TL;DR: The authors' 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, thus providing compelling evidence for organismal relationships.
Abstract: 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. 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. 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.
TL;DR: A phylogenetic analysis of 16 accessions representing six species of Apodanthaceae using the mitochondrial matR gene and the nuclear 18S gene concludes that they belong in the Cucurbitales with which they share inferior ovaries, parietal placentation and a dioecious mating system, traits that can now be interpreted as possible synapomorphies of an enlarged order Cucurbanes.
Abstract: Background: Of the c. 450 families of flowering plants, only two are left “unplaced” in the most recent APG classification of angiosperms. One of these is the Apodanthaceae, a clade of c. 19 holoparasitic species in two or three genera occurring in North and South America, Africa, the Near East, and Australia. Because of lateral gene transfer between Apodanthaceae and their hosts it has been difficult to infer the family’s true closest relatives. Results: Here we report a phylogenetic analysis of 16 accessions representing six species of Apodanthaceae from the United States, Chile, Iran, and Australia, using the mitochondrial matR gene and the nuclear 18S gene. Data matrices include 190 matR sequences from up to 95 families in 39 orders of flowering plants and 197 18S sequences from 101 families representing the 16 orders of rosids. Analyses were performed at the nucleotide and at the amino acid level. Both gene trees agree with angiosperm phylogenies found in other studies using more genes. Apodanthaceae and the seven families of the order Cucurbitales form a clade with 100% bootstrap support from matR and 56% from 18 S. In addition, the Apodanthaceae and Cucurbitales matR gene sequences uniquely share two non-synonymous codon changes and one synonymous change, as well as a codon insertion, already found by Barkman et al. (2007). Conclusions: Apodanthaceae belong in the Cucurbitales with which they share inferior ovaries, parietal placentation and a dioecious mating system, traits that are ancestral in Cucurbitales and which can now be interpreted as possible synapomorphies of an enlarged order Cucurbitales. The occurrence of Apodanthaceae in the Americas, Africa, the Near East, and Australia, and their adaptation to distantly related host species in the Fabaceae and Salicaceae suggest a long evolutionary history.
TL;DR: Sister group comparisons so far do not reveal any reproductive traits that evolved (or were lost) concomitant with a parasitic life style, but the lack of wind pollination suggests that this pollen vector may be maladaptive in parasites, perhaps because of host foliage or flowers borne close to the ground.
Abstract: Premise of the study: The most recent reviews of the reproductive biology and sexual systems of parasitic angiosperms were published 17 yr ago and reported that dioecy might be associated with parasitism. We use current knowledge on parasitic lineages and their sister groups, and data on the reproductive biology and sexual systems of Apodanthaceae, to readdress the question of possible trends in the reproductive biology of parasitic angiosperms. Methods: Fieldwork in Zimbabwe and Iran produced data on the pollinators and sexual morph frequencies in two species of Apodanthaceae. Data on pollinators, dispersers, and sexual systems in parasites and their sister groups were compiled from the literature. Key results: With the possible exception of some Viscaceae, most of the ca. 4500 parasitic angiosperms are animal-pollinated, and ca. 10% of parasites are dioecious, but the gain and loss of dioecy across angiosperms is too poorly known to infer a statistical correlation. The studied Apodanthaceae are dioecious and pollinated by nectar- or pollen-foraging Calliphoridae and other fl ies. Conclusions: Sister group comparisons so far do not reveal any reproductive traits that evolved (or were lost) concomitant with a parasitic life style, but the lack of wind pollination suggests that this pollen vector may be maladaptive in parasites, perhaps because of host foliage or flborne close to the ground.
TL;DR: Comparative studies on floral morphology, anatomy, and histology were performed and Berlinianche was studied for the first time in detail and its affinity to Apodanthaceae was confirmed.
Abstract: Comparative studies on floral morphol- ogy, anatomy, and histology were performed to identify shared features of the genera of Apo- danthaceae (Rafflesiales): Apodanthes, Pilostyles, and Berlinianche. Berlinianche was studied for the first time in detail and its affinity to Apodanth- aceae was confirmed. It has a previously unde- scribed hair cushion on the inner perianth organs and inaperturate pollen. Shared features of mem- bers of Apodanthaceae are: unisexual flowers; three (or four) alternating di-/tetra- or tri-/hexa- merous whorls of scales of which the inner one or two correspond to a perianth; a synandrium with pollen sacs typically arranged in two rings; opening by a dehiscence line between the two rings of pollen sacs; large vesicular hairs above the synandrium; a gynoecium with four united carpels; inferior and unilocular ovaries with four parietal placentae, ovules tenuinucellate, anatro- pous with two well developed integuments, ori- ented in various directions; a nectary disk. Apodanthaceae share some special structural features with Malvales.
TL;DR: Using nuclear and mitochondrial DNA sequences of Apodanthaceae, a worldwide family of endoparasites living inside Fabaceae and Salicaceae, several dating approaches are compared and host-age calibrations did not yield well-constrained results, but they may work better in other parasite clades.