear consistent within families: x = 11 (Calycanthaceae); x = 22 (Atherospermataceae and Siparunaceae); x = 19 (Monimiaceae); and x = 12 and 15 (Lauraceae). The Hernandiaceae have diverse numbers: x = 15 (Gyrocarpoideae) and x = 18 and 20 (Hernandioideae). Karyotype analyses showed that Hennecartia, Kibaropsis, and Matthaea (all Monimiaceae) contained two or three sets of four distinct chromosomes in 38 somatic chromosomes, suggesting that 2n = 38 was derived by aneuploid reduction from 2n = 40, a tetraploid of x = 10. In light of the overall framework of phylogenetic relationships in the Laurales, we show that x = 11 is an archaic base number in the order and is retained in the Calycanthaceae, which are sister to the remainder of the order. Polyploidization appears to have occurred from x = 11 to x = 22 in a common clade of the Siparunaceae, Atherospermataceae, and Gomortegaceae (although 2n = 42 in the Gomortegaceae), and aneuploid reduction from x = 11 to x = 10 occurred in a common clade of the Hernandiaceae, Lauraceae, and Monimiaceae. To understand chromosome evolution in the Lauraceae, however, more studies are needed of genera and species of Cryptocaryeae.