Instituto de Investigación en Cambio Global

It has been recognized that cytokinins are plant hormones that influence not only numerous aspects of plant growth, development and physiology, including cell division, chloroplast differentiation and delay of senescence but the interaction with other organisms, including pathogens. Cytokinins are not only produced by plants but are also by other prokaryotic and eukaryotic organism such as bacteria, fungi, microalgae and insects. Notably, cytokinins are produced both by pathogenic and also beneficial microbes and are known to induce resistance in plants against pathogen infections. In this review the contrasting role of cytokinin for the defence and susceptibility of plants against bacterial and fungal pathogen and pest insects is assessed. We also discuss the cross talk of cytokinins with other phytohormones and the underlying mechanism involved in enhancing plant immunity against pathogen infections and explore possible practical applications in crop plant production.

Biological invasions rank amongst the most deleterious components of global change inducing alterations from genes to ecosystems. The genetic characteristics of introduced pools of individuals greatly influence the capacity of introduced species to establish and expand. The recently demonstrated heritability of microbial communities associated to individual genotypes of primary producers makes them a potentially essential element of the evolution and adaptability of their hosts. Here, we characterized the bacterial communities associated to native and non-native populations of the marine green macroalga Caulerparacemosa through pyrosequencing, and explored their potential role on the strikingly invasive trajectory of their host in the Mediterranean. The similarity of endophytic bacterial communities from the native Australian range and several Mediterranean locations confirmed the origin of invasion and revealed distinct communities associated to a second Mediterranean variety of C. racemosa long reported in the Mediterranean. Comparative analysis of these two groups demonstrated the stability of the composition of bacterial communities through the successive steps of introduction and invasion and suggested the vertical transmission of some major bacterial OTUs. Indirect inferences on the taxonomic identity and associated metabolism of bacterial lineages showed a striking consistency with sediment upheaval conditions associated to the expansion of their invasive host and to the decline of native species. These results demonstrate that bacterial communities can be an effective tracer of the origin of invasion and support their potential role in their eukaryotic host's adaptation to new environments. They put forward the critical need to consider the 'meta-organism' encompassing both the host and associated micro-organisms, to unravel the origins, causes and mechanisms underlying biological invasions.

Summary Historical declines of marine megaherbivores have led to a view of seagrass communities structured largely by abiotic disturbance and plant competition. There is, however, growing recognition of the significance of top‐down control through herbivory, on seagrass ecosystem processes, raising the question of how meadows functioned under historically high populations of megaherbivores. We assess the impacts of such intense herbivory on seagrass meadow composition in the Lakshadweep islands (India), where high‐density feeding aggregations of green turtles have persisted for over a decade. We use a series of complementary approaches: (i) natural herbivory exclosures (ii) published data on seagrass composition before and after turtles established (at one atoll: Agatti) and (iii) present species composition along a turtle herbivory gradient over multiple atolls. Long‐term natural exclosures in Agatti indicated that sustained turtle grazing caused clear shifts in species dominance from the long‐lived, higher‐successional Thalassia hemprichii to the relatively short‐lived, pioneering species Cymodocea rotundata (dominant in grazed areas). T. hemprichii was the dominant species c . 20 years ago but is now restricted to areas within exclosures in Agatti, and to the least grazed meadows (<5%) in other atolls. We conducted field experiments to identify possible mechanisms by which herbivory mediated direct or apparent competitive interactions between seagrass species. To verify if grazing reduced growth rates of T. hemprichii in comparison with C. rotundata , we conducted clipping experiments in 1m 2 plots, simulating turtle herbivory on equal shoot proportions of both species. After 4 months, T. hemprichii shoot density showed major declines in clipped vs. control plots, but C. rotundata shoot density remained relatively unaffected. To test whether selective grazing on T. hemprichii facilitated C. rotundata, we established paired seagrass preference experiments. Turtles had clear preferences for T. hemprichii (64%), but also grazed on C. rotundata . Synthesis . Taken together, our results show that high‐impact turtle herbivory changes seagrass composition, precipitating dominance shifts in grazed meadows by mediating direct and apparent competition. Given the crucial role of megaherbivores in seagrass meadow functioning, our results suggest that past meadows may have had natural functional limits to megaherbivore densities that they could sustainably support.

Societal Impact Statement Seagrass ecosystems are of fundamental importance to our planet and wellbeing. Seagrasses are marine flowering plants, which engineer ecosystems that provide a multitude of ecosystem services, for example, blue foods and carbon sequestration. Seagrass ecosystems have largely been degraded across much of their global range. There is now increasing interest in the conservation and restoration of these systems, particularly in the context of the climate emergency and the biodiversity crisis. The collation of 100 questions from experts across Europe could, if answered, improve our ability to conserve and restore these systems by facilitating a fundamental shift in the success of such work. Summary Seagrass meadows provide numerous ecosystem services including biodiversity, coastal protection, and carbon sequestration. In Europe, seagrasses can be found in shallow sheltered waters along coastlines, in estuaries & lagoons, and around islands, but their distribution has declined. Factors such as poor water quality, coastal modification, mechanical damage, overfishing, land‐sea interactions, climate change and disease have reduced the coverage of Europe’s seagrasses necessitating their recovery. Research, monitoring and conservation efforts on seagrass ecosystems in Europe are mostly uncoordinated and biased towards certain species and regions, resulting in inadequate delivery of critical information for their management. Here, we aim to identify the 100 priority questions, that if addressed would strongly advance seagrass monitoring, research and conservation in Europe. Using a Delphi method, researchers, practitioners, and policymakers with seagrass experience from across Europe and with diverse seagrass expertise participated in the process that involved the formulation of research questions, a voting process and an online workshop to identify the final list of the 100 questions. The final list of questions covers areas across nine themes: Biodiversity & Ecology; Ecosystem services; Blue carbon; Fishery support; Drivers, Threats, Resilience & Response; Monitoring & Assessment; Conservation & Restoration; Governance, Policy & Management; and Communication. Answering these questions will fill current knowledge gaps and place European seagrass onto a positive trajectory of recovery.

Abies nordmanniana is used for Christmas tree production but poor seed germination and slow growth represent challenges for the growers. We addressed the plant growth promoting potential of root-associated bacteria isolated from A. nordmanniana. Laboratory screenings of a bacterial strain collection yielded several Bacillus and Paenibacillus strains that improved seed germination and produced indole-3-acetic acid. The impact of three of these strains on seed germination, plant growth and growth-related physiological parameters was then determined in greenhouse and field trials after seed inoculation, and their persistence was assessed by 16S rRNA gene-targeted bacterial community analysis. Two strains showed distinct and significant effects. Bacillus sp. s50 enhanced seed germination in the greenhouse but did not promote shoot or root growth. In accordance, this strain did not increase the level of soluble hexoses needed for plant growth but increased the level of storage carbohydrates. Moreover, strain s50 increased glutathione reductase and glutathione-S-transferase activities in the plant, which may indicate induction of systemic resistance during the early phase of plant development, as the strain showed poor persistence in the root samples (rhizosphere soil plus root tissue). Paenibacillus sp. s37 increased plant root growth, especially by inducing secondary root formation, under in greenhouse conditions, where it showed high persistence in the root samples. Under these conditions, it further it increased the level of soluble carbohydrates in shoots, and the levels of starch and non-structural carbohydrates in roots, stem and shoots. Moreover, it increased the chlorophyll level in the field trial. These findings indicate that this strain improves plant growth and vigor through effects on photosynthesis and plant carbohydrate reservoirs. The current results show that the two strains s37 and s50 could be considered for growth promotion programs of A. nordmanniana in greenhouse nurseries, and even under field conditions.

Plant-pathogen-interactions have been widely studied, but mostly from the site of the plant secondary defense. Less is known about the effects of pathogen infection on plant primary metabolism. The possibility to transform a fluorescing protein into prokaryotes is a promising phenotyping tool to follow a bacterial infection in plants in a noninvasive manner. In the present study virulent and avirulent Pseudomonas syringae strains were transformed with Green Fluorescent Protein (GFP) to follow the spread of bacteria in vivo by imaging PAM fluorescence and conventional binocular microscopy. The combination of various wavelengths and filters allowed simultaneous detection of GFP transformed bacteria, PAM chlorophyll fluorescence and phenolic fluorescence from pathogen infected plant leaves. The results show that fluorescence imaging allows spatiotemporal monitoring of pathogen spread as well as phenolic- and chlorophyll fluorescence in situ thus providing a novel means to study complex plant-pathogen interactions and relate the responses of primary and secondary metabolism to pathogen spread and multiplication. The study establishes a deeper understanding of imaging data and their implementation into disease screening.

The diel variability in picophytoplankton cell death was analyzed by quantifying the proportion of dead cyanobacteria Prochlorococcus and Synechococcus cells along several in situ diel cycles in the open Mediterranean Sea. During the diel cycle, total cell abundance varied on average 2.8 ± 0.6 and 2.6 ± 0.4 times for Synechococcus and Prochlorococcus populations, respectively. Increasing percentages of dead cells of Prochlorococcus and Synechococcus were observed during the course of the day reaching the highest values around dusk and decreasing as the night progressed, indicating a clear pattern of diel variation in the cell mortality of both cyanobacteria. Diel cycles of cell division were also monitored. The maximum percentage of dead cells (Max % DC) and the G2 + M phase of the cell division occurred within a period of 2 h for Synechoccoccus and 4.5 h for Prochlorococcus, and the lowest fraction of dead cells occurred at early morning, when the maximum number of cells in G1 phase were also observed. The G1 maximum corresponded with the maximal increase in newly divided cells (minimum % dead cells), and the subsequent exposure of healthy daughter cells to environmental stresses during the day resulted in the progressive increase in dying cells, with the loss of these cells from the population when cell division takes place. The discovery of diel patterns in cell death observed revealed the intense dynamics of picocyanobacterial populations in nature.

Heat stress around anthesis is considered to have an increasing impact on wheat yield under the ongoing climate change. However, the effect of high temperatures and their duration on formation of individual yield parameters is still little understood. Within this study, the effect of high temperatures applied during anthesis for 3 and 7 days on yield formation parameters was analysed. The study was conducted in growth chambers under four temperature regimes (daily temperature maxima 26, 32, 35 and 38°C). In the periods preceding and following heat stress regimes the plants were cultivated under ambient weather conditions. The number of grains per spike was reduced under temperatures ≥ 35°C in cv. Bohemia and ≥ 38°C in cv. Tobak. This resulted in a similar response of spike productivity. Thousand grain weight showed no response to temperature regime in cv. Tobak, whereas in cv. Bohemia, a peak response to temperature with maximum at 35°C was observed. The duration of heat stress had only little effect on most yield formation parameters.

Abstract Aim Compare the distribution and composition of temperate Labridae (wrasse) assemblages on shallow water coastal reefs in South‐Western Australia between 2006 and 2015, after a decade characterized by both gradual ocean warming and severe heatwave events. Location South‐Western Australia from Port Gregory to the Recherché Archipelago. Methods Surveys of Labridae fishes were conducted in 2006 and repeated in 2015 across 112 reefs spanning 2,000 km of coastline, using diver‐operated stereo‐video systems (stereo‐DOVs). We used a hierarchical design with seven regions, four locations in each region, four reef sites in each location and twelve transects in each site. Results In 2015, we found an increase in abundance of tropical and subtropical labrid species that were rarely observed in 2006. Three temperate species declined in abundance, which tended to be large and slow growing fish. Twenty‐two labrid species increased in abundance. There was also a discernible poleward shift in 20 of the 25 most abundant and representative labrid species from 2006 to 2015. The labrid community composition was explained predominantly by sea surface temperature (SST), physical reef structure and kelp ( Ecklonia radiata ) cover. Main conclusions Our study reveals that labrid assemblages associated with the shallow water temperate reefs of South‐Western Australia have undergone rapid changes across almost 2,000 km of coastline, with warm‐temperate waters showing the strongest change. However, cool‐temperate waters on the south coast also showed significant changes in the composition of the labrid assemblages. Our findings provide important insights into the effects of warming and habitat loss on warm‐temperate assemblages and the potential trajectory of change for cool‐temperate assemblages under a warmer future.

Necrotic and chlorotic symptoms induced during Pyrenophora teres infection in barley leaves indicate a compatible interaction that allows the hemi-biotrophic fungus Pyrenophora teres to colonise the host. However, it is unexplored how this fungus affects the physiological responses of resistant and susceptible cultivars during infection. To assess the degree of resistance in four different cultivars, we quantified visible symptoms and fungal DNA and performed expression analyses of genes involved in plant defence and ROS scavenging. To obtain insight into the interaction between fungus and host, we determined the activity of 19 key enzymes of carbohydrate and antioxidant metabolism. The pathogen impact was also phenotyped non-invasively by sensor-based multireflectance and -fluorescence imaging. Symptoms, regulation of stress-related genes and pathogen DNA content distinguished the cultivar Guld as being resistant. Severity of net blotch symptoms was also strongly correlated with the dynamics of enzyme activities already within the first day of infection. In contrast to the resistant cultivar, the three susceptible cultivars showed a higher reflectance over seven spectral bands and higher fluorescence intensities at specific excitation wavelengths. The combination of semi high-throughput physiological and molecular analyses with non-invasive phenotyping enabled the identification of bio-signatures that discriminates the resistant from susceptible cultivars.

Magnetically modified Posidonia oceanica sea grass dead biomass was employed as an adsorbent of organic dyes. The adsorption of seven water-soluble organic dyes was characterized using Langmuir adsorption model. The highest calculated maximum adsorption capacity was found for Bismarck brown Y (233.5 mg g-1), while the lowest capacity value was obtained for safranin O (88.1 mg g-1). The adsorption processes followed the pseudo-second-order kinetic model and the thermodynamic studies indicated spontaneous and endothermic adsorption.

Abstract. The Mediterranean Sea is warming faster than the global ocean, with important consequences for organisms and biogeochemical cycles. Warming is a major stressor for key marine benthic macrophytes. However, the effect of warming on marine N2 fixation remains unknown, despite the fact that the high productivity of macrophytes in oligotrophic waters is partially sustained by the input of new nitrogen (N) into the system by N2 fixation. Here, we assess the impact of warming on the N2 fixation rates of three key marine macrophytes: Posidonia oceanica, Cymodocea nodosa, and Caulerpa prolifera. We experimentally measured N2 fixation rates in vegetated and bare sediments at temperatures encompassing current summer mean (25 and 27 ∘C), projected summer mean (29 and 31 ∘C), and projected summer maximum (33 ∘C) seawater surface temperatures (SSTs) by the end of the century under a scenario of moderate greenhouse gas emissions. We found that N2 fixation rates in vegetated sediments were 2.8-fold higher than in bare sediments at current summer mean SST, with no differences among macrophytes. Currently, the contribution of N2 fixation to macrophyte productivity could account for up to 7 %, 13.8 %, and 1.8 % of N requirements for P. oceanica, C. nodosa, and C. prolifera, respectively. We show the temperature dependence of sediment N2 fixation rates. However, the thermal response differed for vegetated sediments, in which rates showed an optimum at 31 ∘C followed by a sharp decrease at 33 ∘C, and bare sediments, in which rates increased along the range of the experimental temperatures. The activation energy and Q10 were lower in vegetated than bare sediments, indicating the lower thermal sensitivity of vegetated sediments. The projected warming is expected to increase the contribution of N2 fixation to Mediterranean macrophyte productivity. Therefore, the thermal dependence of N2 fixation might have important consequences for primary production in coastal ecosystems in the context of warming.

The natural environment has been significantly altered by human activity over the past few decades. There is evidence we are now experiencing the sixth mass extinction, as many species of plants and animals are declining in abundance. We focused on the Orchidaceae because this plant family has experienced one of the biggest reductions in distribution. We investigated patterns in species richness and distribution of orchids, the rate and causes of their decrease and extinction, and factors influencing their occurrence in the Czech Republic and Greece. The key findings are: (i) Method of pollination and type of rooting system are associated with their distributions and they are different in the two countries. We assume that these differences might be due to the difference in the orography, distribution of suitable habitats and types of bedrock in these two countries. (ii) The greatest reduction in distribution was recorded for critically endangered taxa of orchids. The number of sites suitable for orchids in the Czech Republic declined by 8–92%. The most threatened orchid species are Spiranthes spiralis, Anacamptis palustris, Epipogium aphyllum and Goodyera repens. The distribution of orchids in the Czech Republic is mainly determined by the distribution of their habitats. (iii) The most important factor affecting the distribution of Czech orchids in South Bohemia is land cover. And the most important types of habitats (types in KVES) are oak and oak-hornbeam forests and agricultural meadows. Based on this information, it should be possible to improve the management that is crucial for maintaining orchid localities.

ABSTRACT Aim Long‐standing theory predicts that the intensity of biotic interactions increases from high to low latitudes. Studies addressing geographic variation in predation on insect prey have often relied on prey models, which lack many characteristics of live prey. Our goals were to explore global latitudinal patterns of predator attack rates on standardised live insect prey and to compare the patterns in predation on live insects with those on plasticine prey models. Location Global forested areas. Time Period 2021–2023. Major Taxa Arthropods, birds. Methods We measured predation rates in 43 forested locations distributed across five continents from 34.1° S to 69.5° N latitude. At each location, we exposed 20 sets of three bait types, one set per tree. Each set included three live fly larvae (maggots), three live fly puparia and three plasticine models of the puparia. We used glue rings to isolate half of the sets from non‐flying predators. Results Arthropod attack rates on plasticine prey decreased linearly from low to high latitudes, whereas attack rates on maggots had a U shaped distribution, with the lowest predation rates at temperate latitudes and the highest rates at tropical and boreal latitudes. This difference emerged from intensive predator attacks on live maggots, but not on plasticine models, in boreal sites. Site‐specific attack rates of arthropod predators on live and plasticine prey were not correlated. In contrast, bird attack rates on live maggots and plasticine models were positively correlated, but did not show significant latitudinal changes. Main Conclusions Latitudinal patterns in predation differ between major groups of predators and between types of prey. Poleward decreases in both arthropod and combined arthropod and bird predation on plasticine models do not mirror patterns of predation on our live prey, the latter likely reflecting real patterns of predation risk better than do patterns of attack on artificial prey.

Abies nordmanniana is an economically important tree crop widely used for Christmas tree production. After initial growth in nurseries, seedlings are transplanted to the field. Rhizosphere bacterial communities generally impact the growth and health of the host plant. However, the dynamics of these communities during A. nordmanniana growth in nurseries, and during transplanting, has not previously been addressed. By a 16S rRNA gene amplicon sequencing approach, we characterized the composition and dynamics of bacterial communities in the rhizosphere during early plant growth in field and greenhouse nurseries, and for plants transplanted from the the greenhouse to the field. Moreover, the nitrogen cycling potential of rhizosphere bacteria across plant age was addressed in both nurseries. Overall, a rhizosphere core microbiome of A. nordmanniana, comprising 19.9% of the taxa at genus level, was maintained across plant age, nursery production systems and even during the transplantation of plants from the greenhouse to the field. The core microbiome included the bacterial genera Bradyrhizobium, Burkholderia, Flavobacterium, Pseudomonas, Rhizobium, Rhodanobacter and Sphingomonas which harbor several N-fixing and plant growth-promoting taxa. Never-the-less, both plant age and production system caused significant changes in the rhizosphere bacterial communities. Concerning community composition, the relative abundance of Rhizobiales (genera Rhizobium, Bradyrhizobium and Devosia) was higher in the rhizosphere of field-grown A. nordmanniana, while the relative abundance of Enterobacteriales and Pseudomonadales (genus Pseudomonas) was higher in the greenhouse. Analysis of community dynamics across plant age showed that in the field nursery, the most abundant bacterial orders showed more dynamic changes in their relative abundance in the rhizosphere than in the bulk soil. In the greenhouse, age-dependent dynamics even occurred but affected different taxa than for the field-grown plants. The N-cycling potential of rhizosphere bacterial communities showed an increase of the relative abundance of genes involved in nitrogen fixation and denitrification by plant age. Similarly, the relative abundance of reported nitrogen-fixing or denitrifying bacteria increased by plant age. However, different community structures seemed to lead to an increased potential for nitrogen-fixation and denitrification in the field versus greenhouse nurseries.

Submarine Groundwater Discharge (SGD) delivers nutrients to the coastal sea triggering phytoplankton blooms, eutrophication, and can also serve as a pathway for contaminants. Wastewater treatment plants (WWTP) including injection wells in coastal areas influence coastal aquifers and might impact the composition and magnitude of SGD fluxes. In tourist areas, wastewater treatment may be less efficient and larger in volume during high seasons, potentially impacting nutrient fluxes from SGD and exacerbating environmental impacts. This study analyzes the nutrient transfer from treated wastewater injection in karstic aquifers to the coastal sea via SGD, considering the impacts of tourism seasonality. This study is conducted in Cala Deià, a small cove in the Balearic Islands, a Mediterranean tourist destination. The findings suggest that the seasonality of tourism, leading to variations in the volume of wastewater treated in the WWTP, influences the dynamics of the coastal aquifer. This leads to increased SGD water and nutrient fluxes to the sea in summer, i.e. the peak tourist season. The measured DIN, DIP, and DSi inventories in the cove are much larger in August than in April (3, 10, and 1.5 times higher, respectively) due to higher input of nutrients in summer due to SGD impacted by the WWTP. These elevated nutrient flows can support algal blooms in the cove, compromising water quality for local swimmers and tourists. Indeed, in August, shoreline stations exhibited eutrophic Chl-a concentrations, with peaks reaching approximately 4 mg Chl-a L−1. These elevated levels suggest the presence of an algal bloom during the survey. The anthropogenic origin of SGD-driven nutrients is traced in seawater and seagrass meadows, as evidenced by high ∂15N signatures indicative of polluted areas. Thus, the high pressure exerted on coastal areas by tourism activities increased the magnitude of SGD nutrient fluxes, thereby threatening coastal ecosystems and the services they provide.

Recently, there has been a significant increase in number of land cover maps available to researchers and they are now more commonly used. The broad variety requires some system for determining the differences between maps and for estimating their applicability for specific research purposes. We focused on comparing land cover maps from the point of view of how the land cover categories used characterize potentially suitable habitats for species. This comparison includes only freely available global land cover projects with resolutions from 1 km to 10 m. The criteria chosen were temporal and spatial resolution, number of classes and map precision. To demonstrate the differences, two areas of different sizes were always chosen. Our results reveal that maps can significantly vary in their estimates of different types of land cover, even at the same spatial resolution. Results also revealed that one type of vegetation in this area is poorly recorded in all land cover maps. Copernicus CGLS-LC100 and ESA CCI-LC maps appear to be the most suitable for evaluating potentially suitable habitats.

Grape phenolics are considered to have a significant impact on wine quality, with their quantity and dynamics being strongly influenced by environmental conditions. We investigated the effect of ambient sunlight on the temporal dynamics of phenolics in cv. Chardonnay under field conditions during the 2012 ripening season, from véraison until harvest. The phenolic profiles of the grapes were monitored at predefined time intervals by using high-pressure liquid chromatography (HPLC) and spectrophotometric (OD) analysis. The obtained concentrations were correlated with the average sunlight intensities preceding the sampling. No significant correlations were found between the hydroxycinnamic acid, hydroxybenzoic acid and stilbene content, in contrast with the strong relationships found between: (i) OD280 nm, (ii) catechin, (iii) flavanols and (iv) total polyphenols and the fluctuating dose of ambient sunlight reaching the grapes throughout the ripening season. The light-dependent dynamics of several main phenolic compounds in cv. Chardonnay during the ripening period could help to establish correlation models that increase the applicability of meteorological data in the assessment of optimal phenolic ripeness in modern viticulture.

High Resolution Image Download MS PowerPoint Slide Two main substances are responsible for the pigmentation of the eggshells of birds and reptiles: the bluish-green tone comes from biliverdin (BV), and protoporphyrin IX (PP) gives brown hues. BV and PP can form complexes with metal cations. The main objective of this investigation is to carry out a theoretical study that analyzes the interaction of metal cations (Cu 2+, Ag 2+, Au 2+, Cd 2+, Zn 2+, and Hg 2+ ) with BV and PP. The divalent metal ions of Cu and Ag are selected to have compounds with the same charge. Density functional theory (DFT) calculations were used to investigate the antiradical capacity of these systems and to obtain ultraviolet–visible (UV–vis) spectra to analyze color modifications. Antiradical capacity is one of the mechanisms that prevents oxidative stress. The antibacterial capacity was investigated through the formation of triplet states. From our results, we can conclude that metal cations interacting with BV and PP affect the electron donor–acceptor properties of the systems, modify coloration of the eggshell, and increase the photoactivating capacity of pigments, which is related to their antimicrobial action. Electron transfer is an important mechanism of antioxidant defense. These results provide useful information on both the influence of contaminants such as heavy metals on the antimicrobial capacity of natural pigments and the signaling value of eggshell coloration.

We investigated the factors affecting group sizes and population composition of chital (Axis axis), sambar (Rusa unicolor), hog deer (Axis porcinus), northern red muntjac (Muntiacus vaginalis), wild boar (Sus scrofa) and gaur (Bos gaurus) in the Chitwan National Park in southern Nepal. The study revealed that mean group sizes were the largest for chital (winter: 13.76 and summer: 11.01), followed by wild boar (winter: 6.89 and summer: 8.51), hog deer (winter: 5.52 and summer: 6.66), gaur (winter: 4.36 and summer: 5.81), sambar (winter: 1.86 and summer: 2.45) and muntjac (winter: 1.44 and summer: 1.46). The age and sex ratio of ungulates were biased towards females in all species. This study found the highest proportion of young individuals in wild boar and the lowest in gaur. Habitat structure, presence of predators and human disturbances strongly affected group size of ungulates in this area. Larger groups of ungulates were found in less disturbed open areas with high predatory pressure, while smaller groups and solitary individuals were found in highly disturbed forest areas. We recommend that management of human disturbances inside the park and regular monitoring of the changes in the demography of ungulate populations will improve long-term conservation strategies in this park. Journal of Institute of Science and TechnologyVolume 22, Issue 2, January 2018, Page: 156-167