Aerial Laser Scanning Data as a Source of Terrain Modeling in a Fluvial Environment: Biasing Factors of Terrain Height Accuracy.

Airborne light detection and ranging (LiDAR) scanning is a commonly used technology for representing the topographic terrain. As LiDAR point clouds include all surface features present in the terrain, one of the key elements for generating a digital terrain model (DTM) is the separation of the ground points. In this study, we intended to reveal the efficiency of different denoising approaches and an easy-to-use ground point classification technique in a floodplain with fluvial forms. We analyzed a point cloud from the perspective of the efficiency of noise reduction, parametrizing a ground point classifier (cloth simulation filter, CSF), interpolation methods and resolutions. Noise filtering resulted a wide range of point numbers in the models, and the number of points had moderate correlation with the mean accuracies (r = -0.65, p < 0.05), indicating that greater numbers of points had larger errors. The smallest differences belonged to the neighborhood-based noise filtering and the larger cloth size (5) and the smaller threshold value (0.2). The most accurate model was generated with the natural neighbor interpolation with the cloth size of 5 and the threshold of 0.2. These results can serve as a guide for researchers using point clouds when considering the steps of data preparation, classification, or interpolation in a flat terrain.

Integrated Control of Apple Scab and Powdery Mildew in an Organic Apple Orchard by Combining Potassium Carbonates with Wettable Sulfur, Pruning, and Cultivar Susceptibility.

In a 4-year study in a whole-field sanitized organic apple orchard, the effectiveness of nine fungicide treatments, including potassium mono- and bicarbonate and their combinations with wettable sulfur, were evaluated for scab and powdery mildew control on two cultivars with different susceptibility to scab and powdery mildew, under two pruning treatments. The whole-field sanitation practice was performed by removal of infected fallen leaves. Treatment effects on phytotoxicity and yield were also determined. Pruning significantly reduced leaf scab incidence but only on the more scab-susceptible Idared. Pruning significantly reduced mildew incidence in most years and on both cultivars but the more mildew-susceptible Jonathan showed significantly higher mildew incidence than Idared. Among products approved for organic production, the best scab control was achieved with a potassium mono- or bicarbonate treatment combined with wettable sulfur, except for Jonathan in 2011 on leaf and in 2014 on both leaf and fruit, and for Idared in 2013 on fruit. The best mildew control was also achieved with potassium mono- or bicarbonate treatments combined with wettable sulfur, with exceptions on shoots of Idared in 2011 and 2013 and on fruit of Idared in 2012. Leaf phytotoxicity was significantly higher in all potassium carbonate treatments compared with untreated plots, except for Idared in 2012, while fruit russet in these treatments did not differ significantly from the untreated plots. However, phytotoxicity values of all carbonate treatments were significantly lower than the lime sulfur treatment in most years on both cultivars. Yield of the potassium mono- or bicarbonate treatments combined with wettable sulfur was significantly higher than the untreated plots in the pruned treatments for both cultivars in all years. The integrated control approach designed for organic disease management against the two pathogens is discussed.

Toward an Integrated Use of Biological Control by Cladosporium cladosporioides H39 in Apple Scab (Venturia inaequalis) Management.

Apple scab, caused by Venturia inaequalis, is the most important disease in apple production, reducing yield and quality of fruit. Control of apple scab in commercial orchards currently depends on multiple applications of fungicides. The potential of the antagonistic isolate Cladosporium cladosporioides H39, originating from a sporulating colony of V. inaequalis, to control apple scab development was tested in eight trials during 2 years in orchards in Eperjeske (Hungary), Dabrowice (Poland), and Bavendorf (Germany) planted with different cultivars. Treatments were conducted as calendar sprays or after infection periods. Additional trials in an orchard in Randwijk (The Netherlands) focused on the effect of timing of antagonist application before or after infection periods. The overall results of the field trials consistently showed-for the first time-that stand-alone applications of the antagonist C. cladosporioides H39 can reduce apple scab in leaves and fruit. This was demonstrated in an organic growing system as well as in conventional orchards by spray schedules applied during the primary or the summer season. In both systems, the same control levels could be reached as with common fungicide schedules. Efficacies reached 42 to 98% on leaf scab incidence and 41 to 94% on fruit scab. The antagonist was also effective if applied one or even several days (equivalent to approximately 300 to 2,000 degree h) after infection events in several field trials and a trial conducted in Randwijk with single-spray applications at different intervals before or after infection events. Better understanding of the biology of the antagonist will help to further exploit its use in apple scab control.

Optimal conditions for mycelial growth of Schizosaccharomyces japonicus cells in liquid medium: it enables the molecular investigation of dimorphism.

The non-pathogenic dimorphic fission yeast, Schizosaccharomyces japonicus, could be a suitable model organism for investigation of the genetic background of mycelial growth, as it has a haploid chromosome set and its genome is sequenced. Since earlier results have suggested that its morphological transition required solid substrates, but molecular biological experiments would require hyphae production in a liquid medium, we wanted to find circumstances which would enable hyphae production in liquid media. Several external conditions were investigated, but the strongest inducer was fetal bovine serum (FBS). Its positive effect could be hampered by heat and was dependent on pH, temperature and concentration of the serum. Other protein-containing compounds, such as peptone and bovine serum albumin or amino acids, proved to be ineffective or weak. Generally, the uninduced and induced mycelial growth of Sz. japonicus could be improved by lower external pH and higher temperature.

Genetic diversity in Monilinia laxa populations in stone fruit species in Hungary.

The objectives of this study were firstly, to determine the genetic diversity of Monilinia laxa isolates from Hungary, using the PCR-based inter-simple sequence repeat (ISSR) and randomly amplified polymorphic DNA (RAPD) technique; secondly, to prepare genetic diversity groups based on the dendrograms; and finally, to select some relevant isolates to study their fungicide sensitivity. 55 and 77 random amplified polymorphic ISSR and RAPD markers, of which 23 and 18 were polymorphic and 32 and 59 monomorphic, respectively, were used to assess the genetic diversity and to study the structure of M. laxa populations in Hungary. 27 isolates out of 57 ones were confirmed as M. laxa from several orchards (subpopulations) in three geographical regions, in various inoculum sources and in various hosts, were used. 10 fungicides and 12 isolates selected from genetic diversity groups based on the ISSR dendrograms were used to determine the fungicide sensitivity of the selected isolates. The analysis of population structure revealed that genetic diversity within locations, inoculum sources and host (H(S)) accounted for 99 % of the total genetic diversity (H(T)), while genetic diversity among locations, inoculum sources and host represented only 1 %. The relative magnitude of gene differentiation between subpopulations (G(ST)) and the estimate of the number of migrants per generation (Nm) averaged 0.005-0.009 and 53.9-99.2, respectively, for both ISSR and RAPD data set. The results obtained in dendrograms were in accordance with the gene diversity analysis. Grouping of isolates in the dendrograms was irrespective of whether they came from the same or different geographical locations. There was no relationship between clustering among isolates from inoculum sources and hosts. In the fungicide sensitivity tests, five isolates out of 12 were partly insensitive to boscalid+piraclostrobin, cyprodinil, fenhexamid or prochloraz. Obtained results in genetic diversity of M. laxa populations are discussed together with implications for the management of brown rot.

Impacts of N-P-K-Mg Fertilizer Combinations on Tree Parameters and Fungal Disease Incidences in Apple Cultivars with Varying Disease Susceptibility.

Adequate mineral fertilization helps to ensure optimal tree growth, fruit development, and predictable yield of apple trees. This 7-year study (2016-2022) aims to investigate the effect of nitrogen (N), phosphorus (P), potassium (K), and magnesium (Mg) fertilizer combinations (NP, NPK, NPKMg, and control) on eight parameters (trunk cross-sectional area-TCSA; fruit yield-FY; number of fruit per tree-FNT; crop load-CL; fruit diameter-FD; fruit weight-FW; fruit scab incidence-FSI; and powdery mildew incidence on shoot-PMIS) on the cultivars (cvs) 'Golden Reinders' (disease susceptible) and 'Pinova' (scab and mildew tolerant). In the 7-year period, TCSA values continuously increased for both cultivars over the years. Fertilizer treatments showed significant differences on TCSA but the effect varied greatly annually among fertilizer treatments. Fertilizer treatments had increasing effects on FY and FNT in 2018 and 2022, on CL in 2018, on FD in 2018 and 2019, and on FW in 2016 and 2018 in both cultivars compared to the control treatment. FSI values were the lowest in the NPKMg treatment for cv. 'Golden Reinders' in 2016, 2017, and 2022; for cv. 'Pinova' in 2016; PMIS values for cv. 'Golden Reinders' in 2017, 2018, 2021, and 2022; and for cv. 'Pinova' in 2018. Correlation and regression analyses revealed strong and significant (p = 0.05) relationships between FNT versus (vs.) TCSA, FNT vs. FY, FW vs. TCSA, CL vs. FY, FW vs. FD, and FSI vs. FW. In conclusion, our study showed that multiyear application of fertilizer combinations can successfully increase TCSA and yield parameters as well as reduce fungal disease incidences, especially on the disease-susceptible cultivar in sandy soil with moderate fertility, under Central-European continental climate conditions.

Effect of Sanitation Treatments on Leaf Litter Density and Leaf Spot Incidence in Integrated and Organic Sour Cherry Orchards.

A 3-year study was conducted to determine the effect of five sanitation treatments on leaf litter density (LLD), leaf spot incidence, and percent defoliation on two cultivars ('Újfehértói fürtös' and 'Érdi botermo') in two sour cherry orchards: one managed by integrated pest management principles with conventional fungicides and the other managed organically. The following sanitation treatments were compared: sprays of urea or lime sulfur in autumn, removing fallen leaves after leaf fall, straw mulch cover in late winter, sprays of urea or lime sulfur followed by mulch cover, removing fallen leaves followed by mulch cover, and a nonsanitized control. In both orchards and all years, LLD decreased by 2 to 28% in all treatment plots from early December to mid-May. LLD reduction was two to four times higher in the organic orchard compared with the integrated orchard. All treatments, except the lime sulfur or urea treatment alone, resulted in significant (P < 0.05) reduction of LLD in both the integrated and organic orchards compared with nonsanitized plots. Only leaf removal alone or in combination with mulch significantly (P < 0.05) reduced cherry leaf spot incidence and percent defoliation (by 11 to 70% and 15 to 72%, respectively) compared with nonsanitized plots. The application of these sanitation treatments in orchard management practices is discussed.

Comparison and Intercorrelation of Various Bentonite Products for Oenological Properties, Elemental Compositions, Volatile Compounds and Organoleptic Attributes of White Wine.

Bentonite fining is one of the generally applied wine-making technological elements that may seriously affect wine components. The aim of this study was (i) to investigate the effect of 21 bentonite products on eight oenological parameters, 19 elements, 21 volatile organic compounds (VOCs) and 10 organoleptic properties of white wine; and (ii) to quantify intercorrelations among the parameters separately for each of the four quality attributes. Among oenological parameters, sugar, acidity, malic-, lactic-, citric acid and total phenol contents were significant among several bentonite products. The amounts of elements were the lowest in the control wine treatments (with exceptions of, e.g., Ni and Cu); and these values were significantly different from several bentonite products. The relative presence of the VOCs was above 100% for most VOCs, but it was below 100% for 1-propanol, 4-amino-1,5-pentandioic acid and butane-dioic acid, and diethyl ester in all treatments. For organoleptic parameters, the values of clearness, colour, flavour intensity and taste persistency was the lowest in the control wine treatment, while the values of flavour character, flavour quality, taste intensity, taste character, and overall harmony were the highest for the bentonite products of AP, EBE, M-SA, EBE, EBE, respectively. Results of correlation and factor analyses showed strong intercorrelative effects of bentonite fining on the four quality attributes. In conclusion, this study can help in the proper choice of a specific bentonite product in relation to complexity effects of bentonite fining.

Temporal Dynamics of Incidence of Shot Hole Disease Affected by Training Systems and Cultivar Susceptibilities in an Integrated Plum Orchard.

Shot hole disease (SHD) can cause severe epidemics in plum orchards, depending on cultivar susceptibility and training system; however, the combined effect on the progress of temporal disease and on the possible reduction in SHD in the disease management was not investigated. The aim of this 3-year study was (i) to monitor and analyze the temporal dynamics of SHD progress under four training systems (4 × 1.5, 4 × 2, 5 × 2.5 and 6 × 3 m) and on four plum cultivars ('Cacanska lepotica', 'Bluefre', 'Stanley' and 'President') in an integrated plum orchard; (ii) to identify those time periods when training system and cultivar combinations can reduce the disease development. Both SHD incidences and the area under the disease progress curves (AUDPC) were significantly affected by the training system, cultivar and year. Plum cultivars with high or mid-high susceptibility to SHD showed continuous SHD development from May to November, while cultivars with low susceptibility to SHD showed no symptoms until mid-summer and then progressed slowly until November. High (4 × 1.5 m) vs. low (6 × 3 m) density training systems reduced SHD incidence and AUDPC consistently for three cultivars ('Cacanska lepotica', 'Stanley' and 'President') in September, October and November, compared to the high-density training system. Only cv. 'Bluefre' showed no effect either on disease incidence or AUDPC, due to very high disease incidences in all training systems from September to November. In conclusions, combinations of training system and cultivar can significantly reduce SHD incidence, which may be successfully used as a part of the integrated pest management approach during the establishment new plantations.

Tree species composition mapping with dimension reduction and post-classification using very high-resolution hyperspectral imaging.

Tree species' composition of forests is essential in forest management and nature conservation. We aimed to identify the tree species structure of a floodplain forest area using a hyperspectral image. We proposed an efficient novel strategy including the testing of three dimension reduction (DR) methods: Principal Component Analysis, Minimum Noise Fraction (MNF) and Indipendent Component Analysis with five machine learning (ML) algorithms (Maximum Likelihood Classifier, Support Vector Classification, Support Vector Machine, Random Forest and Artificial Neural Network) to find the most accurate outcome; altogether 300 models were calculated. Post-classification was applied by combining the multiresolution segmentation and filtering. MNF was the most efficient DR technique, and at least 7 components were needed to gain an overall accuracy (OA) of > 75%. Forty-five models had > 80% OAs; MNF was 43, and the Maximum Likelihood was 19 times among these models. Best classification belonged to MNF with 10 components and Maximum Likelihood classifier with the OA of 83.3%. Post-classification increased the OA to 86.1%. We quantified the differences among the possible DR and ML methods, and found that even > 10% worse model can be found using popular standard procedures related to the best results. Our workflow calls the attention of careful model selection to gain accurate maps.

NODULIN HOMEOBOX is required for heterochromatin homeostasis in Arabidopsis.

Arabidopsis NODULIN HOMEOBOX (NDX) is a nuclear protein described as a regulator of specific euchromatic genes within transcriptionally active chromosome arms. Here we show that NDX is primarily a heterochromatin regulator that functions in pericentromeric regions to control siRNA production and non-CG methylation. Most NDX binding sites coincide with pericentromeric het-siRNA loci that mediate transposon silencing, and are antagonistic with R-loop structures that are prevalent in euchromatic chromosomal arms. Inactivation of NDX leads to differential siRNA accumulation and DNA methylation, of which CHH/CHG hypomethylation colocalizes with NDX binding sites. Hi-C analysis shows significant chromatin structural changes in the ndx mutant, with decreased intrachromosomal interactions at pericentromeres where NDX is enriched in wild-type plants, and increased interchromosomal contacts between KNOT-forming regions, similar to those observed in DNA methylation mutants. We conclude that NDX is a key regulator of heterochromatin that is functionally coupled to het-siRNA loci and non-CG DNA methylation pathways.

Characterization of three-dimensional spatial aggregation and association patterns of brown rot symptoms within intensively mapped sour cherry trees.

BACKGROUND AND AIMS: Characterization of spatial patterns of plant disease can provide insights into important epidemiological processes such as sources of inoculum, mechanisms of dissemination, and reproductive strategies of the pathogen population. Whilst two-dimensional patterns of disease (among plants within fields) have been studied extensively, there is limited information on three-dimensional patterns within individual plant canopies. Reported here are the detailed mapping of different symptom types of brown rot (caused by Monilinia laxa) in individual sour cherry tree (Prunus cerasus) canopies, and the application of spatial statistics to the resulting data points to determine patterns of symptom aggregation and association. METHODS: A magnetic digitizer was utilized to create detailed three-dimensional maps of three symptom types (blossom blight, shoot blight and twig canker) in eight sour cherry tree canopies during the green fruit stage of development. The resulting point patterns were analysed for aggregation (within a given symptom type) and pairwise association (between symptom types) using a three-dimensional extension of nearest-neighbour analysis. KEY RESULTS: Symptoms of M. laxa infection were generally aggregated within the canopy volume, but there was no consistent pattern for one symptom type to be more or less aggregated than the other. Analysis of spatial association among symptom types indicated that previous year's twig cankers may play an important role in influencing the spatial pattern of current year's symptoms. This observation provides quantitative support for the epidemiological role of twig cankers as sources of primary inoculum within the tree. CONCLUSIONS: Presented here is a new approach to quantify spatial patterns of plant disease in complex fruit tree canopies using point pattern analysis. This work provides a framework for quantitative analysis of three-dimensional spatial patterns within the finite tree canopy, applicable to many fields of research.

Temporal Patterns and Inter-Correlations among Physical and Antioxidant Attributes and Enzyme Activities of Apricot Fruit Inoculated with Monilinia laxa under Salicylic Acid and Methyl Jasmonate Treatments under Shelf-Life Conditions.

Monilinia laxa causes serious postharvest damage on apricot fruits under shelf-life storage conditions. Plant elicitors of methyl jasmonate (MeJA) and salicylic acid (SA) can reduce this damage, and their research can explain the background of the plant defense physiological processes in M. laxa-infected fruits. The aims of this study were: (i) to evaluate the effect of various concentrations of MeJA and SA on brown rot incidence (BRI) and lesion diameter (LD) of apricot fruits; (ii) to measure the temporal patterns for the effect of 0.4 mmol L-1 MeJA and 2 mmol L-1 SA treatments on BRI, LD and seven fruit measures (fruit firmness (FF), lignin content (LC), total soluble phenol content (TSPC), total antioxidant capacity (TAC) and enzyme activities of PAL, POD and SOD) in treatments of M. laxa-inoculated versus (vs.) non-inoculated fruits over an eight-day shelf-life storage period; and (iii) to determine inter-correlations among the seven fruit measures for MeJA and SA treatments. Both MeJA and SA significantly reduced BRI and LD. LC, FF, TAC, TSPC, as well as SOD and PAL activities in the MeJA and SA treatments were higher than the water-treated control in most assessment days and both inoculation treatments. In both inoculation treatments, the activity of POD in the SA-treated fruits was higher than MeJA-treated and control fruits at all dates. In MeJA vs. SA and inoculated vs. non-inoculated treatments, six variable pairs (FF vs. TSPC, FF vs. TAC, TAC vs. PAL, PAL vs. POD, PAL vs. SOD, and POD vs. SOD) showed significant inter-correlation values. Principal component analyses explained 96% and 93% of the total variance for inoculated and non-inoculated treatments, respectively. In inoculated treatments, both PC1 and PC2 explained 41% of the total variance and correlated with FF, TSPC and TAC and with PAL, SOD and POD, respectively. In non-inoculated treatments, PC1 and PC2 explained 49% and 44% of the total variance and correlated with LC, PAL, POD and SOD and with FF, TSPC and TAC, respectively. It can be concluded that MeJA and SA are useful in the practice to enhance the plant defense system against brown rot by reducing fungal growth and by improving physical and antioxidant attributes (FF, LC, TAC and TSPC) and the activity of defense-related enzymes (PAL, POD and SOD) in apricot fruits during shelf-life storage conditions.

A new selective medium for the recovery and enumeration of Monilinia fructicola, M. fructigena, and M. laxa from stone fruits.

Isolation of Monilinia spp. from stone and pome fruit surfaces is difficult due to the presence of several fast-growing fungal species such as Rhizopus, Alternaria, and Penicillium spp. Therefore, a new selective medium (acidified potato dextrose agar [pH 3.6] amended with fosetyl-aluminum [fosetyl-AL] at 500 microg/ml) (APDA-F500) was developed for the recovery of Monilinia propagules. The antifungal agents fosetyl-Al, dichloran, ammonium molybdate, and 2-deoxy-D-glucose (2-dD-glucose) were tested in potato dextrose agar (PDA) for their selective activity against Monilinia fructicola and seven common fungal contaminants of peach, including Alternaria alternata, Aspergillus niger, Colletotrichum acutatum, Gilbertella persicaria, Penicillium expansum, Phomopsis amygdali, and Rhizopus stolonifer. Dichloran, ammonium molybdate, and 2-dD-glucose inhibited spore germination and mycelial growth of all test fungi, including M. fructicola, at comparable levels. Fosetyl-Al added to PDA (PDA-F) at 500 or 1,000 microg/ml did not inhibit germination of any of the fungi but had a strong effect on mycelial growth of six of eight test fungi at 1,000 microg/ml, with the exceptions being R. stolonifer and M. fructicola. Germination and mycelial growth of M. fructicola were least affected on APDA-F500 compared with the other test fungi. On APDA-F500 at pH 3.2 and 3.6, germination of M. fructicola was not inhibited but mycelial growth was reduced by 54.2 and 24.2%, respectively. In all, 17 M. fructicola, 6 M. fructigena, and 6 M. laxa isolates collected from different geographic locations and diverse hosts were evaluated for their germination and mycelial growth on APDA-F500 (at pH 3.6). Germination was not inhibited for any isolate and relative mycelial growth was 45.8 to 83.3%. Field-grown peach fruit from South Carolina and Hungary and plum fruit from Hungary were used to test the selectivity of APDA-F500 for the recovery of three Monilinia spp. compared with PDA-F500 and Monilinia selective medium (MSM) previously developed for Monilinia spp. detection. Percent recovery of M. fructicola from South Carolinian peach fruit was highest on APDA-F500 (0, 17, and 69% in June, July, and August, respectively) compared with PDA-F500 (0, 3.5, and 50%, respectively) and MSM (0, 0, and 6.8%, respectively). Moreover, APDA-F500 selectively recovered M. fructigena and M. laxa propagules from the surfaces of Hungarian peach and plum fruit. Our results indicate that APDA-F500 is a useful medium for selective isolation and enumeration of the three most common Monilinia spp. attacking stone fruits worldwide.

The benefits of combining elemental sulfur with a DMI fungicide to control Monilinia fructicola isolates resistant to propiconazole.

BACKGROUND: Management of demethylation inhibitor (DMI) fungicide resistance in Monilinia fructicola (G. Winter) Honey is a priority in peach orchards of the southeastern United States, but DMI fungicides are still an important component of antiresistance strategies in view of the few effective alternatives. The goal of this study was to investigate potential benefits of a sulfur/propiconazole mixture for the control of propiconazole-resistant isolates. RESULTS: The mixture provided the best control for propiconazole-resistant isolates, regardless of protective or curative application timings, or the presence or absence of fruit injury. Propiconazole-resistant isolates developed disease on detached fruit after protective or curative applications of propiconazole or its mixture with sulfur, but protective applications of the mixture significantly reduced (P = 0.05) disease symptoms compared with the individual compounds. Additive to slightly synergistic effects were observed for the mixture in protective treatments of peaches inoculated with propiconazole-resistant isolates. CONCLUSION: The results suggest that the addition of elemental sulfur to a DMI fungicide is likely to be a relatively inexpensive means to improve brown rot control in peach production areas where reduced sensitivity to DMI fungicides is suspected but has not led to noticeable control failure.

Interactions of retinoids with the ABC transporters P-glycoprotein and Breast Cancer Resistance Protein.

Retinoids - derivatives of vitamin A - are important cell permeant signaling molecules that regulate gene expression through activation of nuclear receptors. P-glycoprotein (Pgp) and ABCG2 are plasma membrane efflux transporters affecting the tissue distribution of numerous structurally unrelated lipophilic compounds. In the present work we aimed to study the interaction of the above ABC transporters with retinoid derivatives. We have found that 13-cis-retinoic acid, retinol and retinyl-acetate inhibited the Pgp and ABCG2 mediated substrate transport as well as the substrate stimulated ATPase activity of these transporters. Interestingly, 9-cis-retinoic acid and ATRA (all-trans retinoic acid), both are stereoisomers of 13-cis-retinoic acid, did not have any effect on the transporters' activity. Our fluorescence anisotropy measurements revealed that 13-cis-retinoic acid, retinol and retinyl-acetate selectively increase the viscosity and packing density of the membrane. Thus, the mixed-type inhibition of both transporters by retinol and ABCG2 by 13-cis-retinoic acid may be the collective result of direct interactions of these retinoids with the substrate binding site(s) and of indirect interactions mediated by their membrane rigidifying effects.

A single active catalytic site is sufficient to promote transport in P-glycoprotein.

P-glycoprotein (Pgp) is an ABC transporter responsible for the ATP-dependent efflux of chemotherapeutic compounds from multidrug resistant cancer cells. Better understanding of the molecular mechanism of Pgp-mediated transport could promote rational drug design to circumvent multidrug resistance. By measuring drug binding affinity and reactivity to a conformation-sensitive antibody we show here that nucleotide binding drives Pgp from a high to a low substrate-affinity state and this switch coincides with the flip from the inward- to the outward-facing conformation. Furthermore, the outward-facing conformation survives ATP hydrolysis: the post-hydrolytic complex is stabilized by vanadate, and the slow recovery from this state requires two functional catalytic sites. The catalytically inactive double Walker A mutant is stabilized in a high substrate affinity inward-open conformation, but mutants with one intact catalytic center preserve their ability to hydrolyze ATP and to promote drug transport, suggesting that the two catalytic sites are randomly recruited for ATP hydrolysis.

Secondary metabolites in fungus-plant interactions.

Fungi and plants are rich sources of thousands of secondary metabolites. The genetically coded possibilities for secondary metabolite production, the stimuli of the production, and the special phytotoxins basically determine the microscopic fungi-host plant interactions and the pathogenic lifestyle of fungi. The review introduces plant secondary metabolites usually with antifungal effect as well as the importance of signaling molecules in induced systemic resistance and systemic acquired resistance processes. The review also concerns the mimicking of plant effector molecules like auxins, gibberellins and abscisic acid by fungal secondary metabolites that modulate plant growth or even can subvert the plant defense responses such as programmed cell death to gain nutrients for fungal growth and colonization. It also looks through the special secondary metabolite production and host selective toxins of some significant fungal pathogens and the plant response in form of phytoalexin production. New results coming from genome and transcriptional analyses in context of selected fungal pathogens and their hosts are also discussed.

FSRD: fungal stress response database.

Adaptation to different types of environmental stress is a common part of life for today's fungi. A deeper understanding of the organization, regulation and evolution of fungal stress response systems may lead to the development of novel antifungal drugs and technologies or the engineering of industrial strains with elevated stress tolerance. Here we present the Fungal Stress Response Database (http://internal.med.unideb.hu/fsrd) aimed to stimulate further research on stress biology of fungi. The database incorporates 1985 fungal stress response proteins with verified physiological function(s) and their orthologs identified and annotated in 28 species including human and plant pathogens, as well as important industrial fungi. The database will be extended continuously to cover other fully sequenced fungal species. Our database, as a starting point for future stress research, facilitates the analysis of literature data on stress and the identification of ortholog groups of stress response proteins in newly sequenced fungal genomes. Database URL: http://internal.med.unideb.hu/fsrd

Metschnikowia Species Share a Pool of Diverse rRNA Genes Differing in Regions That Determine Hairpin-Loop Structures and Evolve by Reticulation.

Modern taxonomy of yeasts is mainly based on phylogenetic analysis of conserved DNA and protein sequences. By far the most frequently used sequences are those of the repeats of the chromosomal rDNA array. It is generally accepted that the rDNA repeats of a genome have identical sequences due to the phenomenon of sequence homogenisation and can thus be used for identification and barcoding of species. Here we show that the rDNA arrays of the type strains of Metschnikowia andauensis and M. fructicola are not homogenised. Both have arrays consisting of diverse repeats that differ from each other in the D1/D2 domains by up to 18 and 25 substitutions. The variable sites are concentrated in two regions that correspond to back-folding stretches of hairpin loops in the predicted secondary structure of the RNA molecules. The substitutions do not alter significantly the overall hairpin-loop structure due to wobble base pairing at sites of C-T transitions and compensatory mutations in the complementary strand of the hairpin stem. The phylogenetic and network analyses of the cloned sequences revealed that the repeats had not evolved in a vertical tree-like way but reticulation might have shaped the rDNA arrays of both strains. The neighbour-net analysis of all cloned sequences of the type strains and the database sequences of different strains further showed that these species share a continuous pool of diverse repeats that appear to evolve by reticulate evolution.