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Host-to-parasite gene transfer in flowering plants: phylogenetic evidence from Malpighiales.

Horizontal gene transfer (HGT) between sexually unrelated species has recently been documented for higher plants, but mechanistic explanations for HGTs have remained speculative. We show that a parasitic relationship may facilitate HGT between flowering plants. The endophytic parasites Rafflesiaceae are placed in the diverse order Malpighiales. Our multigene phylogenetic analyses of Malpighiales show that mitochondrial (matR) and nuclear loci (18S ribosomal DNA and PHYC) place Rafflesiaceae in Malpighiales, perhaps near Ochnaceae/Clusiaceae. Mitochondrial nad1B-C, however, groups them within Vitaceae, near their obligate host Tetrastigma. These discordant phylogenetic hypotheses strongly suggest that part of the mitochondrial genome in Rafflesiaceae was acquired via HGT from their hosts.

Cell Nucleus↗

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.

DNA, Plant↗

The genome sequence of Euphorbia lathyris L., 1753 (Malpighiales: Euphorbiaceae).

We present a genome assembly of Euphorbia lathyris (caper spurge; Streptophyta; Magnoliopsida; Malpighiales; Euphorbiaceae). The genome sequence has a total length of 1 020.39 megabases. Most of the assembly (99.88%) is scaffolded into 10 chromosomal pseudomolecules. The mitochondrial sequences have lengths of 886.36, 41.02, 86.31 and 32.73 kilobases and the plastid genome assembly has a length of 163.75 kilobases. Gene annotation of this assembly on Ensembl identified 26 361 protein-coding genes. This assembly was generated as part of the Darwin Tree of Life project, which produces reference genomes for eukaryotic species found in Britain and Ireland.

Euphorbia lathyris↗

The genome sequence of Hypericum androsaemum L., 1753 (Malpighiales: Hypericaceae).

We present a genome assembly of Hypericum androsaemum (Tutsan; Streptophyta; Magnoliopsida; Malpighiales; Hypericaceae). The genome sequence has a total length of 384.54 megabases. Most of the assembly (99.96%) is scaffolded into 20 chromosomal pseudomolecules. The mitochondrial sequences have lengths of 138.63, 75.57, 98.15 and 103.38 kilobases and the plastid genome assembly has a length of 165.0 kilobases. This assembly was generated as part of the Darwin Tree of Life project, which produces reference genomes for eukaryotic species found in Britain and Ireland.

Hypericum androsaemum↗

Explosive radiation of Malpighiales supports a mid-cretaceous origin of modern tropical rain forests.

Fossil data have been interpreted as indicating that Late Cretaceous tropical forests were open and dry adapted and that modern closed-canopy rain forest did not originate until after the Cretaceous-Tertiary (K/T) boundary. However, some mid-Cretaceous leaf floras have been interpreted as rain forest. Molecular divergence-time estimates within the clade Malpighiales, which constitute a large percentage of species in the shaded, shrub, and small tree layer in tropical rain forests worldwide, provide new tests of these hypotheses. We estimate that all 28 major lineages (i.e., traditionally recognized families) within this clade originated in tropical rain forest well before the Tertiary, mostly during the Albian and Cenomanian (112-94 Ma). Their rapid rise in the mid-Cretaceous may have resulted from the origin of adaptations to survive and reproduce under a closed forest canopy. This pattern may also be paralleled by other similarly diverse lineages and supports fossil indications that closed-canopy tropical rain forests existed well before the K/T boundary. This case illustrates that dated phylogenies can provide an important new source of evidence bearing on the timing of major environmental changes, which may be especially useful when fossil evidence is limited or controversial.

Biological Evolution↗

Female flower and cupule structure in Balanopaceae, an enigmatic rosid family.

The Balanopaceae, whose flowers were poorly known, have, in the past, been variously allocated to the Fagales, Euphorbiaceae, Salicales or other hamamelids and rosids (these groups being in Fagales, Malpighiales and Saxifragales, according to the Angiosperm Phylogeny Group). This paper attempts a clarification based on flower morphology. Female flowers and cupules were studied in Balanops vieillardii, young fruits in B. australiana. The cupules are simple involucres of bracts which are spirally arranged (according to a Fibonacci pattern) on the floral axis preceding the flower. They contrast with the complicated cupules of Fagaceae which consist of a condensed cymose ramification system of axes of several orders around the flower. Flowers appear later than most of the cupular bracts, in contrast to Fagaceae. In addition to a terminal flower there may be several smaller lateral flowers in the axil of cupular bracts, each surrounded by its own small cupule. The female flowers do not have a perianth. They consist of two to three large carpels. At anthesis, the ovary is completely septate; the syncarpous part (ovary and lower style) is completely symplicate. The carpels are free for most of their length, with the free parts once, twice or three times bifurcate, in contrast to simple in Fagales. The stigmatic surface covers the ventral side of each stigmatic branch and at the margins also spreads to the dorsal side. The stigma is wet and secretion appears holocrine. The two ovules per carpel are collateral and axile in early development. However, at anthesis they appear one above the other, because in one ovule the funicle greatly elongates. As the ovary elongates only above the placenta, the ovules appear basal at anthesis. The ovules are (weakly) crassinucellar, bitegmic (not unitegmic), anatropous, and intermediate between apotropous and epitropous (not apotropous). The ovules are mature at anthesis, in contrast to Fagales. In mature ovules the upper part of the nucellus disintegrates, and a weakly differentiated endothelium is present in the inner integument. The morphological results of this study support a position of Balanopaceae in Malpighiales, and not Fagales or other orders, and are thus in accordance with recent molecular results based on chloroplast rbcL sequences data. However, within Malpighiales, as opposed to molecular results, Balanopaceae agree more with Euphorbiaceae s.l. than with Dichapetalaceae/Trigoniaceae and Chrysobalanaceae/Euphroniaceae.

Flowers↗

Phylogenetic relationships of the enigmatic angiosperm family Podostemaceae inferred from 18S rDNA and rbcL sequence data.

The phylogenetic relationships of some angiosperm families have remained enigmatic despite broad phylogenetic analyses of rbcL sequences. One example is the aquatic family Podostemaceae, the relationships of which have long been controversial because of major morphological modifications associated with their aquatic habit. Podostemaceae have variously been associated with Piperaceae, Nepenthaceae, Polygonaceae, Caryophyllaceae, Scrophulariaceae, Rosaceae, Crassulaceae, and Saxifragaceae. Two recent analyses of rbcL sequences suggest a possible sister-group relationship of Podostemaceae to Crassulaceae (Saxifragales). However, the branch leading to Podostemaceae was long, and use of different outgroups resulted in alternative placements. We explored the phylogenetic relationships of Podostemaceae using 18S rDNA sequences and a combined rbcL + 18S rDNA matrix representing over 250 angiosperms. In analyses based on 18S rDNA data, Podostemaceae are not characterized by a long branch; the family consistently appears as part of a Malpighiales clade that also includes Malpighiaceae, Turneraceae, Passifloraceae, Salicaceae, Euphorbiaceae, Violaceae, Linaceae, Chrysobalanaceae, Trigoniaceae, Humiriaceae, and Ochnaceae. Phylogenetic analyses based on a combined 18S rDNA + rbcL data set (223 ingroup taxa) with basal angiosperms as the outgroup also suggest that Podostemaceae are part of a Malpighiales clade. These searches swapped to completion, and the shortest trees showed enhanced resolution and increased internal support compared to those based on 18S rDNA or rbcL alone. However, when Gnetales are used as the outgroup, Podostemaceae appear with members of the nitrogen fixing clade (e.g., Elaeagnaceae, Ulmaceae, Rhamnaceae, Cannabaceae, Moraceae, and Urticaceae). None of the relationships suggested here for Podostemaceae receives strong bootstrap support. Our analyses indicate that Podostemaceae are not closely allied with Crassulaceae or with other members of the Saxifragales clade; their closest relatives, although still uncertain, appear to lie elsewhere in the rosids.

DNA, Ribosomal↗

Molecular phylogenetics of Phyllanthaceae inferred from five genes (plastid atpB, matK, 3'ndhF, rbcL, and nuclear PHYC).

Phyllanthaceae are a pantropical family of c. 2000 species for which circumscription is believed to be coincident with subfamily Phyllanthoideae of Euphorbiaceae sensu lato (Malpighiales) excluding Putranjivaceae. A phylogenetic study of the family using DNA sequence data has delivered largely congruent results from the plastid atpB, matK, ndhF, rbcL, and the nuclear PHYC. Our analyses include sampling from 54 of 59 genera, representing all tribes and subtribes of Phyllanthoideae. The family falls into two major clades characterized by inflorescence and leaf anatomical features. Several traditional taxonomic groupings were retrieved with minor modifications, but most clades recovered are considerably different from previous non-molecular based ideas of relationships. The enigmatic genus Dicoelia and the geographically disjunct genus Lingelsheimia are shown to be embedded in Phyllanthaceae. The taxonomic status of Leptopus diplospermus (=Chorisandrachne) and the debated placement of Andrachne ovalis have been clarified, and Protomegabaria and Richeriella are newly placed. Paraphyly of Cleistanthus and Phyllanthus is confirmed, having three and four other genera embedded, respectively. Petalodiscus is also paraphyletic, including all other Malagassian Wielandieae.

Base Sequence↗

Mitochondrial DNA sequences reveal the photosynthetic relatives of Rafflesia, the world's largest flower.

All parasites are thought to have evolved from free-living ancestors. However, the ancestral conditions facilitating the shift to parasitism are unclear, particularly in plants because the phylogenetic position of many parasites is unknown. This is especially true for Rafflesia, an endophytic holoparasite that produces the largest flowers in the world and has defied confident phylogenetic placement since its discovery >180 years ago. Here we present results of a phylogenetic analysis of 95 species of seed plants designed to infer the position of Rafflesia in an evolutionary context using the mitochondrial gene matR (1,806 aligned base pairs). Overall, the estimated phylogenetic tree is highly congruent with independent analyses and provides a strongly supported placement of Rafflesia with the order Malpighiales, which includes poinsettias, violets, and passionflowers. Furthermore, the phylogenetic placement of Mitrastema, another enigmatic, holoparasitic angiosperm with the order Ericales (which includes blueberries and persimmons), was obtained with these data. Although traditionally classified together, Rafflesia and Mitrastema are only distantly related, implying that their endoparasitic habits result from convergent evolution. Our results indicate that the previous significant difficulties associated with phylogenetic placement of holoparasitic plants may be overcome by using mitochondrial DNA so that a broader understanding of the origins and evolution of parasitism may emerge.

DNA, Mitochondrial↗

Phylogenetic inference in Rafflesiales: the influence of rate heterogeneity and horizontal gene transfer.

BACKGROUND: The phylogenetic relationships among the holoparasites of Rafflesiales have remained enigmatic for over a century. Recent molecular phylogenetic studies using the mitochondrial matR gene placed Rafflesia, Rhizanthes and Sapria (Rafflesiaceae s. str.) in the angiosperm order Malpighiales and Mitrastema (Mitrastemonaceae) in Ericales. These phylogenetic studies did not, however, sample two additional groups traditionally classified within Rafflesiales (Apodantheaceae and Cytinaceae). Here we provide molecular phylogenetic evidence using DNA sequence data from mitochondrial and nuclear genes for representatives of all genera in Rafflesiales. RESULTS: Our analyses indicate that the phylogenetic affinities of the large-flowered clade and Mitrastema, ascertained using mitochondrial matR, are congruent with results from nuclear SSU rDNA when these data are analyzed using maximum likelihood and Bayesian methods. The relationship of Cytinaceae to Malvales was recovered in all analyses. Relationships between Apodanthaceae and photosynthetic angiosperms varied depending upon the data partition: Malvales (3-gene), Cucurbitales (matR) or Fabales (atp1). The latter incongruencies suggest that horizontal gene transfer (HGT) may be affecting the mitochondrial gene topologies. The lack of association between Mitrastema and Ericales using atp1 is suggestive of HGT, but greater sampling within eudicots is needed to test this hypothesis further. CONCLUSIONS: Rafflesiales are not monophyletic but composed of three or four independent lineages (families): Rafflesiaceae, Mitrastemonaceae, Apodanthaceae and Cytinaceae. Long-branch attraction appears to be misleading parsimony analyses of nuclear small-subunit rDNA data, but model-based methods (maximum likelihood and Bayesian analyses) recover a topology that is congruent with the mitochondrial matR gene tree, thus providing compelling evidence for organismal relationships. Horizontal gene transfer appears to be influencing only some taxa and some mitochondrial genes, thus indicating that the process is acting at the single gene (not whole genome) level.

Bayes Theorem↗

Systematic affinities of Rhizophoraceae and Anisophylleaceae, and intergeneric relationships within Rhizophoraceae, based on chloroplast DNA, nuclear ribosomal DNA, and morphology.

A cladistic analysis of sequences from the chloroplast gene rbcL was used to determine the systematic affinities of Rhizophoraceae and Anisophylleaceae. This analysis rejects close relationships of Rhizophoraceae with Celastraceae or Elaeocarpaceae, suggested previously, and identifies Erythroxylaceae as sister group within the Malpighiales, supported by several morphological and anatomical characters. Our molecular results also indicate that Anisophylleaceae are nested within Cucurbitales. Although this placement is novel, this affinity is also well supported by shared morphological characters. Tribal and generic relationships within Rhizophoraceae are evaluated with a combination of six molecular data sets (rbcL, atpB-rbcL intergenic spacer, trnL-trnF intergenic spacer, ITS1, ITS2, and 5.8S) and a morphological data set. These relationships are compared with results from previous morphological cladistic analyses. Against the background of the molecular results, we briefly discuss the evolution of morphological characters traditionally used for tribal subdivision as well as characters presumably significant for adaptation to mangrove habitats, namely, aerial stilt roots and vivipary.

Journal Article↗