Palynology of the perigoniate Aroideae: Zamioculcas,Gonatopus and Stylochaeton (Araceae)

The pollen of the perigoniate Aroideae sensu Mayo et al. (1997) ( Zamioculcas Schott, Gonatopus Hook. f. ex Engl. and Stylochaeton Lepr.) differs ultrastructurally from that of the aperigoniate Aroideae in several important exine and aperture characters. The almost identical zona-aperturate pollen of Zamioculcas and Gonatopus has outside the aperture an elaborated, thick ectexine, while the aperture consists of a thin, but continuous ectexine and a thick, lamellate endexine. In contrast, the omniaperturate pollen of Stylochaeton has a thin, not clearly stratified ectexine and a thin, heterogeneous endexine below. However, the zona-aperturate pollen of Zamioculcas and Gonatopus deviates significantly from the superficially similar zona-aperturate pollen of the unrelated Monstereae (e. g., Monstera Adans., Amydrium Schott): in the apertures of Monstera or Amydrium both the thin, but continuous ectexine and the lamellate endexine, which are typical features for Zamioculcas and Gonatopus , are absent. The palynological data underline not only the present classification of Zamioculcas , Gonatopus and of Stylochaeton into two tribes (Zamioculcadeae and Stylochaetoneae) and the differences of both tribes from the other Aroideae, but show also significant deviations in the respective zona-aperturate condition in Monstereae (Monsteroideae) and Zamioculcadeae (Aroideae).     

Evolutionary dynamics of chloroplast genomes in subfamily Aroideae (Araceae)

Aroideae is the largest and most diverse subfamily of the plant family Araceae. Despite its agricultural and horticultural importance, the genomic resources are sparse for this subfamily. Here, we report de novo assembled and fully annotated chloroplast genomes of 13 Aroideae species. The quadripartite chloroplast genomes (size range of 158,177–170,037 bp) are comprised of a large single copy (LSC; 75,594–94,702 bp), a small single copy (SSC; 12,903–23,981 bp) and a pair of inverted repeats (IRs; 25,266–34,840 bp). Notable gene rearrangements and IRs contraction / expansions were found for Anchomanes hookeri and Zantedeschia aethiopica. Codon usage, amino acid frequencies, oligonucleotide repeats, GC contents, and gene features revealed similarities among the 13 species. The number of oligonucleotide repeats was uncorrelated with genome size or phylogenetic position of the species. Phylogenetic analyses corroborated the monophyly of Aroideae but were unable to resolve the positions of Calla and Schismatoglottis.     

Schottarum (Schismatoglottideae: Araceae) substantiated based on combined nuclear and plastid DNA sequences

Recent studies on Schismatoglottideae have resulted in the recognition of four new monophyletic genera, the resurrection of two additional genera following clarification of their monophyly, and the publication of many taxonomically novel species. However, generic boundaries among some parts of Schismatoglottideae remain unclear owing to several reasons: (1) more taxa are being revealed through our on-going fieldwork, now expanded to previously unsampled localities on Borneo; (2) established occurrence of a high level of homoplasies among the morphological characteristics hitherto used to delimitate genera; and (3) gene regions used in previous studies contradicted some of current taxonomic placements. Among the unsolved groups from previous studies a clade comprising Schismatoglottis sarikeensis and S. josefii needs further investigation. Therefore, phylogenetic analyses were carried out to investigate the position of these two species using the nuclear region, internal transcribed spacer and combined plastid regions: trnL intron and trnL-F intergenic spacer, coding matK+partial 3′ trnK, intergenic spacer trnH-psbA. A total of 23 accessions representing 16 taxa of Schismatoglottideae and Philonotieae were included in the study. Phylogenetic analyses of a total 4,658 bp combined dataset using parsimony, maximum likelihood, and Bayesian methods revealed that S. sarikeensis and S. josefii do not belong to Schismatoglottis, and therefore are transferred to Schottarum (≡Hottarum sarikeense ≡ Schismatoglottis sarikeense). Flowering mechanism, pollination strategy, and fruitset of S. sarikeense are also presented in the paper.     

Patterns and causes of incongruence between plastid and nuclear Senecioneae (Asteraceae) phylogenies

One of the longstanding questions in phylogenetic systematics is how to address incongruence among phylogenies obtained from multiple markers and how to determine the causes. This study presents a detailed analysis of incongruent patterns between plastid and ITS/ETS phylogenies of Tribe Senecioneae (Asteraceae). This approach revealed widespread and strongly supported incongruence, which complicates conclusions about evolutionary relationships at all taxonomic levels. The patterns of incongruence that were resolved suggest that incomplete lineage sorting (ILS) and/or ancient hybridization are the most likely explanations. These phenomena are, however, extremely difficult to distinguish because they may result in similar phylogenetic patterns. We present a novel approach to evaluate whether ILS can be excluded as an explanation for incongruent patterns. This coalescence-based method uses molecular dating estimates of the duration of the putative ILS events to determine if invoking ILS as an explanation for incongruence would require unrealistically high effective population sizes. For four of the incongruent patterns identified within the Senecioneae, this approach indicates that ILS cannot be invoked to explain the observed incongruence. Alternatively, these patterns are more realistically explained by ancient hybridization events.     

Reasons and consequences of the lack of a sporopollenin ektexine in Aroideae (Araceae)

The ultrastructure of pollen walls in Araceae is characterized by the absence of a stable sporopollenin outer exine layer in subfamily Aroideae, and by the presence of several distinctive pollen characters typical for the other aroid subfamilies. This article discusses if and to which extent such distinctive pollen characters are mirrored in various classifications of Araceae, basing either on morphological or on molecular data. Accordingly, the pollen characters perfectly reflect the actual subfamily classification, and also recent arrangements of clades in trees basing on molecular data. The actual subfamilies appear no longer eurypalynous, but now strictly stenopalynous. Aside from the (settled) classification problem the fundamental question is addressed why do Aroideae lack an elaborated sporopollenin ektexine. Possible pollination biology benefits, deriving from an absence of an elaborated sporopollenin ektexine in Aroideae, are presented and discussed. Compared with all other subfamilies the most advanced and by far largest subfamily Aroideae has lost several crucial characters and simultaneously acquired corresponding opposed characters, amongst others a non-sporopollenin exine layer and an unusual thick and spongy endexine. Taken together, losses and acquisitions are interpreted as a major paradigm shift in Araceae evolution, which took place according to the fossil record probably in the Paleogene.     

Complex evolution in Arundinarieae (Poaceae: Bambusoideae): incongruence between plastid and nuclear GBSSI gene phylogenies

The monophyly of tribe Arundinarieae (the temperate woody bamboos) has been unequivocally recovered in previous molecular phylogenetic studies. In a recent phylogenetic study, 10 major lineages in Arundinarieae were resolved based on eight non-coding plastid regions, which conflicted significantly with morphological classifications both at the subtribal and generic levels. Nevertheless, relationships among and within the 10 lineages remain unclear. In order to further unravel the evolutionary history of Arundinarieae, we used the nuclear GBSSI gene sequences along with those of eight plastid regions for phylogenetic reconstruction, with an emphasis on Chinese species. The results of the plastid analyses agreed with previous studies, whereas 13 primary clades revealed in the GBSSI phylogeny were better resolved at the generic level than the plastid phylogeny. Our analyses also revealed many inconsistencies between the plastid DNA and the nuclear GBSSI trees. These results implied that the nuclear genome and the plastid genome had different evolutionary trajectories. The patterns of incongruence suggested that lack of informative characters, incomplete lineage sorting, and/or hybridization (introgression) could be the causes. Seven putative hybrid species were hypothesized, four of which are discussed in detail on the basis of topological incongruence, chromosome numbers, morphology, and distribution patterns, and those taxa probably resulted from homoploid hybrid speciation. Overall, our study indicates that the tribe Arundinarieae has undergone a complex evolution.     

Allotetraploid origin and divergence in Eleusine (Chloridoideae, Poaceae): evidence from low-copy nuclear gene phylogenies and a plastid gene chronogram

Background and Aims Eleusine

(Poaceae) is a small genus of the subfamily Chloridoideae exhibiting considerable morphological and ecological diversity in East Africa and the Americas. The interspecific phylogenetic relationships of Eleusine are investigated in order to identify its allotetraploid origin, and a chronogram is estimated to infer temporal relationships between palaeoenvironment changes and divergence of Eleusine in East Africa.

Methods

Two low-copy nuclear (LCN) markers, Pepc4 and EF-1α, were analysed using parsimony, likelihood and Bayesian approaches. A chronogram of Eleusine was inferred from a combined data set of six plastid DNA markers (ndhA intron, ndhF, rps16-trnK, rps16 intron, rps3, and rpl32-trnL) using the Bayesian dating method.

Key Results

The monophyly of Eleusine is strongly supported by sequence data from two LCN markers. In the cpDNA phylogeny, three tetraploid species (E. africana, E. coracana and E. kigeziensis) share a common ancestor with the E. indica–E. tristachya clade, which is considered a source of maternal parents for allotetraploids. Two homoeologous loci are isolated from three tetraploid species in the Pepc4 phylogeny, and the maternal parents receive further support. The A-type EF-1α sequences possess three characters, i.e. a large number of variations of intron 2; clade E-A distantly diverged from clade E-B and other diploid species; and seven deletions in intron 2, implying a possible derivation through a gene duplication event. The crown age of Eleusine and the allotetraploid lineage are 3·89 million years ago (mya) and 1·40 mya, respectively.

Conclusions

The molecular data support independent allotetraploid origins for E. kigeziensis and the E. africana–E. coracana clade. Both events may have involved diploids E. indica and E. tristachya as the maternal parents, but the paternal parents remain unidentified. The habitat-specific hypothesis is proposed to explain the divergence of Eleusine and its allotetraploid lineage.     

Incongruent plastid and nuclear DNA phylogenies reveal ancient intergeneric hybridization in Pilosella hawkweeds (Hieracium, Cichorieae, Asteraceae)

Phylogenetic relationships for Hieracium subgen. Pilosella were inferred from chloroplast (trnT-trnL, matK) and nuclear (ITS) sequence data. Chloroplast markers revealed the existence of two divergent haplotype groups within the subgenus that did not correspond to presumed relationships. Furthermore, chloroplast haplotypes of the genera Hispidella and Andryala nested each within one of these groups. In contrast, ITS data were generally in accord with morphology and other evidence and were therefore assumed to reflect the true phylogeny. They revealed a sister relationship between Pilosella and Hispidella and a joint clade of Hieracium subgenera Hieracium and Chionoracium (Stenotheca) while genus Andryala represented a third major lineage of the final ingroup cluster. Detailed analysis of trnT-trnL character state evolution along the ITS tree suggested two intergeneric hybridization events between ancestral lineages that resulted in cytoplasmic transfer (from Hieracium/Chionoracium to Pilosella, and from the introgressed Pilosella lineage to Andryala). These chloroplast capture events, the first of which involved a now extinct haplotype, are the most likely explanation for the observed incongruencies between plastid and nuclear DNA markers.     

Close relationship between Asterina and Solasteridae (Asteroidea) supported by both nuclear and mitochondrial gene molecular phylogenies

Phylogenetic relationships among asteroids remain to be extremely controversial in spite of many morphological and molecular studies have been applied to this issue. In the present study, especially focusing on resolving the relationship of Asterina and Solasteridae, we reconstructed the molecular phylogenetic tree of asteroids using nuclear 18S rDNA. A close relationship between Asterina and Solasteridae, which has been suggested from analyses of mitochondrial 12S rDNA and 16S rDNA, is supported here by the nuclear 18S rDNA dataset. The support is even stronger when the sequences of mitochondrial rDNAs and nuclear 18S rDNA are combined as a total dataset. The independent support from both nuclear 18S rDNA and mitochondrial rDNAs strongly argues for a close relationship between the Asterina and Solasteridae.     

The correlation between development of atypical bisexual flowers and phylogeny in the Aroideae (Araceae)

 In the intermediate zone of the inflorescence of genera of Aroideae one can find flowers with male and female characteristics. Until now, two types of developmental sequences of atypical bisexual flowers (ABFs) have been recognized: the Philodendron type and the Cercestis type. In the Philodendron type, bisexual flowers generally consist of functional carpels and staminodes inserted on the same whorl. In the Cercestis type, the gynoecium and stamens are inserted on two different whorls. These different ontogenetic patterns represent two different pathways in the evolution of unisexual flowers in this subfamily. A molecular phylogenetic analysis of 33 genera of Araceae, based on the chloroplast trnL intron and trnL–F intergenic spacer sequences was carried out. We use this phylogenetic analysis and those published by French et al. (1995) and Mayo et al. (1997) to examine the distribution of the two types of ABFs in selected genera. Our results suggest that the two developmental patterns of ABFs in Aroideae sensu Mayo et al. (1997) do not correspond to two separate evolutionary lineages but rather are more or less consistent within clades. Although this new molecular phylogeny does not include all aroid genera, it corroborates in general, at the subfamily level, the molecular analysis of French et al. (1995) based on chloroplast DNA restriction site data and the analysis of Mayo et al. (1997) based on morphological and anatomical data. Received March 15, 2001 Accepted October 11, 2001     

SEM study of surface structures of the spathe in Cryptocoryne and Lagenandra (Araceae: Aroideae: Cryptocoryneae)

SEM studies of the spathe structures in the two closely related genera Cryptocoryne and Lagenandra show differences between the inner and outer surfaces, as well as in cell structures in the various parts of the spathe. The cell structure reveals patterns mat makes it possible to depict homologous structures of the spathe, even though the spathes of the two genera look different. The basal part of the kettle has a mucilage covering of the cells, interpreted as a hitherto unnoticed food source. The cells of the inner surface of the kettle and tube have downward pointing trichomes. On the second day of flowering these collapse and sink into the cell lumen, which is suggested to create a unique lattice-like structure that enables the insects to climb out of the kettle and tube. The cell structure of the flap shows that it is a prolongation and continuation of the spathe tube margin.     

Parallel evolution in eight-barbel loaches of the genus Lefua (Balitoridae, Cypriniformes) revealed by mitochondrial and nuclear DNA phylogenies

The evolutionary history of eight-barbel loaches of the genus Lefua contains important phylogenetic information that will aid in resolution of the faunal formations and evolutionary histories of Japanese and East Asian freshwater fishes. Our sequencing of the mitochondrial D-loop region in a large number of samples allowed construction of the most comprehensive phylogeny of these loaches to date; we demonstrated monophyly of five Lefua species and identified populations of Lufua. sp. and Lefua echigonia. Loaches inhabiting the Tokai region in Japan were morphologically and ecologically indistinguishable from Lefua sp. However, they were included in the L. echigonia lineage. We determined a novel phylogeny by sequencing the nuclear ribosomal S7 subunit and showed that nuclear DNA phylogeny essentially matched the mitochondrial DNA phylogeny. Loaches from the Tokai region were part of the L. echigonia lineage, indicating parallel evolution between Tokai loaches and Lefua sp. in western Japan. We presented the most robust phylogeny to date using concatenated mitochondrial and nuclear sequences. The wealth of molecular information allowed us to speculate on evolutionary processes in the genus Lefua.     

A new genus of Entiminae from North Africa,supported by a phylogenetic analysis (Coleoptera: Curculionidae: Entiminae)

The genus Holcophloeus gen. nov. is here proposed to include Trachyphloeus cruciatus Seidlitz, 1868, and two new species native to North Africa, based on a phylogenetic analysis and an evaluation of the diagnostic characters. The taxonomic position of Holcophloeus in relation to the tribes Trachyphloeini Lacordaire, 1863, and Holcorhinini Desbrochers, 1898, is discussed, and the new genus is attributed to the Holcorhinini. Holcophloeus laurae sp. nov. from south‐eastern Morocco and Holcophloeus weilli sp. nov. from northern Libya are described and illustrated and a key to the species of the new genus is given. The lectotype of Trachyphloeus cruciatus Seidlitz, 1868, is designated. The genus Massimiellus Borovec, 2009, is transferred from Trachyphloeini to Holcorhinini. © 2013 The Linnean Society of London     

Hybridization and polyploidy of an aquatic plant, Ruppia (Ruppiaceae), inferred from plastid and nuclear DNA phylogenies

? Premise of the study: The monogeneric family Ruppiaceae is found primarily in brackish water and is widely distributed on all continents, many islands, and from subartic to tropical zones. Ruppia taxonomy has been confusing because of its simplified morphology yet high phenotypic plasticity and the existence of polyploidy and putative hybrids. This study addresses the current classification of species in the genus, the origin of putative hybrids and polyploids, and the distribution of Ruppia species. ? Methods: Separate molecular phylogenetic analyses using plastid DNA and nuclear-encoded PHYB data sets were performed after chromosome observations. ? Key results: The resultant trees were largely congruent between genomes, but were incongruent in two respects: the first incongruence may be caused by long outgroup branches and their effect on ingroup rooting, and the second is caused by the existence of heterogeneous PHYB sequences for several accessions that may reflect several independent hybridization events. Several morphological species recognized in previous taxonomic revisions appear paraphyletic in plastid DNA and PHYB trees. ? Conclusions: Given the molecular phylogenies, and considering chromosome number and morphology, three species and one species complex comprising six lineages were discerned. A putative allotriploid, an allotetraploid, and a lineage of hybrid origin were identified within the species complex, and a hybrid was found outside the species complex, and their respective putative parental taxa were inferred. With respect to biogeography, a remarkably discontinuous distribution was identified in two cases, for which bird-mediated seed dispersal may be a reasonable explanation.     

Parallel evolution of glucosinolate biosynthesis inferred from congruent nuclear and plastid gene phylogenies

The phytochemical system of mustard-oil glucosides (glucosinolates) accompanied by the hydrolytic enzyme myrosinase (beta-thioglucosidase), the latter usually compartmented in special myrosin cells, characterizes plants in 16 families of angiosperms. Traditional classifications place these taxa in many separate orders and thus imply multiple convergences in the origin of this chemical defense system. DNA sequencing of the chloroplast rbcL gene for representatives of all 16 families and several putative relatives, with phylogenetic analyses by parsimony and maximum likelihood methods, demonstrated instead a single major clade of mustard-oil plants and one phylogenetic outlier. In a further independent test, DNA sequencing of the nuclear 18S ribosomal RNA gene for all these exemplars has yielded the same result, a major mustard-oil clade of 15 families (Akaniaceae, Bataceae, Brassicaceae, Bretschneideraceae, Capparaceae, Caricaceae, Gyrostemonaceae, Koeberliniaceae, Limnanthaceae, Moringaceae, Pentadiplandraceae, Resedaceae, Salvadoraceae, Tovariaceae, and Tropaeolaceae) and one outlier, the genus Drypetes, traditionally placed in Euphorbiaceae. Concatenating the two gene sequences (for a total of 3254 nucleotides) in a data set for 33 taxa, we obtain robust support for this finding of parallel origins of glucosinolate biosynthesis. From likely cyanogenic ancestors, the "mustard oil bomb" was invented twice.     

Pollination mutualism between a new species of the genus Colocasiomyia de Meijere (Diptera: Drosophilidae) and Steudnera colocasiifolia (Araceae) in Yunnan, China

A new species of the genus Colocasiomyia de Meijere (Diptera: Drosophilidae) was discovered from inflorescences of Steudnera colocasiifolia K. Koch (Araceae) in Yunnan, China. The new species is described as Colocasiomyia steudnerae Takenaka and Toda, sp. nov., and we investigated the reproductive ecology of both the fly and the plant species. This fly species reproduces in the inflorescences/infructescences of the plant, and depends almost throughout its entire life cycle on the host plant. The fly species is the most abundant flower visitor for S. colocasiifolia and behaves intimately with the flowering events, suggesting that it is the unique and most efficient pollinator for the host plant. Bagging (insect‐exclusion) treatment of inflorescences resulted in no fruits. These findings strongly suggest that intimate pollination mutualism has evolved between the fly and the host plant, as are known in other Colocasiomyia flies and Araceae plants. One notable feature of this system is that the new species almost monopolizes the host‐plant inflorescence as a visitor, without any cohabiting Colocasiomyia species. In comparison to other cases where two Colocasiomyia species share the same inflorescence and infructescence of Araceae host plants for reproduction by separating their breeding niches microallopatrically between the staminate (upper male‐flower) and the pistillate (lower female‐flower) regions on the spadix, C. steudnerae exhibits a mixture of stamenicolous and pistillicolous breeding habits.