Sean W. Graham

Summary: Complex traits often evolve by co-opting pre-existing genetic components. Orthologs of the transcriptional co-factor SEED DORMANCY 4 (SDR4) are found across seed-bearing and seed-free land plants. In flowering plants, loss-of-function mutants of sdr4 orthologs in rice and Arabidopsis exhibit opposite phenotypes in seed germination, a trait regulated by the phytohormone abscisic acid (ABA). We uncovered a major gene duplication event that closely preceded the origin of seed plants, by using a structure-informed molecular phylogenetic analysis. This duplication gave rise to the paralogous SDR4 and DIG/DIL clades, which are broadly retained across the seed plants. Investigating the role of SDR4 homologs in the common liverwort Marchantia polymorpha , a seed-free model organism with only a single SDR4-like homolog (MpSDR4), may provide insights into how SDR4 evolved to regulate dormancy and germination in other land plants. Loss-of-function mutants of Mp sdr4 and transcriptome profiling showed that MpSDR4 modulates ABA sensitivity, consistent with the idea that SDR4 subsequently evolved to regulate hormone responses related to seed germination, with additional functional divergence between monocots and eudicots. Furthermore, Mp sdr4 mutants exhibit elevated growth of gemma cups, contrasting the roles of SDR4 orthologs in asexual reproduction in liverwort with the regulation of seed traits in flowering plants.

We use genomic and morphological data to show that three distinct bamboo species, Khoomengia honbaensis, Sasa hainanensis and an undescribed bamboo species from Guangxi belong to the small bamboo subtribe Hsuehochloinae of tribe Arundinarieae, which previously was represented by only a single species, Hsuehochloa calcarea. Here we provide detailed morphological and phylogenomic evidence supporting this expansion of the subtribe. We employ various approaches to analyze nuclear and plastid phylogenomic data sets obtained using a deep genome skimming (DGS) survey. All variant analyses strongly support the re-circumscription of Hsuehochloinae to include these three additional taxa, a finding that is also corroborated by multiple probable morphological synapomorphies. We establish a new genus, Guihaiochloa, to accommodate the undescribed southern Chinese bamboo, which differs from the other three species in having a raceme- or panicle-like unit of inflorescence of synflorescence with 3–6 spikelets. We provide both phylogenomic and morphological evidence for the transfer of the Chinese species Sasa hainanensis to the recently described Vietnamese genus Khoonmengia. However, at a broader level, using coalescent methods for inferring relationships among the subtribes still leads to poorly supported relationships among Hsuehochloinae, Ampelocalaminae and a clade comprising the remaining subtribes within Arundinarieae. We show that incomplete lineage sorting is likely a major source of this phylogenetic uncertainty in higher-order Arundinarieae bamboo phylogeny, which is likely the result of extremely short divergent times between the two closely related subtribes and a clade comprising the rest of Arundinarieae.

Premise: The inclusion of heterotrophic plants in broader systematic studies is challenging due to substantial morphological modification, gene loss, and accelerated substitution rates following photosynthesis loss. In particular, inclusion of very long branch (VLB) taxa with highly elevated rates may lead to phylogenomic misinference. Methods: We explored how rapidly evolving plastomes behave in model‐based phylogenetic inferences in the yam order Dioscoreales, which experienced convergent photosynthesis losses. Our taxon sampling included mycoheterotrophic lineages displaying a broad range of substitution rates and all photosynthetic genera. Results: When only moderately rapidly evolving heterotrophs are included, relationships within Dioscoreales are congruent with recent mitochondrial analysis, and generally strongly supported. Photosynthetic Stenomeris (represented by only three genes) is moderately supported as sister to Dioscoreaceae. Notably, Burmanniaceae and Thismiaceae, two mycoheterotrophic families including highly reduced plastomes, are strongly supported as distantly related within Dioscoreales. Constrained analysis rejects combining Thismiaceae with Burmanniaceae, conflicting with current angiosperm classification. Including a subset of VLB mycoheterotrophs in phylogenetic inference results in localized or broad reduction in branch support—substantially so for Afrothismiaceae, a family placing in disparate locations in Dioscoreales (and beyond) in variant analyses, with no ability to differentiate among its alternative placements. Conclusions: Inclusion of highly rate‐elevated taxa in analyses of Dioscoreales phylogeny can depress branch support. Inferred relationships in the order are otherwise congruent with studies based on mitochondrial data; the family‐level classification needs updating. The extraordinarily rapidly evolving family Afrothismiaceae places inconsistently in diverse plastid analyses, pointing to probable analytical limits for plastid‐based phylogenomic analyses of VLB heterotrophs.

Summary: The vast majority of land plants transfer part of the organic carbon they produce by photosynthesis to arbuscular mycorrhizal (AM) fungi inside their root cells; the fungi in turn help plants to take up nutrients and water from the soil. This carbon can subsequently be acquired from mycorrhizal fungi by rare nonphotosynthetic ‘mycoheterotrophic’ plants that tap into the same fungal network. However, recent findings suggest that carbon uptake from AM fungi may exist among some green plants too. If so, this would qualify them as partial mycoheterotrophs (mixotrophs) rather than as pure autotrophs. Here, we discuss the evolutionary, ecophysiological, morphological, genetic, and environmental evidence for the existence and prevalence of this trait. We conclude that there is strong, albeit indirect, evidence for its existence, although its taxonomic distribution remains to be determined. This knowledge gap currently prevents us from inferring the magnitude of AM partial mycoheterotrophy in terrestrial ecosystems, which in turn severely limits our understanding of its role in plant establishment and survival and in ecosystem structure and function.

Premise: Plastid-based data sets continue to play a major role in our understanding of early flowering-plant relationships, although organellar genomes of major lineages outside the monocots and eudicots remain under-sampled. A tendency of mitochondrial RNA-edit sites to mislead phylogenetic analysis in mixed transcriptomic/genomic data sets needs attention in angiosperm-wide studies, which only rarely consider mitochondrial genomes. Methods: We compared mitochondrial- vs. plastid-based phylogenomic inferences, examined the effect of removing putative RNA-edit sites from mitochondrial data, and performed combined organellar analysis (plastid plus filtered mitochondrial genomes). We expanded taxon sampling for multiple angiosperm lineages for phylogenomic analysis using both organellar genomes, representing several poorly sampled lineages (in particular Degeneriaceae, Trimeniaceae) with smaller (few-gene) data sets. Results: Plastid-based inferences recover well-supported relationships that align with and build upon previous studies, and recover well-supported internal relationships for two ANA-grade families (Hydatellaceae, Trimeniaceae) sampled for nearly all species. By contrast, unfiltered mitochondrial inferences of angiosperm phylogeny are generally poorly supported, and recover anomalous relationships compared to plastid-based inferences. However, removing putative mitochondrial RNA-edit sites dramatically reduces inter-organellar conflict and improves overall branch support. Conclusions: We accounted for phylogenomic discordance between the two organellar genomes regarding overall angiosperm-wide relationships and filled in taxonomic gaps (poorly sampled lineages). Removing RNA edit sites substantially improves congruence in interorganellar inferences by effectively correcting a systematic bias in mitochondrial data. Uncertain relationships persist among five major mesangiosperm lineages in plastid-based inferences, but a clade comprising Chloranthales, Ceratophyllales and eudicots is well supported by filtered mitochondrial data.

Angiosperms are the cornerstone of most terrestrial ecosystems and human livelihoods^1,2. A robust understanding of angiosperm evolution is required to explain their rise to ecological dominance. So far, the angiosperm tree of life has been determined primarily by means of analyses of the plastid genome^3,4. Many studies have drawn on this foundational work, such as classification and first insights into angiosperm diversification since their Mesozoic origins^5–7. However, the limited and biased sampling of both taxa and genomes undermines confidence in the tree and its implications. Here, we build the tree of life for almost 8,000 (about 60%) angiosperm genera using a standardized set of 353 nuclear genes⁸. This 15-fold increase in genus-level sampling relative to comparable nuclear studies⁹ provides a critical test of earlier results and brings notable change to key groups, especially in rosids, while substantiating many previously predicted relationships. Scaling this tree to time using 200 fossils, we discovered that early angiosperm evolution was characterized by high gene tree conflict and explosive diversification, giving rise to more than 80% of extant angiosperm orders. Steady diversification ensued through the remaining Mesozoic Era until rates resurged in the Cenozoic Era, concurrent with decreasing global temperatures and tightly linked with gene tree conflict. Taken together, our extensive sampling combined with advanced phylogenomic methods shows the deep history and full complexity in the evolution of a megadiverse clade.

Summary: Afrothismia is a genus of non-photosynthetic mycoheterotrophs from the forests of continental tropical Africa. Multiple phylogenetic inferences using molecular data recover the genus as sister to a clade comprising mycoheterotrophic Thismiaceae and the photosynthetic family Taccaceae, contrary to earlier placements of Afrothismia and Thismiaceae within Burmanniaceae. Morphological support for separating Afrothismia from the rest of Thismiaceae has depended on the zygomorphic flowers of Afrothismia (although some species of Thismia are also zygomorphic), and their clusters of root tubers, each with a terminal rootlet. The number of described species of Afrothismia has recently increased from four to 16, with seven more species as yet undescribed; these discoveries have added morphological characters that support its distinction from Thismiaceae. Most notably, the ovary in Afrothismia has a single stalked placenta, and circumscissile fruits from which seeds are exserted by placental elevation (vs in Thismiaceae, three placentas, a deliquescing fruit lid, and seeds not exserted). Afrothismia stamens are inserted in the lower part of the floral tube, where they are attached to the stigma, and individual flowers are subtended by a single large dorsal bract. In contrast, in Thismiaceae, stamens are inserted at the mouth of the tube, free of and distant from the stigma, and each flower is subtended by a loose whorl of (2 –) 3 (– 4) bracts. Here we formally characterise Afrothismiaceae and review what is known of its development, seed germination, interactions with mycorrhizal Glomeromycota, biogeography, phylogeny and pollination biology. All but one (Afrothismia insignis; Vulnerable) of the 13 species assessed on the IUCN Red List of Threatened Species are either Endangered or Critically Endangered; one species (A. pachyantha) is considered extinct.

Premise: Bryophytes form a major component of terrestrial plant biomass, structuring ecological communities in all biomes. Our understanding of the evolutionary history of hornworts, liverworts, and mosses has been significantly reshaped by inferences from molecular data, which have highlighted extensive homoplasy in various traits and repeated bursts of diversification. However, the timing of key events in the phylogeny, patterns, and processes of diversification across bryophytes remain unclear.Methods: Using the GoFlag probe set, we sequenced 405 exons representing 228 nuclear genes for 531 species from 52 of the 54 orders of bryophytes. We inferred the species phylogeny from gene tree analyses using concatenated and coalescence approaches, assessed gene conflict, and estimated the timing of divergences based on 29 fossil calibrations.Results: The phylogeny resolves many relationships across the bryophytes, enabling us to resurrect five liverwort orders and recognize three more and propose 10 new orders of mosses. Most orders originated in the Jurassic and diversified in the Cretaceous or later. The phylogenomic data also highlight topological conflict in parts of the tree, suggesting complex processes of diversification that cannot be adequately captured in a single gene‐tree topology.Conclusions: We sampled hundreds of loci across a broad phylogenetic spectrum spanning at least 450 Ma of evolution; these data resolved many of the critical nodes of the diversification of bryophytes. The data also highlight the need to explore the mechanisms underlying the phylogenetic ambiguity at specific nodes. The phylogenomic data provide an expandable framework toward reconstructing a comprehensive phylogeny of this important group of plants.

Parasitic plants have evolved to be subtly or severely dependent on host plants to complete their life cycle. To provide new insights into the biology of parasitic plants in general, we assembled genomes for members of the sandalwood order Santalales, including a stem hemiparasite (Scurrula) and two highly modified root holoparasites (Balanophora) that possess chimaeric host–parasite tubers. Comprehensive genome comparisons reveal that hemiparasitic Scurrula has experienced a relatively minor degree of gene loss compared with autotrophic plants, consistent with its moderate degree of parasitism. Nonetheless, patterns of gene loss appear to be substantially divergent across distantly related lineages of hemiparasites. In contrast, Balanophora has experienced substantial gene loss for the same sets of genes as an independently evolved holoparasite lineage, the endoparasitic Sapria (Malpighiales), and the two holoparasite lineages experienced convergent contraction of large gene families through loss of paralogues. This unprecedented convergence supports the idea that despite their extreme and strikingly divergent life histories and morphology, the evolution of these and other holoparasitic lineages can be shaped by highly predictable modes of genome reduction. We observe substantial evidence of relaxed selection in retained genes for both hemi- and holoparasitic species. Transcriptome data also document unusual and novel interactions between Balanophora and host plants at the host–parasite tuber interface tissues, with evidence of mRNA exchange, substantial and active hormone exchange and immune responses in parasite and host.