Evergreen broad-leaved forests (EBLFs) are widely distributed in East Asia and play a vital role in ecosystem stability. The occurrence of these forests in East Asia has been a subject of debate across various disciplines. In this study, we explored the occurrence of East Asian EBLFs from a paleobotanical perspective. By collecting plant fossils from four regions in East Asia, we have established the evolutionary history of EBLFs. Through floral similarity analysis and paleoclimatic reconstruction, we have revealed a diverse spatio-temporal pattern for the occurrence of EBLFs in East Asia. The earliest occurrence of EBLFs in southern China can be traced back to the middle Eocene, followed by southwestern China during the late Eocene-early Oligocene. Subsequently, EBLFs emerged in Japan during the early Oligocene and eventually appeared in central-eastern China around the Miocene. Paleoclimate simulation results suggest that the precipitation of wettest quarter (PWetQ, mm) exceeding 600 mm is crucial for the occurrence of EBLFs. Furthermore, the heterogeneous occurrence of EBLFs in East Asia is closely associated with the evolution of the Asian Monsoon. This study provides new insights into the occurrence of EBLFs in East Asia.

Angiosperms experienced one of the most remarkable radiations of land plants and are now the dominant autotrophs in terrestrial ecosystems. Recent phylogenomic studies based on large-scale data from plastid, mitochondrial, or nuclear transcriptomes/genomes and increased taxon sampling have provided unprecedent resolution into the phylogeny of flowering plants. However, owing to ancient rapid radiations, the interrelationships among the five lineages of Mesangiospermae, the vast majority of angiosperms, remain contentious. Here we show that, although plastid and mitochondrial genomes lack sufficient phylogenetic signal for resolving deeper phylogeny, the relationships among five mesangiosperm lineages can be confidently resolved under better-fitting models using genome-scale data. According to our Bayesian cross-validation and model test in a maximum likelihood framework, site-heterogeneous models (e.g., CAT-GTR + G4, LG + C20 + F + G) outperform site-homogeneous or partition models often used in previous studies. Under site-heterogeneous models, the approximately unbiased test favored our preferred tree recovered from various datasets: Ceratophyllales (coontails) are robustly recovered as sister to monocots, and they together are sister to the clade comprising magnoliids, Chloranthales, and eudicots. Our phylogenomic analyses resolve the last enigma of the deeper phylogeny of angiosperms and emphasize the efficacy of modeling compositional heterogeneity in resolving rapid radiations of plants.

Saussurea is one of the largest and most rapidly evolving genera within the Asteraceae, comprising approximately 520 species from the Northern Hemisphere. A comprehensive infrageneric classification, supported by robust phylogenetic trees and corroborated by morphological and other data, has not yet been published. For the first time, we recovered a well-resolved nuclear phylogeny of Saussurea consisting of four main clades, which was also supported by morphological data. Our analyses show that ancient hybridization is the most likely source of deep cytoplasmic-nuclear conflict in Saussurea, and a phylogeny based on nuclear data is more suitable than one based on chloroplast data for exploring the infrageneric classification of Saussurea. Based on the nuclear phylogeny obtained and morphological characters, we proposed a revised infrageneric taxonomy of Saussurea, which includes four subgenera and 13 sections. Specifically, 1) S. sect. Cincta, S. sect. Gymnocline, S. sect. Lagurostemon, and S. sect. Strictae were moved from S. subg. Saussurea to S. subg. Amphilaena, 2) S. sect. Pseudoeriocoryne was moved from S. subg. Eriocoryne to S. subg. Amphilaena, and 3) S. sect. Laguranthera was moved from S. subg. Saussurea to S. subg. Theodorea.

Dryopteridaceae are the largest fern family and include nearly 20% of extant fern diversity, with 24 currently recognized genera. Recognition and delineation of genera within this family have varied greatly. The three-subfamily classification of Dryopteridaceae was based primarily on molecular phylogenetic relationships but lacked morphological evidence, and the phylogenetic relationships of the subfamilies and genera of Dryopteridaceae are only partially resolved. A comprehensive and robust phylogeny is urgently needed. The heterogeneous morphology of the current members of Dryopteridaceae makes the family and its subfamilies difficult to define by single morphological characteristics or even character combinations. We carried out phylogenetic analyses to reconstruct a highly supported phylogeny of Dryopteridaceae. Our analyses recovered 24 strongly supported clades grouped into seven major clades of Dryopteridaceae. Seven morphological characters including habit, rhizome shape, frond morphology, rachis-costae architecture, appendages on stipe base and lamina, and soral arrangement were found to be informative for identifying different major clades and clades in Dryopteridaceae. Based on phylogenetic reconstruction and morphological analysis, we presented an updated infra-familial classification of Dryopteridaceae with seven subfamilies and 24 genera including four newly proposed subfamilies (Ctenitidoideae, Lastreopsidoideae, Pleocnemioideae, and Polystichopsidoideae). Morphological character combinations of each subfamily are summarized, and a key is provided. Most genera sensu PPG I are recognized, with Stigmatopetris reclassified into Dryopteridoideae and Arthrobotrya considered a synonym of Teratophyllum. A new genus Pseudarachniodes is introduced. This revised classification will serve as a foundational framework for future investigations on taxonomy, biogeography, and diversification of the most species-rich Dryopteridaceae in ferns.

Understanding the ecological adaptation of tree species can not only reveal the evolutionary potential but also benefit biodiversity conservation under global climate change. Quercus is a keystone genus in Northern Hemisphere forests, and its wide distribution in diverse ecosystems and long evolutionary history make it an ideal model for studying the genomic basis of ecological adaptations. Here we used a newly sequenced genome of Quercus gilva, an evergreen oak species from East Asia, with 18 published Fagales genomes to determine how Fagaceae genomes have evolved, identify genomic footprints of ecological adaptability in oaks in general, as well as between evergreen and deciduous oaks. We found that oak species exhibited a higher degree of genomic conservation and stability, as indicated by the absence of large-scale chromosomal structural variations or additional whole-genome duplication events. In addition, we identified expansion and tandem repetitions within gene families that contribute to plant physical and chemical defense (e.g., cuticle biosynthesis and oxidosqualene cyclase genes), which may represent the foundation for the ecological adaptation of oak species. Circadian rhythm and hormone-related genes may regulate the habits of evergreen and deciduous oaks. This study provides a comprehensive perspective on the ecological adaptations of tree species based on phylogenetic, genome evolutionary, and functional genomic analyses.

Rapid growth is an innovative trait of woody bamboos that has been widely studied. However, the genetic basis and evolution of this trait are poorly understood. Taking advantage of genomic resources of 11 representative bamboos at different ploidal levels, we integrated morphological, physiological, and transcriptomic datasets to investigate rapid growth. In particular, these bamboos include two large-sized and a small-sized woody species, compared with a diploid herbaceous species. Our results showed that gibberellin A1 was important for the rapid shoot growth of the world's largest bamboo, Dendrocalamus sinicus, and indicated that two gibberellins (GAs)-related genes, KAO and SLRL1, were key to the rapid shoot growth and culm size in woody bamboos. The expression of GAs-related genes exhibited significant subgenome asymmetry with subgenomes A and C demonstrating expression dominance in the large-sized woody bamboos while the generally submissive subgenomes B and D dominating in the small-sized species. The subgenome asymmetry was found to be correlated with the subgenome-specific gene structure, particularly UTRs and core promoters. Our study provides novel insights into the molecular mechanism and evolution of rapid shoot growth following allopolyploidization in woody bamboos, particularly via subgenome asymmetry. These findings are helpful for understanding of how polyploidization in general and subgenome asymmetry in particular contributed to the origin of innovative traits in plants.

Exploring the worldwide patterns of endemism and the processes that lead to the formation of high-endemism centers is crucial in biogeography. This study examines the geographic distribution and ecological influences on the endemism of liverworts across 390 regions worldwide. We assess phylogenetic endemism and relative phylogenetic endemism in relation to eleven environmental factors, which represent current and Quaternary climate variations, as well as topographic and environmental heterogeneity. Areas with higher endemism in liverworts tend to have higher temperatures, precipitation, and environmental heterogeneity, but lower temperature seasonality and lesser impacts from Quaternary climate changes. Regions exhibiting notably high endemism are predominantly found in tropical Asia, Madagascar, eastern Australia, and the Andes, while those with notably low endemism are generally in temperate Eurasia and North America, parts of Africa, and eastern South America. Centers of neo-endemism are mainly in southern Africa, whereas centers of paleo-endemism are in southern South America, tropical Asia, and New Zealand. Environment variability is a more significant predictor of phylogenetic endemism than current climate conditions, which are themselves more predictive than variables related to Quaternary climate changes. Nevertheless, these three types of explanatory variables combined explain only about one-third of the variance in phylogenetic endemism.

Latitudinal patterns of tree β-diversity reveal important insights into the biogeographical processes that influence forest ecosystems. Although previous studies have extensively documented β-diversity within relatively small spatial extents, the potential drivers of β-diversity along latitudinal gradients are still not well understood at larger spatial extents. In this study, we determined whether tree β-diversity is correlated with latitude in forests of southeastern China, and if so, what ecological processes contribute to these patterns of tree β-diversity. We specifically aimed to disentangle the relative contributions from interspecific aggregation and environmental filtering across various spatial extents. We delineated regional communities comprising multiple nearby national forest inventory (NFI) plots around random focal plots. The number of NFI plots in a regional community served as a surrogate for spatial extent. We also used a null model to simulate randomly assembled communities and quantify the deviation (β-deviation) between observed and expected β-diversity. We found that β-diversity decreased along a latitudinal gradient and that this pattern was clearer at larger spatial extents. In addition, latitudinal patterns of β-deviation were explained by the degree of species spatial aggregation. We also identified environmental factors that drive β-deviation in these forests, including precipitation, seasonality, and temperature variation. At larger spatial extents, these environmental variables explained up to 84% of the β-deviation. Our results reinforce that ecological processes are scale-dependent and collectively contribute to the β-gradient in subtropical forests. We recommend that conservation efforts maintain diverse forests and heterogeneous environments at multiple spatial extents to mitigate the adverse effects of climate change.

Understanding the underlying mechanisms driving species assembly along elevational gradients is of great interest in ecology and biogeography. The Himalaya is one of the world’s hotspots of biodiversity, and the elevational gradient of the central Himalaya in Nepal is one of the longest elevational gradients in the world. Mosses are important constituents of vegetation in the Himalaya, and knowledge of geographic patterns and ecological causes of phylogenetic structure of mosses along elevational gradients in the Himalaya is critical to understanding the assembly of plant diversity in general, and moss diversity in particular, in the Himalaya. Here, we investigate the relationships of phylogenetic structure metrics reflecting different evolutionary depths with elevation and climatic variables representing mean temperature and precipitation conditions, climate extremes, and climate seasonality for mosses distributed along an elevational gradient spanning about 5000 m in the central Himalaya, Nepal. For a given climatic variable, different metrics of phylogenetic structure show different spatial and climatic patterns, but all phylogenetic metrics standardized for species richness show that phylogenetic dispersion in moss assemblages tend to increase with increasing elevation and decreasing temperature. The standardized effect size of mean pairwise distance of moss assemblages shows a triphasic (zig-zag) pattern, which is generally consistent with the triphasic patterns previously found in angiosperms and ferns along the same elevational gradient. Our study shows that temperature-related variables and climate seasonality variables are more important drivers of phylogenetic dispersion in mosses in Nepal, compared with precipitation-related variables and climate extreme variables, respectively.

Patterns and drivers of species-genetic diversity correlations (SGDCs) have been broadly examined across taxa and ecosystems and greatly deepen our understanding of how biodiversity is maintained. However, few studies have examined the role of canopy structural heterogeneity, which is a defining feature of forests, in shaping SGDCs. Here, we determine what factors contribute to α- and β-species-genetic diversity correlations (i.e., α- and β-SGDCs) in a Chinese subtropical forest. For this purpose, we used neutral molecular markers to assess genetic variation in almost all adult individuals of the dominant tree species, Lithocarpus xylocarpus, across plots in the Ailaoshan National Natural Reserve. We also quantified microhabitat variation by quantifying canopy structure heterogeneity with airborne laser scanning on 20 1-ha subtropical forest plots. We found that species α-diversity was negatively correlated with genetic α-diversity. Canopy structural heterogeneity was positively correlated with species α-diversity but negatively correlated with genetic α-diversity. These contrasting effects contributed to the formation of a negative α-SGDC. Further, we found that canopy structural heterogeneity increases species α-diversity and decreases genetic α-diversity by reducing the population size of target species. Species β-diversity, in contrast, was positively correlated with genetic β-diversity. Differences in canopy structural heterogeneity between plots had non-linear parallel effects on the two levels of β-diversity, while geographic distance had a relatively weak effect on β-SGDC. Our study indicates that canopy structural heterogeneity simultaneously affects plot-level community species diversity and population genetic diversity, and species and genetic turnover across plots, thus driving α- and β-SGDCs.

DNA barcoding has been extensively used for species identification. However, species identification of mixed samples or degraded DNA is limited by current DNA barcoding methods. In this study, we use plant species in Juglandaceae to evaluate an assembly-free reads accurate identification (AFRAID) method of species identification, a novel approach for precise species identification in plants. Specifically, we determined (1) the accuracy of DNA barcoding approaches in delimiting species in Juglandaceae, (2) the minimum size of chloroplast dataset for species discrimination, and (3) minimum amount of next generation sequencing (NGS) data required for species identification. We found that species identification rates were highest when whole chloroplast genomes were used, followed by taxon-specific DNA barcodes, and then universal DNA barcodes. Species identification of 100% was achieved when chloroplast genome sequence coverage reached 20% and the original sequencing data reached 500,000 reads. AFRAID accurately identified species for all samples tested after 500,000 clean reads, with far less computing time than common approaches. These results provide a new approach to accurately identify species, overcoming limitations of traditional DNA barcodes. Our method, which uses next generation sequencing to generate partial chloroplast genomes, reveals that DNA barcode regions are not necessarily fixed, accelerating the process of species identification.

Single-stranded DNA-binding proteins (SSBs) play essential roles in the replication, recombination and repair processes of organellar DNA molecules. In Arabidopsis thaliana, SSBs are encoded by a small family of two genes (SSB1 and SSB2). However, the functional divergence of these two SSB copies in plants remains largely unknown, and detailed studies regarding their roles in the replication and recombination of organellar genomes are still incomplete. In this study, phylogenetic, gene structure and protein motif analyses all suggested that SSB1 and SSB2 probably diverged during the early evolution of seed plants. Based on accurate long-read sequencing results, ssb1 and ssb2 mutants had decreased copy numbers for both mitochondrial DNA (mtDNA) and plastid DNA (ptDNA), accompanied by a slight increase in structural rearrangements mediated by intermediate-sized repeats in mt genome and small-scale variants in both genomes. Our findings provide an important foundation for further investigating the effects of DNA dosage in the regulation of mutation frequencies in plant organellar genomes.

Evolutionary transitions from sexual to asexual reproduction should have significant influences on genetic divergence and polymorphism at the genome level. Plant lineages with diverse reproductive systems provide opportunities to investigate this question using comparative approaches and studies of molecular evolution. We investigated evidence for differences among the transcriptomes of 19 Dioscorea species (wild yams) with diverse reproductive systems. These included sexual species, those that propagate primarily by bulbils, and those with mixed sexual and asexual reproductive modes. We examined how transitions between these reproductive systems affected between-species divergence and within-species polymorphism. Primarily asexual species exhibited a reduced efficacy of natural selection and accumulation of deleterious mutations for both divergence and polymorphism. In contrast, species with mixed reproductive strategies involving both seed and clonal reproduction showed no evidence of an increased fixation of harmful mutations at the divergence level, while an accumulation of genetic load present in polymorphism was evident. Our study indicates that the genetic consequences of evolutionary transitions from sexual to predominantly clonal reproduction is likely to depend on both the duration and extent of asexuality occurring in populations.

Plants and their interaction partners offer unparalleled views of evolutionary ecology. Nectar larceny, entailing nectar extraction without pollinating, is thought to be an example of a harmful, antagonistic behavior, but the precise consequences of floral larceny on plant reproductive success remain contentious. We conducted a comprehensive meta-analysis of 153 studies across 120 plant species, using 14 moderators to assess the effects of floral larceny on plant reproductive success and examine the key moderators. We found that floral larceny negatively impacts flower traits, pollinator visitation, pollen deposition, and fruit set, while having a neutral effect on critical female fitness indicators, such as seed set and seed quality, as well as on male fitness. By altering pollinator behavior, floral larceny may reduce geitonogamy, potentially enhancing genetic diversity. Additionally, factors such as pollinator type, plant mating system, and pollen limitation were identified as key moderators of these effects. Our analysis reveals an ultimately neutral effect of floral larceny on plant reproductive success, with potential benefits in certain contexts. These findings suggest that floral larceny plays a complex and multifaceted role within plant-pollinator interactions, facilitating the evolutionary stability and coexistence of floral larcenists and host plants.

Alpine plants possess unique traits to adapt alpine environments. Whether leaf trait relationships of alpine plants can be captured by the two trait dimensions of organ size and resource economics is unknown. We hypothesized that, beyond the trait dimensions of leaf size and resource economics, non-structured carbohydrates (NSC) would reflect a dimension of cold-tolerance in alpine plants. To test this hypothesis, we measured 12 leaf traits critical to leaf construction and growth in 143 species across 7 sites ranging from alpine steppes to alpine meadows along an environmental gradient on the Tibetan Plateau. Furthermore, a cold resistance experiment was conducted at one of these sites to estimate the lethal temperature causing 50% frost damage (LT₅₀) of 11 alpine species. The majority of variations in 12 leaf traits of alpine plants were captured by three trait axes, in which leaf carbon (LCC) and NSC (including leaf starch; LSC and leaf soluble sugars; LSS) were clustered in a new dimension (PC3) beyond leaf size and structure, and resource economics. Although LCC, LSC and LSS all showed negative correlations with mean annual temperature, a significant negative correlation was only found between LSS and LT₅₀. It indicated that PC3 was able to reflect the cold-tolerance of alpine plants to some extent, in which LSS was the most critical trait. The storage and transformation of NSC under stressful conditions could reflect a dimension of long-term metabolic adaptation and cold-tolerance, which is an extension of the resource-utilization strategy beyond construction cost and growth.