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We present a genome assembly from an individual Ilex aquifolium (the English holly; Eudicot; Magnoliopsida; Aquifoliales; Aquifoliaceae). The genome sequence is 800.0 megabases in span. Most of the assembly is scaffolded into 20 chromosomal pseudomolecules. The assembled mitochondrial and plastid genomes have lengths of 538.43 kilobases and 157.52 kilobases in length, respectively.

In this study, we investigated the environmental factors associated with the establishment of invasive English holly (Hex aquifolium) and the spatiotemporal trajectories of its potential spread. We collected georeferenced presence and absence and demographic data of holly between 2011 and 2014 in Saint Edward State Park, Washington. We analyzed them through a spatially explicit framework that combines multiple logistic regression, cellular automata, and bivariate quadratic modeling. Results showed that the presence of holly is influenced by the proximity to evergreen coniferous vegetation and forest edge, distance to streams, forest structure, and slope direction. Our study confirmed that local environmental conditions, species characteristics, and vegetative and dispersal mechanisms play a significant role in determining the rate and success of holly establishment across the different phases of invasion. Invasive species management plans should consider both species characteristics and their habitat and the interactions among them when planning long-term strategies for control and eradication. [ABSTRACT FROM AUTHOR]

The iconic tree species,Arbutus menziesii(Pacific madrona, madrone arbutus), has been in decline in the Pacific Northwest of North America for the past 40 years. It is thought that the fungal pathogenNeofusicoccum arbutihas contributed to the decline of this tree species. In recent years, there have been reports of declining arbutus in the coastal region of southern British Columbia, Canada. We conducted intensive sampling in a park with severely affected arbutus trees to determine the cause and prevalence of decline. The majority of arbutus trees sampled in this study had cankers associated withN. arbutiinfection. We also sought to determine ifN. arbutihas additional hosts that could act as a reservoir for this pathogen. Six new hosts ofN. arbutiwere identified, and a seventh was confirmed; these hosts spanned four taxonomic orders and includedAmelanchier alnifolia, Cytisus scoparius(confirmed),Gaultheria shallon, Ilex aquifolium, Rosasp.,Sorbus sitchensisandSpiraea douglasii. Recovery ofNeofusicoccum arbutifrom both symptomatic and asymptomatic plant tissues of these hosts indicates that it potentially has a broad host range. It remains to be established ifN. arbutiis also pathogenic to these hosts. These results highlight the importance of monitoring for this pathogen and assessing the extent of its geographic distribution and its ability to colonize various hosts. [ABSTRACT FROM AUTHOR]

The present study assessed the effects of climatic conditions on radial growth and functional anatomical traits, including ring width, vessel size, vessel frequency and derived variables, i.e., potential hydraulic conductivity and xylem vulnerability to cavitation in Ilex aquifolium L. trees using long-term tree-ring time series obtained at two climatically contrasting sites, one mesic site in Switzerland (CH) and one drought-prone site in Italy (ITA). Relationships were explored by examining different xylem traits, and point pattern analysis was applied to investigate vessel clustering. We also used generalized additive models and bootstrap correlation functions to describe temperature and precipitation effects. Results indicated modified radial growth and xylem anatomy in trees over the last century; in particular, vessel frequency increased markedly at both sites in recent years, and all xylem traits examined, with the exception of xylem cavitation vulnerability, were higher at the CH mesic compared with the ITA drought site. A significant vessel clustering was observed at the ITA site, which could contribute to an enhanced tolerance to drought-induced embolism. Flat and negative relationships between vessel size and ring width were observed, suggesting carbon was not allocated to radial growth under conditions which favored stem water conduction. Finally, in most cases results indicated that climatic conditions influenced functional anatomical traits more substantially than tree radial growth, suggesting a crucial role of functional xylem anatomy in plant acclimation to future climatic conditions. [ABSTRACT FROM AUTHOR]

We present a new hierarchical strategy for fine-grained categorization. Standard, fully automated systems report a single estimate of the category, or perhaps a ranked list, but have non-neglible error rates for most realistic scenarios, which limits their utility. Instead, we propose a semi-automated system which outputs a it confidence set (CS)-a variable-length list of categories which contains the true one with high probability (e.g., a 99 % CS). Performance is then measured by the expected size of the CS, reflecting the effort required for final identification by the user. The implementation is based on a hierarchical clustering of the full set of categories. This tree of subsets provides a graded family of candidate CS's containing visually similar categories. There is also a learned discriminant score for deciding between each subset and all others combined. Selection of the CS is based on the joint score likelihood under a Bayesian network model. We apply this method to determining the species of a plant from an image of a leaf against either a uniform or natural background. Extensive experiments are reported. We obtain superior results relative to existing methods for point estimates for scanned leaves and report the first useful results for natural images at the expense of asking the user to initialize the process by identifying specific landmarks. [ABSTRACT FROM AUTHOR]

Between 2010 and 2013 several English holly trees showing defoliation, leaf and twig blight were found in natural ecosystems in Sardinia and Corsica. A Phytophthora species was consistently isolated from necrotic tissues of leaves and twigs, and bleeding cankers on stems. Isolates obtained were identified as Phytophthora ilicis, on the basis of morphological features, colony patterns and growth rates at different temperatures. Identity was confirmed by sequence analyses of the ITS, β-tubulin and cox1 gene regions. Two different genotypes within P. ilicis were detected. Pathogenicity of P. ilicis isolates was verified by inoculating freshly cut twigs of English holly. Phytophthora ilicis was originally recorded on English holly in western USA, and to date has been found only in cool temperate regions in northern America and, more recently, in Europe. This is the first report of P. ilicis on English holly in the Mediterranean region. Two additional Phytophthora species were isolated from other trees and shrubs species growing together with English holly, including P. bilorbang from Alnus glutinosa leaves and P. pseudosyringae from rhizosphere soil samples of A. glutinosa and Castanea sativa, and from symptomatic leaves of Hedera helix. [ABSTRACT FROM AUTHOR]

Despite the recognized importance of indirect plant-plant interactions for community structure, we still need to improve our current knowledge on how their outcomes are consistent in space and time, as well as reciprocal between participating species. These caveats are especially relevant in the case of indirect interactions mediated by animals, whose behavior may show high variability. We studied consistency and reciprocity of frugivore-mediated interactions between fleshy-fruited trees. For three years we examined the influence of crop size and neighborhood characteristics (con- and heterospecific fruit abundance and forest cover) on frugivory rates on Crataegus monogyna and Ilex aquifolium, two coexisting species in the secondary forests of the Cantabrian range that share a guild of frugivorous birds. Crop size and neighborhood characteristics influenced frugivory on C. monogyna and I. aquifolium. Both con- and heterospecific fruit abundance affected frugivory, evidencing the occurrence of indirect interactions between trees, although the strength and sign of these effects varied between tree species as well as across years within species. By showing complex temporal patterns in the consistency and reciprocity of indirect interactions, this study emphasizes the need for multispecific, long-term studies to assess the actual contribution of animal-mediated plant-plant indirect interactions to community dynamics. [ABSTRACT FROM AUTHOR]

Seed dispersal at large scales strongly influences plant population dynamics. Still, ecologists have rarely measured seed dispersal at relevant scales, and the role of habitat types in affecting seed dispersal at long distances remains unexplored. We studied seed dispersal of Ilex aquifolium and Crataegus monogyna in northern Spain, hypothesizing that seeds would be recovered at higher rates and at longer distances (LDD) at habitats with fleshy-fruited trees, compared to habitats with other tree types or at open habitats. We tracked seeds in eight landscapes by enriching trees with 15N isotopes at the center of landscapes, and then detected 15N-marked seeds by sampling at distances of up to 700 m. We found that seeds arrive in greater densities and at longer distances in habitats with trees, particularly fleshy-fruited types, producing different LDD probabilities for each habitat. Results also show a disproportional arrival of seeds in habitats similar to those of mother plants, which should affect seed establishment and the genetic diversity of plant neighborhoods. Findings reveal the strong dependence of seed dispersal on the existing templates that guide the movements of avian dispersers in heterogeneous landscapes and also suggest that LDD above tree lines and beyond hard habitat edges can be difficult. [ABSTRACT FROM AUTHOR]

Nursery catalogues have recently been used as a method for tracing the introduction history of invasive plants. Information on species, cultivars, plant size and price can help quantify historical changes in propagule pressure. Propagule pressure of invasive ornamentals has to be combined with climatic data to understand shifting distribution patterns of (potentially) invasive plants. In this study the spread of Ilex aquifolium L. cultivars was investigated at the edge of its distribution margin in Denmark. Danish nursery catalogues from 1841-2007 were studied to 1) reconstruct the introduction history of potentially invasive ornamental genotypes of I. aquifolium in Denmark; 2) explore potential factors that could explain naturalization of the species, i.e. nursery location, plant size, hardiness, price and year; 3) investigate whether or not marketing has been more common inside the historical range of the species in western Denmark; and 4) test the hypothesis that marketing of this frost-sensitive species has increased parallel with higher temperatures in the past 100 years. Price, plant size, hardiness, location, and year were recorded for a total of 3213 entries. Through time, increasing prices and the availability of frost-hardy cultivars were the most significant factors contributing to the sale of I. aquifolium in Denmark. The number of nurseries per area, and hence propagule pressure, was higher in eastern Denmark, outside the natural range of the species. The proportion of very frost-hardy cultivars increased with milder winter temperatures. The main conclusion from this combined nursery catalogue and climate study are that nurseries in an indirect way play an important role in increasing the propagule pressure of potentially invasive ornamentals and thus can affect distribution of native species. [ABSTRACT FROM AUTHOR]

Rapid range expansions are becoming more prevalent, especially as climate continues to change. The escape of ornamental plants within their native range represents a significant, but often overlooked component of this process. Few studies have focused on the role of ornamental plants in range expansions using molecular markers to identify the possibility of mixed native and cultivar populations. The purpose of the present study was to determine the genetic variation of a native woody plant with ornamental conspecifics at the edge of its distribution. We selected Ilex aquifolium L. (English holly) which grows naturally in Denmark but is spreading eastward in Scandinavia, possibly due to a combination of climate change and introduction of more frost-tolerant cultivars. We sampled 187 individuals from older and recently established populations, and cultivated I. aquifolium throughout Denmark, and compared them using nuclear SSR and AFLP. The overall results showed no structure or clustering of plants from the historical or the expanding range, or of wild or cultivated plants. The only clusters found were for a group of cultivated hybrid I. aquifolium. The samples represented four genetic groupings, indicating either hybridization between cultivated and wild I. aquifolium or replacement of the latter by cultivars. Thus, ornamental genotypes of I. aquifolium contribute to the northeastern range expansion of the species and eventual invasiveness within its native range. [ABSTRACT FROM AUTHOR]

Abstract: Dispersal limitations contribute to shaping plant distribution patterns and thus are significant for biodiversity conservation and urban ecology. In fleshy-fruited plants, for example, any preference of frugivorous birds affects dispersal capacities of certain fruit species. We conducted a removal experiment with fruits of Ilex aquifolium, a species that is currently expanding its range margin in northern Europe in response to climate change. The species is also a popular ornamental tree and naturalization has been observed in many parts of its range. Fruits of native I. aquifolium and of three cultivars were offered to birds at the expanding range margin in urban habitats in eastern Denmark. The four fruit types were removed at different rates and red fruits were preferred over a yellow cultivar. Small fruit diameter was positively related to fruit removal, and removal was faster under tree canopies compared with open habitats. The preference for red cultivars compared with native I. aquifolium may contribute to naturalization and potential invasion of garden escapes. Preferential foraging under closed canopies indicates trees and shrubs as recruitment foci for fleshy-fruited plants in urban landscapes. The results should be included in urban forestry and planting of potentially invasive ornamental species. [Copyright &y& Elsevier]

Abstract: Animal population density has been related to resource patch size through various hypotheses such as those derived from island biogeography and resource concentration theory. This theoretical framework can be also applied to plant–herbivore interactions, and it can be modified by the sex of the host-plant, and density-dependent relationships. Leaf-miners are specialised herbivores that leave distinct traces on infested leaves in the form of egg scars, mines, signs of predation and emergence holes. This allows the life cycle of the insect to be reconstructed and the success at the different stages to be estimated. The main stages of the leaf-miner Phytomyza ilicis were recorded in eleven populations of the evergreen host Ilex aquifolium in Denmark. Survival rates were calculated and related to population size, sex of the host plant, and egg and mine densities. Host population size was negatively related to leaf-miner prevalence, with larger egg and mine densities in small populations. Percentage of eggs hatching and developing into mines, and percentage of adult flies emerging from mines also differed among host populations, but were not related to population size or host cover. Feeding punctures left by adults were marginally more frequent on male plants, whereas egg scars and mines were more common on females. Overall survival rate from egg stage to adult emergence was higher on female plants. Egg density was negatively correlated with hatching, while mine density was positively correlated with emergence of the larvae. The inverse effects of host population size were not in line with predictions based on island biogeography and resource concentration theory. We discuss how a thorough knowledge of the immigration behaviour of this fly might help to understand the patterns found. [Copyright &y& Elsevier]

Habitat fragmentation increases seed dispersal limitation across the landscape and may also affect subsequent demographic stages such as seedling establishment. Thus, the development of adequate plans for forest restoration requires an understanding of mechanisms by which fragmentation hampers seed delivery to deforested areas and knowledge of how fragmentation affects the relationship between seed-deposition patterns and seedling establishment. We evaluated the dispersal and recruitment of two bird-dispersed, fleshy-fruited tree species (Crataegus monogyna and Ilex aquifolium ) in fragmented secondary forests of northern Spain. Forest fragmentation reduced the probability of seed deposition for both trees because of decreased availability of woody perches and fruit-rich neighborhoods for seed dispersers, rather than because of reductions in tree cover by itself. The effects of fragmentation went beyond effects on the dispersal stage in Crataegus because seedling establishment was proportional to the quantities of bird-dispersed seeds arriving at microsites. In contrast, postdispersal mortality in Ilex was so high that it obscured the seed-to-seedling transition. These results suggest that the effects of fragmentation are not necessarily consistent across stages of recruitment across species. Habitat management seeking to overcome barriers to forest recovery must include the preservation, and even the planting, of fleshy-fruited trees in the unforested matrix as a measure to encourage frugivorous birds to enter into open and degraded areas. An integrative management strategy should also explicitly consider seed-survival expectancies at microhabitats to preserve plant-population dynamics and community structure in fragmented landscapes. [ABSTRACT FROM AUTHOR]

Abstract: Analyzing spatial patterns in plant communities may provide insights in the importance of different processes for community assembly and dynamics. We applied techniques of spatial point pattern analysis to data from a fully mapped plot of a temperate forest community (Corylus avellana, Crataegus monogyna, Fagus sylvatica, Ilex aquifolium and Taxus baccata) in North-western Spain to conduct a community wide assessment of the type and frequency of intra and interspecific spatial association patterns. We first explored the overall intra and interspecific patterning, and then classified the types of association patterns at various neighbourhoods. By conditioning on the larger scale pattern we then explored small-scale (0–15m) intraspecific and interspecific patterns. Association patterns varied from strong positive association at small scales to, as a by-product, repulsion at intermediate scales. Surprisingly, there were no negative associations at small scales, but trees were arranged in multi-species clumps, up to 2.5m in diameter and comprising a few individuals. Ilex and Corylus, the understorey species, were frequently involved in the clumps, showing positive small-scale association with the other tree species. Our analyses highlighted that animal mediated seed dispersal, interspecific facilitation and perturbation processes may operate successively to shape tree distributional patterns, although their relative importance vary among species. Given the complexity of the patterns described and the current threats to some of the species studied, directed experiments in the field are needed to further elucidate some of the hypotheses derived. [Copyright &y& Elsevier]

An approach integrating phytosociological and stand structure surveys with the predictive modelling of species distribution was applied to analyse the spatial distribution and dynamics of the Apennine beech forests with Taxus and Ilex, a high conservation priority forest habitat in Europe. The homogeneity of the habitat was tested trough a Mann-Whitney test between beech woods with Taxus and those with Ilex with respect to climatic, topographic, structural and environmental parameters: the former have proven to be more microthermic, mesophilous and characterised by a closer canopy. Five statistical models were compared to analyse the relationship between bioclimatic parameters and Taxus and Ilex spatial distribution: Regression Tree Analysis, the most efficient model, has shown that the distribution of Taxus is influenced by precipitation variables, while Ilex is mainly influenced by temperature variables. This model highlighted that Ilex has a potential area that surrounds, at lower altitudes, that of Taxus. A stepwise multiple regression analysis has been applied to identify the factors influencing the regeneration of the two species: beside climatic parameters, Taxus regeneration is negatively influenced by soil nitrate concentration (an indicator of livestock disturbance) while Ilex is negatively influenced by beech forest cover. Traditional management practices seem to have an effect on the regeneration of the two species: frequent cuts favour the regeneration of Ilex, reducing the forest cover and allowing more light penetration, while Taxus, less resistant to grazing livestock, is confined to more inaccessible places. The multiple approach has proven to be useful for the elaboration of two differentiated conservation strategies for the two beech forest types. [ABSTRACT FROM AUTHOR]

The objective of this study was to examine antibacterial, antifungal, and antimycobacterial properties of Ilex aquifolium L. (Aquifoliaceae) growing in Turkey. The ethanol, ethyl acetate, chloroform, and n-hexane extracts prepared from the leaves of I. aquifolium were tested against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Enterobacter aerogenes, Proteus vulgaris, Salmonella typhimurium, and Candida albicans for antibacterial and antifungal evaluation using the microdilution broth susceptibility assay. In addition, antimycobacterial activity of the crude extracts of I. aquifolium was evaluated by microplate Alamar blue assay. The results showed that the extracts tested, except n-hexane, possessed moderate antibacterial and antifungal activity varying from 62.5 to 250 μg/mL. On the other hand, the ethanol extract of the leaves exhibited a minimum inhibitory concentration value (MIC) against Mycobacterium tuberculosis H37Ra strain of 200 μg/mL. [ABSTRACT FROM AUTHOR]

The aim of the study was to determine whether there are genetic variations in growth and leaf phenology (flushing and senescence) among populations of six woody species (Abies alba Mill., Acer pseudoplatanus L., Fagus sylvatica L., Fraxinus excelsior L., Ilex aquifolium L., and Quercus petraea (Matt.) Liebl.) along altitudinal gradients, using a common-garden experiment. We found (i) significant differences in phenology and growth among provenances for most species and (ii) evidence that these among-population differences in phenology were related to the annual temperature at the provenance sites for ash, beech, and oak. It is noteworthy that along the same climatic gradient, species can exhibit opposing genetic clines: beech populations from high elevations flushed earlier than those from low elevations, whereas we observed the opposite trend for ash and oak. For most species, significant altitudinal clines for growth were also revealed. Finally, we highlighted the fact that both phenology timing and growth rate were highly consistent from year to year. The results demonstrated that despite the proximity of the populations in their natural area, differences in altitude led to genetic differentiation in their phenology and growth. These adaptive capacities acting along a natural climatic gradient could allow populations to cope with current climate change. [ABSTRACT FROM AUTHOR]

High infection rates of European holly ( Ilex aquifolium) feeder roots by an unknown root-knot nematode were found in a holly forest at Arévalo de la Sierra (Soria province) in northern Spain. Holly trees infected by the root-knot nematode showed some decline and low growth. Infected feeder roots were distorted and showed numerous root galls of large (8–10 mm) to moderate (2–3 mm) size. Morphometry, esterase and malate dehydrogenase electrophoretic phenotypes and phylogenetic trees of sequences within the ribosomal DNA (rDNA) demonstrated that this nematode species differs clearly from other previously described root-knot nematodes. Studies of host-parasite relationships showed a typical susceptible reaction in naturally infected European holly plants, but did not reproduce on a number of cultivated plants, including tomato, grapevine, princess-tree and olive. The species is described here, illustrated and named as Meloidogyne silvestris n. sp. The new root-knot nematode can be morphologically distinguished from other Meloidogyne spp. by: (i) roundish perineal pattern, dorsal arch low, with fine, sinuous cuticle striae, lateral fields faintly visible; (ii) female excretory pore level with stylet knobs, or just anterior to them, EP/ST ratio about 0·8; (iii) second-stage juveniles with hemizonid located 1 to 2 annuli anterior to excretory pore and short, sub-digitate tail; and (iv) males with lateral fields composed of four incisures, with areolated outer bands. Phylogenetic trees derived from maximum parsimony analysis based on 18S, ITS1-5·8S-ITS2 and D2–D3 of 28S rDNA showed that M. silvestris n. sp. can be differentiated from all described root-knot nematode species, and it is clearly separated from other species with resemblance in morphology, such as M. ardenensis, M. dunensis and M. lusitanica. [ABSTRACT FROM AUTHOR]

The pollen content of eleven honey samples from ten different apiaries in the Baixa Limia – Serra do Xurés Nature Reserve and other honey commercialised by the cooperative as “Mel do Xurés” (north‐west Spain) was subjected to quantitative and qualitative melissopalynological analysis. The quantitative analysis found that ten samples belonged to Maurizio's Class III and one to Class IV. According to the qualitative analysis, four samples were classified as unifloral honey with Erica , four samples as multifloral honey with Erica pollen as the principal component and three samples as multifloral honey with Cytisus ‐type pollen and Erica as the principal component pollen. The pollen spectra differ between the diverse honeys analysed, with a common denominator being Erica and Cytisus ‐type pollen being abundant in all. For the rest of the samples, the pollen spectra were mainly the same, but with different relative percentages among secondary elements. Thus, either as a secondary or an important element, 91% of the honeys contained Quercus , 82% Castanea sativa Miller, 45% Rubus, 36% Cistus and 27% Lithodora prostrate ( Loisel) Griseb,. In particular, we record for the first time the presence of Ribes and Ilex aquifolium L. pollen in Spanish honeys as an important minor or minor pollen component. [ABSTRACT FROM AUTHOR]

Shade tolerance, plastic phenotypic response to light and sensitivity to photoinhibition were studied in holly (Ilex aquifolium L.) seedlings transported from the field to a greenhouse and in adult trees in the field. All plants were growing in, or originated from, continental Mediterranean sites in central Spain. Seedlings tolerated moderate but not deep shade. Mortality was high and growth reduced in 1% sunlight. Survival was maximal in 12% sunlight and minimal in full sunlight, although the relative growth rate of the seedlings surviving in high light was similar to that of plants in moderate shade. Maximum photochemical efficiency at predawn was significantly lower in sun plants than in shade plants in the field, revealing chronic photoinhibition that was most pronounced in winter. Plasticity in response to available light varied according to the variable studied, being low for photosynthetic capacity and stomatal conductance, and high for specific leaf area, root:shoot ratio and leaf area ratio, particularly in seedlings. Differences in water relations and hydraulic features between sun and shade plants in the field were marginal. High water potential at the turgor loss point of field-grown plants suggested that holly is sensitive to drought during both the seedling and the adult stage. Low relative growth rates in both high and low light with low physiological plasticity in response to light indicate the existence of a stress-tolerance mechanism. We conclude that holly is a facultative understory plant in areas of oceanic and relatively mild climate, but an obligate understory plant in dry continental areas such as the study site. The impact of abandonment of traditional management practices and climate change on these Mediterranean populations is discussed. [ABSTRACT FROM AUTHOR]

Abstract Variation in the chloroplast genome of Ilex aquifolium (English holly), a dioecious evergreen tree native to south, west and central Europe, was analysed using polymerase chain reaction–restriction fragment length polymorphisms (PCR-RFLPs) and microsatellites. Differentiation between populations was high (G [sub ST] = 0.595) and evidence for phylogeographical structure was detected (N [sub ST] = 0.697, significantly higher than G [sub ST] ). Two chloroplast lineages were inferred originating from putative glacial refugia in southern Europe (Iberia, Italy and possibly the Balkans). The G [sub ST] value was higher than reported for endozoochorous hermaphrodite species and for anemochorous dioecious species investigated over a similar geographical scale. It appears that dioecy has contributed to strong differentiation between refugia and that this has been maintained following postglacial recolonization as a result of limited seed flow. Palynological records for I. aquifolium are poor, thus these results give an important insight into patterns of glacial isolation and postglacial recolonization of this species. [ABSTRACT FROM AUTHOR]

Two procedures were tested for micropropagation of Ilex aquifolium (English holly), one in which shoots proliferated on solid medium and another one using liquid medium. Different growth regulator treatments and supports were analyzed for optimizing in vitro rooting, showing that indolebutyric acid and agar or cellulose plugs gave the best results. The surival percentage of successfully in vitro or ex vitro rooted plants did not differ significantly between the best treatments. However, the efficiency with ex vitro rooting was 80%, while for in vitro rooting, the final efficiency was 64%. The results show that a correct manipulation of Murashige's stage II of micropropagation and eliminating or decreasing stage III are useful tools to reduce the requirements of acclimatization. [ABSTRACT FROM AUTHOR]

The article focuses on the study on the biological flora of the British Isles, specifically the ilex aquifolium L. Ilex aquifolium L were normally dioecious, with no subspecies, and are native, and widespread in Great Britain. Physical attributions include green twigs, with grey bark, alternate leaves, and could grow up to 23 meters in height. Previous studies suggested that ilex aquiform L, or English holly, were not seen in areas where temperature falls below negative 0.5 degree celsius. Holly flowers, particularly from mature hollies, are abundant in most seasons, but the quantity of the fruits may vary from year to year.

The article presents information about Arbutus unedo at Killarney lakes in Ireland. It was noted that the Arbutus unedo is usually found on cliffs, open rocky slopes, limestones and margins of oakwoods. It was found that the plant is successful in these kinds of areas with the help of regeneration of seeds. Arbutus unedo on limestones likely growth with Taxus baccata and Ilex aquifolium. It was asserted that heavy output of seeds is needed in order for plants to survive in areas that the Arbutus unedo grows.

The market demand for interesting varieties and cultivars of Ilex aquifolium necessitates the exploration and sustainable exploitation of plant individuals thriving in nature without human care. In this work, an effort was made to develop a simple and reliable protocol for vegetative propagation of I. aquifolium plants, with desirable-for-market characteristics, grown in a mountain area of Halkidiki, Greece, and at the same time to proceed with their genetic identification using molecular markers. From these plants, new plants were vegetatively produced which afterward were used as stock plants for providing the needed shoot cuttings for the experiments of rooting and leaves for their genetic analysis. Factors studied in formulating a propagation protocol included the season of cutting collection and the application of 0.2% 1-naphthaleneacetic acid (NAA), as well as the type of shoot cuttings (terminal, subterminal) and the application of auxin. It was found that application of NAA was crucial for rooting response and number of roots formed, whereas the season effect was not significant on rooting. Terminal cuttings treated with 0.2% NAA exhibited the highest rooting percentage (100%) and formed abundant roots (25.7) compared to subterminal ones. All rooted cuttings, after being potted and transferred to acclimatization greenhouse, were successfully hardened. In the spring of the next year, the produced plants blossomed abundantly and formed fruits (bright red berries) presenting their characteristic ornamental appearance that was maintained until Christmas. For the identification procedure, the genotypic profile of the stock plants was also investigated by inter-simple sequence repeat (ISSR) genetic analysis, revealing that they were genetically the same both among themselves and when compared with a certified I. aquifolium 'Agrifoglio Commune' individual, but they differed genetically from I. aquifolium 'Argentea Marginata' and I. aquifolium 'Hellas'. [ABSTRACT FROM AUTHOR]