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 Phytotoxic Tryptoquialanines Produced In Vivo by Penicillium digitatum Are Exported in Extracellular Vesicles

mBio ◽  
2021 ◽  
Vol 12 (1) ◽  
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 ◽  
Content Type ◽  
Green Mold ◽  
Host Pathogen ◽  
Citrus Seeds

ABSTRACT Penicillium 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. IMPORTANCE During the postharvest period, citrus fruits can be affected by phytopathogens such as Penicillium digitatum, which causes green mold disease and is responsible for up to 90% of total citrus losses. Chemical fungicides are widely used to prevent 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.

Inactivation of Penicillium digitatum and Penicillium italicum under In Vitro and In Vivo Conditions by Using UV-C Light

2013 ◽  
Vol 76 (10) ◽  
pp. 1761-1766 ◽  
Author(s):  
GÜLTEN TİRYAKİ GÜNDÜZ ◽  
FIKRET PAZIR
Keyword(s):  
Potato Dextrose Agar ◽  
Penicillium Digitatum ◽  
Citrus Fruits ◽  
Penicillium Italicum ◽  
Inoculation Methods ◽  
Penetration Ability ◽  
Working Distance ◽  
Uv C

In this study, the effects of UV-C on two of the main wound pathogens of citrus fruits, Penicillium digitatum and Penicillium italicum, were investigated with different inoculation methods in vitro and on oranges. P. digitatum and P. italicum spores were inoculated onto the surface of potato dextrose agar or oranges using spread, spot, wound, and piercing inoculation methods. UV-C treatment for 1 min from a working distance of 8 cm reduced the numbers of P. italicum and P. digitatum by about 3.9 and 5.3 log units, respectively, following spread inoculation under in vitro conditions. Significant reductions were obtained after 1-min UV-C treatments of the tested fungi following inoculation using the spread and spot methods. With inoculation by the wound and piercing methods, the tested spores were not inactivated completely even after 10- and 20-min treatment times, respectively. The application of UV-C (7.92 kJ m−2) on oranges reduced the percentage of oranges infected at least threefold compared with the rate of infection in the untreated control samples. UV-C irradiation could effectively inactivate spores of P. italicum and P. digitatum inoculated by the spread plate and spot inoculation methods under in vitro and in vivo conditions. On the other hand, because of the low penetration ability of UV-C light, the tested fungi were not completely inactivated following inoculation with the wound and piercing methods. UV-C treatment has potential for use in surface decontamination of citrus fruits.

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 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 Screening Methods for Isolation of Biocontrol Epiphytic Yeasts against Penicillium digitatum in Lemons

2021 ◽  
Vol 7 (3) ◽  
pp. 166
Author(s):  
Martina María Pereyra ◽  
Mariana Andrea Díaz ◽  
Fabricio Fabián Soliz-Santander ◽  
Anja Poehlein ◽  
Friedhelm Meinhardt ◽  
...  
Keyword(s):  
Action Spectrum ◽  
Penicillium Digitatum ◽  
Rrna Genes ◽  
Screening Methods ◽  
Dual Culture ◽  
Citrus Fruits ◽  
Postharvest Diseases ◽  
Yeast Strains

Worldwide, the green rot caused by Penicillium digitatum is one of the most aggressive postharvest diseases of lemons. Searching for sustainable alternatives to chemical fungicides, epiphytic yeasts as potential biocontrol agents were isolated from citrus fruits using a tailor-made selective medium. For disclosing their antagonistic potential against P. digitatum, obtained isolates were subjected to direct screening methods, both in vitro and in vivo. In the course of the primary in vitro screening that comprised dual culture assays, 43 yeast strains displaying antagonistic activities against the pathogen were selected. Subsequently, such strains were subjected to an in vivo screening that consisted of a microscale test, allowing the selection of six yeast strains for further analysis. In the final screening using macroscale in vivo tests, three strains (AcL2, AgL21, and AgL2) displaying the highest efficiencies to control P. digitatum were identified. The protection efficiencies in lemons were 80 (AcL2), 76.7 (AgL21), and 75% (AgL2). Based on sequence analysis of the PCR amplified D1/D2 domains of the 26S rRNA genes, they were identified as representatives of the species Clavispora lusitaniae. Interestingly, the strains exhibited a broad action spectrum among citrus fruits as they were also able to combat the green mold disease in grapefruit and two orange varieties. The direct screening methods applied in this study favored the recovery of efficient candidates for application as biological control agents to combat fungal infestations of citrus fruits.

Associated microflora of medicinal ferns: biotechnological potentials and possible applications

2016 ◽  
Vol 5 (03) ◽  
pp. 4927 ◽  
Author(s):  
Shubhi Srivastava ◽  
Paul A. K.
Keyword(s):  
Secondary Metabolites ◽  
Growth Promotion ◽  
Biological Activities ◽  
Hydrolytic Enzymes ◽  
In Vitro Production ◽  
Wide Range ◽  
Negative Effect ◽  
Bacteria And Fungi

Plant associated microorganisms that colonize the upper and internal tissues of roots, stems, leaves and flowers of healthy plants without causing any visible harmful or negative effect on their host. Diversity of microbes have been extensively studied in a wide variety of vascular plants and shown to promote plant establishment, growth and development and impart resistance against pathogenic infections. Ferns and their associated microbes have also attracted the attention of the scientific communities as sources of novel bioactive secondary metabolites. The ferns and fern alleles, which are well adapted to diverse environmental conditions, produce various secondary metabolites such as flavonoids, steroids, alkaloids, phenols, triterpenoid compounds, variety of amino acids and fatty acids along with some unique metabolites as adaptive features and are traditionally used for human health and medicine. In this review attention has been focused to prepare a comprehensive account of ethnomedicinal properties of some common ferns and fern alleles. Association of bacteria and fungi in the rhizosphere, phyllosphere and endosphere of these medicinally important ferns and their interaction with the host plant has been emphasized keeping in view their possible biotechnological potentials and applications. The processes of host-microbe interaction leading to establishment and colonization of endophytes are less-well characterized in comparison to rhizospheric and phyllospheric microflora. However, the endophytes are possessing same characteristics as rhizospheric and phyllospheric to stimulate the in vivo synthesis as well as in vitro production of secondary metabolites with a wide range of biological activities such as plant growth promotion by production of phytohormones, siderophores, fixation of nitrogen, and phosphate solubilization. Synthesis of pharmaceutically important products such as anticancer compounds, antioxidants, antimicrobials, antiviral substances and hydrolytic enzymes could be some of the promising areas of research and commercial exploitation.

A Systematic Review on Extracellular Vesicles-Enriched Fat Grafting: A Shifting Paradigm

2020 ◽  
Author(s):  
Mohammad Ghiasloo ◽  
Laura De Wilde ◽  
Kashika Singh ◽  
Patrick Tonnard ◽  
Alexis Verpaele ◽  
...  
Keyword(s):  
Systematic Review ◽  
Graft Survival ◽  
Extracellular Vesicles ◽  
Retention Rate ◽  
Fat Grafting ◽  
Retention Rates ◽  
Fat Graft ◽  
Cochrane Central Register

Abstract Background Recent evidence confirms that mesenchymal stem cells (MSCs) facilitate angiogenesis mainly through paracrine function. Extracellular vesicles (EVs) are regarded as key components of the cell secretome, possessing functional properties of their source cells. Subsequently, MSC-EVs have emerged as a novel cell-free approach to improve fat graft retention rate. Objectives To provide a systematic review of all studies reporting the use of MSC-EVs to improve graft retention rate. Methods A systematic search was undertaken using the Embase, PubMed and the Cochrane Central Register of Controlled Trials databases. Outcome measures included donor/receptor organism of the fat graft, study model, intervention groups, evaluation intervals, EV research data, in vitro and in vivo results. Results Of the total 1717 articles, 62 full-texts were screened. Seven studies reporting on 294mice were included. Overall, EV treated groups showed higher graft retention rates compared to untreated groups. Notably, retention rate was similar following EV- and MSC-treatment. In addition to reduced inflammation, graft enrichment with EVs resulted in early revascularization and better graft integrity. Interestingly, hypoxic preconditioning of MSCs improved their beneficial paracrine effects and led to a more proangiogenic EV population, as observed by both in vitro and in vivo results. Conclusions MSC-EVs appear to offer an interesting cell-free alternative to improve fat graft survival. While their clinical relevance remains to be determined, it is clear that not the cells, but their secretome is essential for graft survival. Thus, a paradigm shift from cell-assisted lipotransfer towards ‘secretome-assisted lipotransfer’ is well on its way.

scholarly journals Plant-Derived Extracellular Vesicles: Current Findings,Challenges, and Future Applications

Membranes ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 411
Author(s):  
Nader Kameli ◽  
Anya Dragojlovic-Kerkache ◽  
Paul Savelkoul ◽  
Frank R. Stassen
Keyword(s):  
Human Health ◽  
Extracellular Vesicles ◽  
Preliminary Results ◽  
In Vivo Experiments ◽  
Health And Disease ◽  
Conducting Research ◽  
Protocol Standardization ◽  
Insight Into

In recent years, plant-derived extracellular vesicles (PDEVs) have gained the interest of many experts in fields such as microbiology and immunology, and research in this field has exponentially increased. These nano-sized particles have provided researchers with a number of interesting findings, making their application in human health and disease very promising. Both in vitro and in vivo experiments have shown that PDEVs can exhibit a multitude of effects, suggesting that these vesicles may have many potential future applications, including therapeutics and nano-delivery of compounds. While the preliminary results are promising, there are still some challenges to face, such as a lack of protocol standardization, as well as knowledge gaps that need to be filled. This review aims to discuss various aspects of PDEV knowledge, including their preliminary findings, challenges, and future uses, giving insight into the complexity of conducting research in this field.

scholarly journals Lipoproteins Are Responsible for the Pro-Inflammatory Property of Staphylococcus aureus Extracellular Vesicles

2021 ◽  
Vol 22 (13) ◽  
pp. 7099
Author(s):  
Pradeep Kumar Kopparapu ◽  
Meghshree Deshmukh ◽  
Zhicheng Hu ◽  
Majd Mohammad ◽  
Marco Maugeri ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Inflammatory Responses ◽  
Surface Proteins ◽  
Host Cells ◽  
Immune Stimulation ◽  
Domains Of Life ◽  
Major Surface ◽  
Staphylococcal Aureus

Staphylococcal aureus (S. aureus), a Gram-positive bacteria, is known to cause various infections. Extracellular vesicles (EVs) are a heterogeneous array of membranous structures secreted by cells from all three domains of life, i.e., eukaryotes, bacteria, and archaea. Bacterial EVs are implied to be involved in both bacteria–bacteria and bacteria–host interactions during infections. It is still unclear how S. aureus EVs interact with host cells and induce inflammatory responses. In this study, EVs were isolated from S. aureus and mutant strains deficient in either prelipoprotein lipidation (Δlgt) or major surface proteins (ΔsrtAB). Their immunostimulatory capacities were assessed both in vitro and in vivo. We found that S. aureus EVs induced pro-inflammatory responses both in vitro and in vivo. However, this activity was dependent on lipidated lipoproteins (Lpp), since EVs isolated from the Δlgt showed no stimulation. On the other hand, EVs isolated from the ΔsrtAB mutant showed full immune stimulation, indicating the cell wall anchoring of surface proteins did not play a role in immune stimulation. The immune stimulation of S. aureus EVs was mediated mainly by monocytes/macrophages and was TLR2 dependent. In this study, we demonstrated that not only free Lpp but also EV-imbedded Lpp had high pro-inflammatory activity.

Sign in / Sign up

Export Citation Format

Share Document