Proteome Analysis of Walnut Bacterial Blight Disease H D Sagawa, Cíntia; de A B Assis, Renata; Zaini, Paulo A ...
International journal of molecular sciences,
10/2020, Letnik:
21, Številka:
20
Journal Article
Recenzirano
Odprti dostop
The interaction between the plant host, walnut (
; Jr), and a deadly pathogen (
pv.
417; Xaj) can lead to walnut bacterial blight (WB), which depletes walnut productivity by degrading the nut ...quality. Here, we dissect this pathosystem using tandem mass tag quantitative proteomics. Walnut hull tissues inoculated with Xaj were compared to mock-inoculated tissues, and 3972 proteins were identified, of which 3296 are from Jr and 676 from Xaj. Proteins with differential abundance include oxidoreductases, proteases, and enzymes involved in energy metabolism and amino acid interconversion pathways. Defense responses and plant hormone biosynthesis were also increased. Xaj proteins detected in infected tissues demonstrate its ability to adapt to the host microenvironment, limiting iron availability, coping with copper toxicity, and maintaining energy and intermediary metabolism. Secreted proteases and extracellular secretion apparatus such as type IV pilus for twitching motility and type III secretion effectors indicate putative factors recognized by the host. Taken together, these results suggest intense degradation processes, oxidative stress, and general arrest of the biosynthetic metabolism in infected nuts. Our results provide insights into molecular mechanisms and highlight potential molecular tools for early detection and disease control strategies.
Plant secretome studies highlight the importance of vascular plant defense proteins against pathogens. Studies on Pierce's disease of grapevines caused by the xylem-limited bacterium
(
) have ...detected proteins and pathways associated with its pathobiology. Despite the biological importance of the secreted proteins in the extracellular space to plant survival and development, proteome studies are scarce due to methodological challenges. Prosit, a deep learning neural network prediction method is a powerful tool for improving proteome profiling by data-independent acquisition (DIA). We explored the potential of Prosit's in silico spectral library predictions to improve DIA proteomic analysis of vascular leaf sap from grapevines with Pierce's disease. The combination of DIA and Prosit-predicted libraries increased the total number of identified grapevine proteins from 145 to 360 and
proteins from 18 to 90 compared to gas-phase fractionation (GPF) libraries. The new proteins increased the range of molecular weights, assisted in the identification of more exclusive peptides per protein, and increased identification of low-abundance proteins. These improvements allowed identification of new functional pathways associated with cellular responses to oxidative stress, to be investigated further.
The Xanthomonadaceae family consists of species of non-pathogenic and pathogenic γ-proteobacteria that infect different hosts, including humans and plants. In this study, we performed a comparative ...analysis using 69 fully sequenced genomes belonging to this family, with a focus on identifying proteins enriched in phytopathogens that could explain the lifestyle and the ability to infect plants. Using a computational approach, we identified seven phytopathogen-enriched protein families putatively secreted by type II secretory system: PheA (CM-sec), LipA/LesA, VirK, and four families involved in N-glycan degradation, NixE, NixF, NixL, and FucA1. In silico and phylogenetic analyses of these protein families revealed they all have orthologs in other phytopathogenic or symbiotic bacteria, and are involved in the modulation and evasion of the immune system. As a proof of concept, we performed a biochemical characterization of LipA from Xac306 and verified that the mutant strain lost most of its lipase and esterase activities and displayed reduced virulence in citrus. Since this study includes closely related organisms with distinct lifestyles and highlights proteins directly related to adaptation inside plant tissues, novel approaches might use these proteins as biotechnological targets for disease control, and contribute to our understanding of the coevolution of plant-associated bacteria.
Following photosynthesis, sucrose is translocated to sink organs, where it provides the primary source of carbon and energy to sustain plant growth and development. Sugar transporters from the SWEET ...(sugar will eventually be exported transporter) family are rate-limiting factors that mediate sucrose transport across concentration gradients, sustain yields, and participate in reproductive development, plant senescence, stress responses, as well as support plant-pathogen interaction, the focus of this study. We identified 25
genes in the walnut genome and distinguished each by its individual gene structure and pattern of expression in different walnut tissues. Their chromosomal locations,
-acting motifs within their 5' regulatory elements, and phylogenetic relationship patterns provided the first comprehensive analysis of the
gene family of sugar transporters in walnut. This family is divided into four clades, the analysis of which suggests duplication and expansion of the SWEET gene family in
. In addition, tissue-specific gene expression signatures suggest diverse possible functions for
genes. Although these are commonly used by pathogens to harness sugar products from their plant hosts, little was known about their role during
pv.
(
) infection. We monitored the expression profiles of the
genes in different tissues of "Chandler" walnuts when challenged with pathogen
417 and concluded that SWEET-mediated sugar translocation from the host is not a trigger for walnut blight disease development. This may be directly related to the absence of type III secretion system-dependent transcription activator-like effectors (TALEs) in
417, which suggests different strategies are employed by this pathogen to promote susceptibility to this major aboveground disease of walnuts.
Walnut blight is a significant above-ground disease of walnuts caused by
pv.
(Xaj). The secreted form of chorismate mutase (CM), a key enzyme of the shikimate pathway regulating plant immunity, is ...highly conserved between plant-associated beta and gamma proteobacteria including phytopathogens belonging to the Xanthomonadaceae family. To define its role in walnut blight disease, a dysfunctional mutant of chorismate mutase was created in a copper resistant strain Xaj417 (XajCM). Infections of immature walnut
(Jr) fruit with XajCM were hypervirulent compared with infections with the wildtype Xaj417 strain. The in vitro growth rate, size and cellular morphology were similar between the wild-type and XajCM mutant strains, however the quantification of bacterial cells by dPCR within walnut hull tissues showed a 27% increase in XajCM seven days post-infection. To define the mechanism of hypervirulence, proteome analysis was conducted to compare walnut hull tissues inoculated with the wild type to those inoculated with the XajCM mutant strain. Proteome analysis revealed 3296 Jr proteins (five decreased and ten increased with FDR ≤ 0.05) and 676 Xaj417 proteins (235 increased in XajCM with FDR ≤ 0.05). Interestingly, the most abundant protein in Xaj was a polygalacturonase, while in Jr it was a polygalacturonase inhibitor. These results suggest that this secreted chorismate mutase may be an important virulence suppressor gene that regulates Xaj417 virulence response, allowing for improved bacterial survival in the plant tissues.
Xanthomonas citri subsp. citri (Xac) is the causal agent of citrus canker. A proteomic analysis under in planta infectious and non-infectious conditions was conducted in order to increase our ...knowledge about the adaptive process of Xac during infection.
For that, a 2D-based proteomic analysis of Xac at 1, 3 and 5 days after inoculation, in comparison to Xac growth in NB media was carried out and followed by MALDI-TOF-TOF identification of 124 unique differentially abundant proteins. Among them, 79 correspond to up-regulated proteins in at least one of the three stages of infection. Our results indicate an important role of proteins related to biofilm synthesis, lipopolysaccharides biosynthesis, and iron uptake and metabolism as possible modulators of plant innate immunity, and revealed an intricate network of proteins involved in reactive oxygen species adaptation during Plants` Oxidative Burst response. We also identified proteins previously unknown to be involved in Xac-Citrus interaction, including the hypothetical protein XAC3981. A mutant strain for this gene has proved to be non-pathogenic in respect to classical symptoms of citrus canker induced in compatible plants.
This is the first time that a protein repertoire is shown to be active and working in an integrated manner during the infection process in a compatible host, pointing to an elaborate mechanism for adaptation of Xac once inside the plant.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
The COVID-19 pandemic has caused a severe global health and economic crisis, with significant consequences for human mortality and morbidity. Therefore, there is an urgent need for more studies on ...the immune response to SARS-CoV-2 infection, both to enhance its effectiveness and prevent its deleterious effects. This study presents the chronology of antibodies during six months after infection in hospitalized patients and the kinetics of serum soluble mediators of the cellular response triggered by SARS-CoV-2. Samples and clinical data from 330 patients hospitalized at the Hospital da Baleia in Belo Horizonte, Brazil, who were suspected of having COVID-19, were collected at the time of hospitalization and during 6 months after infection. The immune response was analyzed by enzyme-linked immunosorbent assay (ELISA) and flow cytometry. There was a significant difference in IgM specific antibody titers from the 7th to 60th days after infection between COVID-19 negative and positive patients. Soon after 60 days after infection, antibody levels started to reduce, becoming similar to the antibody levels of the COVID-19 negative patients. IgG specific antibodies started to be detectable after 9 days of infection and antibody levels were comparatively higher in positive patients as soon as after 7 days. Furthermore, IgG levels remained higher in these patients during the complete period of 180 days after infection. The study observed similar antibody profiles between different patient groups. The soluble systemic biomarkers evaluated showed a decrease during the six months after hospitalization, except for CCL11, CXCL8, CCL3, CCL4, CCL5, IL-6, IFN-g, IL-17, IL-5, FGF-basic, PDGF, VEGF, G-CSF, and GM-CSF. The results indicate that IgM antibodies are more prominent in the early stages of infection, while IgG antibodies persist for a longer period. Additionally, the study identified that patients with COVID-19 have elevated levels of biomarkers after symptom onset, which decrease over time.
The objective of this study was to evaluate extender supplemented with melatonin and freezing curves on the antioxidant enzyme activity, peroxidation lipid and sperm characteristics of cryopreserved ...Brycon orbignyanus milt. Males (n = 16) and females (n = 5) were hormonally induced with two doses (0.5 mg and 5.0 mg kg−1) of carp pituitary extract, and their gametes were collected by light abdominal massage. The fresh milt was diluted at a ratio of 1:4 (milt:extender) in the following solutions: (Control) 10% methyl glycol (MG) + 5% Beltsville thawing solution; (M1) Control + 1 mM melatonin; and (M2) Control + 2 mM melatonin. The freezing curves were C1 (automated freezer) and C2 (dry shipper for 24 h). After each curve was recorded, the straws were transferred to a liquid nitrogen container until the analyses were performed. The samples were thawed in a water bath (60 °C, 8 s) and evaluated using the Sperm Class Analyzer software for the parameters total motility, progressive motility, curvilinear velocity, straight line velocity, mean displacement velocity, straightness, and linearity. The following were also measured: motility time, vitality, morphology, oxidative stress (lipid peroxidation, activity of catalase and superoxide dismutase, quantification of nitric oxide), and fertilization and hatching rates. The data were analyzed within R by one-way analysis of variance and Tukey's test for comparison of means (p < 0.05). A significant difference (p < 0.05) was observed between the solutions in vitality, morphology, motility, and fertilization rate, the solutions with melatonin having the best values. Total motility, progressive motility, and motility time were significantly different. Among oxidative stress markers, only lipid peroxidation and superoxide dismutase activity showed an effect of the curve × solution interaction (p < 0.05), the solutions with melatonin yielding the lowest values. The fertilization and hatching rates were also higher under the melatonin treatments, regardless of the curve. Melatonin 2 mM and slow curve are indicated for the cryopreservation of fish species sperm as it led to the slowest detrimental spermatozoa effects and better fertilization and hatching rates.
•Thawed semen of B. orbignyanus is improved with the addition of melatonin associated with cryoprotectant methylglycol.•The slow freezing curve used influenced the sperm quality of Brycon orbignyanus.•Melatonin 2 mM and slow freezing curve provide the best sperm response based in lipid peroxidation and SOD activity.•Melatonin 2 mM is indicated for the freezing of B. orbignianus sperm, contributing to the germplasm banks of the species.
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