scholarly journals Biocontrol of Penicillium digitatum on Postharvest Citrus Fruits by Pseudomonas fluorescens

2018 ◽  
Vol 2018 ◽  
pp. 1-10 ◽  
Author(s):  
Zhirong Wang ◽  
Mengyao Jiang ◽  
Kewei Chen ◽  
Kaituo Wang ◽  
Muying Du ◽  
...  
Keyword(s):  
Pseudomonas Fluorescens ◽  
Fold Increase ◽  
Penicillium Digitatum ◽  
Bacterial Suspension ◽  
Citrus Peel ◽  
Green Mold ◽  
Induce Resistance ◽  
Growth And Reproduction

The effectiveness of the bacteria antagonist Pseudomonas fluorescens to control green mold caused by Penicillium digitatum on oranges (Citrus sinensis Osbeck, cv. Jincheng) and the possible modes of action were evaluated. Whether in vitro or in vivo, treatments with cell-free autoclaved cultures or culture filtrate had limited capacity to suppress P. digitatum, while P. digitatum was significantly inhibited by bacterial fluid (P. fluorescens in the nutrient broth liquid medium) and bacterial suspension (P. fluorescens in sterile distilled water) with living cells. There was a positive relationship between the concentration of P. fluorescens in bacterial suspension and its biological efficacy. In addition, P. fluorescens was effective when applied preventatively but not when applied curatively. In the inoculated wounds, the population of P. fluorescens was an approximately 28- and 34-fold increase after being incubated at 20°C for 8 d and at 4°C for 16 d, respectively, and P. digitatum could effectively stimulate the growth and reproduction of P. fluorescens. Moreover, P. fluorescens was able to inhibit spore germination and germ tube elongation of P. digitatum as well as induce resistance on citrus peel by increasing the chitinase (CHI) activity and advancing the activities peaks of β-1,3-glucanase (GLU), peroxidase (POD), and phenylalanine ammonia lyase (PAL). All of these results support the potential application of P. fluorescens against green mold on postharvest citrus.

scholarly journals Evaluation of the synergy between Schwanniomyces vanrijiae and propolis in the control of Penicillium digitatum on lemons

2021 ◽  
Vol 31 (1) ◽  
Author(s):  
Kamal A. M. Abo-Elyousr ◽  
Adel D. Al-Qurashi ◽  
Najeeb M. Almasoudi
Keyword(s):  
Fungal Pathogen ◽  
Mycelial Growth ◽  
Penicillium Digitatum ◽  
In Vitro Experiments ◽  
Combined Application ◽  
Green Mold ◽  
Ethanolic Extracts ◽  
Production Losses

Abstract Background Green mold disease on citrus caused by Penicillium digitatum is the most serious and destructive disease. It is causing 90% of production losses during post-harvest handling. Results In this study, the activity of seven yeast isolates from lemons against P. digitatum, a fungal pathogen that causes the green mold disease in lemons, was isolated and examined. In vitro experiments showed that isolate three significantly reduced pathogen growths and were later identified as Schwanniomyces vanrijiae. In addition, 3% ethanolic extracts of propolis (EEP) caused a strong mycelial growth inhibition with inhibition halos of 1.4 cm. The use of S. vanrijiae treatments to protect lemon fruits from green mold has been reported (55%); however, reports describing the application of EEP are limited (40%). Thus, the effectiveness of the combination of S. vanrijiae and 3% EEP in an antagonistic mixture for protecting lemon fruits from P. digitatum was examined. EEP and S. vanrijiae treatments were applied alone and in combination in both in vitro and in vivo conditions. The combined application of 3% EEP + S. vanrijiae on lemon fruits significantly reduced the severity and incidence of green mold (80 and 93.7%, respectively) with much higher efficacy than either treatment alone. Lemon fruits treated with both S. vanrijiae and 3% EEP showed increased levels of antioxidants, peroxidase (POD), polyphenol oxidase (PPO), and phenol than the untreated control. Conclusion The results indicated that the combination of S. vanrijiae + 3% EEP can strongly protect lemon fruits from green mold compared with the sole application of either bioagent.

scholarly journals Phytotoxic tryptoquialanines produced in vivo by Penicillium digitatum are exported in extracellular vesicles

2020 ◽  
Author(s):  
Jonas Henrique Costa ◽  
Jaqueline Moraes Bazioli ◽  
Luidy Darllan Barbosa ◽  
Pedro Luis Theodoro dos Santos Júnior ◽  
Flavia C. G. Reis ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
Extracellular Vesicles ◽  
Metabolic Response ◽  
Penicillium Digitatum ◽  
Citrus Fruits ◽  
Green Mold ◽  
Host Pathogen ◽  
Citrus Seeds

ABSTRACTPenicillium digitatum is the most aggressive pathogen of citrus fruits. Tryptoquialanines are major indole alkaloids produced by P. digitatum. It is unknown if tryptoquialanines are involved in the damage of citrus fruits caused by P. digitatum. To investigate the pathogenic roles of tryptoquialanines, we initially asked if tryptoquialanines could affect the germination of Citrus sinensis seeds. Exposure of the citrus seeds to tryptoquialanine A resulted in a complete inhibition of germination and an altered metabolic response. Since this phytotoxic effect requires the extracellular export of tryptoquialanine A, we investigated the mechanisms of extracellular delivery of this alkaloid in P. digitatum. We detected extracellular vesicles (EVs) released by P. digitatum both in culture and during infection of citrus fruits. Compositional analysis of EVs produced during infection revealed the presence of a complex cargo, which included tryptoquialanines and the mycotoxin fungisporin. The EVs also presented phytotoxicity activity in vitro, and caused damage to the tissues of citrus seeds. Through molecular networking, it was observed that the metabolites present in the P. digitatum EVs are produced in all of its possible hosts. Our results reveal a novel phytopathogenic role of P. digitatum EVs and tryptoquialanine A, implying that this alkaloid is exported in EVs during plant infection.IMPORTANCEDuring the post-harvest period, citrus fruits can be affected by phytopathogens such as Penicillium digitatum, which causes the green mold disease and is responsible for up to 90 % of the total citrus losses. Chemical fungicides are widely used to prevent the green mold disease, leading to concerns about environmental and health risks. To develop safer alternatives to control phytopathogens, it is necessary to understand the molecular basis of infection during the host-pathogen interaction. In the P. digitatum model, the virulence strategies are poorly known. Here, we describe the production of phytotoxic extracellular vesicles (EVs) by P. digitatum during the infection of citrus fruits. We also characterized the secondary metabolites in the cargo of EVs and found in this set of molecules an inhibitor of seed germination. Since EVs and secondary metabolites have been related to virulence mechanisms in other host-pathogen interactions, our data are important for the comprehension of how P. digitatum causes damage to its primary hosts.

scholarly journals Methylation of rRNA as a host defense against rampant group II intron retrotransposition

Mobile DNA ◽  
2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Justin M. Waldern ◽  
Dorie Smith ◽  
Carol Lyn Piazza ◽  
E. Jake Bailey ◽  
Nicholas J. Schiraldi ◽  
...  
Keyword(s):  
Insertional Mutagenesis ◽  
Fold Increase ◽  
Group Ii Intron ◽  
In Vivo Experiments ◽  
Cellular Factors ◽  
Group Ii ◽  
Self Splicing ◽  
Native Host

Abstract Background Group II introns are mobile retroelements, capable of invading new sites in DNA. They are self-splicing ribozymes that complex with an intron-encoded protein to form a ribonucleoprotein that targets DNA after splicing. These molecules can invade DNA site-specifically, through a process known as retrohoming, or can invade ectopic sites through retrotransposition. Retrotransposition, in particular, can be strongly influenced by both environmental and cellular factors. Results To investigate host factors that influence retrotransposition, we performed random insertional mutagenesis using the ISS1 transposon to generate a library of over 1000 mutants in Lactococcus lactis, the native host of the Ll.LtrB group II intron. By screening this library, we identified 92 mutants with increased retrotransposition frequencies (RTP-ups). We found that mutations in amino acid transport and metabolism tended to have increased retrotransposition frequencies. We further explored a subset of these RTP-up mutants, the most striking of which is a mutant in the ribosomal RNA methyltransferase rlmH, which exhibited a reproducible 20-fold increase in retrotransposition frequency. In vitro and in vivo experiments revealed that ribosomes in the rlmH mutant were defective in the m3Ψ modification and exhibited reduced binding to the intron RNA. Conclusions Taken together, our results reinforce the importance of the native host organism in regulating group II intron retrotransposition. In particular, the evidence from the rlmH mutant suggests a role for ribosome modification in limiting rampant retrotransposition.

scholarly journals THE RATE OF BACTERICIDAL ACTION OF PENICILLIN IN VITRO AS A FUNCTION OF ITS CONCENTRATION, AND ITS PARADOXICALLY REDUCED ACTIVITY AT HIGH CONCENTRATIONS AGAINST CERTAIN ORGANISMS

1948 ◽  
Vol 88 (1) ◽  
pp. 99-131 ◽  
Author(s):  
Harry Eagle ◽  
A. D. Musselman
Keyword(s):  
Bacterial Species ◽  
Fold Increase ◽  
Bactericidal Effect ◽  
Penicillin G ◽  
Bacterial Suspension ◽  
Maximal Effect ◽  
Maximal Rate ◽  
Effective Level ◽  
High Concentrations

1. The concentrations of penicillin G which (a) reduced the net rate of multiplication, (b) exerted a net bactericidal effect, and (c) killed the organisms at a maximal rate, have been defined for a total of 41 strains of α- and ß-hemolytic streptococci, Staphylococcus aureus and Staphylococcus albus, Diplococcus pneumoniae, and the Reiter treponoma. 2. The concentration which killed the organisms at a maximal rate was 2 to 20 times the minimal effective level ("sensitivity" as ordinarily defined). With some organisms, even a 32,000-fold increase beyond this maximally effective level did not further increase the rate of its bactericidal effect. However, with approximately half the strains here studied (all 4 strains of group B ß-hemolytic streptococci, 4 of 5 group C strains, 5 of 7 strains of Streptococcus fecalis, 2 of 4 other α-hemolytic streptococci, and 4 of 9 strains of staphylococci), when the concentration of penicillin was increased beyond that optimal level, the rate at which the organisms died was paradoxically reduced rather than increased, so that the maximal effect was obtained only within a relatively narrow optimal zone. 3. There were marked differences between bacterial species, and occasionally between different strains of the same species, not only with respect to the effective concentrations of penicillin, but also with respect to the maximal rate at which they could be killed by the drug in any concentration. Although there was a rough correlation between these two factors, there were many exceptions; individual strains affected only by high concentrations of penicillin might nevertheless be killed rapidly, while strains sensitive to minute concentrations might be killed only slowly. 4. Within the same bacterial suspension, individual organisms varied only to a minor degree with respect to the effective concentrations of penicillin. They varied strikingly, however, in their resistance to penicillin as measured by the times required to kill varying proportions of the cells.

Abstract 41: Explant-Derived Cells from Chronic Heart Failure Activated Endothelial-Mesenchymal Transition and Attenuated Pluripotency Genes Expression

2012 ◽  
Vol 111 (suppl_1) ◽  
Author(s):  
Liudmila Zakharova ◽  
Hikmet Nural ◽  
Mohamed A Gaballa
Keyword(s):  
Gene Expression ◽  
Progenitor Cells ◽  
Smooth Muscle Actin ◽  
Fold Increase ◽  
Gene Expressions ◽  
Mesenchymal Transition ◽  
Cell Outgrowth ◽  
Pluripotency Genes

Cardiac progenitor cells are generated from atria explants; however the cellular origin and the mechanisms of cell outgrowth are unclear. Using transgenic tamoxifen-induced Willms tumor 1 (Wt1)-Cre/ERT and Cre-activated GFP reporter mice, we found approximately 40% of explant-derived cells and 74% of explant-derived c-Kit+ cells originated from the epicardium. In atria from sham hearts, Wt1+ cells were located in a thin epicardial layer, while c-Kit+ cells were primarily found within both the sub-epicardium and the myocardium, albeit at low frequency. No overlap between c-Kit+ and Wt1+ cells was observed, suggesting that epicardial Wt1+ cells do not express c-Kit marker in vivo, but more likely the c-Kit marker was acquired in culture. Compared with 4 days in culture, at day 21 we observed 7 folds increase in Snail gene expression; 32% increase in α-smooth muscle actin (SMA) marker, and 30% decrease in E-cadherin marker, suggesting that the explant-derived cells underwent epithelial to mesenchymal transition (EMT) in vitro. Cell outgrowths released TGF-β (1036.4 ± 1.18 pm/ml) and exhibited active TGF-β signaling, which might triggered the EMT. Compared to shams, CHF cell outgrowths exhibited elevated levels of EMT markers, SMA (49% vs. 34%) and Snail (2 folds), and reduced level of Wt1 (11% vs. 22%). In addition, CHF cell outgrowths had two folds increase in Pai1 gene expression, a direct target of TGF-β signaling. In c-Kit+ cells derived from CHF explants, Nanog gene expression was 4 folds lower and Sox 2 was 2 folds lower compared with cells from shams. Suppression of EMT in cell outgrowth increased the percentage of c-Kit+ and Wt1+ cells by 17%, and 15%, respectively. Also suppression of EMT in c-Kit+ cells resulted in 4 folds increase in Nanog and 3 fold increase in Sox2 gene expressions. Our results showed that CHF may further exuberates EMT while diminishes the re-activation of pluripotency genes. Thus, EMT modulation in CHF is a possible strategy to regulate both the yield and the pluripotency of cardiac-explant-derived progenitor cells.

FKBP51 Modulates Hippocampal Size and Function in Post-translational Regulation of Parkin

2021 ◽  
Author(s):  
Bin Qiu ◽  
Zhaohui Zhong ◽  
Shawn Righter ◽  
Yuxue Xu ◽  
Jun Wang ◽  
...  
Keyword(s):  
Hippocampal Neurons ◽  
Translational Regulation ◽  
Disease Onset ◽  
Fold Increase ◽  
Knock Out ◽  
Fk506 Binding Protein ◽  
Protein Levels ◽  
And Function

Abstract FK506-binding protein 51 (encoded by Fkpb51) has been associated with stress-related mental illness. To identify its function, we studied the morphological consequences of Fkbp51 deletion. Artificial Intelligence-assist morphological analysis identified that Fkbp51 knock-out (KO) mice possess more elongated CA and DG but shorter in height in coronal section when compared to WT. Primary cultured Fkbp51 KO hippocampal neurons were shown to exhibit larger dendritic outgrowth than wild-type (WT) controls, pharmacological manipulation experiments suggest that this may occur through regulation of microtubule-associated protein. Both in vitro primary culture and in vivo labeling support that FKBP51 regulates microtubule-associated protein expression. Furthermore, in the absence of differences in mRNA expression, Fkbp51 KO hippocampus exhibited decreases in βIII-tubulin, MAP2, and Tau protein levels, but a greater than 2.5-fold increase in Parkin protein. Overexpression and knock-down FKBP51 demonstrated that FKBP51 negatively regulates Parkin in a dose-dependent and ubiquitin-mediated manner. These results indicate a potential novel post-translational regulatory of Parkin by FKBP51 and significance of their interaction on disease onset.

scholarly journals Albumin Enhances Caspofungin Activity against Aspergillus Species by Facilitating Drug Delivery to Germinating Hyphae

2015 ◽  
Vol 60 (3) ◽  
pp. 1226-1233 ◽  
Author(s):  
Petros Ioannou ◽  
Aggeliki Andrianaki ◽  
Tonia Akoumianaki ◽  
Irene Kyrmizi ◽  
Nathaniel Albert ◽  
...  
Keyword(s):  
Drug Delivery ◽  
Cell Culture ◽  
Antifungal Agents ◽  
Fold Increase ◽  
Culture Conditions ◽  
The Other ◽  
Related Factors ◽  
Content Type

The modestin vitroactivity of echinocandins againstAspergillusimplies that host-related factors augment the action of these antifungal agentsin vivo. We found that, in contrast to the other antifungal agents (voriconazole, amphotericin B) tested, caspofungin exhibited a profound increase in activity against variousAspergillusspecies under conditions of cell culture growth, as evidenced by a ≥4-fold decrease in minimum effective concentrations (MECs) (P= 0. 0005). Importantly, the enhanced activity of caspofungin againstAspergillusspp. under cell culture conditions was strictly dependent on serum albumin and was not observed with the other two echinocandins, micafungin and anidulafungin. Of interest, fluorescently labeled albumin bound preferentially on the surface of germinatingAspergillushyphae, and this interaction was further enhanced upon treatment with caspofungin. In addition, supplementation of cell culture medium with albumin resulted in a significant, 5-fold increase in association of fluorescently labeled caspofungin withAspergillushyphae (P< 0.0001). Collectively, we found a novel synergistic interaction between albumin and caspofungin, with albumin acting as a potential carrier molecule to facilitate antifungal drug delivery toAspergillushyphae.

scholarly journals Augmented expansion of Treg cells from healthy and autoimmune subjects via adult progenitor cell co-culture

2020 ◽  
Author(s):  
JL Reading ◽  
VD Roobrouck ◽  
CM Hull ◽  
PD Becker ◽  
J Beyens ◽  
...  
Keyword(s):  
T Cell ◽  
Treg Cell ◽  
Regulatory T Cell ◽  
Cellular Therapy ◽  
Ex Vivo ◽  
Fold Increase ◽  
Adoptive Cellular Therapy ◽  
T Cell Therapy

AbstractRecent clinical experience has demonstrated that adoptive regulatory T cell therapy is a safe and feasible strategy to suppress immunopathology via induction of host tolerance to allo- and autoantigens. However, clinical trials continue to be compromised due to an inability to manufacture a sufficient Treg cell dose. Multipotent adult progenitor cells (MAPCⓇ) promote regulatory T cell differentiation in vitro, suggesting they may be repurposed to enhance ex vivo expansion of Tregs for adoptive cellular therapy. Here, we use a GMP compatible Treg expansion platform to demonstrate that MAPC cell-co-cultured Tregs (MulTreg) exhibit a log-fold increase in yield across two independent cohorts, reducing time to target dose by an average of 30%. Enhanced expansion is linked with a distinct Treg cell-intrinsic transcriptional program, characterized by diminished levels of core exhaustion (BATF, ID2, PRDM1, LAYN, DUSP1), and quiescence (TOB1, TSC22D3) related genes, coupled to elevated expression of cell-cycle and proliferation loci (MKI67, CDK1, AURKA, AURKB). In addition, MulTreg display a unique gut homing (CCR7lo β7hi) phenotype and importantly, are more readily expanded from patients with autoimmune disease compared to matched Treg lines, suggesting clinical utility in gut and/or Th1-driven pathology associated with autoimmunity or transplantation. Relative to expanded Tregs, MulTreg retain equivalent and robust purity, FoxP3 TSDR demethylation, nominal effector cytokine production and potent suppression of Th1-driven antigen specific and polyclonal responses in vitro and xeno graft vs host disease (xGvHD) in vivo. These data support the use of MAPC cell co-culture in adoptive Treg therapy platforms as a means to rescue expansion failure and reduce the time required to manufacture a stable, potently suppressive product.

scholarly journals Respuesta de biosurfactantes producidos por Pseudomonas fluorescens para el control de la gota de la papa Phytophthora infestans (Mont) de Bary, bajo condiciones controlada.

2010 ◽  
Vol 11 (1) ◽  
pp. 21
Author(s):  
Hugo F. Rivera ◽  
Erika P. Martínez ◽  
Jairo A. Osorio ◽  
Edgar Martínez
Keyword(s):  
Late Blight ◽  
Phytophthora Infestans ◽  
Pseudomonas Fluorescens ◽  
Negative Impact ◽  
Potato Late Blight ◽  
Pathogen Resistance ◽  
Production Costs ◽  
In Vitro Assays

<p>Phytophthora infestans (Mont.) de Bary, agente causal de la gota de la papa, es considerado la principal limitante de la producción de este cultivo en Colombia. El control habitual del patógeno se realiza con fungicidas de tipo sistémico, que incrementan los costos de producción, pueden inducir la resistencia del patógeno y tiene un impacto negativo en el ambiente. Por tanto, se llevó a cabo este estudio con el propósito de buscar alternativas amigables con el ambiente, que hagan parte de un paquete tecnológico eficaz de control. Dos cepas nativas de Psedomonas fluorescens (039T y 021V), provenientes de cultivos de papa, fueron evaluadas contra P. infestans. Las suspensiones bacterianas y los biosurfactantes parcialmente purificados (BPP), producidos por éstas (obtenidos en medio mínimo de sales con querosén), fueron aplicados sobre foliolos desprendidos en ensayos in vitro y experimentos in vivo en plantas de papa, en condiciones controladas en casa de malla. Los resultados demostraron la capacidad que tienen los biosurfactantes y las suspensiones bacterianas para controlar al patógeno, ya que el BPP 039T logró reducir el nivel de severidad de la enfermedad en 79,9% in vitro y 38,5% in vivo, mientras que el BPP 021V redujo en 78,7% in vitro y 30,2% in vivo. Las suspensiones bacterianas redujeron el nivel de severidad en 72,4% (039T) y 66,1% (021V) en las evaluaciones in vitro y 35% en los experimentos in vivo. Los resultados de esta investigación muestran el potencial que tienen los biosurfactantes para el control de la gota en Colombia.</p><p> </p><p><strong>Evaluation of Biosurfactants Produced by Pseudomonas fluorescens for Potato Late Blight Control (Phytophthora infestans (Mont) de Bary) Under Controlled Conditions</strong></p><p>Phytophthora infestans (Mont.) de Bary, causal agent of potato late blight is considered the main limiting pathogen for the production of this crop in Colombia. The usual control of the disease has been performed with systemic fungicides which increase production costs, can induce pathogen resistance and have a negative impact on the environment. Therefore, this study was carried out in order to find effective and environmentally friendly control alternatives for potato late blight. Two Pseudomonas fluorescens native strains (039T and 021V) isolated from potato crops were evaluated against P. infestans. Bacterial suspensions (obtained from minimal salts medium added with kerosene) and partially purified biosurfactants (BPP) were applied on detached leaflets for in vitro assays and on potato plants in greenhouse, for in vivo assays and the measure of inhibitory effect of the disease was assessed. The results showed the ability of P. fluorescens biosurfactants and bacterial suspensions to control the pathogen. BPP 039T was able to reduce the level of severity disease by 79.9% in vitro and 38.5% in vivo, whereas BPP 021V decreased 78.7% in vitro and 30.2% in vivo. Bacterial suspensions reduced the severity level in 72.4% (039T) and 66.1% (021V) in vitro assessments and 35% in the in vivo experiment. These results show the potential of P. fluorescens biosurfactants to control the potato late blight in Colombia.</p>

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