Saša Stefanović

Monophyly of the Convolvulaceae and circumscription of their major lineages based on DNA sequences of multiple chloroplast loci.

Convolvulaceae, a large family of worldwide distribution, exhibit a rich diversity of morphological characteristics and ecological habitats. Previous efforts to systematize this diversity without a cladistic phylogenetic framework have disagreed on the circumscription of the family as well as tribal composition and relationship. In order to circumscribe the family and assess the relationships among its major lineages, a broad data set was constructed containing representatives of all ten recognized tribes of Convolvulaceae plus representatives of putatively related families within Asteridae. This is done by using four chloroplast regions: rbcL, atpB, psbE-J operon, and trnL-trnF intron/spacer. The results indicate that Convolvulaceae are monophyletic and sister to Solanaceae. Two of the three groups that have been proposed previously as separate families, Cuscuta and Dichondreae, are nested within the Convolvulaceae in this analysis, and the third, Humbertia, is the sister to all other members of the family. The exact position of Cuscuta could not be ascertained, but some alternatives were rejected with confidence. The study identified several distinct monophyletic groups, some of which correspond to earlier taxonomic treatments. Close relationships of tribes Hildebrandtieae with Cresseae and Ipomoeeae with Argyreieae (forming Echinoconieae) were confirmed. The polyphyly of Merremieae, Convolvuleae, Poraneae, and Erycibeae is first identified in this study.

Loss of all plastid ndh genes in Gnetales and conifers: extent and evolutionary significance for the seed plant phylogeny

The exact phylogenetic position of Gnetales, a small, highly modified group of gymnosperms with an accelerated rate of molecular evolution, is one of the most challenging issues for seed plant systematics. Recent results from entire plastid genome (ptDNA) sequencing revealed the absence of the entire suite of plastid ndh genes in several species of Gnetales and the pine family (Pinaceae) potentially highlighting a major structural feature linking these two groups—concerted loss of all plastid genes for the NADH dehydrogenase complex. However, the precise extent of ndh gene loss in gymnosperms has not been surveyed. Using a slot-blot hybridization method, we probed all 11 ndh genes in 162 species from 70 of 85 gymnosperm genera. We find that all ndh genes are absent across Gnetales and Pinaceae, but not in any other group of gymnosperms. This feature represents either a major synapomorphy for a clade consisting of these two lineages or, less likely, a convergent loss. Our survey substantially extends previous inferences based on more limited sampling and, if the former evolutionary interpretation is correct, it provides additional support for the contentious “gnepine” hypothesis, which places Gnetales as sister to Pinaceae.

Classification of Convolvulaceae: A Phylogenetic Approach

Abstract Because recent molecular studies, based on multiple data sets from all three plant genomes, have indicated mutually congruent, well-resolved, and well-supported relationships within Convolvulaceae (the morning-glory family), a formal reclassification of this family is presented here. Convolvulaceae, a large family of worldwide distribution, exhibiting a rich diversity of morphological characteristics and ecological habitats, are now circumscribed within twelve tribes. A key to these tribes of Convolvulaceae is offered. The group of spiny-pollen bearing Convolvulaceae (forming “Echinoconiae”) and tribe Cuscuteae are retained essentially in their traditional sense, Cresseae are circumscribed with only minor modifications, Convolvuleae and Erycibeae are recognized in a restricted sense, while Dichondreae and Maripeae are expanded. Also, to produce a tribal taxonomy that better reflects phylogenetic relationships, the concept of Poraneae is abandoned as artificial, three new tribes are recognized (Aniseieae, Cardiochlamyeae, and Jacquemontieae), and a new tribal status is proposed for the Malagasy endemic Humbertia (Humbertieae). “Merremieae” are tentatively retained even though the monophyly of this tribe is not certain. In addition to the formal classification, we provide clade name definitions for the family as well as for most of the clades recognized presently as tribes. Also, five well-supported clades that are not assigned formal ranks are recognized and their names defined. The reevaluation of traditional taxonomic characters reveals that many homoplasious characters were emphasized in previous classifications, resulting in formal recognition of non-monophyletic groups. Putative morphological synapomorphies for many clades discovered through molecular cladistic analyses are discussed. However, the morphology of several clades that are well-supported by DNA evidence remains poorly understood, creating further challenges for future studies in Convolvulaceae.

Horizontal acquisition of multiple mitochondrial genes from a parasitic plant followed by gene conversion with host mitochondrial genes

BackgroundHorizontal gene transfer (HGT) is relatively common in plant mitochondrial genomes but the mechanisms, extent and consequences of transfer remain largely unknown. Previous results indicate that parasitic plants are often involved as either transfer donors or recipients, suggesting that direct contact between parasite and host facilitates genetic transfer among plants.ResultsIn order to uncover the mechanistic details of plant-to-plant HGT, the extent and evolutionary fate of transfer was investigated between two groups: the parasitic genus Cuscuta and a small clade of Plantago species. A broad polymerase chain reaction (PCR) survey of mitochondrial genes revealed that at least three genes (atp1, atp6 and matR) were recently transferred from Cuscuta to Plantago. Quantitative PCR assays show that these three genes have a mitochondrial location in the one species line of Plantago examined. Patterns of sequence evolution suggest that these foreign genes degraded into pseudogenes shortly after transfer and reverse transcription (RT)-PCR analyses demonstrate that none are detectably transcribed. Three cases of gene conversion were detected between native and foreign copies of the atp1 gene. The identical phylogenetic distribution of the three foreign genes within Plantago and the retention of cytidines at ancestral positions of RNA editing indicate that these genes were probably acquired via a single, DNA-mediated transfer event. However, samplings of multiple individuals from two of the three species in the recipient Plantago clade revealed complex and perplexing phylogenetic discrepancies and patterns of sequence divergence for all three of the foreign genes.ConclusionsThis study reports the best evidence to date that multiple mitochondrial genes can be transferred via a single HGT event and that transfer occurred via a strictly DNA-level intermediate. The discovery of gene conversion between co-resident foreign and native mitochondrial copies suggests that transferred genes may be evolutionarily important in generating mitochondrial genetic diversity. Finally, the complex relationships within each lineage of transferred genes imply a surprisingly complicated history of these genes in Plantago subsequent to their acquisition via HGT and this history probably involves some combination of additional transfers (including intracellular transfer), gene duplication, differential loss and mutation-rate variation. Unravelling this history will probably require sequencing multiple mitochondrial and nuclear genomes from Plantago.See Commentary: http://www.biomedcentral.com/1741-7007/8/147.

Relationships Among Phaseoloid Legumes Based on Sequences from Eight Chloroplast Regions

Abstract Generic level relationships in phaseoloid legumes have received much attention using chloroplast DNA markers. However, despite this attention not all relationships are yet well-resolved. This study includes trnL-F sequences from across a wide sample of phaseoloid legumes as well as seven additional chloroplast DNA loci (rbcL, atpB, trnK/matK, rpl2, clpP, rps16, and ycf4) analyzed separately and in combination. Together, these data provide support for many relationships generally consistent with, but only weakly supported, in earlier studies. Some major discordant phylogenetic results were found in our separate analyses; for example, ycf4 sequences group Glycine and Teramnus with strong support; however, the combined analysis of the remaining seven loci found incongruent groupings (Glycine and Psoraleeae genera; Teramnus and Amphicarpaea) also with strong support. Network analysis of ycf4 revealed that the conflicting signal (relative to the other seven loci) came from first and second codon positions. These positions also showed significant rate acceleration, together indicating that selection driving convergent molecular evolution is the likely cause of the signal in ycf4, rather than shared history. The major clades within the phaseoloid legumes supported by our analysis are discussed.

Population genetic structure of diploid sexual and polyploid apomictic hawthorns (Crataegus; Rosaceae) in the Pacific Northwest

Polyploidy and gametophytic apomixis are two important and associated processes in plants. Many hawthorn species are polyploids and can reproduce both sexually and apomictically. However, the population genetic structure of these species is poorly understood. Crataegus douglasii is represented exclusively by self‐compatible tetraploid pseudogamous apomicts across North America, whereas Crataegus suksdorfii found in the Pacific Northwest is known to include self‐incompatible diploid sexuals as well as polyploid apomicts. We compare population structure and genetic variability in these two closely related taxa using microsatellite and chloroplast sequence markers. Using 13 microsatellite loci located on four linkage groups, 251 alleles were detected in 239 individuals sampled from 15 localities. Within‐population multilocus genotypic variation and molecular diversity are greatest in diploid sexuals and lowest in triploid apomicts. Apart from the isolation of eastern North American populations of C. douglasii, there is little evidence of isolation by distance in this taxon. Genetic diversity in western populations of C. douglasii suggests that gene flow is frequent, and that colonization and establishment are often successful. In contrast, local populations of C. suksdorfii are more markedly differentiated. Gene flow appears to be limited primarily by distance in diploids and by apomixis and self‐compatibility in polyploids. We infer that apomixis and reproductive barriers between cytotypes are factors that reduce the frequency of gene flow among populations, and may ultimately lead to allopatric speciation in C. suksdorfii. Our findings shed light on evolution in woody plants that show heterogeneous ploidy levels and reproductive systems.

Testing the phylogenetic position of a parasitic plant (Cuscuta, Convolvulaceae, asteridae): Bayesian inference and the parametric bootstrap on data drawn from three genomes.

Previous findings on structural rearrangements in the chloroplast genome of Cuscuta (dodder), the only parasitic genus in the morning-glory family, Convolvulaceae, were attributed to its parasitic life style, but without proper comparison to related nonparasitic members of the family. Before molecular evolutionary questions regarding genome evolution can be answered, the phylogenetic problems within the family need to be resolved. However, the phylogenetic position of parasitic angiosperms and their precise relationship to nonparasitic relatives are difficult to infer. Problems are encountered with both morphological and molecular evidence. Molecular data have been used in numerous studies to elucidate relationships of parasitic taxa, despite accelerated rates of sequence evolution. To address the question of the position of the genus Cuscuta within Convolvulaceae, we generated a new molecular data set consisting of mitochondrial (atpA) and nuclear (RPB2) genes, and analyzed these data together with an existing chloroplast data matrix (rbcL, atpB, trnL-F, and psbE-J), to which an additional chloroplast gene (rpl2) was added. This data set was analyzed with an array of phylogenetic methods, including Bayesian analysis, maximum likelihood, and maximum parsimony. Further exploration of data was done by using methods of phylogeny hypothesis testing. At least two nonparasitic lineages are shown to diverge within the Convolvulaceae before Cuscuta. However, the exact sister group of Cuscuta could not be ascertained, even though many alternatives were rejected with confidence. Caution is therefore warranted when interpreting the causes of molecular evolution in Cuscuta. Detailed comparisons with nonparasitic Convolvulaceae are necessary before firm conclusions can be reached regarding the effects of the parasitic mode of life on patterns of molecular evolution in Cuscuta.

Delimitation of major lineages within Cuscuta subgenus Grammica (Convolvulaceae) using plastid and nuclear DNA sequences

Subgenus Grammica, the largest and most diverse group in the parasitic genus Cuscuta, includes 130 species distributed primarily throughout the New World, with Mexico as its center of diversity. To circumscribe the subgenus and assess the relationships among its major lineages, we conducted the first phylogenetic study of Grammica using plastid trnL-F and nrITS sequences from a wide taxonomic sampling covering its morphological, physiological, and geographical diversity. With the exception of one species belonging elsewhere, the subgenus was found to be monophyletic. The results further indicate the presence of 15 well-supported major clades within Grammica. Some of those lineages correspond partially to earlier taxonomic treatments, but the majority of groups are identified in this study for the first time. The backbone relationships among major clades, however, remain weakly supported or unresolved in some cases. The phylogenetic results indicate that the fruit dehiscence character is homoplastic, thus compromising its value as a major taxonomic and evolutionary feature. While several striking cases of long-distance dispersal are inferred, vicariance emerges as the most dominant biogeographical pattern for Cuscuta. Species placed within one of the clades with a predominantly South American distribution are hypothesized to have substantially altered plastid genomes.

Plastid genome evolution in mycoheterotrophic Ericaceae

Unlike parasitic plants, which are linked to their hosts directly through haustoria, mycoheterotrophic (MHT) plants derive all or part of their water and nutrients from autothrophs via fungal mycorrhizal intermediaries. Ericaceae, the heather family, are a large and diverse group of plants known to form elaborate symbiotic relationships with mycorrhizal fungi. Using PHYA sequence data, we first investigated relationships among mycoheterotrophic Ericaceae and their close autotrophic relatives. Phylogenetic results suggest a minimum of two independent origins of MHT within this family. Additionally, a comparative investigation of plastid genomes (plastomes) grounded within this phylogenetic framework was conducted using a slot-blot Southern hybridization approach. This survey encompassed numerous lineages of Ericaceae with different life histories and trophic levels, including multiple representatives from mixotrophic Pyroleae and fully heterotrophic Monotropeae and Pterosporeae. Fifty-four probes derived from all categories of protein coding genes typically found within the plastomes of flowering plants were used. Our results indicate that the holo-mycoheterotrophic Ericaceae exhibit extensive loss of genes relating to photosynthetic function and expression of the plastome but retain genes with possible functions outside photosynthesis. Mixotrophic taxa tend to retain most genes relating to photosynthetic functions but are varied regarding the plastid ndh gene content. This investigation extends previous inferences that the loss of the NDH complex occurs prior to becoming holo-heterotrophic and it shows that the pattern of gene losses among mycoheterotrophic Ericaceae is similar to that of haustorial parasites. Additionally, we identify the most desirable candidate species for entire plastome sequencing.

Molecular Reappraisal of Relationships Between Crataegus and Mespilus (Rosaceae, Pyreae)—Two Genera or One?

Abstract Mespilus and Crataegus are sister genera in Rosaceae tribe Pyreae. Mespilus has been seen to comprise not only the medlar, Mespilus germanica, of western Eurasia but also the Arkansas, U.S.A. endemic, Mespilus canescens. Crataegus, on the other hand, consists of 140–200 species found throughout the northern hemisphere. Diagnoses of these two genera rely on morphological features of leaves, flowers and fruits. However, character states supposed to be diagnostic of Mespilus occur in species of Crataegus. We used two nuclear (ribosomal ITS and LEAFY intron2) and four intergenic chloroplast DNA regions (trnS-trnG, psbA-trnH, trnH-rpl2, and rpl20-rps12) to estimate the phylogeny of Mespilus and Crataegus. Maximum parsimony, maximum likelihood, and Bayesian analyses all corroborate the sister group relationship between Crataegus and Mespilus, and Crataegus brachyacantha sister to the rest of Crataegus. However, incongruence between chloroplast and nuclear data supports the hypothesis of a hybrid origin for Mespilus canescens, with Crataegus brachyacantha or its ancestor as the maternal parent. Accordingly, we (1) restrict Crataegus section Brevispinae to Crataegus brachyacantha (2) distinguish the Arkansas endemic as a nothospecies; (3) describe a new section and a new nothosection within Crataegus to contain the former species of Mespilus and Crataemespilus; and (4) make two new combinations under Crataegus.