RESUMO
Copper nanoparticles (Cu-NPs) have shown great antifungal activity against phytopathogenic fungi, making them a promising and affordable alternative to conventional fungicides. In this study, we evaluated the antifungal activity of Cu-NPs against Fusarium kuroshium, the causal agent of Fusarium dieback, and this might be the first study to do so. The Cu-NPs (at different concentrations) inhibited more than 80% of F. kuroshium growth and were even more efficient than a commercial fungicide used as a positive control (cupric hydroxide). Electron microscopy studies revealed dramatic damage caused by Cu-NPs, mainly in the hyphae surface and in the characteristic form of macroconidia. This damage was visible only 3 days post inoculation with used treatments. At a molecular level, the RNA-seq study suggested that this growth inhibition and colony morphology changes are a result of a reduced ergosterol biosynthesis caused by free cytosolic copper ions. Furthermore, transcriptional responses also revealed that the low- and high-affinity copper transporter modulation and the endosomal sorting complex required for transport (ESCRT) are only a few of the distinct detoxification mechanisms that, in its conjunction, F. kuroshium uses to counteract the toxicity caused by the reduced copper ion.
RESUMO
Fusarium kuroshium is a novel member of the Ambrosia Fusarium Clade (AFC) that has been recognized as one of the symbionts of the invasive Kuroshio shot hole borer, an Asian ambrosia beetle. This complex is considered the causal agent of Fusarium dieback, a disease that has severely threatened natural forests, landscape trees, and avocado orchards in the last 8 years. Despite the interest in this species, the molecular responses of both the host and F. kuroshium during the infection process and disease establishment remain unknown. In this work, we established an in vitro pathosystem using Hass avocado stems inoculated with F. kuroshium to investigate differential gene expression at 1, 4, 7 and 14 days post-inoculation. RNA-seq technology allowed us to obtain data from both the plant and the fungus, and the sequences obtained from both organisms were analyzed independently. The pathosystem established was able to mimic Fusarium dieback symptoms, such as carbohydrate exudation, necrosis, and vascular tissue discoloration. The results provide interesting evidence regarding the genes that may play roles in the avocado defense response to Fusarium dieback disease. The avocado data set comprised a coding sequence collection of 51,379 UniGenes, from which 2,403 (4.67%) were identified as differentially expressed. The global expression analysis showed that F. kuroshium responsive UniGenes can be clustered into six groups according to their expression profiles. The biologically relevant functional categories that were identified included photosynthesis as well as responses to stress, hormones, abscisic acid, and water deprivation. Additionally, processes such as oxidation-reduction, organization and biogenesis of the cell wall and polysaccharide metabolism were detected. Moreover, we identified orthologues of nucleotide-binding leucine-rich receptors, and their possible action mode was analyzed. In F. kuroshium, we identified 57 differentially expressed genes. Interestingly, the alcohol metabolic process biological category had the highest number of upregulated genes, and the enzyme group in this category may play an important role in the mechanisms of secondary metabolite detoxification. Hydrolytic enzymes, such as endoglucanases and a pectate lyase, were also identified, as well as some proteases. In conclusion, our research was conducted mainly to explain how the vascular tissue of a recognized host of the ambrosia complex responds during F. kuroshium infection since Fusarium dieback is an ambrosia beetle-vectored disease and many variables facilitate its establishment.
RESUMO
It is known that some nutrients can have both negative and positive effects on some populations of insects. To test this, the Logrank test and the Interval Overlap Test were evaluated for two crop cycles (February-May and May-August) of the 7705 tomato hybrid, and the effect on the psyllid, Bactericera cockerelli (Sulc.) (Hemiptera: Triozidae), was examined under greenhouse conditions. Tomato plants were in polythene bags and irrigated with the following solutions: T1-Steiner solution, T2-Steiner solution with nitrogen reduced to 25%, T3-Steiner solution with potassium reduced to 25%, and T4-Steiner solution with calcium reduced to 25%. In the Logrank test, a significant difference was found when comparing the survival parameters of B. cockerelli generated from the treatment cohorts: T1-T2; T1-T3; T1-T4; T2-T3; and T3-T4, while no significant differences were found in the T2-T4 comparison in the February-May cycle. In the May-August cycle, significant differences were found when comparing the survival parameters generated from the treatment cohorts: T1-T2; T1-T3; and T1-T4, while no significant differences were found in the T2-T3; T2-T4; and T3-T4 comparisons of survival parameters of B. cockerelli fed with the 7705 tomato hybrid. Also, the Interval Overlap Test was done on the treatment cohorts (T1, T2, T3, and T4) in the February-May and May-August cycles. T1 and T2 compare similarly in both cycles when feeding on the treatments up to 36 d. Similarly, in T1 and T3, the behavior of the insect is similar when feeding on the treatments up to 40 and 73 d, respectively. Comparisons T2-T3 and T2-T4 are similar when feeding on both treatments up to 42, 38 and 37, 63 d, respectively. Finally, the T3-T4 comparison was similar when feeding in both treatments up to 20 and 46 d, respectively.
