Interactions of natural products with β-lactoglobulins, members of the lipocalin family

Natural products constitute an important source of new drugs. The bioavailability of the drugs depends on their absorption, distribution, metabolism and elimination. To achieve good bioavailability, the drug must be soluble in water, stable in the gastrointestinal tract and palatable. Binding proteins may improve the solubility of drug compounds, masking unwanted properties, such as bad taste, bitterness or toxicity, transporting or protecting these compounds during processing and storage. The focus of this thesis was to study the interactions, including ligand binding and the effect of pH and temperature, of bovine and reindeer β-lactoglobulin (βLG) with such compounds as retinoids, phenolic compounds as well as with compounds from plant extracts, and to investigate the transport properties of the βLG-ligand complex. To examine the binding interactions of different ligands to βLG, new methods were developed. The fluorescence binding method for the evaluation of ligand binding to βLG was miniaturized from a quartz cell to a 96-well plate. A method of ultrafiltration sampling combined with high-performance liquid chromatography was developed to assess the binding of compounds from extracts. The interactions of phenolic compounds or retinoids and βLG were investigated using the 96-well plate method. The majority of flavones, flavonols, flavanones and isoflavones and all of the retinoids included were shown to bind to bovine and reindeer βLG. Phenolic compounds, contrary to retinol, were not released at acidic pH. Those results suggest that βLG may have more binding sites, probably also on the surface of βLG. An extract from Camellia sinensis (L.) O. Kunze (black tea), Urtica dioica L. (nettle) and Piper nigrum (black pepper) were used to evaluate whether βLG could bind compounds from plant extracts. Piperine from P. nigrum was found to bind tightly and rutin from U. dioica weakly to βLG. No components from C. sinensis bound to βLG in our experiment. The uptake and membrane permeation of bovine and reindeer βLG, free and bound with retinol, palmitic acid and cholesterol, were investigated using Caco-2 cell monolayers. Both bovine and reindeer βLG were able to cross the Caco-2 cell membrane. Free and βLG-bound retinol and palmitic acid were transported equally, whereas cholesterol could not cross the Caco-2 cell monolayer free or bound to βLG. Our results showed that βLG can bind different natural product compounds, but cannot enhance transport of retinol, palmitic acid or cholesterol through Caco-2 cells. Despite this, βLG, as a water-soluble binding protein, may improve the solubility of natural compounds, possibly protecting them from early degradation and transporting some of them through the stomach. Furthermore, it may decrease their bad or bitter taste during oral administration of drugs or in food preparations. βLG can also enhance or decrease the health benefits of herbal teas and food preparations by binding compounds from extracts.

Agroforestry systems for sustainable livelihoods and improved land management in the East Usambara Mountains, Tanzania

Forest destruction for agriculture continues to be a major threat to the rich biological diversity in the East Usambara Mountains in the north-eastern corner of Tanzania. The highest ratio of endemic plant and animal species found on 100 km2 anywhere in the world is depending on the remaining natural forests. Forests are vitally important for the local population in many different ways, and nationally they are an important source of water and hydroelectricity. The soils, of low fertility and mostly acidic Ferrasols, mainly have the nutrients in the topsoil. After clear-cutting, the soils soon become poor when the topsoil is eroded. High-value cardamom is nowadays unsustainably cultivated in the natural forests of the East Usambaras. The general aim was to study the possibilities to develop new profitable and sustainable agroforestry systems for the benefit of the local people that could contribute to relieving the pressure on the remaining natural forests in the East Usambara Mountains. Results from a spice crop agroforestry trial, established in cooperation with a local farmer, showed a clear advantage of intercropping cardamom (Elettaria cardamomum) and black pepper (Piper nigrum) with trees, especially with Grevillea robusta. The nitrogen fixing tree species Gliricidia sepium also improved the nitrogen and organic matter content of the soil over levels found in the natural forest. With improved agroforestry methods for spice production the households generated as much as13 times the net income obtained with traditional forest cultivation practices. There are thus sustainable and profitable ways to cultivate spices as cash crops in well-managed homegardens. However, the farmers need stable markets, access to credit and comprehensive extension services. The soil fertility depletion should be reversed with organic manure application and an enabling policy environment for the smallholder-farming sector. Strong farmers organisations and equal rights to resources and decision-making are needed. Organic spices have an increasing demand, and their export would be profitable for these farmers. What is, however, most needed for a change is a political will of a government that understands the importance of agricultural and forestry development for poverty reduction.

Effect of phenolic-rich plant materials on protein and lipid oxidation reactions

The antioxidant activity of natural plant materials rich in phenolic compounds is being widely investigated for protection of food products sensitive to oxidative reactions. In this thesis plant materials rich in phenolic compounds were studied as possible antioxidants to prevent protein and lipid oxidation reactions in different food matrixes such as pork meat patties and corn oil-in water emulsions. Loss of anthocyanins was also measured during oxidation in corn oil-in-water emulsions. In addition, the impact of plant phenolics on amino acid level was studied using tryptophan as a model compound to elucidate their role in preventing the formation of tryptophan oxidation products. A high-performance liquid chromatography (HPLC) method with ultraviolet and fluorescence detection (UV-FL) was developed that enabled fast investigation of formation of tryptophan derived oxidation products. Byproducts of oilseed processes such as rapeseed (Brassica rapa L.), camelina (Camelina sativa) and soy meal (Glycine max L.) as well as Scots pine bark (Pinus sylvestris) and several reference compounds were shown to act as antioxidants toward both protein and lipid oxidation in cooked pork meat patties. In meat, the antioxidant activity of camelina, rapeseed and soy meal were more pronounced when used in combination with a commercial rosemary extract (Rosmarinus officinalis). Berry phenolics such as black currant (Ribes nigrum) anthocyanins and raspberry (Rubus idaeus) ellagitannins showed potent antioxidant activity in corn oil-in-water emulsions toward lipid oxidation with and without β-lactoglobulin. The antioxidant effect was more pronounced in the presence of β-lactoglobulin. The berry phenolics also inhibited the oxidation of tryptophan and cysteine side chains of β-lactoglobulin. The results show that the amino acid side chains were oxidized prior the propagation of lipid oxidation, thereby inhibiting fatty acid scission. In addition, the concentration and color of black currant anthocyanins decreased during the oxidation. Oxidation of tryptophan was investigated in two different oxidation models with hydrogen peroxide (H2O2) and hexanal/FeCl2. Oxidation of tryptophan in both models resulted in oxidation products such as 3a-hydroxypyrroloindole-2-carboxylic acid, dioxindolylalanine, 5-hydroxy-tryptophan, kynurenine, N-formylkynurenine and β-oxindolylalanine. However, formation of tryptamine was only observed in tryptophan oxidized in the presence of H2O2. Pine bark phenolics, black currant anthocyanins, camelina meal phenolics as well as cranberry proanthocyanidins (Vaccinium oxycoccus) provided the best antioxidant effect toward tryptophan and its oxidation products when oxidized with H2O2. The tryptophan modifications formed upon hexanal/FeCl2 treatment were efficiently inhibited by camelina meal followed by rapeseed and soy meal. In contrast, phenolics from raspberry, black currant, and rowanberry (Sorbus aucuparia) acted as weak prooxidants. This thesis contributes to elucidating the effects of natural phenolic compounds as potential antioxidants in order to control and prevent protein and lipid oxidation reactions. Understanding the relationship between phenolic compounds and proteins as well as lipids could lead to the development of new, effective, and multifunctional antioxidant strategies that could be used in food, cosmetic and pharmaceutical applications.

Climate change and crop yields in the Indian Cardamom Hills, 1978-2007 CE

In this study we analyzed climate and crop yields data from Indian cardamom hills for the period 1978-2007 to investigate whether there were significant changes in weather elements, and if such changes have had significant impact on the production of spices and plantation crops. Spatial and temporal variations in air temperatures (maximum and minimum), rainfall and relative humidity are evident across stations. The mean air temperature increased significantly during the last 30 years; the greatest increase and the largest significant upward trend was observed in the daily temperature. The highest increase in minimum temperature was registered for June (0.37A degrees C/18 years) at the Myladumpara station. December and January showed greater warming across the stations. Rainfall during the main monsoon months (June-September) showed a downward trend. Relative humidity showed increasing and decreasing trends, respectively, at the cardamom and tea growing tracts. The warming trend coupled with frequent wet and dry spells during the summer is likely to have a favorable effect on insect pests and disease causing organisms thereby pesticide consumption can go up both during excess rainfall and drought years. The incidence of many minor pest insects and disease pathogens has increased in the recent years of our study along with warming. Significant and slight increases in the yield of small cardamom (Elettaria cardamomum M.) and coffee (Coffea arabica), respectively, were noticed in the recent years.; however the improvement of yield in tea (Thea sinensis) and black pepper (Piper nigrum L.) has not been seen in our analysis.

The role of macrophytes in the economy of the lower Sondu-Miriu river of Lake Victoria

This paper highlights the results of a 12 month's ecological study of macrophytes and their role in the economy of the lower Sondu-Miriu river of Lake Victoria. A total of 34 species of aquatic macrophytes were identified. These were grouped as emergent, floating leaved, free floating and submersed. The major community type identified were dominated by Cladium jamaicanse (Crantz) Kurk., Cyperus paprus (L.) and Cyperus latifolius (Poir). The study observed an over dependence on macrophytes by the local community for the supply of bedding, grazing fields, fuel, roofing and other building materials. The white (heart) of Typha shoot, Nile cabbage (Pistia stratiotes and a legume solanum nigrum traditionally known as "osuga" are used as vegetables by local community. The only source of protein in this arid region is the fish caught in the swamps and birds which inhabit the swamp. The swamp provides the only source of green pasture in the dry season for the animals. An export oriented economy has developed involving mat making and basket weaving with exports reaching as far as Japan. The study proposes a rational and sustainable exploitation of the littoral zone to support the adjacent rural community.

镉超积累植物的生态特征及污染耐性机理分析

在已发现的超积累植物中，镉超积累植物种类较少，对其重金属耐性及超积累机理的研究还未有突破性进展，这些植物金属耐性机理的描述对于镉污染土壤的植物修复研究是非常有意义的。本研究以镉超积累植物龙葵（Solanum nigrum L.）、球果蔊菜（Rorippa globosa (Turcz.) Thell.）为研究对象，以茄子（Solanum melongena L.）和风花菜（Rorippa islandica (Oeder.) Borb.）为对照植物，描述了镉超积累植物污染耐性的生态特征，系统探讨了镉超积累植物的耐性机制。 （1）龙葵较茄子有更高的镉耐性及镉积累性；球果蔊菜较风花菜有更高的镉耐性及镉积累性，球果蔊菜茎部受抑制现象是实现其特异吸收和积累重金属的适应性反应，这种地上部营养物质的重新分配现象可能是球果蔊菜作为超积累植物独特的生态特征。 （2）龙葵根系中SOD和CAT活性及叶片中POD和CAT活性的协同作用在缓解镉积累引起的膜脂过氧化损伤中发挥着积极作用，球果蔊菜叶片SOD和CAT的协同作用在缓解镉积累引起的膜脂过氧化损伤中也发挥重要作用，可以推断龙葵、球果蔊菜体内抗氧化酶防御系统在其镉耐性中扮演重要的角色。 （3）镉的污染胁迫均能明显促进龙葵和球果蔊菜体内脯氨酸的积累，脯氨酸在其镉耐性及积累性中扮演着重要角色，其中叶片脯氨酸可能发挥着更大的作用。 （4）龙葵叶片中柠檬酸和乙酸含量的动态变化与其镉积累有关，球果蔊菜叶片中苹果酸和酒石酸含量的动态变化与其镉积累有关，有机酸可能在龙葵及球果蔊菜镉耐性及积累性中起到一定的作用，但高含量的有机酸并不是植物镉耐性和超积累性的主要原因。 （5）植物螯合肽可以作为植物镉超积累特征的生物指标，但植物螯合肽的合成并非镉超积累植物耐性的主要机制。

植物修复镉-多环芳烃污染土壤强化措施研究

本论文以镉超富集植物龙葵(Solanum nigrum L.)为材料，研究化学强化剂对龙葵修复镉-多环芳烃复合污染的作用，并对筛选出的强化剂在不同浓度水平镉-多环芳烃复合污染土壤的适用性进行研究，以期为镉-多环芳烃复合污染土壤的植物修复强化技术提供理论指导。 （1）采用室内盆栽试验的方法，研究外源半胱氨酸、谷氨酸、甘氨酸及其复合处理对超富集植物龙葵修复镉-多环芳烃复合污染土壤的影响。试验表明，各浓度水平的单因素处理对植物株高及地上部干重并没有显著影响（p>0.05），而复合处理则具有显著的促进作用(p<0.05)。各处理能够显著增强龙葵对镉的吸收和富集能力，其中半胱氨酸处理使重金属提取率分别达到1.80%及1.83%，与对照相比提高1.80及1.83倍。研究发现甘氨酸、谷氨酸及半胱氨酸复合处理能够促进镉及多环芳烃在土壤环境中的去除作用，且0.3mmol•kg-1甘氨酸+0.3mmol•kg-1谷氨酸+0.3mmol•kg-1半胱氨酸处理效果最佳，与对照相比龙葵地上部Cd含量增加2.26倍，土壤多环芳烃总量的去除率提高5.46倍。 （2）采用室内盆栽试验的方法，研究EDTA、水杨酸、TW80及其复合处理对超富集植物龙葵修复镉-多环芳烃复合污染土壤的影响。研究发现，单因素EDTA处理对龙葵具有很强的植物毒性作用，显著降低了龙葵地上部干重(p<0.05)。水杨酸及TW80处理促进了土壤中多环芳烃的生物降解作用，其中以0.9mmol•kg-1水杨酸处理效果最佳，芴、苯并(a)蒽、 、苯并(b)荧蒽、苯并(a)芘及苯并(g,h,i) 的降解率分别达到52.3、35.1、60.7、54.5、69.3及68.8％。此外，研究结果显示水杨酸对龙葵地上部提取镉总量具有很好的促进作用，即水杨酸处理能够促进污染物镉和多环芳烃的在土壤环境中的去除作用。 同样，复合处理0.1mmol•kg-1EDTA+0.5mmol•kg-1TW80和0.5mmol•kg-1EDTA+0.3mmol•kg-1水杨酸对以上两种污染物的去除均有很好的促进作用，表明在化学强化剂的协助条件下利用超富集植物修复重金属和有机复合污染土壤是可行的。 （3）采用室外盆栽试验的方法，研究化学强化剂0.1mmol•kg-1EDTA、0.9mmol•kg-1半胱氨酸、0.9mmol•kg-1水杨酸、0.3mmol•kg-1TW80及其复合处理对不同类型的镉和苯并(a)芘土壤修复适用性进行研究。研究发现，高浓度镉和苯并(a)芘对其在土壤环境中的去除产生抑制作用，其抑制强度随浓度的升高而增强。研究结果显示，不同类型的污染土壤对化学强化剂的需求不同，当土壤投加5mg•kg-1Cd时，EDTA+半胱氨酸及EDTA处理对龙葵吸收镉具有显著的促进作用，而土壤镉投加浓度为15mg•kg-1Cd时，仅EDTA+半胱氨酸处理的促进效果最佳；当土壤投加1mg•kg-1Bap时，水杨酸+TW80处理能够显著增强Bap的降解作用，而土壤投加2mg•kg-1Bap时，TW80处理的强化效果最佳；当土壤投加5mg•kg-1Cd和1mg•kg-1Bap时，水杨酸+TW80+EDTA处理对土壤中两种污染物去除均有很好的促进作用，而土壤投加15mg•kg-1Cd和2mg•kg-1Bap时，半胱氨酸+TW80处理效果最佳。

东北茶藨属的分类研究

本文在前人工作的基础上，对薄弱茶藨属植物进行了植物形态分类、叶柄解剖结构及种子脂肪酸成分的综合研究。结果表明：1、刺李属应与茶藨属合并，作为茶藨属下的一个分类等级。２、东北茶藨属分为2亚属、8组，共有14种、2亚种、8变种。其中将R. pauciflorum Turcz. ex Ledeb.降为R. nigrum L.的亚种；将R. palczewskii A. Pojark和R. Liouanum Kitag。分别作为R. spicatum Robs的亚种和变种，将R. repens Bar降为R. triste Pall的变种。3、本属植物叶柄维管束类型为三隙维管束。其维管束形状可以作为划分亚属的依据。2、γ-亚麻酸是本属的特征性成分。本文还详细描述了每个种的形态分类学特征。地理分布和文献考证，并附有每个种的叶片外形图，叶柄横切面解剖图及照片，花部外形图及解剖图。以及分种检索表。

Triterpenoids from the mangrove plant Hibiscus tiliaceus

Three new triterpenoids with the rarely occurring nigrum skeleton, namely (20E)-22-hydroxynigrum-20-en-3-one (1), 21 beta-hydroxynigrum-22(29)-en-3-one (2), and 21 alpha-hydroxynigrum-22(29)-en-3-one (3), were isolated from the mangrove plant Hibiscus tiliaceus. Additionally, five known triterpenoids including friedelin (4),12-oleanen-3 beta-ol (5), 3 beta-hydroxy-12-oleanen-28-oic acid (6),20(29)-lupen-3 beta,28-diol (7). and cucurbita-5,23-dien-3 beta,25-diol (8) were also isolated and identified. The latter structures were elucidated by a detailed NMR and MS analyses, as well as by comparison with reported literature data.

Seringueira em sistemas agroflorestais.

Consorciacao com culturas de ciclo curto. consorciacao com culturas perenes. seringueira x cafeeiro. sistemas temporarios. Consorciacao do cafeeiro durante a formacao do seringal. Substituicao de cafezal em fim de ciclo por seringal. Sistemas permanentes. Seringal em renques no cafezal. Arborizacao do cafezal com seringueiras. Seringueira x cacaueira. Cacaueira x seringais velhos e desfolhados. Novos plantios consorciados de seringueira x cacaueiro. Seringueira x citros. Substituicao de pomares citricos decadentes por seringais. Consorciacao de citros durante a formacao do eringal. Consorciacao permanente de seringueira x citros. Seringueira x pimenteira-do-reino. Seringueira x palmeiras e plantas menos exigentes de luz. Seringueira x guaranazeiro. Seringueir-ras como componentes de quintais agroflorestais.

Identificação dos principais fungos das sementes de trigo.

Procedimento para identificação dos fungos das sementes de trigo; Descrição diagnostica dos principais fungos das sementes de trigo; Sclerotium Tode; Rhizoctonia DC; Chaetomium Kunze; Pleospora Rabenh; Sporobolomyces Kluy. & Niel; Rhodotorula Harrison; Phoma Sacc.; Septoria tritici Rob; Stagonospora nodorum (Berk.) Cas. & Germ.; Stagonospora avenae (Frank) Bisset f. sp. triticae; Colletotrichum graminicola (Ces.) Wilson; Fusarium tricinctum (Corda) Sacc; Fusarium moniliforme Sheldon; Fusarium avenaceum (Fr.) Sacc; Fusarium acuminatum Ell. & Kellerm; Fusarium equiseti (Corda) Sacc.; Fusarium graminearum Schw.; Mucor Micheli; Rhizopus Ehrenb; Aspergillus Link.; Penicillium Link.; Alternaria Nees; Epicoccum Link; Cladosporium Link; Nigrospora Zimm; Curvularia Boedijn; Drechslera tritici-repentis (Died.) Drech; Bipolaris sorokiniana (Sacc. in Sorok.) Shoem; Chave sistemática dos principais fungos de sementes de trigo; Ilustrações dos principais fungos encontrados em sementes de trigo.

Identificação dos principais fungos das sementes de trigo.

Procedimento para identificação dos fungos das sementes de trigo; Descrição diagnostica dos principais fungos das sementes de trigo; Sclerotium Tode; Rhizoctonia DC; Chaetomium Kunze; Pleospora Rabenh; Sporobolomyces Kluy. & Niel; Rhodotorula Harrison; Phoma Sacc.; Septoria tritici Rob; Stagonospora nodorum (Berk.) Cas. & Germ.; Stagonospora avenae (Frank) Bisset f. sp. triticae; Colletotrichum graminicola (Ces.) Wilson; Fusarium tricinctum (Corda) Sacc; Fusarium moniliforme Sheldon; Fusarium avenaceum (Fr.) Sacc; Fusarium acuminatum Ell. & Kellerm; Fusarium equiseti (Corda) Sacc.; Fusarium graminearum Schw.; Mucor Micheli; Rhizopus Ehrenb; Aspergillus Link.; Penicillium Link.; Alternaria Nees; Epicoccum Link; Cladosporium Link; Nigrospora Zimm; Curvularia Boedijn; Drechslera tritici-repentis (Died.) Drech; Bipolaris sorokiniana (Sacc. in Sorok.) Shoem; Chave sistemática dos principais fungos de sementes de trigo; Ilustrações dos principais fungos encontrados em sementes de trigo.

The role of the seed bank in recovery of temperate heath and blanket bog following wildfires

Questions - Are the germinable seed banks of upland heath and blanket bog reduced following wildfires? Are some species at particular risk? Do the impacts of wildfires on seed banks differ between heathlands and blanket bog?

Location - Northern Ireland, United Kingdom.

Methods - Vegetation surveys and seed bank sampling were conducted in 2012 at burned and unburned areas within six upland sites where large wildfires had occurred during spring 2011. Differences in seedling abundance, species richness and Jaccard similarity indices between burned and unburned areas were compared using GLMMs. Differences in the community composition were examined using pRDA.

Results - In total, 24 of the 51 species in the vegetation were detected in the germinable seed bank. Species richness and the abundance of seedlings other than Calluna vulgaris were lower in areas where wildfires had occurred. Species composition of both germinable seed banks and vegetation differed between burned and unburned areas within sites; with negative associations between burned areas and some key indicator species including Drosera rotundifolia, Eriophorum vaginatum, Empetrum nigrum, Narthecium ossifragum and Trichophorum germanicum. We did not find any evidence of significant interactions between burning and habitat, suggesting that wildfires had similar impacts on each species regardless of the habitat in which they occurred.

Conclusions - This study differs from other UK studies in that it examines impacts of wildfires at sites that have not been previously intensively managed by burning. In particular, we highlight potential impacts on N. ossifragum and D. rotundifolia, which are key components of the upland flora and, to our knowledge, were not present in previous UK studies.

Potential Impact of Climate Change on Fungal Distributions: Analysis of 2 Years of Contrasting Weather in the UK

The impact of climate change on fungal growth and spore production is less well documented than for allergenic pollen grains, although similar implications for respiratory tract diseases in humans occur. Fungal spores are commonly described as either “dry” or “wet” according to the type of weather associated with their occurrence in the air. This study examined the distribution of selected fungal spores (Alternaria spp., Cladosporium spp., Didymella spp., Epicoccum spp., Leptosphaeria spp. and rusts) occurring in the West Midlands of UK during 2 years of contrasting weather. Spore specimens were collected using a 7-day volumetric air sampler and then analysed with the aid of light microscopy. Distributions of spores were then studied using normality tests and Mann–Whitney U test, while relationships with meteorological parameters were investigated using Spearman’s rank test and angular-linear correlation for wind direction analysis. Our results showed that so-called wet spores were more sensitive to the weather changes showing statistically significant differences between the 2 years of study, in contrast to “dry” spores. We predict that in following years we will observe accelerated levels in allergenic fungal spore production as well as changes in species diversity. This study could be a starting point to revise the grouping system of fungal spores as either “dry” or “wet” types and their response to climate change

Species distribution models reveal apparent competitive and facilitative effects of a dominant species on the distribution of tundra plants

Abiotic factors are considered strong drivers of species distribution and assemblages. Yet these spatial patterns are also influenced by biotic interactions. Accounting for competitors or facilitators may improve both the fit and the predictive power of species distribution models (SDMs). We investigated the influence of a dominant species, Empetrum nigrum ssp. hermaphroditum, on the distribution of 34 subordinate species in the tundra of northern Norway. We related SDM parameters of those subordinate species to their functional traits and their co-occurrence patterns with E. hermaphroditum across three spatial scales. By combining both approaches, we sought to understand whether these species may be limited by competitive interactions and/or benefit from habitat conditions created by the dominant species. The model fit and predictive power increased for most species when the frequency of occurrence of E. hermaphroditum was included in the SDMs as a predictor. The largest increase was found for species that 1) co-occur most of the time with E. hermaphroditum, both at large (i.e. 750 m) and small spatial scale (i.e. 2 m) or co-occur with E. hermaphroditum at large scale but not at small scale and 2) have particularly low or high leaf dry matter content (LDMC). Species that do not co-occur with E. hermaphroditum at the smallest scale are generally palatable herbaceous species with low LDMC, thus showing a weak ability to tolerate resource depletion that is directly or indirectly induced by E. hermaphroditum. Species with high LDMC, showing a better aptitude to face resource depletion and grazing, are often found in the proximity of E. hermaphroditum. Our results are consistent with previous findings that both competition and facilitation structure plant distribution and assemblages in the Arctic tundra. The functional and co-occurrence approaches used were complementary and provided a deeper understanding of the observed patterns by refinement of the pool of potential direct and indirect ecological effects of E. hermaphroditum on the distribution of subordinate species. Our correlative study would benefit being complemented by experimental approaches.