An increasing abundance of Aphanizomenon ovalisporum in water bodies from diverse world regions has been reported in the last few years, with the majority of the isolated strains producing the toxin cylindrospermopsin (CYN), leading to a rise in ecological and health risks. The understanding of CYN synthesis is crucial in the control of CYN production. An amidinotransferase (AMDT) seems to be the first enzyme involved in the synthesis of CYN. In this study, we have cloned and overexpressed the aoaA gene from the constitutive CYN producer A. ovalisporum UAM‐MAO. The recombinant purified AoaA was characterized, confirming that it is an l‐arginine:glycine AMDT. It shows an optimal activity between 32 and 37°C, at pH from 8 to 9. The activity exhibits a mixed (ping‐pong/sequential) kinetic mechanism, and is inhibited by the reaction product guanidine acetate (GAA) in a noncompetitive manner. Mg²⁺ stimulates AoaA activity while Co²⁺ and Mn²⁺ inhibit it. AoaA conserves the critical residues of the catalytic site and substrate specificity of AMDTs, as the previously reported AMDT from Cylindrospermopsis raciborskii Cyr. Both proteins can be included in a new group of prokaryotic AMDTs involved in CYN production.

Research and regulatory efforts in toxicology are increasingly focused on the development of suitable non-animal methodologies for human health risk assessment. In this work we used human intestinal Caco-2 and HT29/MTX cell lines to address the potential risks of mixtures of the emerging contaminants tetrabromobisphenol A (TBBPA) and commercial polystyrene nanoparticles (PSNPs). We employed different in vitro settings to evaluate basal cytotoxicity through three complementary endpoints (metabolic activity, plasmatic, and lysosomal membrane integrity) and the induction of the oxidative stress and DNA damage responses with specific endpoints. Although no clear pattern was observed, our findings highlight the predominant impact of TBBPA in the combined exposures under subcytotoxic conditions and a differential behavior of the Caco-2 and HT29/MTX co-culture system. Distinctive outcomes detected with the mixture treatments include reactive oxygen species (ROS) increases, disturbances of mitochondrial inner membrane potential, generation of alkali-sensitive sites in DNA, as well as significant changes in the expression levels of relevant DNA and oxidative stress related genes.

Arginine (Arg) and glycine (Gly) seem to be the only substrates accepted by the amidinotransferase that catalyze the first step of the synthesis pathway of the cyanotoxin cylindrospermopsin (CYN), leading to guanidinoacetate (GAA). Here, the effect of these amino acids on the production of CYN in cultures of the cylindrospermopsin-producing strain, Aphanizomenon ovalisporum UAM-MAO, has been studied. Arg clearly increased CYN content, the increment appearing triphasic along the culture. On the contrary, Gly caused a decrease of CYN, observable from the first day on. Interestingly, the transcript of the gene ntcA, key in nitrogen metabolism control, was also enhanced in the presence of Arg and/or Gly, the trend of the transcript oscillations being like that of aoa/cyr. The inhibitory effect of Gly in CYN production seems not to result from diminishing the activity of genes considered involved in CYN synthesis, since Gly, as Arg, enhance the transcription of genes aoaA-C and cyrJ. On the other hand, culture growth is affected by Arg and Gly in a similar way to CYN production, with Arg stimulating and Gly impairing it. Taken together, our data show that the influence of both Arg and Gly on CYN changes seems not to be due to a specific effect on the first step of CYN synthesis; it rather appears to be the result of changes in the physiological cell status.

Ditylenchus dipsaci is a plant-parasitic nematode with a great economic impact on bulbous crops, including garlic (Allium sativum L.), and is distributed worldwide, particularly in the Mediterranean region. Traditionally, garlic was a rainfed crop in Spain, but irrigated areas have increased during the last few decades. However, the expected climatic conditions, with longer and more intense droughts, will make it necessary to reduce the water supply to garlic crops. This poses the urgent need to select garlic cultivars more tolerant to water scarcity and that are also more resistant to plant pathogenic organisms. The aim of this work was to analyze the influence of water stress on the host response of garlic plants to D. dipsaci. The specific objectives were to evaluate the level of nematode infestation in plants from four garlic genotypes treated with a reduced irrigation regime and compare them with those of control plants not subjected to water stress. The observed results were correlated with changes in the bulb and root development, as well as in the physiological parameters (total chlorophyll concentration and proline accumulation). The effects were different depending on whether the plants were subjected to water stress before or after nematode inoculation, as well as whether the water stress was continuous or discontinuous. Garlic inter-cultivar variability also affected the obtained results.

Ruta graveolens L. has been described as possessing antifungal and nematicidal activity. Among the bioactive compounds present in this plant, alkaloids and furanocoumarins have attracted considerable attention. The aim of this study was to evaluate the in vitro biological activity of extracts from rue enriched in bioactive compounds against Fusarium oxysporum, F. circinatum, and Meloidogyne incognita, and to correlate the chemical profile of the extracts with their biological activities. Six extracts with contrasting chemical profiles, obtained by pressurized liquid extraction and supercritical fluid extraction using green solvents, were selected for biological evaluation. The highest F. oxysporum growth inhibition was achieved with the extracts enriched in fatty acids and furanocoumarins at concentrations of 4, 8, and 16 mg/mL, while for F. circinatum, the highest growth inhibition was obtained using the extract enriched in terpenes at 16 mg/mL; moreover, the six extracts evaluated caused mortality in M. incognita. Therefore, enriched extracts of R. graveolens might be considered as an alternative for pathogen control on economically important crops such as potatoes, tomatoes, and onions, among others. Correlations between biological activities and chemical compositions suggest the importance of fatty acids against F. oxysporum, fatty acids and terpenes against F. circinatum, and alkaloids, coumarins, and furanocoumarins for M. incognita.