Grass Subfamily Research Articles

Phytolith spectra of the monotypic grass Danthonidium gammiei (Bhide) Hubb in Hook (Danthonioideae) from Northern Western Ghats, India: Taxonomic and environmental significance of its distinctive Bilobates

Grass Silica Short Cell Phytoliths (GSSCPs) are regarded as a potential adjunctive tool for grass subfamily identification owing to their definite morphology and affinity towards certain grass subfamilies. In the present study phytolith spectra of the monotypic genus Danthonidium gammiei (Bhide) Hubb in Hook, collected from northern Western Ghats, Maharashtra is studied in detail for the first time. This grass profusely produces Bilobates along with other GSSCPs and Elongate types. A thorough study is undertaken to find out potential of the Danthonidium gammiei Bilobates in distinguishing this grass from other major Bilobate producing grass sub-families like Panicoideae and Aristidoideae. The possible relationship between Bilobates specific for Danthonidium gammiei and the intensity of rainfall in northern Western Ghats is also discussed here. Statistical analyses (PCA, CA, and CCA) also establish the potential of Bilobate phytoliths in identifying Danthonioideae subfamily and discriminating different rainfall zones of the region.

Some unique anatomical scaling relationships among genera in the grass subfamily Pooideae.

Members of the grass family Poaceae have adapted to a wide range of habitats and disturbance regimes globally. The cellular structure and arrangements of leaves can help explain how plants survive in different climates, but these traits are rarely measured in grasses. Most studies are focussed on individual species or distantly related species within Poaceae. While this focus can reveal broad adaptations, it is also likely to overlook subtle adaptations within more closely related groups (subfamilies, tribes). This study, therefore, investigated the scaling relationships between leaf size, vein length area (VLA) and vessel size in five genera within the subfamily Pooideae. The scaling exponent of the relationship between leaf area and VLA was -0.46 (±0.21), which is consistent with previous studies. In Poa and Elymus, however, minor vein number and leaf length were uncorrelated, whereas in Festuca these traits were positively correlated (slope = 0.82±0.8). These findings suggest there are broad-scale and fine-scale variations in leaf hydraulic traits among grasses. Future studies should consider both narrow and broad phylogenetic gradients.

Taxonomic and environmental significance of Poaceae and Cyperaceae phytoliths from the Northern Territory, Australia

Phytoliths are a valuable tool for reconstructing past grassy ecosystems. However, they are not commonly utilised as a palaeoecological proxy in Australia due to a lack of modern reference material from plants, particularly grasses. This study analyses phytolith reference material from 49 grass and 4 sedge species from the Northern Territory, Australia. This is used to develop a detailed classification scheme and multivariate morphospace analysis to examine taxonomic and ecological patterns of grass silica short cell phytolith (GSSCP) production within the Poaceae (grass) family. The results of detrended correspondence analysis show that grass subfamilies, tribes, and genera in this region can be differentiated through phytolith assemblages, and that these assemblages reflect the environmental conditions associated with various grass taxa. The developed detailed classification system and examination of co-occurrence patterns reduces redundancy of phytolith morphotypes across subfamilies, improving taxonomic and palaeoecological resolution of phytolith assemblages. Finally, based on the results of this study, a detailed scheme is presented for palaeoecological reconstructions in grassy ecosystems of northern Australia.

Genome-wide analysis and expression of the aquaporin gene family in Avena sativa L.

Oat (Avena sativa L.) belongs to the early maturity grass subfamily of the Gramineae subfamily oats (Avena) and has excellent characteristics, such as tolerance to barrenness, salt, cold, and drought. Aquaporin (AQP) proteins belong to the major intrinsic protein (MIP) superfamily, are widely involved in plant growth and development, and play an important role in abiotic stress responses. To date, previous studies have not identified or analyzed the AsAQP gene family system, and functional studies of oat AQP genes in response to drought, cold, and salt stress have not been performed. In this study, AQP genes (AsAQP) were identified from the oat genome, and various bioinformatics data on the AQP gene family, gene structure, gene replication, promoters and regulatory networks were analyzed. Quantitative real-time PCR technology was used to verify the expression patterns of the AQP gene family in different oat tissues under different abiotic stresses. In this study, a total of 45 AQP genes (AsAQP) were identified from the oat reference genome. According to a phylogenetic analysis, 45 AsAQP were divided into 4 subfamilies (PIP, SIP, NIP, and TIP). Among the 45 AsAQP, 23 proteins had interactions, and among these, 5AG0000633.1 had the largest number of interacting proteins. The 20 AsAQP genes were expressed in all tissues, and their expression varied greatly among different tissues and organs. All 20 AsAQP genes responded to salt, drought and cold stress. The NIP subfamily 6Ag0000836.1 gene was significantly upregulated under different abiotic stresses and could be further verified as a key candidate gene. The findings of this study provide a comprehensive list of members and their sequence characteristics of the AsAQP protein family, laying a solid theoretical foundation for further functional analysis of AsAQP in oats. This research also offers valuable reference for the creation of stress-tolerant oat varieties through genetic engineering techniques.

Chloridoideae grass phytoliths can distinguish two different rainfall zones: A case study from northern parts of Western Ghats, India

Grass silica short cell phytoliths (GSSCPs), which are profusely produced within the grasses are often subfamily-specific and hence may act as one of the reliable plant proxies in deciphering eco-climatic conditions of an area. To assess how reliably grass phytolith morphs can capture the signal of rainfall variations of an area and to overcome the bias caused due to the redundancy and the multiplicity of GSSCPs, some Chloridoid grasses and soil samples from Maharashtra part of the Western Ghats along the west-east rainfall gradient are studied. The study reveals that Saddle are the dominant morphotype produced by the members of this grass subfamily irrespective of the rainfall variations. Surface soil phytolith assemblage revealed that Saddle morphs are inversely proportional to rainfall i.e., they increase with decrease in rainfall and vice versa. Based on this data, regions with MAP >2000 mm and <2000 mm could be separated in the northern Western Ghats. The results of the CCA and PCA also corroborated these observations.

A genome assembly for Orinus kokonorica provides insights into the origin, adaptive evolution and further diversification of two closely related grass genera

Ancient whole-genome duplication (WGD) or polyploidization is prevalent in plants and has played a crucial role in plant adaptation. However, the underlying genomic basis of ecological adaptation and subsequent diversification after WGD are still poorly understood in most plants. Here, we report a chromosome-scale genome assembly for the genus Orinus (Orinus kokonorica as representative) and preform comparative genomics with its closely related genus Cleistogenes (Cleistogenes songorica as representative), both belonging to a newly named subtribe Orininae of the grass subfamily Chloridoideae. The two genera may share one paleo-allotetraploidy event before 10 million years ago, and the two subgenomes of O. kokonorica display neither fractionation bias nor global homoeolog expression dominance. We find substantial genome rearrangements and extensive structural variations (SVs) between the two species. With comparative transcriptomics, we demonstrate that functional innovations of orthologous genes may have played an important role in promoting adaptive evolution and diversification of the two genera after polyploidization. In addition, copy number variations and extensive SVs between orthologs of flower and rhizome related genes may contribute to the morphological differences between the two genera. Our results provide new insights into the adaptive evolution and subsequent diversification of the two genera after polyploidization.

Soil surface grass phytolith morphotypes across bioclimatic gradients and biomes at about latitude 29°S in South Africa

This study examines the distribution patterns of grass subfamilies and their diagnostic silica phytoliths along a transect running approximately at latitude 28° S in South Africa. The transect encompasses 77 surface samples collected along moisture and temperature gradients, across 6 biomes and 39 ecozones, and spanning elevations between near-sea level and over 2253 m. This study tests the relation between the distribution and density of grass subfamilies and the proportions of their diagnostic grass silica short cell phytoliths (GSSCP) representing them, and how these proportions correlate with five climatic variables (mean temperature of the wettest quarter, annual precipitation, precipitation of the wettest quarter, precipitation variability, and the aridity index). The results suggest that C3-grass diagnostic GSSCP morphotypes increase with a decline in temperature in the wettest quarter, occurring in areas receiving mostly winter rain or at high elevations in areas of summer rain. Of the C4 grass subfamilies, the Panicoideae increase with summer precipitation and low precipitation variability, while the Aristidoideae-Stipagrostis the opposite. The distribution of Chloridoideae depends on complex changes in all climatic variables, suggesting aridity gradients only in the eastern half of the transect. In view of the data, this study reviews the usefulness of the phytolith humidity-aridity index (Iph) and the water stress index (Fs) and proposes the Panicoideae index (PI) and Stipagrostis index (SI) as indicator of aridity and high precipitation variability in southern Africa.

Leaf transformation for efficient random integration and targeted genome modification in maize and sorghum

Transformation in grass species has traditionally relied on immature embryos and has therefore been limited to a few major Poaceae crops. Other transformation explants, including leaf tissue, have been explored but with low success rates, which is one of the major factors hindering the broad application of genome editing for crop improvement. Recently, leaf transformation using morphogenic genes Wuschel2 (Wus2) and Babyboom (Bbm) has been successfully used for Cas9-mediated mutagenesis, but complex genome editing applications, requiring large numbers of regenerated plants to be screened, remain elusive. Here we demonstrate that enhanced Wus2/Bbm expression substantially improves leaf transformation in maize and sorghum, allowing the recovery of plants with Cas9-mediated gene dropouts and targeted gene insertion. Moreover, using a maize-optimized Wus2/Bbm construct, embryogenic callus and regenerated plantlets were successfully produced in eight species spanning four grass subfamilies, suggesting that this may lead to a universal family-wide method for transformation and genome editing across the Poaceae.

Independent recruitment of FRUITFULL-like transcription factors in the convergent origins of vernalization-responsive grass flowering

Flowering in response to low temperatures (vernalization) has evolved multiple times independently across angiosperms as an adaptation to match reproductive development with the short growing season of temperate habitats. Despite the context of a generally conserved flowering time network, evidence suggests that the genes underlying vernalization responsiveness are distinct across major plant clades. Whether different or similar mechanisms underlie vernalization-induced flowering at narrower (e.g., family-level) phylogenetic scales is not well understood. To test the hypothesis that vernalization responsiveness has evolved convergently in temperate species of the grass family (Poaceae), we carried out flowering time experiments with and without vernalization in several representative species from different subfamilies. We then determined the likelihood that vernalization responsiveness evolved through parallel mechanisms by quantifying the response of Pooideae vernalization pathway FRUITFULL (FUL)-like genes to extended periods of cold. Our results demonstrate that vernalization-induced flowering has evolved multiple times independently in at least five grass subfamilies, and that different combinations of FUL-like genes have been recruited to this pathway on several occasions.

Genomic insights into the origin, adaptive evolution, and herbicide resistance of Leptochloa chinensis, a devastating tetraploid weedy grass in rice fields

Chinese sprangletop (Leptochloa chinensis), belonging to the grass subfamily Chloridoideae, is one of the most notorious weeds in rice ecosystems. Here, we report a chromosome-scale reference genome assembly and a genomic variation map of the tetraploid L. chinensis. The L. chinensis genome is derived from two diploid progenitors that diverged ∼10.9 million years ago, and its two subgenomes display neither fractionation bias nor overall gene expression dominance. Comparative genomic analyses reveal substantial genome rearrangements in L. chinensis after its divergence from the common ancestor of Chloridoideae and, together with transcriptome profiling, demonstrate the important contribution of tetraploidization to the gene sources for the herbicide resistance of L. chinensis. Population genomic analyses of 89 accessions from China reveal that L. chinensis accessions collected from southern/southwestern provinces have substantially higher nucleotide diversity than those from the middle and lower reaches of the Yangtze River, suggesting that L. chinensis spread in China from the southern/southwestern provinces to the middle and lower reaches of the Yangtze River. During this spread, L. chinensis developed significantly increased herbicide resistance, accompanied by the selection of numerous genes involved in herbicide resistance. Taken together, our study generated valuable genomic resources for future fundamental research and agricultural management of L. chinensis, and provides significant new insights into the herbicide resistance as well as the origin and adaptive evolution of L. chinensis.

Phylotranscriptomics Resolves the Phylogeny of Pooideae and Uncovers Factors for Their Adaptive Evolution.

Adaptation to cool climates has occurred several times in different angiosperm groups. Among them, Pooideae, the largest grass subfamily with ∼3,900 species including wheat and barley, have successfully occupied many temperate regions and play a prominent role in temperate ecosystems. To investigate possible factors contributing to Pooideae adaptive evolution to cooling climates, we performed phylogenetic reconstruction using five gene sets (with 1,234 nuclear genes and their subsets) from 157 transcriptomes/genomes representing all 15 tribes and 24 of 26 subtribes. Our phylogeny supports the monophyly of all tribes (except Diarrheneae) and all subtribes with at least two species, with strongly supported resolution of their relationships. Molecular dating suggests that Pooideae originated in the late Cretaceous, with subsequent divergences under cooling conditions first among many tribes from the early middle to late Eocene and again among genera in the middle Miocene and later periods. We identified a cluster of gene duplications (CGD5) shared by the core Pooideae (with 80% Pooideae species) near the Eocene–Oligocene transition, coinciding with the transition from closed to open habitat and an upshift of diversification rate. Molecular evolutionary analyses homologs of CBF for cold resistance uncovered tandem duplications during the core Pooideae history, dramatically increasing their copy number and possibly promoting adaptation to cold habitats. Moreover, duplication of AP1/FUL-like genes before the Pooideae origin might have facilitated the regulation of the vernalization pathway under cold environments. These and other results provide new insights into factors that likely have contributed to the successful adaptation of Pooideae members to temperate regions.

The role of plants in the formation of species-specific features in grass flies (Diptera, Chloropidae, Meromyza).

In the current manuscript, we present the results of comparative analysis of seven species of Meromyza flies in the “variegata” cluster and of the evolutionary close species M.inornata, based the following criteria: 1) 14 external key features; 2) shape and area of the anterior processes of postgonites; 3) mtDNA CO1 region and 4) host plant diversity data. We could demonstrate the primary role of host plants in species formation inside genus Meromyza and calculated the timing of the divergence of M.inornata and the species of “variegata” cluster. Based on our estimates of evolution rate for mtDNA CO1 gene, we could conclude that that divergence of herbs happened before the speciation of grass flies Meromyza. Meromyza species, close to the ancestral species of the cluster, are adapted to the wide range of host plants. We revealed the most informative variables h1, S and Plant analysing data with the following statistical methods: linear discriminant analysis - LDA, regularised discriminant analysis - RDA, flexible discriminant analysis – FDA and probabilistic neural network - PNN. The highest classification accuracy was achieved using PNN (99%) and the lowest when using LDA (95.8%). When the Plant trait was excluded, the classification accuracy decreased by 14%. We revealed the significant trends in size change of the anterior process of the postgonite amongst studies species. This morphological structure is an element of male reproductive apparatus critical for the restriction of interspecies mating. We determined three branches of speciation in the “variegata” cluster and five trends in the evolution of this cluster, based on the external morphological features. We showed that M.variegata and especially M.mosquensis, the species closest to the ancestral haplotype, have the largest number of features typical of those of M.inornata. Based on the external features and the area of the anterior process of the postgonite, we reconstructed the phylogenetic position of M.elbergi in the cluster. In accordance with the obtained outcomes, we could conclude that the distribution, species diversity and the adaptation of the grass flies to narrow oligophagy were directly connected to host plant diversity. The adaptation to different host plants could be the main factor in divergence of grass flies and their evolution started later than the diversification in the Pooideae subfamily of grasses.

Phylogenetic, ecological and intraindividual variability patterns in grass phytolith shape.

Grass silica short cell (GSSC) phytoliths appear to be the most reliable source of fossil evidence for tracking the evolutionary history and paleoecology of grasses. In recent years, modern techniques that quantitatively assess phytolith shape variation have widened opportunities for the classification of grass fossil phytoliths. However, phylogenetic, ecological and intraindividual variability patterns in phytolith shape remain largely unexplored. The full range of intraindividual phytolith shape variation [3650 two-dimensional (2-D) outlines] from 73 extant grass species, 48 genera, 18 tribes and eight subfamilies (particularly Pooideae) was analysed using geometric morphometric analysis based on semi-landmarks spanning phytolith outlines. The 2-D phytolith shape is mainly driven by deep-time diversification of grass subfamilies. There is distinct phytolith shape variation in early-diverging lineages of Pooideae (Meliceae, Stipeae). The amount of intraindividual variation in phytolith shape varies among species, resulting in a remarkable pattern across grass phylogeny. The phylogenetic pattern in phytolith shape was successfully revealed by applying geometric morphometrics to 2-D phytolith shape outlines, strengthening the potential of phytoliths to track the evolutionary history and paleoecology of grasses. Geometric morphometrics of 2-D phytolith shape is an excellent tool for analysis requiring large numbers of phytolith outlines, making it useful for quantitative palaeoecological reconstruction.

The Streptochaeta Genome and the Evolution of the Grasses.

In this work, we sequenced and annotated the genome of Streptochaeta angustifolia, one of two genera in the grass subfamily Anomochlooideae, a lineage sister to all other grasses. The final assembly size is over 99% of the estimated genome size. We find good collinearity with the rice genome and have captured most of the gene space. Streptochaeta is similar to other grasses in the structure of its fruit (a caryopsis or grain) but has peculiar flowers and inflorescences that are distinct from those in the outgroups and in other grasses. To provide tools for investigations of floral structure, we analyzed two large families of transcription factors, AP2-like and R2R3 MYBs, that are known to control floral and spikelet development in rice and maize among other grasses. Many of these are also regulated by small RNAs. Structure of the gene trees showed that the well documented whole genome duplication at the origin of the grasses (ρ) occurred before the divergence of the Anomochlooideae lineage from the lineage leading to the rest of the grasses (the spikelet clade) and thus that the common ancestor of all grasses probably had two copies of the developmental genes. However, Streptochaeta (and by inference other members of Anomochlooideae) has lost one copy of many genes. The peculiar floral morphology of Streptochaeta may thus have derived from an ancestral plant that was morphologically similar to the spikelet-bearing grasses. We further identify 114 loci producing microRNAs and 89 loci generating phased, secondary siRNAs, classes of small RNAs known to be influential in transcriptional and post-transcriptional regulation of several plant functions.

TaWRKY10 transcription factor is a novel jasmonic acid signalling regulator involved in immunity against Septoria tritici blotch disease in wheat

Septoria tritici blotch (STB), caused by the fungus Zymoseptoria tritici, is currently the main threat to worldwide wheat production in temperate climates. Understanding the genetic mechanisms that underpin the Z. tritici–wheat interaction will be crucial for generating new control strategies against STB. Plant hormones are essential regulators of growth and immunity. In particular, jasmonic acid (JA) plays a central role in defence against necrotrophic plant pathogens, but its role in mediating immunity against Z. tritici is largely unknown. Here we identify the transcription factor TaWRKY10 that is taxonomically restricted to the grass subfamily Pooideae as a novel regulator of JA responses in wheat. We demonstrate by using virus‐induced gene silencing that silencing TaWRKY10 leads to increased resistance against Z. tritici with an earlier onset of necrotic symptoms, but with reduced pathogen sporulation. Wheat plants silenced for TaWRKY10 show an up‐regulated JA response. Transcriptional profiling of TaWRKY10 knock‐down plants indicates that it is a negative regulator of the JA receptor TaCOI1 gene expression. Our data indicate that TaWRKY10 down‐regulates JA responses, contributing to the susceptibility of wheat to Z. tritici. We postulate that manipulating TaWRKY10 may provide a strategy to boost STB resistance in wheat.

A 313 plastome phylogenomic analysis of Pooideae: Exploring relationships among the largest subfamily of grasses

In this study, we analyzed 313 plastid genomes (plastomes) of Poaceae with a focus on expanding our current knowledge of relationships among the subfamily Pooideae, which represented over half the dataset (164 representatives). In total, 47 plastomes were sequenced and assembled for this study. This is the largest study of its kind to include plastome-level data, to not only increase sampling at both the taxonomic and molecular levels with the aim of resolving complex and reticulate relationships, but also to analyze the effects of alignment gaps in large-scale analyses, as well as explore divergences in the subfamily with an expanded set of 14 accepted grass fossils for more accurate calibrations and dating. Incorporating broad systematic assessments of Pooideae taxa conducted by authors within the last five years, we produced a robust phylogenomic reconstruction for the subfamily, which included all but two supergeneric taxa (Calothecinae and Duthieeae).We further explored how including alignment gaps in plastome analyses oftentimes can produce incorrect or misinterpretations of complex or reticulate relationships among taxa of Pooideae. This presented itself as consistently changing relationships at specific nodes for different stripping thresholds (percentage-based removal of gaps per alignment column). Our summary recommendation for large-scale genomic plastome datasets is to strip alignment columns of all gaps to increase pairwise identity and reduce errant signal from poly A/T bias. To do this we used the “mask alignment” tool in Geneious software. Finally, we determined an overall divergence age for Pooideae of roughly 84.8 Mya, which is in line with, but slightly older than most recent estimates.

Cross-reactivity in Skin Prick Test Results of Members Within Pooideae Subfamily.

ObjectiveMolecular similarities of grass pollen antigens have led to the view that cross-reactivity exists within members of the Pooideae subfamily of grasses. This has resulted in testing for only the most antigenically representative member of Pooideae, Timothy grass (Phleum pratense), despite little literature to support the claim that Phleum is the most representative member or that in vitro cross-reactivity correlates with in vivo cross-reactivity. The aim of the study was to determine if patients with allergic rhinitis symptoms and positive skin prick test results to meadow fescue (Festuca pratensis) also have positive results to Timothy grass.Study DesignRetrospective cross-sectional study.SettingTertiary care center in middle Missouri.MethodsA retrospective chart review identified patients ≥12 years old with a diagnosis of allergic rhinitis who underwent skin prick testing between March 2016 and July 2018, by using a search with CPT code 95004 (Current Procedural Terminology). Positive skin prick test results were based on wheal produced ≥3 mm than the negative control.ResultsAfter review of 2182 charts, 1587 patients met criteria to test for Phleum and Festuca. In total, 1239 patients had a positive result for Phleum or Festuca. Of these, 479 (38.6%) tested positive for Festuca alone, while 342 (27.6%) and 418 (33.7%) tested positive for Phleum alone and Phleum+Festuca, respectively.ConclusionClinical cross-reactivity among Pooideae members may not be as complete as traditionally thought. P pratense may not be the most antigenically representative subfamily member, and other grasses may need to be included in skin prick testing.

Phytolith records of flourishing early Holocene Pooideae linked to an 8.2 ka cold event in subtropical China

The grass subfamily Pooideae originated in a temperate niche during the late Cretaceous; it is the largest Poaceae subfamily, consisting of almost 4,000 species, which are distributed worldwide. Pooideae responses to climate changes at different time scales, and different ecological zones are thus important in understanding Poaceae evolutionary processes and their relationship with climate change. In the study described in this article, we reconstructed Pooideae variability during the early Holocene, as inferred by a phytolith sequence from the Lower Yangtze in subtropical China. The phytolith assemblage was marked by three increases in Pooideae phytoliths, dated to ca 8.4–8.0, 7.8–7.6, and 7.4–7.2 ka BP (before present, 1950 AD), with each representing pronounced increases in Pooideae extent and distribution. All these increases were within age ranges that agreed well with the timing of weak Asian Monsoon events, at 8.2, 7.7, and 7.3 ka BP. The first Pooideae flourishing period in subtropical China was the most significant, lasting for approximately four centuries and being characterized by a double peak, which equated with an event at 8.2 ka. This suggested that cold and/or dry conditions—which occurred over a period of several hundred years and were linked to weakening of the Asian monsoon—probably caused Pooideae to flourish in the Lower Yangtze region. Comparison of two diagnostic trapezoid phytolith types—namely wavy and wavy narrow—which showed different changes between ca 8.4 and 8.0 ka BP, suggested that they responded differently to the climate change represented by the 8.2 ka event. Our phytolith records have provided not only new data clarifying the detailed Pooideae response to the 8.2 ka event but also a reliable index for past cold climates in subtropical China.

Palaeovegetation changes recorded in Palaeolake Olduvai OGCP Core 2A (2.09–2.12 Ma) Naibor Soit Formation Olduvai Gorge, Tanzania

For five decades Olduvai Gorge has been a key site to reconstruct and understand the relationship between environmental and landscape conditions and use of affordances by early African hominin populations. Following the first Olduvai Gorge Coring project (OGCP) during 2014, a multiproxy microbiological analysis, which includes phytoliths, pollen, diatoms, sponge spicules and chrysophyte cysts, was undertaken on samples collected from various borehole cores. The aim of the study is to better understand palaeoenvironmental and palaeovegetation conditions and changes through time and their relationship to hominin presence and evolution. This study details the first palaeobotanical and palaeoenvironmental study of Borehole 2A at Olduvai Gorge. It represents the as yet oldest known sedimentary sequence in the Olduvai Basin for a portion of the pre-Bed I Naibor Soit Formation; this is a unit that is not accessible in any natural exposures in Olduvai Gorge, and has only recently been encountered by drilling. Here we present the results from a particularly phytolith-prone portion between ~2.09 Ma and 2.12 Ma.Phytolith results indicate a savannah environment dominated by grasses, where Poaceae were a key component and where the C3 Pooideae grasses were mostly dominant, alternating with C4 grasses. Oscillations between grass subfamilies, C3 Pooideae, C4 Chloridoideae, and C4 Panicoideae to lesser degrees, indicate five substantial climatic shifts, varying between more humid and arid conditions. Associated with phytoliths, freshwater indicators such as diatoms, sponge spicules and chrysophyte cysts were also identified, suggesting the presence of wetlands in the lake catchment area. Pollen is extremely rare in the sediments but when present, comprises fungal spores and Poaceae pollen, thus supporting the wetland and grassland reconstructions, respectively. These results offer for the first time, a whole picture of the palaeovegetation and associated palaeoenvironments for this pre-Bed I period. Together with previous results from other areas and chronological periods, they improve our understanding of the evolution and adaptation of early hominins and their close relationship to the surrounding landscape.

Increased above-ground resource allocation is a likely precursor for independent evolutionary origins of annuality in the Pooideae grass subfamily.

Semelparous annual plants flower a single time during their 1-yr life cycle, investing much of their energy into rapid reproduction. By contrast, iteroparous perennial plants flower multiple times over several years, and partition their resources between reproduction and persistence. To which extent evolutionary transitions between life-cycle strategies are internally constrained at the developmental, genetic and phylogenetic level is unknown. Here we study the evolution of life-cycle strategies in the grass subfamily Pooideae and test if transitions between them are facilitated by evolutionary precursors. We integrate ecological, life-cycle strategy and growth data in a phylogenetic framework. We investigate if growth traits are candidates for a precursor. Species in certain Pooideae clades are predisposed to evolve annuality from perenniality, potentially due to the shared inheritance of specific evolutionary precursors. Seasonal dry climates, which have been linked to annuality, were only able to select for transitions to annuality when the precursor was present. Allocation of more resources to above-ground rather than below-ground growth is a candidate for the precursor. Our findings support the hypothesis that only certain lineages can respond quickly to changing external conditions by switching their life-cycle strategy, likely due to the presence of evolutionary precursors.