From invasive species stand to species-rich grassland: Long-term changes in plant species composition during Solidago invaded site restoration.

Biological invasions degrade ecosystems, negatively affecting human well-being and biodiversity. Restoration of invaded agricultural ecosystems is among specific goals of European Union Biodiversity Strategy. Successful restoration of invaded lands is a long-term process that requires monitoring to assess the effects of interventions. Here, we present the results of a long-term experiment (8 years) on restoration of semi-natural grassland on abandoned arable field overgrown by invasive Solidago species (S. gigantea and S. canadensis). We examined effect of different invaders removal methods (rototilling, turf stripping, herbicide application) and seed application practices (commercial seed mixture, fresh hay) on changes in species composition and taxonomic diversity of restored vegetation. Our results showed a positive effect of grassland restoration on taxonomic diversity and species composition, manifested by a decrease in Solidago cover and an increase in cover and richness of target graminoids and forbs characteristic of grassland. The seed source had a longer lasting and still observable effect on the vegetation composition than the Solidago removal treatments, which ceased to differ significantly in their influence after the first few years. Applying fresh hay as a seed source increased the cover of grassland species such as Arrhenatherum elatius and Poa pratensis. For commercial seed mixture, we observed the high cover of Lolium perenne and Schedonorus pratensis (introduced with seed mixture) at the beginning and the slow decrease along the experiment course. The most striking effect was the fresh hay with herbicide application, which resulted in the lowest Solidago cover and the highest cover of target graminoids. Nonetheless, with years the non-chemical methods, including no treatment, gives comparable to herbicide effectiveness of restoration. Overall, during the experiment, alpha diversity increased, while beta and gamma diversity reached a species maximum in the third year, and then decreased. In conclusion, this study gives guidance to successful restoration of species-rich grasslands on sites invaded by Solidago. It should be emphasised that short-term effect differ considerably from long-term outputs, especially highlighting the importance of seed source, as well as effectiveness of environmentally friendly methods such as regular mowing to control the invader.

Seed limitation interacts with biotic and abiotic factors to constrain novel species' impact on community biomass and richness.

Seed limitation can narrow down the number of coexisting plant species, limit plant community productivity, and also constrain community responses to changing environmental and biotic conditions. In a 10-year full-factorial experiment of seed addition, fertilisation, warming and herbivore exclusion, we tested how seed addition alters community richness and biomass, and how its effects depend on seed origin and biotic and abiotic context. We found that seed addition increased species richness in all treatments, and increased plant community biomass depending on nutrient addition and warming. Novel species, originally absent from the communities, increased biomass the most, especially in fertilised plots and in the absence of herbivores, while adding seeds of local species did not affect biomass. Our results show that seed limitation constrains both community richness and biomass, and highlight the importance of considering trophic interactions and soil nutrients when assessing novel species immigrations and their effects on community biomass.

Tipping the balance: The role of seed density, abiotic filters, and priority effects in seed-based wetland restoration.

Sowing native seeds is a common approach to reintroduce native plants to degraded systems. However, this method is often overlooked in wetland restoration despite the immense global loss of diverse native wetland vegetation. Developing guiding principles for seed-based wetland restoration is critical to maximize native plant recovery, particularly in previously invaded wetlands. Doing so requires a comprehensive understanding of how restoration manipulations, and their interactions, influence wetland plant community assembly. With a focus on the invader Phragmites australis, we established a series of mesocosm experiments to assess how native sowing density, invader propagule pressure, abiotic filters (water and nutrients), and native sowing timing (i.e., priority effects) interact to influence plant community cover and biomass in wetland habitats. Increasing the density of native seeds yielded higher native cover and biomass, but P. australis suppression with increasing sowing densities was minimal. Rather, community outcomes were largely driven by invader propagule pressure: P. australis densities of ≤500 seeds/m2 maintained high native cover and biomass. Low-water conditions increased the susceptibility of P. australis to dominance by native competitors. Early sowing of native seeds showed a large and significant benefit to native cover and biomass, regardless of native sowing density, suggesting that priority effects can be an effective restoration manipulation to enhance native plant establishment. Given the urgent wetland restoration need combined with the limited studies on seed-based wetland restoration, these findings provide guidance on restoration manipulations that are grounded in ecological theory to improve seed-based wetland restoration outcomes.

Reducing dispersal limitation via seed addition increases species richness but not above-ground biomass.

Seed dispersal limitation, which can be exacerbated by a number of anthropogenic causes, can result in local communities having fewer species than they might potentially support, representing a potential diversity deficit. The link between processes that shape natural variation in diversity, such as dispersal limitation, and the consequent effects on productivity is less well known. Here, we synthesised data from 12 seed addition experiments in grassland communities to examine the influence of reducing seed dispersal limitation (from 1 to 60 species added across experiments) on species richness and productivity. For every 10 species of seed added, we found that species richness increased by about two species. However, the increase in species richness by overcoming seed limitation did not lead to a concomitant increase in above-ground biomass production. This highlights the need to consider the relationship between biodiversity and ecosystem functioning in a pluralistic way that considers both the processes that shape diversity and productivity simultaneously in naturally assembled communities.

Experimental dispersal reveals characteristic scales of biodiversity in a natural landscape.

Ecological theory posits that dispersal among habitat patches links local communities and is a key "regional" process that maintains biological diversity. However, manipulations required to experimentally test regional processes are infeasible for most systems, and thus more work is needed to detect the scales at which regional processes manifest and their overall effect on diversity. In a Californian grassland, a hotspot for global biodiversity, we used a seed vacuum to increase dispersal at spatial scales varying from 1 m to 10 km while maintaining a realistic spatial structure of species pools and environmental conditions. We found that dispersal limitation has a profound influence on diversity; species richness increased with the spatial scale of seed mixing, doubling in plots that received seed from large (≥5 km) compared with small (≤5 m) scales. This increase in diversity corresponded to an increase in how well species distributions were explained by environmental conditions, from modest at small scales (R2 = 0.34) to strong at large scales (R2 = 0.52). Responses to the spatial scale of seed mixing were nonlinear, with no differences below 5 m or above 5 km. Nonlinearities were explained by homogeneity of environmental conditions below 5 m and by a lack of additional variation in the species pool above 5 km. Our approach of manipulating natural communities at different spatial scales reveals (i) nonlinear transitions in the importance of environmental sorting and dispersal, and (ii) the negative effects of dispersal limitation on local diversity, consistent with previous research suggesting that large numbers of species are headed toward regional extinction.

Traits linked with species invasiveness and community invasibility vary with time, stage and indicator of invasion in a long-term grassland experiment.

Much uncertainty remains about traits linked with successful invasion - the establishment and spread of non-resident species into existing communities. Using a 20-year experiment, where 50 non-resident (but mostly native) grassland plant species were sown into savannah plots, we ask how traits linked with invasion depend on invasion stage (establishment, spread), indicator of invasion success (occupancy, relative abundance), time, environmental conditions, propagule rain, and traits of invaders and invaded communities. Trait data for 164 taxa showed that invader occupancy was primarily associated with traits of invaders, traits of recipient communities, and invader-community interactions. Invader abundance was more strongly associated with community traits (e.g. proportion legume) and trait differences between invaders and the most similar resident species. Annuals and invaders with high-specific leaf area were only successful early in stand development, whereas invaders with conservative carbon capture strategies persisted long-term. Our results indicate that invasion is context-dependent and long-term experiments are required to comprehensively understand invasions.

Experimental Testing of Dispersal Limitation in Soil Bacterial Communities with a Propagule Addition Approach.

The role of dispersal in the assembly of microbial communities remains contentious. This study tested the importance of dispersal limitation for the structuring of local soil bacterial communities using an experimental approach of propagule addition. Microbes extracted from soil pooled from samples collected at 20 localities across ~ 400 km in a temperate steppe were added to microcosms of local soils at three sites; the microcosms were then incubated in situ for 3 months. We then assessed the composition and diversity of bacterial taxa in the soils using 16S rRNA gene amplicon sequencing. The addition of the regional microbial pool did not cause significant changes in the overall composition or diversity of the total bacterial community, although a very small number of individual taxa may have been affected by the addition treatment. Our results suggest a negligible role of dispersal limitation in structuring soil bacterial communities in our study area.

Limited prospects for future alpine treeline advance in the Canadian Rocky Mountains.

Treeline advance has occurred throughout the twentieth century in mountainous regions around the world; however, local variation and temporal lags in responses to climate warming indicate that the upper limits of some treelines are not necessarily in climatic equilibrium. These observations suggest that factors other than climate are constraining tree establishment beyond existing treelines. Using a seed addition experiment, we tested the effects of seed availability, predation and microsite limitation on the establishment of two subalpine tree species (Picea engelmannii and Abies lasiocarpa) across four treelines in the Canadian Rocky Mountains. The effect of vegetation removal on seedling growth was also determined, and microclimate conditions were monitored. Establishment limitations observed in the field were placed in context with the effects of soil properties observed in a parallel experiment. The seed addition experiment revealed reduced establishment with increasing elevation, suggesting that although establishment within the treeline ecotone is at least partially seed limited, other constraints are more important beyond the current treeline. The effects of herbivory and microsite availability significantly reduced seedling establishment but were less influential beyond the treeline. Microclimate monitoring revealed that establishment was negatively related to growing season temperatures and positively related to the duration of winter snow cover, counter to the conventional expectation that establishment is limited by low temperatures. Overall, it appears that seedling establishment beyond treeline is predominantly constrained by a combination of high soil surface temperatures during the growing season, reduced winter snowpack and unfavourable soil properties. Our study supports the assertion that seedling establishment in alpine treeline ecotones is simultaneously limited by various climatic and nonclimatic drivers. Together, these factors may limit future treeline advance in the Canadian Rocky Mountains and should be considered when assessing the potential for treeline advance in alpine systems elsewhere.

Dispersal and neutral sampling mediate contingent effects of disturbance on plant beta-diversity: a meta-analysis.

A major challenge in ecology, conservation and global-change biology is to understand why biodiversity responds differently to similar environmental changes. Contingent biodiversity responses may depend on how disturbance and dispersal interact to alter variation in community composition (ß-diversity) and assembly mechanisms. However, quantitative syntheses of these patterns and processes across studies are lacking. Using null-models and meta-analyses of 22 factorial experiments in herbaceous plant communities across Europe and North America, we show that disturbance diversifies communities when dispersal is limited, but homogenises communities when combined with increased immigration from the species pool. In contrast to the hypothesis that disturbance and dispersal mediate the strength of niche assembly, both processes altered ß-diversity through neutral-sampling effects on numbers of individuals and species in communities. Our synthesis suggests that stochastic effects of disturbance and dispersal on community assembly play an important, but underappreciated, role in mediating biotic homogenisation and biodiversity responses to environmental change.

When does seed limitation matter for scaling up reforestation from patches to landscapes?

Restoring forest to hundreds of millions of hectares of degraded land has become a centerpiece of international plans to sequester carbon and conserve biodiversity. Forest landscape restoration will require scaling up ecological knowledge of secondary succession from small-scale field studies to predict forest recovery rates in heterogeneous landscapes. However, ecological field studies reveal widely divergent times to forest recovery, in part due to landscape features that are difficult to replicate in empirical studies. Seed rain can determine reforestation rate and depends on landscape features that are beyond the scale of most field studies. We develop mathematical models to quantify how landscape configuration affects seed rain and forest regrowth in degraded patches. The models show how landscape features can alter the successional trajectories of otherwise identical patches, thus providing insight into why some empirical studies reveal a strong effect of seed rain on secondary succession, while others do not. We show that seed rain will strongly limit reforestation rate when patches are near a threshold for arrested succession, when positive feedbacks between tree canopy cover and seed rain occur during early succession, and when directed dispersal leads to between-patch interactions. In contrast, seed rain has weak effects on reforestation rate over a wide range of conditions, including when landscape-scale seed availability is either very high or very low. Our modeling framework incorporates growth and survival parameters that are commonly estimated in field studies of reforestation. We demonstrate how mathematical models can inform forest landscape restoration by allowing land managers to predict where natural regeneration will be sufficient to restore tree cover. Translating quantitative forecasts into spatially targeted interventions for forest landscape restoration could support target goals of restoring millions of hectares of degraded land and help mitigate global climate change.

Species pool size alters species-area relationships during experimental community assembly.

The species pool concept has advanced our understanding for how biodiversity is coupled at local and regional scales. However, it remains unclear how species pool size, the number of species available to disperse to a site, influences community assembly across spatial scales. We provide one of the first studies that assesses diversity across scales after experimentally assembling grassland communities from species pools of different sizes. We show that species pool size causes scale-dependent effects on diversity in grasslands undergoing restoration by altering the shape of the species-area relationship (SAR). Specifically, larger species pools increased the slope of the SAR, but not the intercept, suggesting that dispersal from a larger pool causes species to be more spatially aggregated. This increased aggregation appears to be caused by sampling effects due to fewer individuals arriving per species, rather than stronger species sorting across variation in soil moisture. These scale-dependent effects suggest that studies evaluating species pools at a single, small scale may underestimate their effects, thereby contributing to uncertainty about the importance of regional processes for community assembly and their consequences for ecological restoration.

Seedling recruitment correlates with seed input across seed sizes: implications for coexistence.

Understanding controls on recruitment is critical to predicting community assembly, diversity, and coexistence. Theory posits that at mean fecundity, recruitment of highly fecund small-seeded plants should be primarily microsite limited, which is indicated by a saturating recruitment function. In contrast, species that produce fewer large seeds are more likely to be seed-limited, which is characterized by a linear recruitment function. If these patterns hold in nature, seed predation that disproportionately affects larger-seeded species can limit their establishment. We tested these predictions by comparing recruitment functions among 16 co-occurring perennial forb species that vary by over two orders of magnitude in seed size. We also assessed how postdispersal seed predation by mice influenced recruitment. We added seeds at densities from zero to three times natural fecundity of each species to undisturbed plots and examined spatial variation in recruitment by conducting experiments across 10 grassland sites that varied in productivity and resource availability. Consistent across two replicated years, most species had linear recruitment functions across the range of added seed densities, indicative of seed-limited recruitment. Depending on year, the recruitment functions of only 19-37% of target species saturated near their average fecundity, and this was not associated with seed size. Recruitment was strongly inhibited by rodent seed predation for large-seeded species but not for smaller-seeded species. Proportional recruitment was more sensitive to spatial variation in recruitment conditions across sites for some small-seeded species than for large-seeded species. These results contradict the common belief that highly fecund small-seeded species suffer from microsite-limited recruitment. Rather, they imply that, at least episodically, recruitment can be strongly correlated to plant fecundity. However, proportional recruitment of small-seeded species was inhibited at productive sites to a greater extent than large-seeded species. Results also show that in a system where the dominant granivore prefers larger seeds, low-fecundity large-seeded species can suffer from even greater seed-limited recruitment than would occur in the absence of predators.

Data on different seed harvesting methods used in grassland restoration on ex-arable land.

We present data of the grassland restoration experiment performed in the Bílé Karpaty Mts. (White Carpathians, Czech Republic) in dry species-rich meadows. First we harvested seed material in a preserved source meadow (donor site hereafter) by brush harvesting the vegetation once (B1 hereafter), brush harvesting three times during the season (B3 hereafter), and by cutting green hay (GH hereafter). Then we determined the species composition and seed quantity of the harvested material. Furthermore, we transferred the seeds to an experimental site on ex-arable land (receptor site hereafter), and monitored the development of the meadow communities in the following five years. Data are interpreted in: Á-J. Albert, O. Mudrák, I. Jongepierová, K. Fajmon, I. Frei, M. Sevcíková, J. Klimesová, J. Dolezal, Grassland restoration on ex-arable land by transfer of brush-harvested propagules and green hay. Agriculture, Ecosystems & Environment 272 (2019), 74-82.

Seed incorporation during vinification and its impact on chemical and organoleptic properties in Syzygium cumini wine.

Syzgium cumini (Jamun) is an evergreen tropical tree, its various parts are known for many therapeutic properties. The present work represents the production of wines from jamun fruits using two different native isolates (Saccharomyces cerevisiae - KF551990 and Pichia gummiguttae - MCC 1273) and influence of jamun seeds on the physico-chemical parameters, chromatic properties, phenolic components and sensory attributes of wine. Wine produced was bottle aged for one year. On aging there was a reduction in bitterness and astringency. Aging lead to reduction in monomeric anthocyanin with an increase in co-pigmented and polymeric anthocynins thus affecting the wine color. Anthocyanin analysis in jamun wine indicated petunidin 3,5-diglucoside as the principal anthocyanin. PCA analysis of wine revealed association of young jamun wine with anthocyanin components. PLS analysis exhibited both positive and negative correlation between various attributes indicating sensory perception of jamun wine is affected by overall composition of the wine.

Limits to tree species invasion in pampean grassland and forest plant communities.

Factors limiting tree invasion in the Inland Pampas of Argentina were studied by monitoring the establishment of four alien tree species in remnant grassland and cultivated forest stands. We tested whether disturbances facilitated tree seedling recruitment and survival once seeds of invaders were made available by hand sowing. Seed addition to grassland failed to produce seedlings of two study species, Ligustrum lucidum and Ulmus pumila, but did result in abundant recruitment of Gleditsia triacanthos and Prosopis caldenia. While emergence was sparse in intact grassland, seedling densities were significantly increased by canopy and soil disturbances. Longer-term surveys showed that only Gleditsia became successfully established in disturbed grassland. These results support the hypothesis that interference from herbaceous vegetation may play a significant role in slowing down tree invasion, whereas disturbances create microsites that can be exploited by invasive woody plants. Seed sowing in a Ligustrum forest promoted the emergence of all four study species in understorey and treefall gap conditions. Litter removal had species-specific effects on emergence and early seedling growth, but had little impact on survivorship. Seedlings emerging under the closed forest canopy died within a few months. In the treefall gap, recruits of Gleditsia and Prosopis survived the first year, but did not survive in the longer term after natural gap closure. The forest community thus appeared less susceptible to colonization by alien trees than the grassland. We conclude that tree invasion in this system is strongly limited by the availability of recruitment microsites and biotic interactions, as well as by dispersal from existing propagule sources.