The plant and its associated microbiota constitute a holobiont. Within this framework, the seed endophyte reservoir, shaped through multigenerational selection, exhibits pronounced host specificity, mutualistic potential, and signatures of co-evolution. We hypothesise that this reservoir operates as a 'symbiotic toolbox' forming an 'Anticipated Utility Microbiota' within the holobiont. Upon germination, specific microbes from this toolbox may undergo resuscitation to buffer environmental stresses, thereby influencing plant fitness. Using axenic Vicia sativa seeds, we simulated cold, salinity, and drought stresses and applied 16S rRNA sequencing to track seed symbiont resuscitation. Taxa showing resuscitation across stresses were classified as generalists, whilst those resuscitating under specific stresses were specialists. Microbial inoculants from these taxa were then tested in pots for host growth effects. As expected, distinct resuscitation patterns under different stresses supported the hypothesised seed 'symbiotic toolbox'. We identified 115 generalist amplicon sequence variants (e.g. Methylobacterium, Pantoea, and Sphingomonas) and stress-specific specialists: 60 cold specialists (e.g. Stenotrophomonas and Geobacter), 79 salt specialists (e.g. Leptotrichia), and 13 drought specialists (e.g. Proteobacteria). Strikingly, generalist microbial inoculants consistently promoted seedling growth across stresses, whilst specialist inoculants showed stress-specific efficacy. This study elucidates a holobiont mechanism whereby vertically transmitted seed microbes constitute a 'symbiotic toolbox' that differentially resuscitates under stress, thereby enhancing seedling fitness.

Plants interact closely with arbuscular mycorrhizal (AM) fungi. They allocate photosynthates to AM fungi in exchange for nutrients, thereby influencing plant fitness. Although plant phenotypes result from multiple traits constrained by trade-offs, the effects of AM fungi on plants are often studied using one trait. Plant response to AM fungi therefore needs to be analysed using multiple traits. Four Asteraceae species were inoculated with ten AM fungal strains using a factorial design in the greenhouse. The effect of AM fungal inoculation and their taxonomic identity on plants vegetative and reproductive traits were assessed. The predictability of the effects was evaluated based on AM fungal phylogenetic relatedness. The effect of the inoculation depended on the trait considered. Compared with the non-inoculated control, biomass allocation to roots decreased, while allocations to shoots or reproduction increased depending on the AM fungi or the plant. Significant differences were observed among AM fungi inoculates used, whether looking at separate traits, trade-offs between vegetative or reproductive growth, and the trait syndrome. In Centaurea cyanus, changes in plant phenotype were associated with the phylogenetic distance between AM fungi. These results underline the importance of using multi-trait approaches to understand AM fungal effects on plant phenotype. In accordance with the holobiont concept, the outcomes of this interaction depended on both the host plant and the AM fungus involved. It contributes to a better understanding of the biological effectors shaping the reaction norm (i.e., the range of phenotypic variation of a given host genotype) within the plant holobiont.

Finding more sustainable ways to produce food is a major challenge for humanity in the face of biodiversity extinction and climate change. Consequently, research on the ability of agroecosystems to provide multiple functions is growing. In this regard, the relative importance of organic farming and landscape-scale measures for improving multifunctionality has recently been debated. We investigated the effects of farming system (conventional vs. organic) at field scale, total length of hedgerows in the landscape and their interaction on the multifunctionality of 40 winter cereal fields in Brittany (France). Our multifunctionality assessment integrated 21 indicators of five agroecosystem goods: biodiversity conservation, nutrient cycling and soil structure, pest and disease regulation, food production and socio-economic performance. Many indicators of biodiversity conservation, pest and disease regulation, and socio-economic performance were higher in organic than in conventional systems. However, indicators of nutrient cycling and soil structure did not improve and food production was much lower in organic systems. Total hedgerow length in the landscape had less influence than organic farming on indicators, although we observed positive interactions. Granivorous carabid abundance and semi-net margin were highest in organic fields located in well-preserved hedgerow landscapes. Synthesis and applications. Our study suggests that field-scale organic farming is necessary to promote biodiversity conservation and associated ecological functioning in crop fields, whereas landscape-scale preservation of semi-natural habitats alone is likely insufficient. Preservation of hedgerows in the landscape brings additional ecological and socio-economic benefits for organic systems without compromising agricultural production. More broadly, our results call for more ambitious research into the myriad possible combinations of farming practices and agri-environmental measures at both field and landscape scales, to improve both below-ground and above-ground functioning. Evoluer vers une production agricole durable est un d & eacute;fi majeur pour l'humanit & eacute; face & agrave; l'extinction de la biodiversit & eacute; et au changement climatique. Par cons & eacute;quent, les recherches sur la capacit & eacute; des agro & eacute;cosyst & egrave;mes & agrave; assurer de nombreuses fonctions se multiplient. & Agrave; cet & eacute;gard, l'importance relative de l'agriculture biologique et des mesures paysag & egrave;res pour am & eacute;liorer la multifonctionnalit & eacute; a r & eacute;cemment & eacute;t & eacute; d & eacute;battue. Nous avons & eacute;tudi & eacute; les effets de l'agriculture biologique, de la longueur totale de haies dans le paysage et de leur interaction sur la multifonctionnalit & eacute; de 40 champs de c & eacute;r & eacute;ales d'hiver en Bretagne (France). Notre & eacute;valuation de la multifonctionnalit & eacute; est bas & eacute;e sur 21 indicateurs de cinq biens agro & eacute;cosyst & eacute;miques : la conservation de la biodiversit & eacute;, le cycle des nutriments et la structure du sol, la r & eacute;gulation des bio-agresseurs et des maladies, la production agricole, et la performance socio-& eacute;conomique. De nombreux indicateurs de conservation de la biodiversit & eacute;, de r & eacute;gulation des bio-agresseurs et des maladies et de performance socio-& eacute;conomique & eacute;taient plus & eacute;lev & eacute;s en agriculture biologique qu'en agriculture conventionnelle. Cependant, les indicateurs du cycle des nutriments et de la structure du sol n'& eacute;taient pas am & eacute;lior & eacute;s et la production agricole & eacute;tait beaucoup plus faible en agriculture biologique. La longueur totale de haies dans le paysage avait moins d'influence que l'agriculture biologique sur les indicateurs, bien que nous ayons observ & eacute; des interactions positives. L'abondance des carabes granivores et la marge semi-nette & eacute;taient plus & eacute;lev & eacute;es dans les champs en agriculture biologique situ & eacute;s dans les paysages bocagers les mieux pr & eacute;serv & eacute;s. Synth & egrave;se et applications. Notre & eacute;tude sugg & egrave;re que l'agriculture biologique est n & eacute;cessaire pour promouvoir la conservation de la biodiversit & eacute; et le fonctionnement & eacute;cologique associ & eacute; dans les champs cultiv & eacute;s, tandis que la pr & eacute;servation des habitats semi-naturels & agrave; l'& eacute;chelle du paysage est probablement insuffisante & agrave; elle seule. La pr & eacute;servation des haies dans les paysages apporte des avantages & eacute;cologiques et socio-& eacute;conomiques suppl & eacute;mentaires en agriculture biologique sans compromettre la production agricole. Plus g & eacute;n & eacute;ralement, nos r & eacute;sultats appellent & agrave; une recherche plus ambitieuse sur la myriade de combinaisons possibles de pratiques agricoles et de mesures agro-environnementales & agrave; l'& eacute;chelle du champ et du paysage, afin d'am & eacute;liorer & agrave; la fois le fonctionnement souterrain et a & eacute;rien des agro & eacute;cosyst & egrave;mes.

Agricultural intensification, landscape simplification, and climate change threaten biodiversity and ecosystem services in arable lands. Increasing semi-natural habitats and landscape heterogeneity can mitigate these impacts by providing diverse habitats, resources and modifying climate at the landscape scale. As effective natural enemies in arable lands, carabids play a key role in pest and weed seed regulation and are influenced by field management and landscape. This study hypothesized that field management directly influences carabid communities and weed seed predation, while landscape factors affect them directly and indirectly through air temperature at the landscape scale. We sampled 77 winter cereal fields across 20 landscape windows representing regional landscape heterogeneity and composition. We monitored air temperature, carabid communities, and weed seed predation during two sampling sessions in late spring and early summer 2023. Piecewise Structural Equation Models were built to test for the direct and indirect effects of field-scale factors, landscape and climate at the landscape scale on carabids and weed seed predation. For both sampling sessions, results showed that the amount of semi-natural habitats and landscape heterogeneity primarily influence carabid activity-density and composition, which in turn affect weed seed predation. Grasslands, by providing resources and refuges, favour carabids but also appear linked to higher maximum air temperature, possibly influencing carabid composition via thermotolerance traits. The study highlights the importance of semi-natural habitats and landscape heterogeneity in shaping carabid communities and their ecosystem services in arable fields. Furthermore, for the first time, we have highlighted the potential influence of landscape context on carabids mediated by air temperature, which may affect weed regulation services through seed predation.

Agricultural intensification reduces not only biodiversity in agroecosystems but also key ecosystem functions such as soil fertility. By reintroducing biological diversity in fields, weeds may enhance soil biological fertility through their influence on crop microbiota. However, letting weeds grow in crop fields will depend on weed competitiveness, farmers' perception and acceptance, and on crop management, which influences the occurrence and the abundance of weeds in the field. This study assessed the use of eight weed species to modify wheat plant root endospheric mycobiota, and the applicability of using these plants for mycobiota enrichment in the field. By combining controlled lab experiments and field studies, we demonstrated that weeds act either as a refuge for a high diversity of fungi or as a vector for transferring fungi to the crop, particularly symbionts. Weeds differed in their competitive effect on wheat growth and only three species significantly reduced wheat growth. Interviews with farmers' revealed that weed species were better known and more appreciated in crop fields by organic farmers than by conventional farmers. Floristic surveys confirmed that both weed occurrence and cover were higher in organic fields than in conventional fields. A multicriteria analysis showed that Trifolium repens and V. persica had the highest potential for promoting wheat plant mycobiota. Among the weed species tested, these two are worth considering as auxiliaries to improve soil biological fertility in crop fields. Their use with the goal of selecting appropriate crop mycobiota should be relatively easier in organic farms where weeds are better accepted, whereas their use in conventional farms would require raising farmers' awareness of the benefits of weeds for soil fertility.

Microbes inhabit virtually all river ecosystems, influencing energy flow and playing a key role in global sustainability and climate change. Yet, there is uncertainty about how various taxonomic groups respond to large-scale factors in river networks. We analysed microbial community richness and composition across six European Atlantic catchments using environmental DNA sequencing. Our findings reveal different drivers for diversity and composition: land use is pivotal for eukaryotes, while climate and geology are crucial for prokaryotes. A strong regional influence shapes these communities, with warmer, drier regions (Portugal and France) differing from cooler, wetter ones (Northern Spain, Ireland and the United Kingdom). These patterns suggest potential indicators for global change, such as taxa resistant to temperature increases and water scarcity, or those sensitive to land use changes.

Landscape heterogeneity is known as a major factor of community structure and composition. Whether this effect of the landscape extends at different scales and particularly at the relevant scale for microorganisms remained to be determined. We used the cases produced by aquatic larvae of Trichoptera, which assemble organic or mineral particles, as naturally replicated experimental systems representing structured substrates to determine the effect of landscape structuration on microbial communities. A metabarcoding approach was used to characterise fungal, bacterial and diatom communities on cases produced by six Trichoptera species and related unstructured organic and mineral substrates. The structuration of the particles constituting the cases was also determined as a measure of microscale landscape. Structured substrates harboured communities of diatoms, fungi and bacteria that differed from those found on unstructured substrates. Microbial communities also differed between organic and mineral substrates. We found a higher microbial diversity on structured substrates than on unstructured substrates. The heterogeneity of the microscale landscape also affected bacterial and fungal communities within cases. These results highlight the importance of microscale landscape structuration for microbial diversity and demonstrate that approaches of landscape ecology could be downscaled to the microscale.

Summary Understanding the drivers of assemblages of arbuscular mycorrhizal fungi (AMF) is essential to leverage the benefits of AMF for plant growth and health. Arbuscular mycorrhizal fungi are heterogeneously distributed in space even at small scale. We review the role of plant distribution in driving AMF assemblages (the passenger hypothesis), using a transposition of the conceptual framework of landscape ecology. Because rooting systems correspond to habitat patches with limited carrying capacity that differ in quality due to host‐preference effects, we suggest considering plant communities as mosaics of AMF microhabitats. We review how predictions from landscape ecology apply to plant community effects on AMF, and the existing evidence that tests these predictions. Although many studies have been conducted on the effect of plant compositional heterogeneity on AMF assemblages, they mostly focused on the effect of plant richness, while only a few investigated the effect of configurational heterogeneity, plant connectivity or plant community temporal dynamics. We propose key predictions and future prospects to fill these gaps. Considering plant communities as landscapes extends the passenger hypothesis by including a spatially explicit dimension and its associated ecological processes and may help understand and manipulate AMF assemblages at small spatial scales.

Societal Impact StatementAgricultural intensification is a major driver of biodiversity decline in agrosystems. For instance, it has been shown that conventional farming leads to a decline in soil microbial diversity and triggers a strong selection process, altering the functioning of the whole ecosystem. The present study shows that organic farming increases diversity and affects composition of crop plant microbiota, mostly as a response to field management and soil characteristics. Furthermore, crop plant microbiota influences crop production and resistance to pathogens. Therefore, agricultural practices affect plant performance through microorganism‐mediated changes, which may be important pillars of future sustainable crop production.Summary Agricultural intensification threatens biodiversity, but the effects of intensification on microorganisms are still overlooked despite their role in ecosystem functioning. Microorganisms associated with plants provide many services that affect plant growth and health. Organic farming is expected to strongly affect species composition, richness, and their interactions. We analyzed the effect of the farming system on endophytic microbial assemblages associated with winter wheat plants and plant performance in the field. We collected environmental data through farmer interviews, soil analyses, and plant inventories and analyzed root microbiota at vegetative and flowering stages. Organic farming increased fungal and bacterial diversity associated to wheat plants and affected species composition in most phyla. This effect was mostly due to soil characteristics and field management and a little to plant diversity in the field. Microbial responses were more pronounced at the late developmental stage, likely as a result of accumulative effect of management actions during plant development. Seed production and resistance to pathogens were related to specific phyla that are important for seed production and/or wheat resistance to septoriose. This work advances our understanding of how agricultural practices affect plant performance through microorganism‐mediated changes and supports the use of microorganisms as pillars of sustainable crop production.

Microorganisms associated with plants provide essential functions to their hosts, and therefore affect ecosystem productivity. Agricultural intensification has modified microbial diversity in the soil reservoir and may affect plant microbial recruitment. Weeds develop spontaneously in crop fields, and could influence microorganisms associated with crop plants through a neighbourhood effect. We explore the effect of weed species on crop plant microbiota as potentially auxiliary plants that affect agricultural productivity. We combined field and controlled laboratory studies to analyse the neighbourhood effect of weeds on wheat root endospheric mycobiota and growth. First, we analysed the effect of weed species diversity and identity recorded in the neighbourhood of individual wheat plants on soil and wheat root mycobiota in the field. Second, we used a plant-matrix design in laboratory conditions to test the effect of weed identity (9 weed treatments) and their ability to transmit root mycobiota to wheat roots, and the resulting impact on wheat growth. In contrast to soil mycobiota, we demonstrated that wheat root endospheric mycobiota was influenced by the diversity and identity of weeds developing in their 1 m 2 neighbourhood. Wheat root endospheric microbiota strongly differs in terms of richness and composition depending on the neighbouring weed plant species. Weed species transmitted from 13% to 74% of their root microbiota to wheat roots depending on weed identity in controlled conditions. Synthesis. Weed neighbours modified wheat plant performance, possibly as a result of competitive interactions and changes in microbiota. Our findings suggest that crop root mycobiota was variable and was modulated by their weed neighbourhood. Synergistic effects between mycobiota of crops and weeds could therefore contribute to soil biodiversity and sustainable agriculture.

Bocage landscapes are characterized by a network of hedgerows that delimits arable fields. Such landscapes provide many ecosystem services, including biodiversity conservation, but their effects on weed communities remain largely unknown. Bocage landscapes could affect weed communities through two main processes: plant spillover from hedgerows and increased environmental heterogeneity in arable fields. These bocage effects are also likely to vary between farming systems (conventional vs. organic) due to differences in management practices. We sampled weed communities more than 20 m from field margins in 74 arable fields (37 per farming system). Fields were located along two independent landscape gradients of total length of hedgerows (with or without a shrub layer) and organic farming cover, in Brittany (France). We analysed the effect of 'bocage' (i.e. the density and complexity of hedgerow networks) and farming systems at field and landscape scales on species and functional diversity of weed communities. Further, we used fidelity to non-crop habitats and Ellenberg indicator values to assess the 'plant spillover' and 'environmental heterogeneity' hypotheses, respectively. Weed communities were more diverse and more abundant in organic farming systems. In addition, weed communities were more diverse, but not more abundant, in denser and more complex bocage landscapes. 'Bocage' increased species diversity of weeds, but also community-weighted variance of specific leaf area, plant height and seed mass. Positive effects of 'bocage' on weed diversity were driven by increased environmental heterogeneity rather than spillover of transient species from hedgerows. 'Bocage' effects were independent of farming systems at field and landscape scales. Synthesis and applications. Maintaining diverse weed communities is key to agroecological weed management and biodiversity conservation in agricultural landscapes. Farmers are often concerned that hedgerows harbour competitive plants spreading into field edges, thereby increasing weed pressure. However, our study shows that dense and complex bocage landscapes promote weed diversity in field cores, most likely by increasing environmental heterogeneity. Thus, bocage landscapes could actually enhance ecosystem services provided by weed communities and reduce weed-crop competition.

Microorganisms are associated with all plants, recently leading to the hologenome concept. We reviewed the assembly processes of plant microbiota and analyzed its structure during the emergence of dysbioses. In particular, we discussed the Anna Karenina Principle (AKP) based on Leo Tolstoy's assertion applied to plant microbiota: ‘All healthy microbiota are alike; each disease-associated microbiota is sick in its own way.' We propose the AKP to explain how stochastic processes in plant microbiota assembly due to several external stressors could lead to plant diseases. Finally, we propose the AKP to conceptualize plant dysbioses as a transitory loss of host capacity to regulate its microbiota, implying a loss of function that leads to a reduction of the host's fitness.

The hologenome concept considers the entity formed by a host and its microbiota, the holobiont, as new level of hierarchical organization subject to neutral and selective forces. We used grafted plants to formally evaluate the hologenome concept. We analyzed the root-endosphere microbiota of two independent watermelon and grapevine plant systems, including ungrafted and reciprocal-grafting combinations. Grafted and ungrafted hosts harbor markedly different microbiota compositions. Furthermore, the results indicate a non-random assembly of bacterial communities inhabiting the root endosphere of chimeric plants with interactive effect of both the rootstock and scion on the recruitment of microorganisms. Because chimeric plants did not have a random microbiota, the null hypothesis that holobionts assemble randomly and hologenome concept is an intellectual construction only can be rejected. The study supports the relevance of hologenome as biological level of organization and opens new avenues for a better fundamental understanding of plants as holobionts.

Agriculture intensification led to the gradual destruction of semi-natural habitats and landscape simplification in the North-Western Europe. All these profound changes affected plant assemblages, especially in semi-natural ecosystems such as grasslands. They could have negative effects on the plant diversity and the related ecosystem functions, such as primary productivity. At the landscape scale this biodiversity-productivity relationship still have to be tested in real-world systems. It could have important economic implications concerning fodder production. We used a large-scale sampling design (30 landscape windows - 1 x1 km) to quantify landscape structure variables (semi-natural habitat amounts and landscape heterogeneity), grassland plant taxonomic diversity and functional diversity calculated from multidimensional functional spaces (functional richness, evenness, dispersion), and grassland productivity (mean, temporal stability, spatial variability) derived from satellite image time-series. Taxonomic dissimilarity, functional evenness and functional dispersion of grassland plant assemblages were related to semi-natural habitat amount. Functional evenness and dispersion decreased with grassland amount whereas taxonomic dissimilarity and functional evenness increased with hedgerow amount. Only taxonomic and functional richness depended on landscape heterogeneity, and with a positive relationship. Grassland plant assemblages with a higher functional evenness presented a higher and more stable mean productivity. In addition to the direct effect of landscape heterogeneity on plant productivity, we demonstrated plant trait-mediated effect of landscape structure on mean and temporal stability of grassland productivity at the landscape scale. Landscape management, especially through the conservation of hedgerows can help maintain high and resilient production of fodder in the future as a result of their positive effects on plant functional diversity.

In ever‐changing landscapes, there is increasing evidence that current plant assemblages are shaped by the temporal dynamics of landscape connectivity. So far, attempts to take the temporal dynamics of connectivity into account have only focused on the degree of connectivity at one or several moments in time, but neglected the cumulative effects of temporal changes in connectivity. We investigated the effects of the temporal dynamics of landscape connectivity (i.e. the degree of connectivity, the magnitude and the variability of its temporal changes) over the last seven decades on current woodland plant herbaceous assemblages. The assemblages were described using a taxonomic approach combined with a functional approach based on four traits linked to the colonisation capacity of plant species. The taxonomic diversity of current woodland assemblages did not respond to the degree of connectivity nor to the magnitude and the variability of temporal changes, but the mean and/or the diversity of trait values linked to their colonisation capacity did. Responses, mainly driven by generalist species, were modulated by the type of connectivity trend experienced by woodlands. In woodlands experiencing an upward connectivity trend, high magnitude of temporal changes in connectivity increased the abundance of species that invest mostly in sexual reproduction at the expense of vegetative reproduction whereas the degree of connectivity and the variability of its temporal changes had no effect. In woodlands experiencing a downward connectivity trend, the diversity of seed mass values was independent on the magnitude but decreased with the variability of temporal changes, and increased with the degree of connectivity. Synthesis . Overall, we show that, besides the degree of connectivity, the cumulative effects of decades of changes in connectivity shape woodland plant community assembly by selecting for particular trait values in plant species. This study opens new perspectives for integrating the temporal dynamics of landscape connectivity in the theoretical framework of plant assembly rules. It should also be considered in the development of management strategies to restore and maintain landscape connectivity.

Background and aims Considering the plant microbiota, temporal changes are expected depending on plant development stages and environmental pressures because of modifications in plant requirements and available soil microbial reservoir. Methods Herein, we analyzed the composition of root endosphere microbiota of grafted vine plants using two grapevine cultivars (Merlot and Cabernet-Sauvignon as scion grafted on rootstocks of different clones) as models both sampled in a single vineyard at three dates over a period of two growing years. Results Highly conserved temporal patterns were found in the two cultivars. Intra-annual changes in microbial community composition were recorded whereas convergent microbial communities were observed on the two September dates. In particular, the increase in Actinobacteria and decrease in Glomeromycota in September were interpreted as shifts in the microbiota community patterns related to plant physiological requirements (e.g. water supply). A high proportion of non-random assembly of the root endospheric bacterial community confirmed the deterministic influence of the plant or/and the environment in microbial recruitment over time. The modified normalized stochasticity ratio (MST) showed that deterministic processes of assembly (MST < 50%) were commonplace despite the changes in the root microbiota composition observed among sampling dates. Conclusion Our study suggests an intra-annual rhythm of microbiota shifts, marginally random, with a succession within the root-microbiota endosphere likely governed by active plant filtering. A better knowledge of microbial-recruitment at work, seems important for both fundamental and applied perspectives.

Agricultural intensification has been demonstrated to induce a loss of biodiversity. Despite the key role of symbiotic microorganisms in plant nutrition and protection, the impact of agricultural intensification on these microorganisms is not fully understood. Organic farming and field edges (as semi-natural elements) may promote a higher microbial diversity thanks to lower anthropic disturbance and higher plant diversity. We sampled wheat individuals in pairs of wheat fields (one organic and one conventional) along a distance gradient to the edges (hedgerow vs. grassy), in 20 landscape windows selected along an uncorrelated gradient of organic farming and hedgerow density. We demonstrated that organic farming shaped microbial composition and increased fungal and bacterial richness, while hedgerows had a neutral or negative effect on richness depending on the microbial phyla considered. In contrast to bacteria, fungal communities were heterogeneously distributed within fields, having a higher diversity for some phyla close to field edges. Overall we highlighted that fungi responded more to the field scale while bacteria were more affected by landscape scale. The effect of agricultural intensification on plant microbiota and therefore on the functions provided by microorganisms to the plants has to be considered at a multiple spatial scale-from field to landscape.

Urbanization and agricultural intensification are the main drivers of biodiversity losses through multiple stressors, especially habitat fragmentation, isolation and loss. Designing Blue and Green Infrastructure Networks (BGIN) has been recommended as a potential tool for land-use planning to increase ecosystem services while preserving biodiversity. All municipalities in France are required to perform BGIN planning. This article focuses on the Couesnon watershed (Brittany, France) and the participatory process used to define and analyze five possible pathways of future land-use and land-cover changes that included implementation of BGINs. Impacts on biodiversity were estimated by quantifying the change in landscape connectivity of woodlands, grasslands and wetlands. The effectiveness of BGIN policies was assessed by comparing current landscape connectivity (2018) to those in possible futures. Landscape connectivity referred to functional connectivity for three indicator species (Abax parallelepipedus, Maniola jurtina and Arvicola sapidus) across three landscape features: woodlands, grasslands and wetlands, respectively. Results allowed impacts of urban and agricultural land-use changes to be identified in terms of extent and quality. If BGIN policies were applied effectively to control the expansion of gray infrastructure, they would help increase the area and the quality of grassland and woodland connectivity by no more than 2%. Agricultural land-use and land-cover changes could have more impact on the extent of grassland (−82% to +38%) and wetland (−49% to +47%) connectivity. Current and future trends for hedgerows implied a decrease in woodland connectivity of 9.8–33.8%. Impacts on the quality of landscape connectivity is not proportional with the extent, as a decrease of the latter can have relatively more negative impacts on the former, and inversely. The study highlights that the BGIN strategy can preserve landscape connectivity effectively in urban ecosystems, where human density is higher, but can be threatened by agricultural intensification.