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Virology has gained much attention in recent years due to the COVID-19 pandemic and other recurrent epidemics/pandemics such as Ebola, zika, or now, monkeypox. We have analysed all recent biology schoolbooks for secondary school approved for the Austrian market. Our aim was to find out whether virological content was presented comprehensively, comprehensibly, and in an error-free manner. This also relates to visual representations of viruses, processes related to virology (e.g., replication), and references to daily life. Furthermore, we looked for tasks related to virology that may help students to deepen their newly acquired knowledge and/or to put it into practice, either by hands-on experimentation or transfer tasks. We examined 97 books (76 books for lower secondary and 21 books for upper secondary school). For this analysis, we developed and tested a coding matrix accompanied with a comprehensive coding guide to facilitate reproducible and reliable ratings. Since inter-rater reliability was found to be very high, both the coding matrix and guide can be recommended for further studies in this field. Overall, the virological content provided was free of errors, with the text and illustrations being mostly comprehensive and comprehensible. This was expected, since all Austrian school books must be approved by a governmental commission. However, individual books even for the same target (age) group differed widely in content and detail. In particular, few books clearly differentiated between viruses and bacteria, clarified that viruses are non-living and emphasized that antibiotics are ineffective against viruses. Yet precisely such knowledge is essential to enable students to make scientifically based decisions in health-related situations, especially for the prevention and treatment of viral diseases (e.g., whether to take antibiotics when suffering from a viral disease).

The current COVID-19 pandemic shows how little many people know about viruses Yet apart from COVID-19, the world has observed epidemic spread of another SARS virus, of the Ebola virus, and of the Zika virus during the last two decades The human immunodeficiency virus (HIV) is still one of the most dangerous viruses worldwide Some types of the human papillomavirus (HPV) are the main cause of cervical cancer Cases of measles, also caused by a virus, increase in numbers due to lack of access to or refusal of vaccination Furthermore, there is the widespread belief that viruses are similar to bacteria and may thus be fought off with antibiotics Yet viruses have no metabolism Thus, antibiotics cannot work against them, but may instead cause more harm than help, given side effects such as killing beneficial bacteria (e g , in the intestine) Second, misuse of antibiotics is one key factor in the evolution of antibiotic-resistant bacterial strains - a strong public health issue nowadays This article informs readers what viruses are, how they are distinct from bacteria, how they may have evolved, and how diseases they cause may be prevented Additionally, insights from studies concerning students' virus-related knowledge are summarized [ABSTRACT FROM AUTHOR] Copyright of American Biology Teacher (University of California Press) is the property of University of California Press and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission However, users may print, download, or email articles for individual use This abstract may be abridged No warranty is given about the accuracy of the copy Users should refer to the original published version of the material for the full abstract (Copyright applies to all Abstracts )

Education about virus biology at school is of pivotal interest to raise public awareness concerning means of disease transmission and, thus, methods to prevent infection, and to reduce unnecessary antibiotic treatment due to patient pressure on physicians in case of viral diseases such as influenza. This study aimed at making visible the knowledge of Austrian high school and university students with respect to virus biology, virus structure and health-education issues. The data presented here stem from comprehensive questionnaire analyses, including the task to draw a virus, from a cross-sectional study with 133 grade 7 and 199 grade 10 high school students, and 133 first-year biology and 181 first-year non-biology university students. Analyses were performed both quantitatively and qualitatively. ANOVA revealed a highly significant group effect for total knowledge relating to virus biology and health issues (F(3, 642) = 44.17, p < 0.01, η2p = 0.17). Specific post-hoc tests by means of the Tukey test showed significant differences between all groups (p < .01) with the exception of 1st year non-biology students and grade 10 high school students. Students enrolled in university-level biology outperformed all other groups, even though they had not yet encountered this topic at their courses; part of this phenomenon might be due to their affinity for learning about biological topics. However, even many first-year biology students had a high number of severe misconceptions, e.g., defining a virus as a pro- or eukaryotic cell, or falsely naming malaria as a viral disease. Since there was no significant difference in virus-related knowledge between high schools, virus biology seems to have been taught similarly among the tested schools. However, the majority of participants stated that the virus-related knowledge they had acquired at school was not sufficient. Based on the results presented here we urgently suggest improving and intensifying teaching this topic at school, since virus-related knowledge was by far too fragmentary among many participants. Such lack of health-relevant knowledge may contribute to pressure on physicians by patients to unnecessarily prescribe antibiotics, and possibly lead to potentially dangerous neglect concerning vaccination. The effectiveness of newly developed virus-related teaching units and material could be tested with the instrument used here.

Epilepsy is not a regular topic in many countries’ schools. Thus many people harbor misconceptions about people suffering from this disease. It was our aim to a) examine what grade ten students know and believe about epilepsy, and b) to develop and test a teaching unit to improve their knowledge and attitude. The test group comprised eight grade ten classes from six different Austrian high schools (54 girls and 51 boys aged 14–17), the control group (no intervention) five grade ten classes from the same schools (26 girls and 37 boys aged 14–17). The teaching unit consisted of three 45-min lessons using different methods and material. Changes in knowledge about and attitude towards epilepsy as a result of the intervention were psychometrically assessed in a pre-test intervention post-test design (along with a follow-up assessment two months after the intervention) by means of a questionnaire capturing different facets of epilepsy-related knowledge and attitude. Across all knowledge/attitude domains, students of the test group had a significantly improved knowledge about and a more positive attitude towards epilepsy and people suffering from it after the teaching unit. However, starting levels were different between the five knowledge/attitude domains tested. Medical background knowledge was lowest and consequently associated with the highest increase after the intervention. This study shows that epilepsy-related knowledge of many grade ten high school students is fragmentary and that some harbor beliefs and attitudes which require improvement. Our comprehensive but concise teaching unit significantly increased knowledge about epilepsy and positively influenced attitude towards individuals with epilepsy. Thus we recommend implementing this unit into regular school curricula.

The fungus Boeremia sambuci-nigrae (formerly Phoma sambuci-nigrae) causes the corymb wilt disease in elder (Sambucus nigra L.). Here, we assessed the genetic diversity of fungi within the Phoma-complex which associate with commercially cultivated elder in Styria (Austria). We sampled leaves, corymb stalks and berries of elder trees in one organically and in four conventionally managed orchards. In each orchard an area was left untreated allowing a sampling of fungicide treated and untreated trees. Nineteen taxa in the Phoma-complex were isolated and identified based on ITS and LSU nucDNA sequences. One isolate is closely related to Chaetosphaeronema, whereas the majority of the strains belong to the genera Epicoccum and Boeremia in Didymellaceae. Six isolates are monophyletic with Boeremia sambuci-nigrae. The results indicate a varying effectiveness of fungicide treatment for the different fungal groups investigated. The diverse distribution of isolates among the sampling sites suggests the influence of treatment effects and possibly also of climatic conditions.

Eukaryote genomes contain multiple copies of nuclear ribosomal DNA (nrDNA) harboring both highly conserved and variable regions. This has made nrDNA the most popular genetic marker for phylogenetic studies and the region of choice for barcoding projects. Furthermore, many scientists believe that all copies of nrDNA within one nucleus are practically identical due to concerted evolution. Here, we investigate the model plant species Arabidopsis thaliana for intragenomic variation of the internal transcribed spacer (ITS) region of nrDNA. Based on a modified deep sequencing approach, we provide a comprehensive list of ITS polymorphisms present in the two most widely used accessions of A. thaliana-Col-0 and Ler. Interestingly, we found that some polymorphisms are shared between these genetically very distinct accessions. On the other hand, the high number of accession-specific polymorphisms shows that each accession can be clearly and easily characterized by its specific ITS polymorphism patterns and haplotypes. Network analysis based on the detected haplotypes demonstrates that the study of ITS polymorphism patterns and haplotypes is an extremely powerful tool for population genetics. Using the methods proposed here, it will now be possible to extend the traditionally species-bound barcoding concept to populations.

Cytokinins are phytohormones that are involved in various regulatory processes throughout plant development, but they are also produced by pathogens and known to modulate plant immunity. A novel transgenic approach enabling autoregulated cytokinin synthesis in response to pathogen infection showed that cytokinins mediate enhanced resistance against the virulent hemibiotrophic pathogen Pseudomonas syringae pv tabaci. This was confirmed by two additional independent transgenic approaches to increase endogenous cytokinin production and by exogenous supply of adenine- and phenylurea-derived cytokinins. The cytokinin-mediated resistance strongly correlated with an increased level of bactericidal activities and up-regulated synthesis of the two major antimicrobial phytoalexins in tobacco (Nicotiana tabacum), scopoletin and capsidiol. The key role of these phytoalexins in the underlying mechanism was functionally proven by the finding that scopoletin and capsidiol substitute in planta for the cytokinin signal: phytoalexin pretreatment increased resistance against P. syringae. In contrast to a cytokinin defense mechanism in Arabidopsis (Arabidopsis thaliana) based on salicylic acid-dependent transcriptional control, the cytokinin-mediated resistance in tobacco is essentially independent from salicylic acid and differs in pathogen specificity. It is also independent of jasmonate levels, reactive oxygen species, and high sugar resistance. The novel function of cytokinins in the primary defense response of solanaceous plant species is rather mediated through a high phytoalexin-pathogen ratio in the early phase of infection, which efficiently restricts pathogen growth. The implications of this mechanism for the coevolution of host plants and cytokinin-producing pathogens and the practical application in agriculture are discussed.

The ascomycete Cymadothea trifolii, a member of the Dothideomycetes, is unique among obligate biotrophic fungi in its capability to only partially degrade the host cell wall and in forming an astonishingly intricate interaction apparatus (IA) in its own hyphae, while the attacked host plant cell is triggered to produce a membranous bubble opposite the IA. However, no sequence data are currently available for this species. Based on molecular phylogenetic results obtained from complete SSU and partial LSU data, we show that the genus Cymadothea belongs to the Mycosphaerellaceae (Capnodiales, Dothideomycetes). This is the first report of sequences obtained for an obligate biotrophic member of Mycosphaerellaceae.

Nuclear ribosomal genes in most eukaryotes are present in multiple copies and often used for taxonomic and phylogenetic analyses. We comprehensively examined intragenomic polymorphism levels of three nuclear ribosomal loci for four important plant pathogenic fungi by polymerase chain reaction amplification and cloning. Here, we show that single nucleotide polymorphisms are present in an unexpectedly high amount. This might have implications for studies of fungal evolution, phylogenetics, and population genetics. Furthermore, our work demonstrates that the majority of all ribosomal sequences obtained from one individual and gene is identical to the majority rule consensus sequence of all detected sequence variants. Due to the large number of polymorphisms found and the fact that the polymorphism level differed markedly even between ribosomal genes of one and the same individual, we assume that nuclear ribosomal genes might not always evolve in a strictly concerted manner.

The major autolysin of Staphylococcus aureus (AtlA) and of Staphylococcus epidermidis (AtlE) are well-studied enzymes. Here we created an atlA deletion mutant in S. aureus that formed large cell clusters and was biofilm-negative. In electron micrographs, the mutant cells were distinguished by rough outer cell surface. The mutant could be complemented using the atlE gene from S. epidermidis. To study the role of the repetitive sequences of atlE, we expressed in Escherichia coli the amidase domain encoded by the gene, carrying no repeat regions (amiE) or two repeat regions (amiE-R1,2), or the three repeat regions alone (R1,2,3) as N-terminal His-tag fusion proteins. Only slight differences in the cell wall lytic activity between AmiE and AmiE-R1,2 were observed. The repetitive sequences exhibit a good binding affinity to isolated peptidoglycan and might contribute to the targeting of the amidase to the substrate. AmiE and AmiE-R1,2 have a broad substrate specificity as shown by similar activities with peptidoglycan lacking wall teichoic acid, O-acetylation, or both. As the amidase activity of AtlA and AtlE has not been proved biochemically, we used purified AmiE-R1,2 to determine the exact peptidoglycan cleavage site. We provide the first evidence that the amidase indeed cleaves the amide bond between N-acetyl muramic acid and L-alanine.

Cellular interactions between the ascomycete Cymadothea trifolii and Trifolium repens (white clover) were analyzed using high-pressure freezing and freeze substitution. Cymadothea trifolii, a biotrophic leaf pathogen, forms a unique structure within its own hyphae, presumably for nutrient uptake from its host. This structure, called an interaction apparatus, consists of long, thin, often net-like cisternae surrounded by a membrane continuous with the fungal plasma membrane. The plant plasmalemma opposite the interaction apparatus invaginates to produce a host bubble. The interaction apparatus and host bubble are apoplastic and are linked by a tube with an electron dense sheath that may channel nutrients from the host to the pathogen. Within the tube, the cell walls of host and parasite appear altered. The interaction apparatus and host bubble may be analogous to haustoria in other obligately biotrophic fungi while the electron dense sheath of the tube may be equivalent to the haustorial neckband.

In order to study the mechanisms behind the infection process of the necrotrophic fungus Botrytis cinerea, the subcellular distribution of hydrogen peroxide (H2O2) was monitored over a time frame of 96 h post inoculation (hpi) in Arabidopsis thaliana Col-0 leaves at the inoculation site (IS) and the area around the IS which was defined as area adjacent to the inoculation site (AIS). H2O2 accumulation was correlated with changes in the compartment-specific distribution of ascorbate and glutathione and chloroplast fine structure. This study revealed that the severe breakdown of the antioxidative system, indicated by a drop in ascorbate and glutathione contents at the IS at later stages of infection correlated with an accumulation of H2O2 in chloroplasts, mitochondria, cell walls, nuclei and the cytosol which resulted in the development of chlorosis and cell death, eventually visible as tissue necrosis. A steady increase of glutathione contents in most cell compartments within infected tissues (up to 600% in chloroplasts at 96 hpi) correlated with an accumulation of H2O2 in chloroplasts, mitochondria and cell walls at the AIS indicating that high glutathione levels could not prevent the accumulation of reactive oxygen species (ROS) which resulted in chlorosis. Summing up, this study reveals the intracellular sequence of events during Botrytis cinerea infection and shows that the breakdown of the antioxidative system correlated with the accumulation of H2O2 in the host cells. This resulted in the degeneration of the leaf indicated by severe changes in the number and ultrastructure of chloroplasts (e.g. decrease of chloroplast number, decrease of starch and thylakoid contents, increase of plastoglobuli size), chlorosis and necrosis of the leaves.

There is increasing evidence that pathogens do not only elicit direct defense responses, but also cause pronounced changes in primary carbohydrate metabolism. Cell-wall-bound invertases belong to the key regulators of carbohydrate partitioning and source-sink relations. Whereas studies have focused so far only on the transcriptional induction of invertase genes in response to pathogen infection, the role of post-translational regulation of invertase activity has been neglected and was the focus of the present study. Expression analyses revealed that the high mRNA level of one out of three proteinaceous invertase inhibitors in source leaves of Arabidopsis thaliana is strongly repressed upon infection by a virulent strain of Pseudomonas syringae pv. tomato DC3000. This repression is paralleled by a decrease in invertase inhibitor activity. The physiological role of this regulatory mechanism is revealed by the finding that in situ invertase activity was detectable only upon infection by P. syringae. In contrast, a high invertase activity could be measured in vitro in crude and cell wall extracts prepared from both infected and non-infected leaves. The discrepancy between the in situ and in vitro invertase activity of control leaves and the high in situ invertase activity in infected leaves can be explained by the pathogen-dependent repression of invertase inhibitor expression and a concomitant reduction in invertase inhibitor activity. The functional importance of the release of invertase from post-translational inhibition for the defense response was substantiated by the application of the competitive chemical invertase inhibitor acarbose. Post-translational inhibition of extracellular invertase activity by infiltration of acarbose in leaves was shown to increase the susceptibility to P. syringae. The impact of invertase inhibition on spatial and temporal dynamics of the repression of photosynthesis and promotion of bacterial growth during pathogen infection supports a role for extracellular invertase in plant defense. The acarbose-mediated increase in susceptibility was also detectable in sid2 and cpr6 mutants and resulted in slightly elevated levels of salicylic acid, demonstrating that the effect is independent of the salicylic acid-regulated defense pathway. These findings provide an explanation for high extractable invertase activity found in source leaves that is kept inhibited in situ by post-translational interaction between invertase and the invertase inhibitor proteins. Upon pathogen infection, the invertase activity is released by repression of invertase inhibitor expression, thus linking the local induction of sink strength to the plant defense response.

International audience; Agrobacterium tumefaciens-mediated random insertional mutagenesis was used to investigate pathogenicity determinants in Leptosphaeria maculans. One tagged nonpathogenic mutant, termed m186, is analyzed in detail here. Microscopic analyses of infected plant tissues revealed that m186 is specifically blocked at the invasive growth phase after an unaffected initial penetration stage and is unable to switch to the necrotrophic lifestyle. In addition, m186 exhibits an altered cell wall and seems to be affected in its ability to produce cell-wall-degrading enzymes. The T-DNA insertion occurs in the intergenic region between two head-to-tail genes, leading to a constitutive upregulation of their expression. Complementation experiments showed that only one of these two genes, Lmepi, fully accounts for the mutant phenotype. Bioinformatics and expression analyses along with functional studies suggested that the Lmepi gene encodes for the highly conserved UDP-glucose-4-epimerase, a key enzyme of the Leloir pathway involved in galactose metabolism. For the third time, this study highlights the intimate connection between primary metabolism and pathogenicity in L. maculans. This finding, along with similar data obtained from the related species Stagonospora nodorum, indicates the importance of in planta nutrition for the success of infection of plants by fungi belonging to class Dothideomycete.

International audience; Random insertional mutagenesis was used to investigate pathogenicity determinants in Leptosphaeria maculans. One tagged nonpathogenic mutant, termed m20, was analysed in detail here. The mutant phenotype was investigated by microscopic analyses of infected plant tissues and in vitro growth assays. Complementation and silencing experiments were used to identify the altered gene. Its function was determined by bioinformatics analyses, cell biology experiments and functional studies. The mutant was blocked at the invasive growth phase after an unaffected initial penetration stage, and displayed a reduced growth rate and an aberrant hyphal morphology in vitro. The T-DNA insertion occurred in the intergenic region between two head-to-tail genes, leading to a complex deregulation of their expression. The unique gene accounting for the mutant phenotype was suggested to be the orthologue of the poorly conserved Saccharomyces cerevisiae gpi15, which encodes for one component of the glycosylphosphatidylinositol (GPI) anchor biosynthesis pathway. Consistent with this predicted function, a functional translational fusion with the green fluorescent protein (GFP) was targeted to the endoplasmic reticulum. Moreover, the mutant exhibited an altered cell wall and addition of glucosamine relieved growth defects. It is concluded that the GPI anchor biosynthetic pathway is required for morphogenesis, cell wall integrity and pathogenicity in Leptosphaeria maculans.

Summary • The intercellular ascomycetous pathogen Cymadothea trifolii, causing sooty blotch of clover, proliferates within leaves of Trifolium spp. and produces a complex structure called interaction apparatus (IA) in its own hyphae. Opposite the IA the plant plasmalemma invaginates to form a bubble. Both structures are connected by a tube with an electron-dense sheath. • Using immunocytochemistry on high-pressure frozen and freeze-substituted samples, we examined several plant and fungal cell wall components, including those in new host wall appositions at the interaction site, as well as a fungal polygalacturonase. • Within the tube linking IA and host bubble, labelling was obtained for cellulose and xyloglucan but not for rhamnogalacturonan-I and homogalacturonans. The IA labelled for chitin and β-1,3-glucans, and for a fungal polygalacturonase. Plant wall appositions reacted with antibodies against callose, xyloglucans and rhamnogalacturonan-I. • Cymadothea trifolii partly degrades the host cell wall. Structural elements remain intact, but the pectin matrix is dissolved. A fungal polygalacturonase detected in the IA is probably a key factor in this process. Owing to the presence of chitin and β-1,3-glucans, the IA itself is considered an apoplastic compartment.