Rising temperature modulates pH niches of fen species.

Rising temperatures may endanger fragile ecosystems because their character and key species show different habitat affinities under different climates. This assumption has only been tested in limited geographical scales. In fens, one of the most endangered ecosystems in Europe, broader pH niches have been reported from cold areas and are expected for colder past periods. We used the largest European-scale vegetation database from fens to test the hypothesis that pH interacts with macroclimate temperature in forming realized niches of fen moss and vascular plant species. We calibrated the data set (29,885 plots after heterogeneity-constrained resampling) with temperature, using two macroclimate variables, and with the adjusted pH, a variable combining pH and calcium richness. We modelled temperature, pH and water level niches for one hundred species best characterizing European fens using generalized additive models and tested the interaction between pH and temperature. Fifty-five fen species showed a statistically significant interaction between pH and temperature (adj p Ë‚ .01). Forty-six of them (84%) showed a positive interaction manifested by a shift or restriction of their niche to higher pH in warmer locations. Nine vascular plants and no moss showed the opposite interaction. Mosses showed significantly greater interaction. We conclude that climate significantly modulates edaphic niches of fen plants, especially bryophytes. This result explains previously reported regional changes in realized pH niches, a current habitat-dependent decline of endangered taxa, and distribution changes in the past. A warmer climate makes growing seasons longer and warmer, increases productivity, and may lower the water level. These effects prolong the duration and intensity of interspecific competition, support highly competitive Sphagnum mosses, and, as such, force niches of specialized fen species towards narrower high-pH ranges. Recent anthropogenic landscape changes pose a severe threat to many fen species and call for mitigation measures to lower competition pressure in their refugia.

The bright side of parasitic plants: what are they good for?

Parasitic plants are mostly viewed as pests. This is caused by several species causing serious damage to agriculture and forestry. There is however much more to parasitic plants than presumed weeds. Many parasitic plans exert even positive effects on natural ecosystems and human society, which we review in this paper. Plant parasitism generally reduces the growth and fitness of the hosts. The network created by a parasitic plant attached to multiple host plant individuals may however trigger transferring systemic signals among these. Parasitic plants have repeatedly been documented to play the role of keystone species in the ecosystems. Harmful effects on community dominants, including invasive species, may facilitate species coexistence and thus increase biodiversity. Many parasitic plants enhance nutrient cycling and provide resources to other organisms like herbivores or pollinators, which contributes to facilitation cascades in the ecosystems. There is also a long tradition of human use of parasitic plants for medicinal and cultural purposes worldwide. Few species provide edible fruits. Several parasitic plants are even cultivated by agriculture/forestry for efficient harvesting of their products. Horticultural use of some parasitic plant species has also been considered. While providing multiple benefits, parasitic plants should always be used with care. In particular, parasitic plant species should not be cultivated outside their native geographical range to avoid the risk of their uncontrolled spread and the resulting damage to ecosystems.

Parasites on parasites: hyper-, epi-, and autoparasitism among flowering plants.

All organisms engage in parasitic relations, as either parasites or hosts. Some species may even play both roles simultaneously. Among flowering plants, the most widespread form of parasitism is characterized by the development of an intrusive organ called the haustorium, which absorbs water and nutrients from the host. Despite this functionally unifying feature of parasitic plants, haustoria are not homologous structures; they have evolved 12 times independently. These plants represent ca. 1% of all extant flowering species and show a wide diversity of life histories. A great variety of plants may also serve as hosts, including other parasitic plants. This phenomenon of parasitic exploitation of another parasite, broadly known as hyper- or epiparasitism, is well described among bacteria, fungi, and animals, but remains poorly understood among plants. Here, we review empirical evidence of plant hyperparasitism, including variations of self-parasitism, discuss the diversity and ecological importance of these interactions, and suggest possible evolutionary mechanisms. Hyperparasitism may provide benefits in terms of improved nutrition and enhanced host-parasite compatibility if partners are related. Different forms of self-parasitism may facilitate nutrient sharing among and within parasitic plant individuals, while also offering potential for the evolution of hyperparasitism. Cases of hyperparasitic interactions between parasitic plants may affect the ecology of individual species and modulate their ecosystem impacts. Parasitic plant phenology and disperser feeding behavior are considered to play a major role in the occurrence of hyperparasitism, especially among mistletoes. There is also potential for hyperparasites to act as biological control agents of invasive primary parasitic host species.

Water-stress physiology of Rhinanthus alectorolophus, a root-hemiparasitic plant.

Root-hemiparasitic plants of the genus Rhinanthus acquire resources through a water-wasting physiological strategy based on high transpiration rate mediated by the accumulation of osmotically active compounds and constantly open stomata. Interestingly, they were also documented to withstand moderate water stress which agrees with their common occurrence in rather dry habitats. Here, we focused on the water-stress physiology of Rhinanthus alectorolophus by examining gas exchange, water relations, stomatal density, and biomass production and its stable isotope composition in adult plants grown on wheat under contrasting (optimal and drought-inducing) water treatments. We also tested the effect of water stress on the survival of Rhinanthus seedlings, which were watered either once (after wheat sowing), twice (after wheat sowing and the hemiparasite planting) or continuously (twice and every sixth day after that). Water shortage significantly reduced seedling survival as well as the biomass production and gas exchange of adult hemiparasites. In spite of that drought-stressed and even wilted plants from both treatments still considerably photosynthesized and transpired. Strikingly, low-irrigated plants exhibited significantly elevated photosynthetic rate compared with high-irrigated plants of the same water status. This might relate to biochemical adjustments of these plants enhancing the resource uptake from the host. Moreover, low-irrigated plants did not acclimatize to water stress by lowering their osmotic potential, perhaps due to the capability to tolerate drought without such an adjustment, as their osmotic potential at full turgor was already low. Contrary to results of previous studies, hemiparasites seem to close their stomata in response to severe drought stress and this happens probably passively after turgor is lost in guard cells. The physiological traits of hemiparasites, namely the low osmotic potential associated with their parasitic lifestyle and the ability to withstand drought and recover from the wilting likely enable them to grow in dry habitats. However, the absence of osmotic adjustment of adults and sensitivity of seedlings to severe drought stress demonstrated here may result in a substantial decline of the hemiparasitic species with ongoing climate change.

Mixotrophy in Land Plants: Why To Stay Green?

Mixotrophic plants combine photosynthesis and heterotrophic nutrition. Recent research suggests mechanisms explaining why mixotrophy is so common in terrestrial ecosystems. First, mixotrophy overcomes nutrient limitation and/or seedling establishment constraints. Second, although genetic drift may push mixotrophs to full heterotrophy, the role of photosynthesis in reproduction stabilizes mixotrophy.

Development of 14 microsatellite markers in Odontites vernus s.l. (Orobanchaceae) and cross-amplification in related taxa.

PREMISE OF THE STUDY: Microsatellite primers were developed for the first time in the root hemiparasite herb Odontites vernus (Orobanchaceae). These markers will be useful to investigate the role of polyploidization in the evolution of this diploid-tetraploid complex, as well as the extent of gene flow between different ploidy levels. METHODS AND RESULTS: Fourteen polymorphic and reproducible loci were identified and optimized from O. vernus using a microsatellite-enriched library and 454 Junior sequencing. The set of primers amplified di- to pentanucleotide repeats and showed two to 13 alleles per locus. Transferability was tested in 30 taxa (19 belonging to Odontites and 11 from eight other genera of Orobanchaceae tribe Rhinantheae). CONCLUSIONS: The results indicate the utility of the newly developed microsatellites in O. vernus and several other species, which will be useful for taxon delimitation and conservation genetics studies.

Hydathode trichomes actively secreting water from leaves play a key role in the physiology and evolution of root-parasitic rhinanthoid Orobanchaceae.

BACKGROUND AND AIMS: Root hemiparasites from the rhinanthoid clade of Orobanchaceae possess metabolically active glandular trichomes that have been suggested to function as hydathode trichomes actively secreting water, a process that may facilitate resource acquisition from the host plant's root xylem. However, no direct evidence relating the trichomes to water secretion exists, and carbon budgets associated with this energy-demanding process have not been determined. METHODS: Macro- and microscopic observations of the leaves of hemiparasitic Rhinanthus alectorolophus were conducted and night-time gas exchange was measured. Correlations were examined among the intensity of guttation, respiration and transpiration, and analysis of these correlations allowed the carbon budget of the trichome activity to be quantified. We examined the intensity of guttation, respiration and transpiration, correlations among which indicate active water secretion. KEY RESULTS: Guttation was observed on the leaves of 50 % of the young, non-flowering plants that were examined, and microscopic observations revealed water secretion from the glandular trichomes present on the abaxial leaf side. Night-time rates of respiration and transpiration and the presence of guttation drops were positively correlated, which is a clear indicator of hydathode trichome activity. Subsequent physiological measurements on older, flowering plants indicated neither intense guttation nor the presence of correlations, which suggests that the peak activity of hydathodes is in the juvenile stage. CONCLUSIONS: This study provides the first unequivocal evidence for the physiological role of the hydathode trichomes in active water secretion in the rhinanthoid Orobanchaceae. Depending on the concentration of organic elements calculated to be in the host xylem sap, the direct effect of water secretion on carbon balance ranges from close to neutral to positive. However, it is likely to be positive in the xylem-only feeding holoparasites of the genus Lathraea, which is closely related to Rhinanthus. Thus, water secretion by the hydathodes might be viewed as a physiological pre-adaptation in the evolution of holoparasitism in the rhinanthoid lineage of Orobanchaceae.

Integrating ecology and physiology of root-hemiparasitic interaction: interactive effects of abiotic resources shape the interplay between parasitism and autotrophy.

Root hemiparasites are green photosynthetic plants, which parasitically acquire resources from host xylem. Mineral nutrients and water, two principal below-ground abiotic resources, were assumed to affect the interaction between hemiparasites and their hosts. The shape of these effects and the underlying physiological mechanisms have, however, remained unclear. We conducted a glasshouse experiment with root-hemiparasitic Rhinanthus alectorolophus, in which we manipulated the availability of mineral nutrients and water. Biomass production and Chl fluorescence of the hemiparasites and hosts were recorded, together with proportion of host-derived carbon in hemiparasite biomass. The abiotic resources had profound interactive effects on the performance of both the hemiparasite and the hosts, as well as the balance of above-ground biomass between them. These effects were mainly based on an increase of growth and photosynthetic efficiency under high nutrient concentrations, on the hemiparasite's ability to induce strong water stress on the hosts if water is limiting, and on release of the host from parasitism by simultaneous abundance of both resources. Hemiparasitism is a highly variable interaction, in which environmental conditions affect both the parasitic and autotrophic (and thus competitive) components. A hemiparasite's own photosynthesis plays a crucial role in the assimilation of parasitized mineral resources and their transformation into growth and fitness.

Ploidy-specific symbiotic interactions: divergence of mycorrhizal fungi between cytotypes of the Gymnadenia conopsea group (Orchidaceae).

Polyploidy is widely recognized as a major mechanism of sympatric speciation in plants, yet little is known about its effects on interactions with other organisms. Mycorrhizal fungi are among the most common plant symbionts and play an important role in plant nutrient supply. It remains to be understood whether mycorrhizal associations of ploidy-variable plants can be ploidy-specific. We examined mycorrhizal associations in three cytotypes (2x, 3x, 4x) of the Gymnadenia conopsea group (Orchidaceae), involving G. conopsea s.s. and G. densiflora, at different spatial scales and during different ontogenetic stages. We analysed: adults from mixed- and single-ploidy populations at a regional scale; closely spaced adults within a mixed-ploidy site; and mycorrhizal seedlings. All Gymnadenia cytotypes associated mainly with saprotrophic Tulasnellaceae (Basidiomycota). Nonetheless, both adults and seedlings of diploids and their autotetraploid derivatives significantly differed in the identity of their mycorrhizal symbionts. Interploidy segregation of mycorrhizal symbionts was most pronounced within a site with closely spaced adults. This study provides the first evidence that polyploidization of a plant species can be associated with a shift in mycorrhizal symbionts. This divergence may contribute to niche partitioning and facilitate establishment and co-existence of different cytotypes.

How does elevated grassland productivity influence populations of root hemiparasites? Commentary on Borowicz and Armstrong (Oecologia 2012).

In their recent study, Borowicz and Armstrong (Oecologia 169:783-792, 2012) investigated effects of nutrient availability and competition for light on a perennial root hemiparasite Pedicularis canadensis. Their study showed a reduction of community productivity as a result of hemiparasite infection independently of a clear positive effect of increased nutrients. In contrast, there was a minimal effect of increased competition for light on growth of the parasite. Here, we summarize the available data on the influence of nutrient availability (closely related to productivity) on temperate grassland root hemiparasites thus expanding the discussion presented by Borowicz and Armstrong (Oecologia 169:783-792, 2012). Most studies show that root hemiparasites are highly sensitive to elevated competition for light in productive environments, which is manifested as an increase in mortality coupled to a decrease in population density. Such responses reflect increased mortality of hemiparasite seedlings that are physiologically inefficient in terms of photosynthesis and nutrient acquisition owing to a limited root network and consequently, are highly sensitive to competition for light. However, the susceptibility of hemiparasites to competition for light tends to decrease for individuals that survive the critical seedling stage. Moreover, survivors benefit from elevated nutrient availability, resulting in increased growth and fecundity. Elevated productivity can thus have opposing effects on the survival and growth of hemiparasites depending on life stage. We conclude that the findings by Borowicz and Armstrong (Oecologia 169:783-792, 2012) are not in conflict with this general view that root hemiparasite population ecology is strongly influenced by competition for light in highly productive environments.

Symbiotic germination capability of four Epipactis species (Orchidaceae) is broader than expected from adult ecology.

PREMISE OF THE STUDY: Both abiotic and biotic factors shape species distributions. Orchids produce minute seeds with few nutrient reserves, thus requiring mycorrhizal fungi for germination. Therefore, both environmental conditions and mycorrhizal fungi distribution affect their germination success, but these ecological requirements and their congruence with habitat preferences of adults remain poorly understood. We investigated the importance of these factors during germination in four forest orchid species of the genus Epipactis. METHODS: We sowed seeds of three habitat specialists and one generalist in different forest types at sites harboring adults of at least one of these ecologically diverging species. We analyzed germination pattern and identified mycorrhizal fungi of both seedlings and adults. KEY RESULTS: Habitat conditions had little influence on germination pattern as seedlings grew in more habitats than expected from the adults' ecology. Ectomycorrhizal fungi availability did not limit germination. Suitable mycorrhizal fungi, mostly pezizalean ascomycetes, were recruited in various forest types, though the fungal communities differed according to habitat type. Finally, orchids with divergent ecological preferences shared similar mycorrhizal fungi. CONCLUSIONS: Limited adult distribution contrasted with successful seed germination at diverse sites and indicates existence of niche differentiation between adults and seedlings. Ecological specialization may thus be determined by factors other than mycorrhizal fungi that act later in the ontogeny, perhaps during the transition to above-ground development.

Glycerol-treated nuclear suspensions--an efficient preservation method for flow cytometric analysis of plant samples.

Flow cytometry (FCM) has been widely used in plant science to determine the amount of nuclear DNA, either in absolute units or in relative terms, as an indicator of ploidy. The requirement for fresh material in some applications, however, limits the value of FCM in field research, including plant biosystematics, ecology and population biology. Dried plant samples have proven to be a suitable alternative in some cases (large-scale ploidy screening) although tissue dehydration is often associated with a decrease in the quality of FCM analysis. The present study tested, using time-scale laboratory and in situ field experiments, the applicability of glycerol-treated nuclear suspension for DNA flow cytometry. We demonstrate that plant nuclei preserved in ice-cold buffer + glycerol solution remain intact for at least a few weeks and provide estimates of nuclear DNA content that are highly comparable and of similar quality to those obtained from fresh tissue. The protocol is compatible with both DAPI and propidium iodide staining, and allows not only the determination of ploidy level but also genome size in absolute units. Despite its higher laboriousness, glycerol-preserved nuclei apparently represent the most reliable way of sample preservation for genome size research. We assume that the protocol will provide a vital alternative to other preservation methods, especially when stringent criteria on the quality of FCM analysis are required.

The role of heterotrophic carbon acquisition by the hemiparasitic plant Rhinanthus alectorolophus in seedling establishment in natural communities: a physiological perspective.

• Heterotrophic acquisition of substantial amounts of organic carbon by hemiparasitic plants was clearly demonstrated by numerous studies. Many hemiparasites are, however, also limited by competition for light preventing the establishment of their populations on highly productive sites. • In a growth-chamber experiment, we investigated the effects of competition for light, simulated by shading, on growth and heterotrophic carbon acquisition by the hemiparasite Rhinanthus alectorolophus attached to C(3) and C(4) hosts using analyses of biomass production and stable isotopes of carbon. • Shading had a detrimental effect on biomass production and vertical growth of the hemiparasites shaded from when they were seedlings, while shading imposed later caused only a moderate decrease of biomass production and had no effect on the height. Moreover, shading increased the proportion of host-derived carbon in hemiparasite biomass (up to 50% in shaded seedlings). • These results demonstrate that host-derived carbon can play a crucial role in carbon budget of hemiparasites, especially if they grow in a productive environment with intense competition for light. The heterotrophic carbon acquisition can allow hemiparasite establishment in communities of moderate productivity, helping well-attached hemiparasites to escape from the critical seedling stage.

Image cytometric measurements of diploid, triploid and tetraploid fish erythrocytes in blood smears reflect the true dimensions of live cells.

Comparative image cytometry of erythrocytes of diploid and triploid tench Tinca tinca L. and evolutionary tetraploid sterlet Acipenser ruthenus L. was performed on whole live unstained cells, live cells with stained nuclei and on stained fixed whole cells and their nuclei to test if erythrocyte measurements made from blood smears reflect the true dimensions of live cells. Nuclear area and perimeter were the best ploidy level predictors distinguishing accurately among live and fixed diploid, triploid and tetraploid cells, without significant differences between live and fixed cells within a ploidy level. Redundancy analysis revealed insignificant marginal effect of fixation (explained 2.3% of variation, F = 0.804), whereas the effect of ploidy level was highly significant (explained 50.6% of variation, F = 34.874). The erythrocyte measurements of diploid, triploid and tetraploid fish erythrocytes and their nuclei made from blood smears reflect the true dimensions of live cells, and the fixation procedure did not substantially affect their predictive value for ploidy level determination.

Interactions between hemiparasitic plants and their hosts: the importance of organic carbon transfer.

Hemiparasitic plants display a unique strategy of resource acquisition combining parasitism of other species and own photosynthetic activity. Despite the active photoassimilation and green habit, they acquire substantial amount of carbon from their hosts. The organic carbon transfer has a crucial influence on the nature of the interaction between hemiparasites and their hosts which can oscillate between parasitism and competition for light. In this minireview, we summarize methodical approaches and results of various studies dealing with carbon budget of hemiparasites and the ecological implications of carbon heterotrophy in hemiparasites.

Size determination of Acipenser ruthenus spermatozoa in different types of electron microscopy.

In this study three types of scanning electron microscopes were used for the size determination of spermatozoa of sterlet Acipenser ruthenus - high vacuum scanning electron microscope (SEM, JEOL 6300), environmental scanning electron microscope (ESEM, Quanta 200 FEG), field emission scanning electron microscope (FESEM, JEOL 7401F) with cryoattachment Alto 2500 (Gatan) and transmission electron microscope (TEM, JEOL 1010). The use of particular microscopes was tied with different specimen preparation techniques. The aim of this study was to evaluate to what degree the type of used electron microscope can influence the size of different parts of spermatozoa. For high vacuum SEM the specimen was prepared using two slightly different procedures. After chemical fixation with 2.5% glutaraldehyde in 0.1M phosphate buffer and post-fixation by 1% osmium tetroxide, the specimen was dehydrated by acetone series and dried either by critical point method or by means of t-butylalcohol. For ESEM fresh, unfixed material was used, which was dropped on microscopic copper grids. In FESEM working in cryo-mode the specimen was observed in a frozen state. Ultrathin sections from chemically fixed and Epon embedded specimens were prepared for TEM observation. Distinct parts of sterlet spermatozoa were measured in each microscope and the data obtained was statistically processed. Results confirmed that the classical chemical procedure of specimen preparation for SEM including critical point drying method led to a significant contraction of all measured values, which could deviate up to 30% in comparison with values measured on the fresh chemically untreated specimen in ESEM. Surprisingly sperm dimensions determinated on ultrathin sections by TEM are comparable with values obtained in ESEM or FESEM.

Heterotrophic carbon gain by the root hemiparasites, Rhinanthus minor and Euphrasia rostkoviana (Orobanchaceae).

Hemiparasitic plants gain virtually all mineral nutrients and water from their host plant whilst organic carbon is provided, at least in part, by their own photosynthetic activity, although their rates of assimilation are substantially lower than that found in non-parasitic plants. Hence, hemiparasites must gain at least some of their organic carbon heterotrophically from the host plant. Despite this, heterotrophic carbon gain by root hemiparasites has been investigated only for a few genera. We investigated heterotrophic carbon gain by two root hemiparasites, Rhinanthus minor L. and Euphrasia rostkoviana Hayne (Orobanchaceae), using natural abundance stable isotope (delta(13)C) profiles of both parasites attached to C(3) (wheat) and C(4) (maize) hosts coupled to a linear two-source isotope-mixing model to estimate the percentage of carbon in the parasite that was derived from the host. Both R. minor and E. rostkoviana attached to maize hosts were significantly more enriched in (13)C than those attached to wheat hosts with R. minor becoming more enriched in (13)C than E. rostkoviana. The natural abundance (13)C profiles of both parasites were not significantly different from their wheat hosts, but were less enriched in (13)C than maize hosts. Using a linear two-source isotope-mixing model, we estimated that R. minor and E. rostkoviana adult plants derive c. 50 and 25% of their carbon from their hosts, respectively. In light of these results, we hypothesise that repeatedly observed negative effect of competition for light on hemiparasites acts predominantly in early ontogenetic stages when parasites grow unattached or the abstraction of host nutrients is less effective.

Fine structure and morphology of sterlet (Acipenser ruthenus L. 1758) spermatozoa and acrosin localization.

Ultrastructure of sterlet Acipenser ruthenus L. 1758 sperm was examined by scanning and transmission electron microscopy, which allowed us to use various methods for visualizations of different parts of sterlet spermatozoa. Sperm cells possess a head with a distinct acrosome, a midpiece and a single flagellum surrounded by the flagellar plasma membrane. The average length of the head including the acrosome and the midpiece was estimated as 5.14+/-0.42 microm. Nine to 10 posterolateral projections were derived from the acrosome. Three inter-twining endonuclear canals bounded by membranes traversed the nucleus in its whole length from the acrosome to the implantation fossa. Acrosin was located in all the three parts (acrosome, endonuclear canals and implantation fossa). The proximal and distal centrioles located in the midpiece compacted of nine peripheral triplets of microtubules. One cut of the midpiece contained from two to six mitochondria with area of 215+/-85 nm(2) in average. The flagellum was 42.47+/-1.89 microm in length with typical eukaryotic organization of one central pair and nine peripheral pairs of microtubules. It passed through a cytoplasmic channel in the midpiece, which was formed by an invagination at the plasmalemma. The flagellum gradually developed two lateral extensions of its plasma membrane, so-called "fins". Detected morphological variation can be described by four principal component axes corresponding to groups of individual morphometric characters defined on the sperm structures. Correlations among the characters indicate that the sperms are variable in their shape rather than size. Significant variation among examined fish individuals was found only in flagellum and nucleus length. Comparison between the present and previous studies of morphology of sturgeon spermatozoa confirmed large inter- and/or intra-specific differences that could be of substantial taxonomic value.