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Laboulbeniales (Ascomycota: Laboulbeniomycetes) are obligate ectoparasitic fungi of arthropods with a worldwide distribution. Their effects on host physiology and behaviour as well as their ecology have recently gained wider attention. One aspect that is virtually unknown regarding Laboulbeniales and arthropod-associated fungi in general, is how abiotic factors shape the distribution of these parasites. We used ant- and bat fly-associated Laboulbeniales to study whether climatic elements play a role in the distribution of fungal species. We collected uninfected and Laboulbeniales-infected insects belonging to three species: bat flies Nycteribia schmidlii and Penicillidia conspicua (Diptera: Nycteribiidae) and the ant Myrmica scabrinodis (Hymenoptera: Formicidae). We used climatic variables and performed statistical analyses to explain the distribution of Laboulbeniales infection. Our results show a higher likelihood of Laboulbeniales presence in habitats with low annual mean temperature and humidity, suggesting that climatic elements can considerably shape the distribution of Laboulbeniales species.

Overproduction of toxic cyanobacteria is a type of harmful algal blooms (HABs). The heptapeptide microcystins (MCs) are one of the most common cyanotoxins. There is increasing research concerning the effects of MCs on growth and physiology of vascular plants, however there is a lack of studies on their direct effects on aquatic macrophytes in the real environment. Here we report the occurrence of a MC producing HAB in Lake Bardos, Hungary in 2012 with harmful effects on cytological, histological and biochemical parameters of Ceratophyllum submersum (soft hornwort) plants naturally growing at the blooming site. Blue-Green Sinapis Test (BGST) showed high toxicity of HAB samples. Cell-free water samples contained a significant amount of MCs (7.31 ± 0.17 μg L−1) while C. submersum plants contained 1.01 ± 0.21 μg g DW−1 MCs. Plants showed significant increases of protein content and decreases of anthocyanin content and carotenoid/chlorophyll ratio, indicating physiological stress- as compared to plants from the control (MC free) sampling site of the same water body. Histological and cytological studies showed (i) radial swelling and the abnormal formation of lateral buds at the shoot tip leading to abnormal development; (ii) the fragmentation of nuclei as well as accumulation of phenolics in the nucleus indicating that the HAB induced cell death and stress reactions at the nuclear level. The most relevant effect was the increase of histone H3 phosphorylation in metaphase chromosomes: since MCs are strong inhibitors of protein phosphatases, this alteration is related to the biochemical targets of these toxins. The HAB decreased peroxidase activity, but increased nuclease and protease activities, showing the decreased capacity of plants to face biotic stress and as the cytological changes, the induction of cell death. This study is one of the first to show the complex harmful changes in aquatic plants that co-exist with HABs.

Two new 14-noreudesmane sesquiterpenes, one new phenylpropane heterodimer, caulilexin C, and uvaol were isolated from the 70% MeOH extract of the fruit peel of Elaeagnus rhamnoides. The structures of the compounds were elucidated by HRESIMS and advanced NMR methods. The absolute configuration of (R)-6,9-dihydroxy-1-oxo-14-noreudesm-5,7,9-triene was determined by the TDDFT-ECD method. The new compounds, together with structurally similar naphthalenes (musizin, musizin-8-O-glucoside, torachrysone-8-O-glucoside) and 1,4-naphthoquinone (2-methylstipandrone), isolated previously from Rumex aquaticus, were investigated for their antiviral activity against Herpes simplex virus type 2 (HSV-2) using two different methods. Applying the traditional virus yield reduction test, (R)-6,9-dihydroxy-1-oxo-14-noreudesm-5,7,9-triene, 1-[3-methoxy-4-(2-methoxy-4-(1E)-propenyl-phenoxy)-phenyl]-propane-1,2-diol, and musizin caused a 2.00 log10, 3.49 log10, and 2.33 log10 reduction of HSV-2 yield, respectively, at a concentration of 12.5 μM. 2-Hydroxy-1-methoxy-6,9-dioxo-14-noreudesm-1,3,5(10),7-tetraene exhibited an antiviral effect at concentration of 50 μM only. Similar results were obtained when the qPCR method was used to test the antiviral activity of the compounds.

Chemical investigation of aerial parts of Helichrysum italicum yielded two new pyrone derivatives (1 and 2) along with ten known compounds. The structures of the new compounds were established by 1D and 2D NMR spectra as well as by HRESIMS data. Compound 1 represented a rare dimer of substituted α- and γ-pyrone units. DFT-NMR and TDDFT-ECD calculations were carried out to determine the absolute configuration of 1 but failed, representing the limitation of TDDFT-ECD calculation for the configurational assignment. All compounds were measured for their antibacterial and cytotoxic activity but proved to be inactive.

A new cyclic pentapeptide, cotteslosin C (1), a new aflaquinolone, 22-epi-aflaquinolone B (3), and two new anthraquinones (9 and 10), along with thirty known compounds (2, 4 – 8, 11 – 34) were isolated from a co-culture of the sponge-associated fungus Aspergillus versicolor with Bacillus subtilis. The new metabolites were only detected in the co-culture extract, but not when the fungus was grown under axenic conditions. Furthermore, the co-culture extract exhibited an enhanced accumulation of the known constituents versicolorin B (14), averufin (16), and sterigmatocyctin (19) by factors of 1.5, 2.0, and 4.7, respectively, compared to the axenic fungal culture. The structures of the isolated compounds were elucidated on the basis of 1D and 2D NMR spectra and mass spectrometry as well as by comparison with literature data. The absolute configuration of compounds 3, 9, and 10 was determined by ECD (electronic circular dichroism) analysis aided by TDDFT-ECD (time-dependent density functional theory electronic circular dichroism) calculations. Compounds 15, 18 – 21, and 26 exhibited strong to moderate cytotoxic activity against the mouse lymphoma cell line L5178Y, with IC50 values ranging from 2.0 to 21.2 µM, while compounds 14, 16, 31, 32, and 33 displayed moderate inhibitory activities against several gram-positive bacteria, with MIC values ranging from 12.5 to 50 µM.

Phenyl 2,3,6,2′,3′,6′-hexa-O-acetyl-4′-S-acetyl-1,4,4′-trithio-β- d -maltoside and its tri-, tetra- and pentasaccharide homologues were prepared by completely regio- and stereoselective photoinitiated thiol-ene coupling reactions of 2-acetoxy-glucal and 4-thioglucose type reaction partners. Complex protecting group strategies could be avoided since all thiols and glucals were prepared from a single starting material, phenyl 2,3,6-tri-O-acetyl-4-S-acetyl-1,4-dithio-β- d -glucopyranoside. The method represents a simple alternative to the known syntheses of similar products.

Biomass productivity of shrub willow plants grown in short rotation system can be improved by genetic means or by innovative cropping technologies. In the present study we analyzed the growth and physiological responses of willow plants to plant biostimulators, such as 1-triacontanol (TRIA), a saturated primary alcohol and seaweed extract (Kelpak®). Testing a novel approach, we soaked stem cuttings in TRIA or Kelpak solutions for 48 h before plantation. These treatments enlarged height and diameter of stems, furthermore increased stem and leaf weights in comparison to the water control. In agreement with these greenhouse observations, field tests showed statistically significant enhancement in height and diameter of woody stems harvested in winter. Application of 25% Kelpak solution was the most effective in stimulating all traits including stem weight per plant after pre-planting treatment of cuttings. In an alternative treatment protocol, Kelpak was applied as foliar treatment or in combination between TRIA (10 mg L−1) treatment of cuttings and foliar spray with Kelpak 1 or 2% solutions in the greenhouse. The green pixel numbers revealed variable degrees of stimulation in shoot growth. These treatments resulted in an increase in stem and leaf weights. Improvement of photosynthetic functions was indicated by more efficient electron transport rates (ETRs) of photosystems. An increased nicotinamide and thiamine contents were detected in the leaves of stimulated plants. The present study can serve as a foundation for additional laboratory and field studies optimizing the application of these stimulators in energy plantations.

Four new monoterpenoid bisindole alkaloids, flabellipparicine (1), 19,20-dihydrovobparicine (2), 10′-demethoxy-19,20-dihydrovobatensine D (3), and 3′-(2-oxopropyl)ervahanine A (4), and 10 known monoterpenoid indole alkaloids were isolated from the stems of Tabernaemontana divaricata. All structures were elucidated based on spectroscopic methods, and the absolute configuration of 1 was established using conformational analysis and TDDFT-ECD calculation of selected stereoisomers. Compound 1 represents the first flabelliformide-apparicine-type bisindole alkaloid, in which the flabelliformide-like unit connects to the apparicine-like unit with a C-3-C-22′ bond and an N-1-C-16′ bond to form an uncommon five-membered ring between the two monomers. All alkaloids were evaluated for their cytotoxicity against two human cancer cell lines, MCF-7 and A-549. Compounds 2, 4, and 14 exhibited cytotoxicity against MCF-7 and A-549 with IC50 values in the range of 2 nM to 8 μM. Fil: Cai, You Sheng. Wuhan University; China Fil: Sarotti, Ariel Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina Fil: Zhou, Ting Lan. Wuhan University; China Fil: Huang, Rong. Wuhan University; China Fil: Qiu, Guofu. Wuhan University; China Fil: Tian, Congkui. Hubei University For Nationalities; China Fil: Miao, Ze Hong. Chinese Academy of Sciences; República de China Fil: Mándi, Attila. University of Debrecen; Hungría Fil: Kurtán, Tibor. University of Debrecen; Hungría Fil: Cao, Shugeng. University Of Hawaii At Hilo; Estados Unidos Fil: Yang, Sheng Ping. Wuhan University; China

Two new cryptic 3,4-dihydronaphthalen-(2H)-1-one (1-tetralone) derivatives, aspvanicin A (1) and its epimer aspvanicin B (2), as well as several known cryptic metabolites (3–8), were obtained from the ethyl acetate extract of the co-culture of the endophytic fungus Aspergillus versicolor KU258497 with the bacterium Bacillus subtilis 168 trpC2 on solid rice medium. When A. versicolor was cultured axenically in liquid Wickerham medium supplemented with 3.5% DMSO, an additional three known secondary metabolites (9–11) were isolated that were lacking when the fungus was fermented on rice medium. The structures of the new compounds were elucidated using one- and two-dimensional NMR spectroscopy as well as HRESIMS. The relative and absolute configurations of 1 and 2 were determined by the combination of NMR and electronic circular dichroism (ECD) analysis aided by DFT conformational analysis and TDDFT-ECD calculations. The ECD calculations revealed that although the sign of the blue-shifted overlapping n-π∗ ECD transition follows the helicity rule of cyclic aryl ketones, the calculation of low-energy conformers and ECD spectra was necessary to determine the stereochemistry. All metabolites were assessed for their antibacterial and cytotoxic activities; one of the new diastereomers, compound 2, showed moderate cytotoxic activity against the mouse lymphoma cell line L5178Y.

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We tested the effect of urbanization on air pollution based on the chlorophyll-a content of Celtis occidentalis leaves along an urbanization gradient (urban, suburban and rural areas) in Debrecen, Hungary. Chlorophyll-a content of Celtis occidentalis, Acer campestre, and Corylus avellana were compared to test which species is the most useful to study the effects of urbanization. Furthermore, the effects of washing solutions (distilled water, tap water, and rainwater) on chlorophyll-a content of tree leaves were also tested during sample preparation. Chlorophyll-a was extracted from leaf samples with acetone, and it was measured using a spectrophotometer. Along the urbanization gradient, chlorophyll-a content of C. occidentalis leaves was the lowest in the urban area; thus, this species proved to be an effective indicator of anthropogenic emission load. Differences were not significant among species in the suburban and rural areas, where the level of air pollution was moderate. We found that effects of the washing solutions on chlorophyll-a content did not differ significantly from each other. Thus, tap water can be used safely to clean the leaf surface, without significant influence on chlorophyll-a. Our study demonstrated that the chlorophyll-a content of leaves was a useful indicator to assess the level of air pollution.

Arthropod-parasitic fungi of the order Laboulbeniales are known to exhibit specialization to individual host taxa in most cases. Some species exhibit ecological specificity to multiple, often unrelated hosts in certain microhabitats; and often position specificity to different host body parts. The myrmecophilous Rickia wasmannii (Ascomycota: Laboulbeniales) infects Myrmica species (Hymenoptera: Formicidae) (host specificity), and occasionally other arthropod inquilines inside the ant nest (ecological specificity). An effect of the position of infection on the thallus densities has also been reported. Another determinative factor that may also exist in the Rickia-Myrmica host-parasite system, the chronological age of ant worker hosts, has also been linked to parasite load. Comprehensive studies on the age-related infection intensity, however, are still lacking. Here we investigated whether the level of infection correlates with the age of the M. scabrinodis host consistently. We found that older hosts exhibited higher parasite load, even though the infection level of the different colonies varied widely. The results highlight that the level of R. wasmannii infections are strongly influenced by host individual and host colony factors.

The freshwater cyanobacterium Cylindrospermopsis raciborskii w as investigated for carotenoid composition. Besides β-carotene, echinenone and (9/9′ Z )-echinenone a carotenoid glycoside was found to be the main component. This compound was isolated and subsequently acetylated for structural elucidation. The acetyl derivative was fully characterized by UV–vis, ECD, NMR and HRMS techniques. The detailed 1 H and 13 C NMR chemical shift assignment of the major carotenoid supported the unequivocal identification of (2′ S )-2-hydroxymyxol 2′-α- l -fucoside.

Public awareness has been raised on the importance of natural history and academic collections for science and society in a time when reduced financial support and staff cuts are prevalent. In the field of biology, new species and new interspecies associations are constantly discovered by making use of museum collections, digitalised materials or citizen science programs. In our study, the Myrmica Latreille, 1804 image collection of AntWeb.org was screened for fungal ectoparasites. A total of 397 imaged specimens from 133 species were visually investigated. A single specimen of M. hellenica Finzi, 1926, collected in Greece by U. Sahlberg, showed a conspicuous fungal infection. The parasite was identified using microscopic methods as Rickia wasmannii Cavara, an ectoparasitic fungal species specialised to Myrmica ants. This finding represents a new country record and a new Myrmica species for the host spectrum of R. wasmannii. According to our results, online entomological databases can be screened relatively easily for ectoparasitic fungal infections from new hosts and new regions. However, depending on quality of the insect voucher photos, additional investigation of the material could be needed to confirm the identity of the parasite.

In 2017 the European Goldfinch (Carduelis carduelis) was voted to be the “bird of the year” in Hungary. This is a partially migrant species; most of the European populations are resident, however, its breeding range extends to East from the Sweden-Poland-Moldova axis towards the Yenisei with some of the populations wintering in Kazakhstan and South of Turkey. The European Goldfinch is classified within the Carduelinae subfamily including approximately a hundred species. Several taxonomic changes were introduced in this group during the last fifteen years, however, we still do not understand much of their origin and evolutionary history. My aim in this paper is to collect existing knowledge on the phylogeny and evolution of the Carduelinae finches and their allies, with a particular focus on the European Goldfinch and its closest relatives. Furthermore, here I point out uncertainties in different phylogenetic sources of finches, which careful consideration can be useful in similar evolutionary studies. Finally, I summarise some vision for future research.

We tagged 12 adult Common Cuckoos (Cuculus canorus) with non-platform terminal transmitter (non-PTT) GPS-UHF telemetry at their breeding grounds in Hungary. One male and two female Cuckoos (one of them twice) were again observed in subsequent years, and GPS fixes documented their migration routes to and from Africa, as far south as Namibia. All four routes showed the species-typical clockwise loop migration. Although currently non-PPT GPS tracking with remotely downloadable data as an ornithological method is primarily suitable to map home ranges of birds, it could be a complement to PTT technology in migration research, especially for delivery of higher spatial accuracy.

Gills are major targets for acute metal toxicity in fish, due to their permanent contact with aquatic pollutants. To assess the effects of metals on gills of the Leuciscus cephalus (chub), fish individuals were collected from two sites in the Tur River, Romania, in upstream (site 1) and downstream (site 2) of a metal pollution source. Quantitative and hyperspectral analyses showed that Zn, Sr, and Fe concentrations were significantly higher in gills from site 2 compared with site 1. Malondialdehyde and advanced oxidation protein products levels increased 17 and 28%, respectively, whereas reduced glutathione level diminished significantly in the gills of fish collected from site 2 compared to site 1. The activities of superoxide dismutase, catalase, and glutathione-S-transferase increased significantly at 41, 21, and 28%, respectively. Proliferating cell nuclear antigen (PCNA) protein levels, as well as the amount of DNA damage, were significantly increased for site 2 compared with site 1. The induced oxidative stress generated hyperplasia, hypertrophy, and inflammation in the epithelial cells and apoptosis. Hence, this could suggest that gill cells have tried to counteract the oxidative stress-induced DNA fragmentation by PCNA up-regulation, but the PCNA expression decreased on longer time due to the low level of GSH, resulting in apoptosis.

There are extensive data on effects of antifungal agents on the plant pathogens, especially on Fusariums spp. species. However, investigations on the interaction of chemicals and the treated cultivars are rare. The aim of the study was to test two types of fungicide mixtures, azoxystrobin-propiconazole, and prothioconazole-tebuconazole, which are applied in wheat cultivars intensively, on six fodder maize hybrids that were infected with Fusarium proliferatum in the R1 growth stage in a field trial. The effect of the fungicide treatment was tested on the starch content and antifungal, antioxidant polyphenols of the kernels in the R3–R4 and R6 stage of the cultivars. The level of the fungal presence and the fumonisin concentration of the kernels were increased significantly under the artificial infection. The fumonisin concentration was variable at the R6 stage of the hybrid maize kernels. The treatment with prothioconazole and tebuconazole was found to be suitable when it was done before flowering, while the azoxystrobin-propiconazole treatments were equally successful before and after maize flowering considering the decreasing fumonisin concentration of the kernels. Both fungicide mixtures, when they were applied after maize flowering, affected the starch biosynthesis to the R3–R4 stage significantly. Meanwhile, azoxystrobin-propiconazole also significantly affected the antioxidant flavone/flavanol contents from the R3–R4 stage to the R6 stage.

Background. The Amur sleeper, Perccottus glenii Dybowski, 1877, is the most invasive alien fish species in the native aquatic communities in central Europe. Although the accelerated invasion of P. glenii has been well documented, there is little information on the ecological and growth parameters of non-native populations in this region. The aim of this study was to describe the growth features of the population of P. glenii in a shallow oxbow lake in the Carpathian Basin. Materials and methods. Our study sample consisted of 1239 individuals (628 ♂ + 611 ♀) collected from an oxbow lake near the Tisza River in the same month for three years (2013–2015). The length, weight, age structure, sex-dependent growth rate, and the condition factor of the collected specimens were determined. Results. The sex ratio was 0.49 (♀ ÷ (♀ + ♂)). The standard length and weight of the collected P. glenii specimens ranged from 20.7 to 127.7 mm and from 0.3 to 75.8 g, respectively. The length–weight relations (SL–W) were allometrically negative for the males (W = 3.2 × 10–5SL2.960), females (W = 3.8 × 10–5SL2.921), and both sexes (W = 3.5 × 10–5SL2.940), without significant differences between males and females. According to the length-frequency analysis, five age groups were differentiated. The von Bertalanffy growth models were L∞ = 138.87 mm, k = 0.21, t0 = –0.54 (♂), L∞ = 174.21 mm, k = 0.17, t0 = –0.33 (♀), L∞ = 154.01 mm, k = 0.19, t0 = –0.45 (♂ + ♀). The growth performance parameters were Ф′ = 3.61 (♂), Ф′ = 3.71 (♀), Ф′ = 3.65 (♂ + ♀), respectively. Conclusion. Due to unequal investment in reproduction, there was a significant difference in the growth rate between males and females. The literature data showed that invasive gobies and odontobutids (e.g., P. glenii) exhibit more opportunistic reproduction strategy in newly colonized areas, which may contribute to the invasion success. This strategy (e.g., earlier maturation, longer spawning period, etc.) results in slower growth rate due to energetic trade-off between reproduction and somatic growth. The observed growth rate of invasive Amur sleeper population (especially in the older age groups) was slower than that of native and more established naturalized populations in Eurasia.

Lay Summary We found that innovative great tit females are prone to cuckold their mates. Innovative problem solving can be important to animals for survival and reproduction in nature, yet we found no evidence that females mated to males with poor problem-solving performance compensate for poor male quality by obtaining superior genes for their offspring from other males via cuckoldry. Instead, females’ infidelity increased with their own innovativeness.

Aims Cyanobacteria may have considerable effects on community functioning, mostly because they produce various metabolites that adversely affect other organisms. Here we synthesized existing knowledge about the effects of toxic cyanobacteria on the germination and growth of terrestrial plants. We also aimed to test the chemical effects of a Nostoc (Cyanobacteria) extract on the germination and growth of species of alkali habitats to investigate whether cyanobacteria can alter community structure and diversity via affecting the establishment success of plants. Location Cyanobacterium colonies from the Hortobagy National Park, east Hungary; indoor experiments at the University of Debrecen, Hungary. Methods To review the effects of toxic cyanobacteria on terrestrial plants, we conducted a literature search. To test these effects on native plants, field-collected Nostoc colonies were used to prepare a cell-free water extract, and treatments (watering with Nostoc extract and watering with tap water) were tested on 3 × 100 seeds of nine alkali grassland species. After 5 wk, seedling number, seedling length and fresh and dry weights were measured. Results We collected data on the effects of cyanobacteria on 27 species, but they were mostly focused on crops irrigated with cyanobacteria-containing water, not on floras native to natural ecosystems. In the germination experiment species identity and treatment had a significant effect on almost all variables, but their interaction only affected germination rate and fresh weight. Fresh weight decreased significantly only in the invasive Hordeum jubatum, but germination rate decreased significantly in five species. Conclusions Based on our findings, terrestrial cyanobacterium colonies can affect the establishment success of grassland plants, through which they may be important in determining which species can be incorporated into the community. Thus, cyanobacteria might play an important role in shaping diversity, species composition and the structure of natural plant communities.

We allocated some new types of bait traps using different volatile aromatic compounds in the Velyka Dobron’ Game Reserve (Ukraine, Transcarpathian region) where light trap surveys on Macroheterocera were already carried out since 2009. The phenylacetaldehyde-based traps were especially effective for the species of subfamilies Plusiinae and Helio-thinae, while the members of some other subfamilies (Hadeninae, Noctuinae) were more attracted by the isoamyl-alcohol-based traps. The highest attractivity was observed in case of the mixture containing isoamyl-alcohol, acetic acid and red wine. The highest num-ber of species was observed in mid-July, but the peak of individual numbers appeared in early August. Majority of species of this assemblage was formed by bivoltine species connected to herbaceous food plants. The dominant species of this late summer period was Trachea atriplicis (second brood). It was sharply replaced by the monovoltine Allophyes oxyacanthae in September. The bulk of species of this early autumnal period was formed by monovoltine species with woody-shrubby larval food plants. Biogeographical spectrum of the assemblage was dominated by widely distributed Euro-Siberian species combined with significant presence of Mediterranean and Boreo-continental faunal elements. In the ecological spectrum of the assemblage the species connected to forested habitats are well represented but together with the presence of numerous generalist species.

Eight new hydroquinone derivatives, gliomastins A–D (1–4), 9-O-methylgliomastin C (5), acremonin A 1-O-β-D-glucopyranoside (6), gliomastin E 1-O-β-D-glucopyranoside (7), and 6′-O-acetyl-isohomoarbutin (8), together with seven known analogues were isolated from the marine-derived fungus Gliomastix sp. Their structures were elucidated by extensive spectroscopic analysis including 1D and 2D NMR measurements aided by DFT NMR calculations as well as MS data. TDDFT-ECD and OR calculations were performed to determine the absolute configurations of 1 and the aglycones of 6 and 7. Compound 1 features a novel skeleton, biogenetically derived from a Diels–Alder reaction between derivatives of 11 and 13. Compound 2 represents a rare sulfur-containing alkaloid derived from the known hydroquinone 13. Compounds 1, 10 and 12 showed strong cytotoxicity against the L5178Y mouse lymphoma cell line with IC50 values of 1.8, 1.0 and 1.1 μM, respectively. Compound 3 exhibited moderate antitubercular activity against Mycobacterium tuberculosis with a MIC value of 12.5 μM.

In the present study, a new species of Craniophora Snellen, 1867, C. minuscula sp. n., is described as new to science from Japan, during course of the investigations on the Craniophora pontica -group. The external and genitalia morphology of both sexes is provided including the discrimination of the new species from the closely allied congeners.

In this study, we research ground-dwelling arthropods along four road stretches, which represent the main verge habitats (arid grassland, agricultural area, forest, and wetland) of Hungary. Besides the faunistic investigation of arthropod communities, we described and compared dominance relation and species composition. We identified 83 carabids, 81 spiders, and 4 isopod species. Seven beetle species and one spider species are included in the invertebrate Red List of Hungary. The registered species were dominant on main road verges bordering arid grassland and wetland. Main road verges proved to be a diverse habitat; hence, it is important to carry on more investigation.

The present study has focused on an investigation of the antibacterial effects of Juncus inflexus and the isolation and identification of its active compounds. Eleven phenanthrenes were isolated from a methanolic extract of the roots. Four compounds (jinflexins A-D, 1-4) are new natural products, while seven phenanthrenes [juncuenins A (5), B (6), and D (8), juncusol (7), dehydrojuncuenins A (9) and B (11), and dehydrojuncusol (10)] were isolated for the first time from the plant. Jinflexin D (4) is a dimer with an unprecedented heptacyclic ring system. The absolute configurations of the new compounds were determined by TDDFT-ECD calculations, and their enantiomeric purity was checked by chiral HPLC analysis. Extracts of different polarity (n-hexane, dichloromethane, and ethyl acetate) were evaluated for their antimicrobial effects against methicillin-resistant Staphylococcus aureus, extended-spectrum β-lactamase (ESBL)-producing Citrobacter freundii, Escherichia coli, Enterobacter cloacae, Klebsiella pneumoniae, multiresistant Acinetobacter baumannii, and Pseudomonas aeruginosa. The MIC values of the isolated compounds were determined by a microdilution method. Jinflexin B (2), juncusol (7), juncuenin D (8), and dehydrojuncuenin B (11) showed significant activity (MIC value range 12.5-100 μg/mL) against MRSA strains.

We compared the macro-moth assemblages of four forests from which one is extended on the Ukrainian and three on the Hungarian side of the Bereg lowland, and additionally one in the adjacent part of the Hungarian plain (NE of Carpathian Basin). The data were collected by light trapping during at least in five years in all sites. Numerous faunistically significant and/or rare species were recorded. The species composition, faunal types and ecological components were rather similar in all sites, dominated by widely distributed Euro-Siberian species but with considerable participation of Boreo- Continental and Holo-Mediterranean—West Asiatic species. Considering the ecological connections of the species, the largest part of the assemblage is composed by species of different types of deciduous forests (oak woods and hardwood gallery forests) but also with significant proportion of wetland components (marshy-boggy habitats and tall forb communities) and generalist species. The similarity of the species composition proved to be high in all dominant taxonomical groups (Geometridae, Erebidae and Noctuidae, including the most diverse subfamilies). Several species have high conservation significance.

Information about the diameter and the height of woody species is fundamental to developing growth and yield models in forest stands. Ten nonlinear height–diameter functions were fitted and evaluat...

A chemical investigation of the endophyte Penicillium sp. (strain ZO-R1-1), isolated from roots of the medicinal plant Zingiber officinale, yielded nine new indole diterpenoids (1-9), together with 13 known congeners (10-22). The structures of the new compounds were elucidated by 1D and 2D NMR analysis in combination with HRESIMS data. The absolute configuration of the new natural products 1, 3, and 7 was determined using the TDDFT-ECD approach and confirmed for 1 by single-crystal X-ray determination through anomalous dispersion. The isolated compounds were tested for cytotoxicity against L5178Y, A2780, J82, and HEK-293 cell lines. Compound 1 was the most active metabolite toward L5178Y cells, with an IC50 value of 3.6 μM, and an IC50 against A2780 cells of 8.7 μM. Interestingly, 1 features a new type of indole diterpenoid scaffold with a rare 6/5/6/6/6/6/5 heterocyclic system bearing an aromatic ring C, which is suggested to be important for the cytotoxic activity of this natural product against L5278Y and A2780 cells.

l-Arabinose and d-galactose are the principal constituents of l-arabinogalactan, and also co-occur in other hemicelluloses and pectins. In this work we hypothesized that similar to the induction of relevant glycoside hydrolases by monomers liberated from these plant heteropolymers, their respective catabolisms in saprophytic and phytopathogenic fungi may respond to the presence of the other sugar to promote synergistic use of the complex growth substrate. We showed that these two sugars are indeed consumed simultaneously by Aspergillus nidulans, while l-arabinose is utilised faster in the presence than in the absence of d-galactose. Furthermore, the first two genes of the Leloir pathway for d-galactose catabolism - encoding d-galactose 1-epimerase and galactokinase - are induced more rapidly by l-arabinose than by d-galactose eventhough deletion mutants thereof grow as well as a wild type strain on the pentose. d-Galactose 1-epimerase is hyperinduced by l-arabinose, d-xylose and l-arabitol but not by xylitol. The results suggest that in A. nidulans, l-arabinose and d-xylose - both requiring NADPH for their catabolisation - actively promote the enzyme infrastructure necessary to convert β-d-galactopyranose via the Leloir pathway with its α-anomer specific enzymes, into β-d-glucose-6-phosphate (the starting substrate of the oxidative part of the pentose phosphate pathway) even in the absence of d-galactose.

Co-cultivation of the endophytic fungus Fusarium tricinctum with Streptomyces lividans on solid rice medium led to the production of four new naphthoquinone dimers, fusatricinones A–D (1–4), and a new lateropyrone derivative, dihydrolateropyrone (5), that were not detected in axenic fungal controls. In addition, four known cryptic compounds, zearalenone (7), (−)-citreoisocoumarin (8), macrocarpon C (9) and 7-hydroxy-2-(2-hydroxypropyl)-5-methylchromone (10), that were likewise undetectable in extracts from fungal controls, were obtained from the co-culture extracts. The known antibiotically active compound lateropyrone (6), the depsipeptides enniatins B (11), B1 (12) and A1 (13), and the lipopeptide fusaristatin A (14), that were present in axenic fungal controls and in co-culture extracts, were upregulated in the latter. The structures of the new compounds were elucidated by 1D and 2D NMR spectra as well as by HRESIMS data. The relative and absolute configuration of dihydrolateropyrone (5) was elucidated by TDDFT-ECD calculations.

Chub (Squalius cephalus L. 1758) specimens of three age groups with different types of diet were collected in November 2013 in the River Szamos/Somes, Hungary. The Ca, K, Mg, Na, Cd, Cr, Cu, Fe, Mn, Pb, Sr and Zn concentrations were analyzed in the muscle, gills and liver samples of chub by microwave assisted plasma-atomic emission spectrometry (MP-AES). The Kruskal–Wallis test revealed significant differences among different age groups based on the trace element concentrations in the liver, muscle and gills. The trace element concentration pattern in muscle and liver of different age groups differed, may be according to the different diet types of the groups. Meanwhile no differentiation among the age groups based on the trace element concentration in the gills was observed, probably because the pattern of trace elements in the gills is related to the habitat preference, which does not differ during the life-span of chub. In contrast to expectations, trace element concentrations in juveniles were the highest in most cases, certainly because of their specific diet, relatively fast metabolic rate and inadequately developed detoxification system. Only the copper concentrations in liver increased with fish age. Considerable concentrations of trace elements in the tissues of juveniles were observed in the case of elements whose concentrations in the River Szamos were higher in 2013 than in previous years. According to this phenomenon, trace element patterns in the tissues of juveniles may be good indicators of recent pollution of watercourses.

We report the synthesis of some novel β-lactam hybrids of 2-mercaptobenzothiazole and benzoquinoline. These compounds were synthesized by a [2 + 2]-cycloaddition reaction of imines 8a-d and ketenes derived from substituted acetic acids. The reaction was totally diastereoselective leading exclusively to the formation of cis-β-lactams 10a-m. All products were obtained in good to excellent yields and their structures were established based on IR, 1H NMR, 13C NMR spectral data and elemental analysis. Schiff bases 8a-d and β-lactam hybrids 10a-m were evaluated for antimicrobial activities against six bacterial species. The minimum inhibitory concentration (MIC) values indicate that two of the β-lactams, 10k and 10m, have good activities against the two Gram-negative bacteria, E. coli and P. aeruginosa, while three of the Schiff bases, 8a-c, are active against P. aeruginosa and the Gram-positive pathogen S. aureus. The molecular and cellular basis for these observed antibacterial properties are not determined. Moreover, the five most active compounds showed acceptably low cytotoxicity (less than 25% cell growth inhibition after 72 h of incubation) against the MCF-7 cell line, and below 10% in vitro hemolytic activity at 50 and 200 µM concentrations. These results suggest a need for further inquiry into the reason for why these compounds are bioactive, and as to what their full biological activities and antibiotic potential may be. The cis stereochemistry of β-lactam 10a was confirmed by X-ray crystallographic studies.

Small, cysteine-rich and cationic antifungal proteins from natural sources are promising candidates for the development of novel treatment strategies to prevent and combat infections caused by drug-resistant fungi. However, limited information about their structure and antifungal mechanism hampers their future applications. In the present study, we determined the solution structure, dynamics and associated solvent areas of the Neosartorya (Aspergillus) fischeri antifungal protein NFAP. Genome mining within the genus revealed the presence of orthologous genes in N. fischeri and Neosartorya spathulata, and genes encoding closely related proteins can be found in Penicillium brasiliensis and Penicillium oxalicum. We show that the tertiary structure of these putative proteins can be resolved using the structure of NFAP as reliable template for in silico prediction. Localization studies with fluorescence-labelled protein pointed at an energy-dependent uptake mechanism of NFAP in the sensitive model fungus Neurospora crassa and subsequent cytoplasmic localization coincided with cell-death induction. The presented results contribute to a better understanding of the structure/function relationship of NFAP and related proteins and pave the way towards future antifungal drug development.

An increasing number of infections originating from probiotic use are reported worldwide, with the majority of such cases caused by the yeast Saccharomyces 'boulardii', a subtype of S. cerevisiae. Reliably linking infectious cases to probiotic products requires unequivocal genotyping data, however, these techniques are often time-consuming and difficult to implement in routine diagnostics. This leads to a widespread lack of genetic data regarding the origin of Saccharomyces infections. We propose a quick and reliable PCR-based protocol for the identification of S. 'boulardii' based on a combined analysis of interdelta fingerprinting and microsatellite typing. By applying various typing methods and our proposed method to the clinical yeast collection of a Hungarian hospital we show that probiotic origin is common among clinical Saccharomyces, and that the new multiplex method enables rapid and unequivocal identification of probiotic yeast infections. This method can be applied for the identification of yeast infection sources, helping decisions on probiotic use.

Three new compounds (9-11) were isolated together with eight known analogues from the fungus Pseudallescheria boydii associated with the South China Sea soft coral Sinularia sandensis. The structures of the new compounds were elucidated on the basis of the spectroscopic analysis, and the absolute configurations including the sulfur stereogenic center of a sulfoxide moiety were determined by comparison of experimental ECD spectra to TDDFT/ECD calculations. Epimeric chiral sulfoxides differing in the absolute configuration of the sulfur chirality center could be efficiently distinguished and assigned by comparing the experimental ECD to those of calculations for the sulfur epimers. In the in vitro biotests for osteoclastogenesis effects, compounds 1, 5, 7, and 10 exhibited a stimulatory activity, while compound 3 displayed an inhibitory activity.

The class Laboulbeniomycetes comprises biotrophic parasites associated with arthropods and fungi. Two orders are currently recognized, Pyxidiophorales and Laboulbeniales. Herpomyces is an isolated genus of Laboulbeniales, with species that exclusively parasitize cockroaches (Blattodea). Here, we evaluate 39 taxa of Laboulbeniomycetes with a three-locus phylogeny (nrSSU, ITS, nrLSU) and propose a new order in this class. Herpomycetales accommodates a single genus, Herpomyces, with currently 26 species, one of which is described here based on morphological and molecular data. Herpomyces shelfordellae is found on Shelfordella lateralis cockroaches from Hungary, Poland, and the USA. We also build on the six-locus dataset from the Ascomycota Tree of Life paper (Schoch and colleagues, 2009) to confirm that Laboulbeniomycetes and Sordariomycetes are sister classes, and we apply laboulbeniomyceta as a rankless taxon for the now well-resolved node that describes the most recent common ancestor of both classes.

One of the promising directions of the combined approach is the design of dual-acting antibiotics - heterodimeric structures on the basis of antimicrobial agents of different classes. In this study a novel series of azithromycin-glycopeptide conjugates were designed and synthesized. The structures of the obtained compounds were confirmed using NMR spectroscopy and mass spectrometry data including MS/MS analysis. All novel hybrid antibiotics were found to be either as active as azithromycin and vancomycin against Gram-positive bacterial strains or have superior activity in comparison with their parent antibiotics. One compound, eremomycin-azithromycin conjugate 16, demonstrated moderate activity against Enterococcus faecium and Enterococcus faecalis strains resistant to vancomycin, and equal to vancomycin's activity for the treatment of mice with Staphylococcus aureus sepsis.

The discovery and understanding of the mode of action of new antimicrobial agents is extremely urgent, since fungal infections cause 1.5 million deaths annually. Antifungal peptides and proteins represent a significant group of compounds that are able to kill pathogenic fungi. Based on phylogenetic analyses the ascomycetous, cysteine-rich antifungal proteins can be divided into three different groups: Penicillium chrysogenum antifungal protein (PAF), Neosartorya fischeri antifungal protein 2 (NFAP2) and “bubble-proteins” (BP) produced, for example, by P. brevicompactum. They all dominantly have β-strand secondary structures that are stabilized by several disulfide bonds. The PAF group (AFP antifungal protein from Aspergillus giganteus, PAF and PAFB from P. chrysogenum, Neosartorya fischeri antifungal protein (NFAP)) is the best characterized with their common β-barrel tertiary structure. These proteins and variants can efficiently be obtained either from fungi production or by recombinant expression. However, chemical synthesis may be a complementary aid for preparing unusual modifications, e.g., the incorporation of non-coded amino acids, fluorophores, or even unnatural disulfide bonds. Synthetic variants up to ca. 6–7 kDa can also be put to good use for corroborating structure determination. A short overview of the structural peculiarities of antifungal β-strand disulfide bridged proteins will be given. Here, we describe the structural propensities of some known antifungal proteins from filamentous fungi which can also be prepared with modern synthetic chemistry methods.

BackgroundPhysiological regulators of life history trade-offs need to be responsive to sudden changes of resource availability. When homeostasis is challenged by unpredictable stressors, vertebrates respond through a set of physiological reactions, which can promote organismal survival. Glucocorticoids have been traditionally recognized as one of the main regulators of the physiological stress response, but the role of an evolutionarily more conserved pathway, the hypothalamic-pituitary-somatotropic (HPS) axis producing insulin-like growth factor-1 (IGF-1) has received much less attention. Although IGF-1 is known to affect several life history traits, little is known about its role in the physiological stress response and it has never been studied directly in adult wild animals.MethodsIn this study, we combined field observations with a controlled experiment to investigate how circulating levels of IGF-1 change in response to stress and whether this change is due to concomitant change in glucocorticoids in a free-living songbird, the bearded reedling Panurus biarmicus. We used a standard capture-restraint protocol in field observation, in which we took first and second (stress induced: 15 minutes later) samples. In a follow-up experiment, we used a minimally invasive oral corticosterone manipulation.ResultsWe showed that corticosterone levels significantly increased while IGF-1 levels significantly decreased during capture and handling stress. However, change in corticosterone levels were not related to change in IGF-1 levels. We found that experimentally elevated corticosterone levels did not affect IGF-1 levels.DiscussionOur results are the first to highlight that circulating IGF-1 levels are responsive to stress independently from glucocorticoids and suggest that the HPS axis is an autonomous physiological pathway that may play an important role as regulator of life-history decisions.

(±)-Cryptomeriolide, a pair of racemic bis- seco-abietane diterpenoids, were isolated from the bark of Cryptomeria fortunei. The separation of enantiomers was achieved by using chiral stationary phase HPLC. Their structures including the absolute configuration and conformations in solution and solid state were determined by extensive analysis of spectroscopic data, single-crystal X-ray diffraction, and comparison of calculated and experimental electronic circular dichroism data. A bioinspired one-pot enantiomeric synthesis of 1a and 1b was accomplished via a readily made intermediate orthoquinone from sugiol. All compounds including the synthetic intermediates were assayed for their cytotoxic activities on human cancer cell lines HL-60, A549, and SGC7901.

In this study, we analyzed the expression of the structural genes encoding the five enzymes comprising the Leloir pathway of D-galactose catabolism in the industrial cell factory Penicillium chrysogenum on various carbon sources. The genome of P. chrysogenum contains a putative galactokinase gene at the annotated locus Pc13g10140, the product of which shows strong structural similarity to yeast galactokinase that was expressed on lactose and D-galactose only. The expression profile of the galactose-1-phosphate uridylyl transferase gene at annotated locus Pc15g00140 was essentially similar to that of galactokinase. This is in contrast to the results from other fungi such as Aspergillus nidulans, Trichoderma reesei and A. niger, where the ortholog galactokinase and galactose-1-phosphate uridylyl transferase genes were constitutively expressed. As for the UDP-galactose-4-epimerase encoding gene, five candidates were identified. We could not detect Pc16g12790, Pc21g12170 and Pc20g06140 expression on any of the carbon sources tested, while for the other two loci (Pc21g10370 and Pc18g01080) transcripts were clearly observed under all tested conditions. Like the 4-epimerase specified at locus Pc21g10370, the other two structural Leloir pathway genes - UDP-glucose pyrophosphorylase (Pc21g12790) and phosphoglucomutase (Pc18g01390) - were expressed constitutively at high levels as can be expected from their indispensable function in fungal cell wall formation.

A new distribution record for Prolixandromyces triandrus Santam. (Ascomycota: Laboulbeniales) is presented from the Bukk Mountains in northeastern Hungary, from the host species Velia (Plesiovelia) saulii Tamanini, 1947 (Heteroptera: Veliidae). Hitherto, this fungal parasite had only been observed in the western Mediterranean region and the Macaronesia Archipelago. Prolixandromyces. triandrus seems to be abundant in the reported Hungarian host population. Additionally, ribosomal DNA barcodes for this fungal species are also presented. Incidence of the parasite and potential of the molecular investigations of host-parasite relationships of this ectoparasitic fungus is discussed. A brief review is given of known hosts of P. triandrus and of Laboulbeniales from aquatic/semiaquatic insect hosts in Hungary.

Six new pyridone alkaloids, named penipyridones A–F (1–6), were isolated from the fermentation broth of an Antarctic moss-derived fungus, Penicillium funiculosum GWT2-24. Their structures were elucidated from extensive NMR and MS data. Although they possess the same major chromophore and some of them presented almost mirror ECD spectra, their absolute configurations were found to be uniformly S, as evidenced by X-ray single-crystal diffraction analysis, stereocontrolled total synthesis, and chemical conversions. TDDFT-ECD calculations of compounds 3 and 6 revealed that subtle conformational changes are responsible for the significantly different ECD curves. None of the compounds were cytotoxic (IC50 > 50 μM), while compounds 1, 2, 5, and 7 elicited lipid-lowering activity in HepG2 hepatocytes.

Two new xanthone derivatives, 12-dimethoxypinselin ( 1 ) and 12- O -acetyl-AGI-B4 ( 2 ), as well as two new phenolic bisabolane-type sesquiterpenes, 11,12-dihydroxysydonic acid ( 15 ) and 1-hydroxyboivinianic acid ( 16 ), together with one new alkaloid, scopulamide ( 21 ) and one new α-pyrone derivative, scopupyrone ( 26 ), in addition to twenty-three known compounds ( 3 – 14 , 17 – 20 , 22 – 25 , 27 – 29 ) were isolated from solid rice cultures of the marine-derived fungus Scopulariopsis sp. obtained from the Red Sea hard coral Stylophora sp. All compounds were unambiguously identified through extensive NMR spectroscopic analyses, and by comparison with the literature. Marfey's reaction was performed to determine the absolute configuration of scopulamide ( 21 ) and TDDFT-ECD calculations were used to assign the configuration of AGI-B4 ( 3 ) and scopupyrone ( 26 ). All isolated compounds were evaluated for their cytotoxicity against L5178Y mouse lymphoma cells and the structure-activity relationships were discussed.

In the current study, decomposition of diclofenac, diflunisal, ibuprofen, mefenamic acid and piroxicam was tested using nine identified strains of endophytic and epiphytic fungi (from Ascomycota) adapted to natural products resembling the pharmaceuticals. The strains were isolated from a medicinal plant, Plantago lanceolata leaves. Metabolites were tentatively identified by liquid chromatography – tandem mass spectrometry (LC-MS3). Eighteen of the 45 combinations resulted in significant decrease of the concentration of the NSAIDs in model solutions. The most active strains were Aspergillus nidulans and Bipolaris tetramera, while Epicoccum nigrum and Aspergillus niger showed somewhat less potency. Piroxicam and diclofenac were most resistant to biotransformation, while ibuprofen and mefenamic acid were efficiently metabolized by most strains. Ten metabolites could be tentatively identified, including hydroxy-metabolites of all tested NSAIDs, and a dihydroxy-metabolite of piroxicam. This biotransformation is likely to modify the toxicity and bioaccumulation potential of these pharmaceuticals. The results highlight the applicability of polyphenol-rich dried medicinal plant materials as an excellent source of fungi with high biotransforming potential. The results also suggest more in-depth testing of these fungi for biodegradation processes.

A new cytotoxic viriditoxin derivative, cladosporinone (1), along with the known viriditoxin (2) and two viriditoxin derivatives (3 and 4) were obtained from the fungus C ladosporium cladosporioides isolated from the sediment of a hypersaline lake in Egypt. The structure of the new compound (1) was determined by 1D and 2D NMR measurements as well as by high-resolution ESIMS and electronic circular dichroism spectroscopy. All isolated compounds were studied for their cytotoxicity against the murine lymphoma cell line L5187Y and for their antibiotic activity against several pathogenic bacteria. Viriditoxin (2) was the most active compound in both bioassays. Compound 1 also exhibited strong cytotoxicity against the murine lymphoma cell line L5187Y with an IC50 value of 0.88 μm, whereas its antibiotic activity was weak.

Small passerines are faced with a trade-off when foraging during winter. Increasing energy reserves makes them more vulnerable to predators, while a low level of reserves exposes them to a high risk of starvation. Whether small birds under these circumstances are allowed to reduce their foraging activity under increased predation risk, for example in feeding sites more exposed to predators, remains controversial in former behavioural and ecological researches. In this study, we investigated the foraging activity of free-living Tree Sparrow Passer montanus flocks feeding on an artificial feeding platform. The predation risk perceived by the sparrows was manipulated by placing the platform either close to or far from a bushy shelter. Foraging activity, assessed as cumulative activity of sparrows per unit time on the platform, did not differ between the low-risk and the high-risk conditions and did not significantly change during the day. Feeding efficiency, assessed as pecking rate, was not either reduced under the high-risk condition. Our results suggest that sparrows were forced to feed almost continuously during the day in order to maintain their preferred level of energy reserves. However, several behavioural changes helped sparrows to adopt a safer foraging policy when feeding far from cover, as we found in another study. Altogether, sparrow flocks feeding far from cover decreased the overall foraging time (the time when any sparrow stayed on the platform) by approximately 20% as compared to the near cover condition. A possible way to maintain the same level of foraging activity despite of the reduction in overall foraging time is discussed.

1.Floral deception is widespread in orchids, with more than one third of the species being pollinated this way. The evolutionary success of deceptive orchids is puzzling, as species employing this strategy are thought to have low reproductive success (less flowers yielding fruits) because of low pollination rates. However, direct measurements of total seed production in orchids – which is a better measure of reproductive success – are scarce due to the extremely small size of their seeds. 2.Here, we quantified seed numbers in 1,015 fruits belonging to 48 orchid species from the Pannonian ecoregion (central Europe) and obtained fruit-set and thousand-seed weight data for these species from the literature. We used phylogenetic comparative methods to test the hypothesis that deceptive species should compensate for their lower fruit-set by having either more flowers, larger seeds or more seeds in a fruit. 3.Similarly to previous studies, we found that deceptive orchids have substantially lower fruits-set than nectar-rewarding ones. Also, we found that deceptive species have more seeds in a fruit but not more flowers or larger seeds compared to nectar-rewarding ones. Based on our results, deceptive species compensate for their lower fruit-set by having higher seed numbers per fruit. As a consequence, their seed numbers per shoot do not differ from that of nectar-rewarding ones. 4.Together with other benefits of deceptive pollination (e.g. lower energy expenditure due to the lack of nectar production and higher genetic variability due to decreased probability of geitonogamous pollination), our results can explain why deceptive strategies are so widespread in the orchid family. 5.Synthesis. Our results indicate that deceptive orchids can compensate for their lower fruit-set by having more (but not larger) seeds in a fruit than rewarding species. These findings highlight possible ways in which plants can increase their reproductive success in face of pollinator limitation. We emphasize that fruit-set in itself is an inappropriate measure of the reproductive success of orchids – the total number of seeds per shoot is a much better approximation.