What are the drivers of female success in food-deceptive orchids?

A large suite of floral signals, and environmental and biotic characteristics influence the behavior of pollinators, affecting the female success of food-deceptive orchids. In this study, we examined the many factors shaping the reproductive output of three orchid taxa: Dactylorhiza majalis, D. incarnata var. incarnata, and D. fuchsii. We applied a statistical model to correlate female success (number of fruit sets) with individual characteristics (plant and inflorescence height, number of flowers, and spur length), number of pollinaria removed, flowering time, and density of floral units of co-flowering rewarding plants. Our findings suggested that the broad spectrum of variations in Dactylorhiza's morphological traits, floral display, and flowering phenology within different environmental contexts has a significant impact on their reproductive success. The number of fruits increased with an increase in the number of pollinaria removed in the studied Dactylorhiza taxa. In contrast, a higher number of flowers per inflorescence and higher inflorescences in relation to individual height always decreased fruit set. We observed that low number of co-flowering rewarding plants in populations could affect the Dactylorhiza reproductive output as magnets and competitor plants. The synchronization of flowering, or lack thereof, between Dactylorhiza and rewarding plants can limit reproductive success. This demonstrates that the food deception strategy is multidirectional, and reproductive output can vary considerably both spatially and temporally within the context of this strategy.

The patterns of inbreeding depression in food-deceptive Dactylorhiza orchids.

Introduction: Inbreeding depression (ID) in food-deceptive plants have been reported previously, however, it has not been often proven that selfed seeds germinate better than outbred ones or that selfing affects ID. To resolved these issues, food-deceptive related Dactylorhiza majalis, D. incarnata var. incarnata and D. fuchsii orchids were investigated. Methods: Hand pollination treatments and control pollination were conducted. Fruit set, number of seeds per fruit, seed length, number of well-developed seeds per fruit, and proportion of in vitro asymbiotic germination seeds, were analyzed in relation to inflorescence levels and used as fitness indicators for these orchids. The ID and pollen limitation were measured. Results: The lowest ID (δ = -1.000) was in D. majalis, and present in combination with a high pollen limitation in its populations. D. fuchsii showed higher ID (δ = 0.366), and D. incarnata var. incarnata weak ID (δ = 0.065), although ID varied between its fitness components. The seed number per fruit differed significantly between the treatments and the inflorescence levels in treatments. Discussion: This study emphasizes that the breeding system rather than the flower position on the inflorescence shaped the quality and quantity of reproductive output. The ID and its effect on germination of food-deceptive orchid seeds undoubtedly played an important role.

Performance of integrated sequencing batch reactor (SBR) and reverse osmosis (RO) process for leachate treatment: effect of pH.

Purpose: In this paper the performance and effectiveness of the reverse osmosis (RO) process for the biologically pretreated leachate was investigated. The RO process was carried out separately for two different pH: 8.0 and 9.3. Methods: A general pollution parameters as well as organic and inorganic indicators were determined in raw, biologically pretreated and RO treated leachate. The performance characteristics of the reverse osmosis system were made on the basis of permeate flux, electroconductivity removal rate, concentration factor and efficiency in removal of analyzed parameters. Results: The use of SBR pretreatment had very good efficiency in BOD (97.3%) and ammonia nitrogen (95.4%) removal. The lowest effectivity was observed for chloride (11.6%), boron (3.9%) and TDS (1.2%). Pretreated leachate was subjected to RO system. The normalized average flux was 0.53 (42.3 L/m2·h) for pH = 8.0 and 0.68 (33.5 L/m2·h) for pH = 9.3. The lower membrane fouling at higher pH can be explained by electrostatic repulsion between the negatively charged membrane surface and organic substances. Independently of the process pH, a two-step membrane fouling was observed. The greatest differences in removal rates were observed for boron, which had a higher retention rate at higher pH, and ammonia nitrogen, whose removal rate decreased at higher pH. The obtained permeate pH after RO process was lower than the feed pH in two analyzed value of pH. Conclusions: The higher flux value at pH = 9.3 is result of high content of organic matter in leachate, which is better rejected at higher pH because of higher electrostatic repulsion between organic matter and membrane surface. This indicates that the organic matter content should be taken into account when determining the operating parameters (pH values) of the RO system.

Pollen transfer and patterns of reproductive success in pure and mixed populations of nectariferous Platanthera bifolia and P. chlorantha (Orchidaceae).

Plant species evolution is driven by many factors that have different roles in space and time. Using different field and laboratory methods, we studied reproductive patterns and their determinants in pure and mixed P. bifolia and P. chlorantha populations in different habitats. We also considered the probability of hybridisation between these two species and the role of intra-population processes in maintaining species integrity. Generally, we found a high level of reproductive success in both Platantherans. In both species, male (MRS) and female (FRS) reproductive success depended on floral display, and male reproductive success additionally on population structure. The flower traits were only weakly related to reproductive success. Moths' assemblages varied spatially and temporally, and their diversity and numbers were correlated with MRS in the year, when their abundance was markedly lower. Analysis of patterns of pollen transfer showed that pollen was transported up to 25 m (average 8.2 ± 4.83 m) and showed gene exchange between these two Platanthera species. The germination level of both species was significantly lower than seed viability, although P. bifolia seed germinated with higher frequency than P. chlorantha seeds. We noted differences in viability and germination of seeds developed as an effect of experimental interspecies crossings and those developed from natural pollination. The presence of intermediate ecotypes together with observations of spontaneous interspecies crosses in the field and viability of seeds produced in interspecies crossing suggest that both pre- and postzygotic reproductive barriers are not complete and do not prevent hybrid production.

Spatiotemporal Variations in Seed Set and Pollen Limitation in Populations of the Rare Generalist Species Polemonium caeruleum in Poland.

A vast majority of angiosperms are pollinated by animals, and a decline in the number and diversity of insects often affects plant reproduction through pollen limitation. This phenomenon may be particularly severe in rare plant species, whose populations are shrinking. Here, we examined the variability in factors shaping reproductive success and pollen limitation in red-listed Polemonium caeruleum L. During a 5-year study in several populations of P. caeruleum (7-15, depending on year), we assessed the degree of pollen limitation based on differences in seed set between open-pollinated (control) and hand-pollinated flowers. We analysed the effects of flower visitors, population size, and meteorological data on plant reproductive success and pollen limitation. Our study showed that pollen limitation rarely affected P. caeruleum populations, and was present mainly in small populations. Pollen limitation index was negatively affected by the size of population, visitation frequency of all insects, and when considering the visitation frequency of individual groups, also by honeybee visits. Seed production in control treatment was positively influenced by the population size, average monthly precipitation in June and visits of hoverflies, while visits of honeybees, average monthly temperature in September, and average monthly precipitation in August influenced seed production negatively. As generalist plant P. caeruleum can be pollinated by diverse insect groups, however, in small populations their main visitors, the honeybees and bumblebees, may be less attracted, eventually leading to the disappearance of these populations. In pollination of P. caeruleum managed honeybees may play a dual role: while they are the most frequent and efficient flower visitors, their presence decreases seed set in open-pollinated flowers, which is most probably related to efficient pollen collection by these insects.

Deceptive strategy in Dactylorhiza orchids: multidirectional evolution of floral chemistry.

BACKGROUND AND AIMS: The deception strategies of orchids remain poorly understood, especially in regard to the chemical compounds emitted from their flowers and their interaction with various taxonomic groups of pollinators. We investigated the phylogenetic relationships and compared the variation of floral chemical compounds between food-deceptive Dactylorhiza taxa (D. incarnata var. incarnata and D. incarnata var. ochroleuca, D. fuchsii and D. majalis) from populations in north-eastern Poland. We propose a model of the evolution of deception based on floral chemical signals in this genus. METHODS: A Bayesian approach based on polymorphic plastid DNA (trnL, trnF and psbC-trnK), internal transcribed spacer (ITS) sequences and flow cytometry data was applied to confirm the taxonomic status of the studied orchids. We also identified and classified the pollinators and flower visitors in each Dactylorhiza population to the taxonomic level and compared our results with literature data. The chemical composition of pentane and diethyl ether extracts from the flowers was analysed by gas chromatography-mass spectrometry. Variation of the floral chemical components was visualized by non-metric multidimensional scaling based on Bray-Curtis dissimilarity. KEY RESULTS: The genetic distinctiveness of D. incarnata, D. fuchsii and D. majalis was confirmed. No hybrids between them were found, but the chloroplast DNA (cpDNA), ITS haplotypes and flow cytometry showed genetic similarity between D. incarnata var. incarnata and D. incarnata var. ochroleuca. We determined that Apis mellifera (Hymenoptera) was the only shared pollinator of these taxa. Strangalia attenuata and Alosterna tabacicolor (Coleoptera) and Volucella pellucens and V. bombylans (Hymenoptera) were observed pollinating D. fuchsii. Visualization of the emission rates of the 61 floral chemical compounds detected from pentane extracts (mainly hydrocarbons and aldehydes) and the 51 from diethyl extracts (with abundant groups of benzenoids and non-aromatic acids) strongly differentiated D. incarnata, D. fuchsii and D. majalis, while those of the two varieties of D. incarnata (var. incarnata and var. ochroleuca) were almost identical. CONCLUSIONS: While the genetic data clearly supported the distinct lineages of D. incarnata, D. fuchsii and D. majalis, the patterns of emission of their flower chemical compounds were more complex within the series of shared compounds (alkanes and aldehydes) and taxon-specific compounds (benzenoids and esters). Their floral bouquet can influence the sexual, social and feeding behaviour of pollinators in different ways. We observed that the floral chemical compounds attracted both shared and species-specific pollinators to Dactylorhiza, confirming the multidirectional character of floral chemical signals in these food-deceptive taxa. Reduction of species-specific pollination levels in Dactylorhiza orchid taxa may promote hybridization between them.

Use of the landfill water pollution index (LWPI) for groundwater quality assessment near the landfill sites.

The purpose of the paper is to assess the groundwater quality near the landfill sites using landfill water pollution index (LWPI). In order to investigate the scale of groundwater contamination, three landfills (E, H and S) in different stages of their operation were taken into analysis. Samples of groundwater in the vicinity of studied landfills were collected four times each year in the period from 2004 to 2014. A total of over 300 groundwater samples were analysed for pH, EC, PAH, TOC, Cr, Hg, Zn, Pb, Cd, Cu, as required by the UE legal acts for landfill monitoring system. The calculated values of the LWPI allowed the quantification of the overall water quality near the landfill sites. The obtained results indicated that the most negative impact on groundwater quality is observed near the old Landfill H. Improper location of piezometer at the Landfill S favoured infiltration of run-off from road pavement into the soil-water environment. Deep deposition of the groundwater level at Landfill S area reduced the landfill impact on the water quality. Conducted analyses revealed that the LWPI can be used for evaluation of water pollution near a landfill, for assessment of the variability of water pollution with time and for comparison of water quality from different piezometers, landfills or time periods. The applied WQI (Water Quality Index) can also be an important information tool for landfill policy makers and the public about the groundwater pollution threat from landfill.

The effect of gender and age structure on municipal waste generation in Poland.

In this study the effect of gender and age structure on municipal waste generation was investigated. The data from 10-year period, from 2001 to 2010 year, were taken into consideration. The following parameters of gender and age structure were analyzed: men and woman quantity, female to male ratio, number of working, pre-working and post-working age men/women, number of unemployed men/women. The results have showed a strong correlation of annual per capita waste generation rate with number of unemployed women (r=0.70) and female to male ratio (r=0.81). This indicates that waste generation rate is more depended on ratio of men and women that on quantitative size of each group. Using the regression analysis a model describing the dependence between female to male ratio, number of unemployed woman and waste quantity was determined. The model explains 70% of waste quantity variation. Obtained results can be used both to improve waste management and to a fuller understanding of gender behavior.

Removal of nitrogen compounds from landfill leachate using reverse osmosis with leachate stabilization in a buffer tank.

In this paper, a removal of nitrogen compounds from a landfill leachate during reverse osmosis (RO) was evaluated. The treatment facility consists of a buffer tank and a RO system. The removal rate of N─NH4, [Formula: see text] and [Formula: see text] in the buffer tank reached 14%, 91% and 41%, respectively. The relatively low concentration of organic carbon limits N─NH4 oxidation in the buffer tank. The removal rate for the total organic nitrogen (TON) was 47%. The removal rate in RO was 99% for [Formula: see text], 84.1% for [Formula: see text] and 41% for [Formula: see text]. The accumulation of [Formula: see text] may be the result of a low pH, which before the RO process is reduced to a value of 6.0-6.5. Besides it, the cause for a low removal rate of the [Formula: see text] in the buffer tank and during RO may be free ammonia, which can inhibit the [Formula: see text] oxidation. The removal rates of total inorganic nitrogen and TON in the RO treatment facility were similar being 99% and 98.5%, respectively.

Mineral and organic compounds in leachate from landfill with concentrate recirculation.

The effect of a reverse osmosis concentrate recirculation on the mineral and organic compounds in a landfill leachate was investigated. Investigated was the quality of a leachate from two landfills operated for different periods (a 20-year-old Cell A and a 1-year-old Cell B), where the concentrate was recirculated. Examined were general parameters (conductivity, pH), organic compounds (biochemical oxygen demand (BOD), chemical oxygen demand (COD), total organic nitrogen, BOD/COD), and inorganic compounds (nitrogen ammonia, sulfite, sulfate, cyanide, boron, chloride, ferrous, zinc, chrome, copper). The findings from the first year of investigation showed that over the initial period of recirculation, the concentration of organic compounds (BOD, COD) increased, but after 6 months their values stabilized. It indicates that the concentrate recirculation accelerated organic decomposition, especially in the new landfill Cell. The analysis of inorganic parameters showed that recirculation landfills produce a leachate with a higher concentration of N-NH4, and Cl(-). In case of the old landfill Cell, an increase in B and Fe was also noticeable. These compounds are cyclically washed out from a waste dump and require an additional pretreatment in order to exclude them from recirculation cycle. The increased concentration of Cu, Zn, and Fe was noticed during the initial months of recirculation and in the season of intense atmospheric precipitation in the leachate from both Cells. Higher values of electro conductivity, Cl(-), N-NH4 (+), B, and Fe in the leachate from the old field indicate that the attenuation capacity of this landfill is close to exhaustion.

Assessment of groundwater quality near the landfill site using the modified water quality index.

The purpose of this paper is to assess the groundwater quality near a landfill site using the modified water quality index. A total of 128 groundwater samples were analyzed for pH, electrical conductivity (EC), total organic carbon (TOC), polycyclic aromatic hydrocarbon (PAH), Cd, Pb, Zn, Cu, Cr, and Hg. The analytical results have showed a decreasing trend in concentration for TOC, Cd, Pb, Hg, and Cu and an increasing one for pH, EC, and PAH. The modified water quality index, which was called landfill water pollution index (LWPI), was calculated to quantify the overall water quality near the landfill site. The analysis reveals that groundwater in piezometers close to the landfill is under a strong landfill impact. The LWPI in piezometers ranged from 0.52 to 98.25 with a mean value of 7.99. The LWPI in groundwater from the nearest house wells varied from 0.59 to 0.92. A LWPI value below 1 proves that analyzed water is not affected by the landfill. Results have shown that LWPI is an efficient method for assessing and communicating the information on the groundwater quality near the landfill.