Nitrogen isotopes reveal independent origins of N2-fixing symbiosis in extant cycad lineages.

Cycads are ancient seed plants (gymnosperms) that emerged by the early Permian. Although they were common understory flora and food for dinosaurs in the Mesozoic, their abundance declined markedly in the Cenozoic. Extant cycads persist in restricted populations in tropical and subtropical habitats and, with their conserved morphology, are often called 'living fossils.' All surviving taxa receive nitrogen from symbiotic N2-fixing cyanobacteria living in modified roots, suggesting an ancestral origin of this symbiosis. However, such an ancient acquisition is discordant with the abundance of cycads in Mesozoic fossil assemblages, as modern N2-fixing symbioses typically occur only in nutrient-poor habitats where advantageous for survival. Here, we use foliar nitrogen isotope ratios-a proxy for N2 fixation in modern plants-to probe the antiquity of the cycad-cyanobacterial symbiosis. We find that fossilized cycad leaves from two Cenozoic representatives of extant genera have nitrogen isotopic compositions consistent with microbial N2 fixation. In contrast, all extinct cycad genera have nitrogen isotope ratios that are indistinguishable from co-existing non-cycad plants and generally inconsistent with microbial N2 fixation, pointing to nitrogen assimilation from soils and not through symbiosis. This pattern indicates that, rather than being ancestral within cycads, N2-fixing symbiosis arose independently in the lineages leading to living cycads during or after the Jurassic. The preferential survival of these lineages may therefore reflect the effects of competition with angiosperms and Cenozoic climatic change.

Oldest evidence of abundant C4 grasses and habitat heterogeneity in eastern Africa.

The assembly of Africa's iconic C4 grassland ecosystems is central to evolutionary interpretations of many mammal lineages, including hominins. C4 grasses are thought to have become ecologically dominant in Africa only after 10 million years ago (Ma). However, paleobotanical records older than 10 Ma are sparse, limiting assessment of the timing and nature of C4 biomass expansion. This study uses a multiproxy design to document vegetation structure from nine Early Miocene mammal site complexes across eastern Africa. Results demonstrate that between ~21 and 16 Ma, C4 grasses were locally abundant, contributing to heterogeneous habitats ranging from forests to wooded grasslands. These data push back the oldest evidence of C4 grass-dominated habitats in Africa-and globally-by more than 10 million years, calling for revised paleoecological interpretations of mammalian evolution.

The evolution of hominoid locomotor versatility: Evidence from Moroto, a 21 Ma site in Uganda.

Living hominoids are distinguished by upright torsos and versatile locomotion. It is hypothesized that these features evolved for feeding on fruit from terminal branches in forests. To investigate the evolutionary context of hominoid adaptive origins, we analyzed multiple paleoenvironmental proxies in conjunction with hominoid fossils from the Moroto II site in Uganda. The data indicate seasonally dry woodlands with the earliest evidence of abundant C4 grasses in Africa based on a confirmed age of 21 million years ago (Ma). We demonstrate that the leaf-eating hominoid Morotopithecus consumed water-stressed vegetation, and postcrania from the site indicate ape-like locomotor adaptations. These findings suggest that the origin of hominoid locomotor versatility is associated with foraging on leaves in heterogeneous, open woodlands rather than forests.

The origin and evolution of open habitats in North America inferred by Bayesian deep learning models.

Some of the most extensive terrestrial biomes today consist of open vegetation, including temperate grasslands and tropical savannas. These biomes originated relatively recently in Earth's history, likely replacing forested habitats in the second half of the Cenozoic. However, the timing of their origination and expansion remains disputed. Here, we present a Bayesian deep learning model that utilizes information from fossil evidence, geologic models, and paleoclimatic proxies to reconstruct paleovegetation, placing the emergence of open habitats in North America at around 23 million years ago. By the time of the onset of the Quaternary glacial cycles, open habitats were covering more than 30% of North America and were expanding at peak rates, to eventually become the most prominent natural vegetation type today. Our entirely data-driven approach demonstrates how deep learning can harness unexplored signals from complex data sets to provide insights into the evolution of Earth's biomes in time and space.

The history and challenge of grassy biomes.

Grassy biomes are >20 million years old but are undervalued and under threat today.

High silicon concentrations in grasses are linked to environmental conditions and not associated with C4 photosynthesis.

The uptake and deposition of silicon (Si) as silica phytoliths is common among land plants and is associated with a variety of functions. Among these, herbivore defense has received significant attention, particularly with regard to grasses and grasslands. Grasses are well known for their high silica content, a trait which has important implications ranging from defense to global Si cycling. Here, we test the classic hypothesis that C4 grasses evolved stronger mechanical defenses than C3 grasses through increased phytolith deposition, in response to extensive ungulate herbivory ("C4 -grazer hypothesis"). Despite mixed support, this hypothesis has received broad attention, even outside the realm of plant biology. Because C3 and C4 grasses typically dominate in different climates, with the latter more abundant in hot, dry regions, we also investigated the effects of water availability and temperature on Si deposition. We compiled a large dataset of grasses grown under controlled environmental conditions. Using phylogenetically informed generalized linear mixed models and character evolution models, we evaluated whether photosynthetic pathway or growth condition influenced Si concentration. We found that C4 grasses did not show consistently elevated Si concentrations compared with C3 grasses. High temperature treatments were associated with increased concentration, especially in taxa adapted to warm regions. Although the effect was less pronounced, reduced water treatment also promoted silica deposition, with slightly stronger response in dry habitat species. The evidence presented here rejects the "C4 -grazer hypothesis." Instead, we propose that the tendency for C4 grasses to outcompete C3 species under hot, dry conditions explains previous observations supporting this hypothesis. These findings also suggest a mechanism via which anthropogenic climate change may influence silica deposition in grasses and, by extension, alter the important ecological and geochemical processes it affects.

3D shape analysis of grass silica short cell phytoliths: a new method for fossil classification and analysis of shape evolution.

Fossil grass silica short cell phytoliths (GSSCP) have been used to reconstruct the biogeography of Poaceae, untangle crop domestication history and detect past vegetation shifts. These inferences depend on accurately identifying the clade to which the fossils belong. Patterns of GSSCP shape and size variation across the family have not been established and current classification methods are subjective or based on a 2D view that ignores important 3D shape variation. Focusing on Poaceae subfamilies Anomochlooideae, Pharoideae, Pueliodieae, Bambusoideae and Oryzoideae, we observed in situ GSSCP to establish their orientation and imaged isolated GSSCP using confocal microscopy to produce 3D models. 3D geometric morphometrics was used to analyze GSSCP shape and size. Classification models were applied to GSSCP from Eocene sediments from Nebraska, USA, and Anatolia, Turkey. There were significant shape differences between nearly all recognized GSSCP morphotypes and between clades with shared morphotypes. Most of the Eocene GSSCP were classified as woody bamboos with some distinctive Nebraska GSSCP classified as herbaceous bamboos. 3D morphometrics hold great promise for GSSCP classification. It accounts for the complete GSSCP shape, automates size measurements and accommodates the complete range of morphotypes within a single analytical framework.

Lineage-based functional types: characterising functional diversity to enhance the representation of ecological behaviour in Land Surface Models.

Process-based vegetation models attempt to represent the wide range of trait variation in biomes by grouping ecologically similar species into plant functional types (PFTs). This approach has been successful in representing many aspects of plant physiology and biophysics but struggles to capture biogeographic history and ecological dynamics that determine biome boundaries and plant distributions. Grass-dominated ecosystems are broadly distributed across all vegetated continents and harbour large functional diversity, yet most Land Surface Models (LSMs) summarise grasses into two generic PFTs based primarily on differences between temperate C3 grasses and (sub)tropical C4 grasses. Incorporation of species-level trait variation is an active area of research to enhance the ecological realism of PFTs, which form the basis for vegetation processes and dynamics in LSMs. Using reported measurements, we developed grass functional trait values (physiological, structural, biochemical, anatomical, phenological, and disturbance-related) of dominant lineages to improve LSM representations. Our method is fundamentally different from previous efforts, as it uses phylogenetic relatedness to create lineage-based functional types (LFTs), situated between species-level trait data and PFT-level abstractions, thus providing a realistic representation of functional diversity and opening the door to the development of new vegetation models.

Exploring cycad foliage as an archive of the isotopic composition of atmospheric nitrogen.

Molecular nitrogen (N2 ) constitutes the majority of Earth's modern atmosphere, contributing ~0.79 bar of partial pressure (pN2 ). However, fluctuations in pN2 may have occurred on 107 -109  year timescales in Earth's past, perhaps altering the isotopic composition of atmospheric nitrogen. Here, we explore an archive that may record the isotopic composition of atmospheric N2 in deep time: the foliage of cycads. Cycads are ancient gymnosperms that host symbiotic N2 -fixing cyanobacteria in modified root structures known as coralloid roots. All extant species of cycads are known to host symbionts, suggesting that this N2 -fixing capacity is perhaps ancestral, reaching back to the early history of cycads in the late Paleozoic. Therefore, if the process of microbial N2 fixation records the δ15 N value of atmospheric N2 in cycad foliage, the fossil record of cycads may provide an archive of atmospheric δ15 N values. To explore this potential proxy, we conducted a survey of wild cycads growing in a range of modern environments to determine whether cycad foliage reliably records the isotopic composition of atmospheric N2 . We find that neither biological nor environmental factors significantly influence the δ15 N values of cycad foliage, suggesting that they provide a reasonably robust record of the δ15 N of atmospheric N2 . Application of this proxy to the record of carbonaceous cycad fossils may not only help to constrain changes in atmospheric nitrogen isotope ratios since the late Paleozoic, but also could shed light on the antiquity of the N2 -fixing symbiosis between cycads and cyanobacteria.

Comment on "The global tree restoration potential".

Bastin et al's estimate (Reports, 5 July 2019, p. 76) that tree planting for climate change mitigation could sequester 205 gigatonnes of carbon is approximately five times too large. Their analysis inflated soil organic carbon gains, failed to safeguard against warming from trees at high latitudes and elevations, and considered afforestation of savannas, grasslands, and shrublands to be restoration.

Assembly of modern mammal community structure driven by Late Cretaceous dental evolution, rise of flowering plants, and dinosaur demise.

The long-standing view that Mesozoic mammaliaforms living in dinosaur-dominated ecosystems were ecologically constrained to small size and insectivory has been challenged by astonishing fossil discoveries over the last three decades. By studying these well-preserved early mammaliaform specimens, paleontologists now agree that mammaliaforms underwent ecomorphological diversification during the Mesozoic Era. This implies that Mesozoic mammaliaform communities had ecological structure and breadth that were comparable to today's small-bodied mammalian communities. However, this hypothesis remains untested in part because the primary focus of most studies is on individual taxa. Here, we present a study quantifying the ecological structure of Mesozoic mammaliaform communities with the aim of identifying evolutionary and ecological drivers that influenced the deep-time assembly of small-bodied mammaliaform communities. We used body size, dietary preference, and locomotor mode to establish the ecospace occupation of 98 extant, small-bodied mammalian communities from diverse biomes around the world. We calculated ecological disparity and ecological richness to measure the magnitude of ecological differences among species in a community and the number of different eco-cells occupied by species of a community, respectively. This modern dataset served as a reference for analyzing five exceptionally preserved, extinct mammaliaform communities (two Jurassic, two Cretaceous, one Eocene) from Konservat-Lagerstätten. Our results indicate that the interplay of at least three factors, namely the evolution of the tribosphenic molar, the ecological rise of angiosperms, and potential competition with other vertebrates, may have been critical in shaping the ecological structure of small-bodied mammaliaform communities through time.

Evolution of phytolith deposition in modern bryophytes, and implications for the fossil record and influence on silica cycle in early land plant evolution.

Anecdotal evidence indicating substantial silica accumulation in tissues of bryophytes suggests that silica (phytolith) deposition evolved early on in embryophytes. To test this hypothesis, we conducted the first survey of phytolith content representing the major liverwort, moss and hornwort clades. We also assessed the diagnostic value of bryophyte phytoliths. Silica extracted from bryophyte material through wet-ashing was described, focusing on abundance, classifying taxa as nonproducers, light producers and higher producers; and phytolith morphotypes. Ancestral state reconstruction of these characters was performed for mosses and liverworts using published phylogenies. Phytoliths are present in multiple subclades within liverworts, mosses and hornworts, but these phyla were not ancestrally high silica-producers. Higher deposition occurs in liverworts and mosses with specialized water-conducting cells. We hypothesize that active, high silica accumulation was not ancestral for embryophytes, but became possible in clades with increased water conductance. Phytoliths of diagnostic structures (e.g. pegged rhizoids) could help track bryophyte clades or water conductance evolution in the fossil record.

Contribution of forests to the carbon sink via biologically-mediated silicate weathering: A case study of China.

During silicate weathering, atmospheric carbon dioxide (CO2) is consumed and base cations are released from silicate minerals to form carbonate and bicarbonate ions, which are finally deposited as carbonate complexes. Continental silicate weathering constitutes a stable carbon sink that is an important influence on long-term climate change, as it sequesters atmospheric carbon dioxide at a million-year time scale. Traditionally, CO2 sequestered through silicate weathering is estimated by measuring the flux of the base cations to watersheds. However, plants also absorb considerable amounts of base cations. Plant biomass is often removed from ecosystems during harvesting. The base cations are subsequently released after decomposition of the harvested plant materials, and thereby enhance CO2 consumption related to weathering. Here, we analyze plant biomass storage-harvest fluxes (production and removal of biomass from forests) of base cations in forests across China to quantify the relative contribution of forest trees to the terrestrial weathering-related carbon sink. Our data suggest that the potential CO2 consumption rate for biomass-related silicate weathering (from the combined action of with afforestation/reforestation, controlled harvesting and rock powder amendment) in Chinese forests is 7.9±4.1Tg CO2yr-1. This represents ~34% of the chemical weathering rate in China. Globally, forests may increase CO2 sequestration through biologically-mediated silicate weathering by ~32%.

Comment on "The extent of forest in dryland biomes".

Bastin et al (Reports, 12 May 2017, p. 635) infer forest as more globally extensive than previously estimated using tree cover data. However, their forest definition does not reflect ecosystem function or biotic composition. These structural and climatic definitions inflate forest estimates across the tropics and undermine conservation goals, leading to inappropriate management policies and practices in tropical grassy ecosystems.

Phytolith carbon sequestration in global terrestrial biomes.

Terrestrial biogeochemical carbon (C) sequestration is coupled with the biogeochemical silicon (Si) cycle through mechanisms such as phytolith C sequestration, but the size and distribution of the phytolith C sink remain unclear. Here, we estimate phytolith C sequestration in global terrestrial biomes. We used biome data including productivity, phytolith and silica contents, and the phytolith stability factor to preliminarily determine the size and distribution of the phytolith C sink in global terrestrial biomes. Total phytolith C sequestration in global terrestrial biomes is 156.7±91.6TgCO2yr-1. Grassland (40%), cropland (35%), and forest (20%) biomes are the dominant producers of phytolith-based carbon; geographically, the main contributors are Asia (31%), Africa (24%), and South America (17%). Practices such as bamboo afforestation/reforestation and grassland recovery for economic and ecological purposes could theoretically double the above phytolith C sink. The potential terrestrial phytolith C sequestration during 2000-2099 under such practices would be 15.7-40.5PgCO2, equivalent in magnitude to the C sequestration of oceanic diatoms in sediments and through silicate weathering. Phytolith C sequestration contributes vitally to the global C cycle, hence, it is essential to incorporate plant-soil silica cycling in biogeochemical C cycle models.

Biodiversity and Topographic Complexity: Modern and Geohistorical Perspectives.

Topographically complex regions on land and in the oceans feature hotspots of biodiversity that reflect geological influences on ecological and evolutionary processes. Over geologic time, topographic diversity gradients wax and wane over millions of years, tracking tectonic or climatic history. Topographic diversity gradients from the present day and the past can result from the generation of species by vicariance or from the accumulation of species from dispersal into a region with strong environmental gradients. Biological and geological approaches must be integrated to test alternative models of diversification along topographic gradients. Reciprocal illumination among phylogenetic, phylogeographic, ecological, paleontological, tectonic, and climatic perspectives is an emerging frontier of biogeographic research.

Linked canopy, climate, and faunal change in the Cenozoic of Patagonia.

Vegetation structure is a key determinant of ecosystems and ecosystem function, but paleoecological techniques to quantify it are lacking. We present a method for reconstructing leaf area index (LAI) based on light-dependent morphology of leaf epidermal cells and phytoliths derived from them. Using this proxy, we reconstruct LAI for the Cenozoic (49 million to 11 million years ago) of middle-latitude Patagonia. Our record shows that dense forests opened up by the late Eocene; open forests and shrubland habitats then fluctuated, with a brief middle-Miocene regreening period. Furthermore, endemic herbivorous mammals show accelerated tooth crown height evolution during open, yet relatively grass-free, shrubland habitat intervals. Our Patagonian LAI record provides a high-resolution, sensitive tool with which to dissect terrestrial ecosystem response to changing Southern Ocean conditions during the Cenozoic.

Applying morphometrics to early land plant systematics: a new Leclercqia (Lycopsida) species from Washington State, USA.

PREMISE OF THE STUDY: Early land plant fossils can be challenging to interpret due to their morphological simplicity and often fragmentary nature. Morphometric techniques using commonly preserved characters might increase diagnostic value of such material. To evaluate the utility of morphometrics in assessing morphospecies boundaries in the Devonian, we compared degrees of variation within the cosmopolitan lycopsid genus Leclercqia with that of living relatives (Lycopodium-Spinulum spp.) Of particular interest was determining whether a new morphotype of Leclercqia from the Middle Devonian Chilliwack flora of Washington State fell within or outside the range of variation of previously described species. METHODS: Morphological variation of Leclercqia was assessed across the geographic range of the genus using six vegetative and three reproductive characters. The new morphotype and two previously described species (L. complexa, L. andrewsii) were compared using linear discriminant analysis (LDA). Extant Lycopodium-Spinulum species and variants were similarly analyzed to assess inter- vs. intraspecific variation in living lycopsids. KEY RESULTS: The LDA comparisons of Lycopodium-Spinulum yielded notable morphological disparity between species but substantial overlap between intraspecific variants. Among the fossils, LDA separates the new morphotype, Leclercqia complexa, and L. andrewsii to a similar degree as Lycopodium and Spinulum species. Based on these results and further study, we describe a new species of Leclercqia: Leclercqia scolopendra Benca et Strömberg sp. nov. CONCLUSIONS: Morphometric analyses can aid in informing taxonomic assignment of fragmentary early land plant fossils using readily preserved features, even in the absence of reproductive structures. Applications of this approach to the Chilliwack flora suggest Leclercqia displayed greater morphological variation, taxonomic diversity, and biogeographic extent than previously thought.

Molecular dating, evolutionary rates, and the age of the grasses.

Many questions in evolutionary biology require an estimate of divergence times but, for groups with a sparse fossil record, such estimates rely heavily on molecular dating methods. The accuracy of these methods depends on both an adequate underlying model and the appropriate implementation of fossil evidence as calibration points. We explore the effect of these in Poaceae (grasses), a diverse plant lineage with a very limited fossil record, focusing particularly on dating the early divergences in the group. We show that molecular dating based on a data set of plastid markers is strongly dependent on the model assumptions. In particular, an acceleration of evolutionary rates at the base of Poaceae followed by a deceleration in the descendants strongly biases methods that assume an autocorrelation of rates. This problem can be circumvented by using markers that have lower rate variation, and we show that phylogenetic markers extracted from complete nuclear genomes can be a useful complement to the more commonly used plastid markers. However, estimates of divergence times remain strongly affected by different implementations of fossil calibration points. Analyses calibrated with only macrofossils lead to estimates for the age of core Poaceae ∼51-55 Ma, but the inclusion of microfossil evidence pushes this age to 74-82 Ma and leads to lower estimated evolutionary rates in grasses. These results emphasize the importance of considering markers from multiple genomes and alternative fossil placements when addressing evolutionary issues that depend on ages estimated for important groups.