scholarly journals NADPH Oxidase Regulates the Growth and Pathogenicity of Penicillium expansum

2021 ◽  
Vol 12 ◽  
Author(s):  
Xuemei Zhang ◽  
Yuanyuan Zong ◽  
Di Gong ◽  
Lirong Yu ◽  
Edward Sionov ◽  
...  
Keyword(s):  
Nadph Oxidase ◽  
Radial Growth ◽  
Penicillium Expansum ◽  
Activity Levels ◽  
Lauryl Sulfate ◽  
Ros Accumulation ◽  
Negative Effect ◽  
Colonization Process ◽  
Regulatory Effects ◽  
Apple Fruits

The occurrence of reactive oxygen species (ROS) during the colonization of necrotrophic pathogens attacking fruit is critical during the attack, but its importance in Penicillium expansum remains unclear. This study aimed to determine the regulatory effects of NADPH oxidase (Nox) genes on the growth and pathogenicity of P. expansum in apple fruits. Deletion mutants of ΔPeNoxA, ΔPeNoxR, and ΔPeRacA genes were constructed to determine the contribution to the colonization process. The ΔPeRacA strain had a significant effect on the reduction of growth and pathogenicity, the ΔPeNoxA strain negatively regulated the growth and development of P. expansum and did not show any significant effect on the pathogenicity, and the ΔPeNoxR strain showed no effect on the growth or pathogenicity of P. expansum in the apple fruits. However, analysis of the content of O2– and H2O2 in the mycelium of all the Nox mutants showed a significant reduction, confirming the functionality of Nox mutations. Growth under stress conditions in the presence of Congo red, sodium lauryl sulfate, and H2O2 showed a negative effect on the radial growth of ΔPeNoxA, but a positive effect on radial growth reduction by ΔPeNoxR and ΔPeRacA mutants was shown. Interestingly, the host antioxidant activity levels of superoxide dismutase (SOD) andcatalase (CAT) in the fruits after inoculation with ΔPeNoxA, ΔPeNoxR, and ΔPeRacA mutants declined, suggesting reduced ROS accumulation in the colonized region. These results suggest that PeNoxA, PeNoxR, and PeRacA differentially regulate the growth and pathogenicity of P. expansum by producing ROS, and that PeRacA showed the strongest regulatory effect.

scholarly journals First Report of Penicillium carneum Causing Blue Mold on Stored Apples in Pennsylvania

Plant Disease ◽  
2012 ◽  
Vol 96 (12) ◽  
pp. 1823-1823 ◽  
Author(s):  
K. A. Peter ◽  
I. Vico ◽  
V. Gaskins ◽  
W. J. Janisiewicz ◽  
R. A. Saftner ◽  
...  
Keyword(s):  
Negative Control ◽  
Penicillium Expansum ◽  
Malt Extract ◽  
Conidial Suspension ◽  
Blue Mold ◽  
Separate Species ◽  
First Report ◽  
Golden Delicious ◽  
Apple Fruits ◽  
Β Tubulin

Blue mold decay occurs during long term storage of apples and is predominantly caused by Penicillium expansum Link. Apples harvested in 2010 were stored in a controlled atmosphere at a commercial Pennsylvania apple packing and storage facility, and were examined for occurrence of decay in May 2011. Several decayed apples from different cultivars, exhibiting blue mold symptoms with a sporulating fungus were collected. One isolate recovered from a decayed ‘Golden Delicious’ apple fruit was identified as P. carneum Frisvad. Genomic DNA was isolated, 800 bp of the 3′ end of the β-tubulin locus was amplified using gene specific primers and sequenced (4). The recovered nucleotide sequence (GenBank Accession No. JX127312) indicated 99% sequence identity with P. carneum strain IBT 3472 (GenBank Accession No. JF302650) (3). The P. carneum colonies strongly sporulated and had a blue green color on potato dextrose agar (PDA), Czapek yeast autolysate agar (CYA), malt extract agar (MEA), and yeast extract sucrose agar (YES) media at 25°C after 7 days. The colonies also had a beige color on plate reverse on CYA and YES media. The species tested positive for the production of alkaloids, as indicated by a violet reaction for the Ehrlich test, and grew on CYA at 30°C and on Czapek with 1,000 ppm propionic acid agar at 25°C; all of which are diagnostic characters of this species (2). The conidiophores were hyaline and tetraverticillate with a finely rough stipe. Conida were produced in long columns, blue green, globose, and averaged 2.9 μm in diameter. To prove pathogenicity, Koch's postulates were conducted using 20 ‘Golden Delicious’ apple fruits. Fruits were washed, surface sterilized with 70% ethanol, and placed onto fruit trays. Using a nail, 3-mm wounds were created and inoculated with 50 μl of a 106/ml conidial suspension or water only as a negative control. The fruit trays were placed into boxes and were stored in the laboratory at 20°C for 7 days. The inoculated fruit developed soft watery lesions, with hard defined edges 37 ± 4 mm in diameter. The sporulating fungus was reisolated from infected tissue of all conidia inoculated apples and confirmed to be P. carneum by polymerase chain reaction (PCR) using the β-tubulin locus as described. Water inoculated control apples were symptomless. Originally grouped with P. roqueforti, P. carneum was reclassified in 1996 as a separate species (1). P. carneum is typically associated with meat products, beverages, and bread spoilage and produces patulin, which is not produced by P. roqueforti (1,2). Our isolate of P. carneum was susceptible to the thiabendazole (TBZ) fungicide at 250 ppm, which is below the recommended labeled application rate of 600 ppm. The susceptibility to TBZ suggests that this P. carneum isolate has been recently introduced because resistance to TBZ has evolved rapidly in P. expansum (4). To the best of our knowledge, P. carneum has not previously been described on apple, and this is the first report of P. carneum causing postharvest decay on apple fruits obtained from storage in Pennsylvania. References: (1) M. Boyson et al. Microbiology 142:541, 1996. (2) J. C. Frisvad and R. A. Samson. Stud. Mycol. 49:1, 2004. (3) B. G. Hansen et al. BMC Microbiology 11:202, 2011. (4) P. L. Sholberg et al. Postharvest Biol. Technol. 36:41, 2005.

Antifungal Activity of Eugenol against Penicillium, Aspergillus, and Fusarium Species

2010 ◽  
Vol 73 (6) ◽  
pp. 1124-1128 ◽  
Author(s):  
DANIELA CAMPANIELLO ◽  
MARIA ROSARIA CORBO ◽  
MILENA SINIGAGLIA
Keyword(s):  
Antifungal Activity ◽  
Radial Growth ◽  
Aspergillus Terreus ◽  
Fusarium Species ◽  
Critical Value ◽  
Model System ◽  
Penicillium Expansum ◽  
Emericella Nidulans ◽  
Colony Diameter ◽  
Fungistatic Activity

The antifungal activity of eugenol in a model system against aspergilli (Aspergillus niger, Aspergillus terreus, and Emericella nidulans), penicilli (Penicillium expansum, Penicillium glabrum, and Penicillium italicum), and fusaria (Fusarium oxysporum and Fusarium avenaceum) was investigated. Minimum detection time (time to attain a colony diameter of 1 cm) and the kinetic parameters were evaluated. The effectiveness of the active compound seemed to be strain or genus dependent; 100 mg/liter represented a critical value for P. expansum, P. glabrum, P. italicum, A. niger, and E. nidulans because a further increase of eugenol resulted in fungistatic activity. The radial growth of A. terreus and F. avenaceum was inhibited at 140 mg/liter, and growth of F. oxysporum was completely inhibited at 150 mg/liter.

scholarly journals Biocontrol Efficacy of Antagonist Yeasts to Gray Mold and Blue Mold on Apples and Pears in Controlled Atmospheres

Plant Disease ◽  
2002 ◽  
Vol 86 (8) ◽  
pp. 848-853 ◽  
Author(s):  
Shiping Tian ◽  
Qing Fan ◽  
Yong Xu ◽  
Haibo Liu
Keyword(s):  
Pathogenic Fungi ◽  
Storage Conditions ◽  
Gray Mold ◽  
Penicillium Expansum ◽  
Cryptococcus Albidus ◽  
Blue Mold ◽  
Controlled Atmospheres ◽  
Biocontrol Efficacy ◽  
Significant Difference ◽  
Apple Fruits

Biocontrol capability of the yeasts Trichosporon sp. and Cryptococcus albidus against Botrytis cinerea and Penicillium expansum was evaluated in apple (cv. Golden Delicious) and pear (cv. Jingbai) fruits at 1°C in air and under controlled atmospheres (CA) with 3% O2 + 3% CO2 or 3% O2 + 8% CO2. Trichosporon sp. controlled gray mold and blue mold of apple fruits more effectively than C. albidus (P < 0.05). Apple fruits treated with Trichosporon sp. and C. albidus had a lower incidence of gray mold rot than blue mold rot in the same storage conditions. Biocontrol efficacy of the yeasts for controlling gray mold and blue mold was better in apples than in pears. Populations of the yeasts in drop-inoculated wounds in fruits increased rapidly after 20 days at 1°C both in air and in CA conditions. There was no significant difference in colony diameters of the two pathogens cultured in 0 to 15% CO2 concentrations after 7 days at 20°C, but the colony diameter of both B. cinerea and P. expansum at 20% CO2 was significantly less than in other treatments (P < 0.05). CA with 3% O2 + 8% CO2 inhibited the pathogenic fungi more than CA with 3% O2 + 3% CO2.

Production of Mycotoxins by Penicillium expansum Inoculated into Apples

2008 ◽  
Vol 71 (8) ◽  
pp. 1714-1719 ◽  
Author(s):  
MITSURU WATANABE
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
High Performance ◽  
Hplc Analysis ◽  
Penicillium Expansum ◽  
Liquid Chromatography Mass Spectrometry ◽  
Blue Mold ◽  
Positive Ion ◽  
Uv And Fluorescence Detection ◽  
Apple Fruits

We investigated the production of mycotoxins in apple fruits inoculated with spores of 40 strains of apple blue mold, Penicillium expansum. Patulin and citrinin contents in the extracts from apples stored at 25°C for 12 days after inoculation were determined by high-performance liquid chromatography (HPLC) analysis with UV and fluorescence detection. Patulin and citrinin were produced by 90% (36) and 80% (32) of the 40 strains, indicating that P. expansum is a consistent producer of these mycotoxins. The patulin content in the extracts was substantially higher than the citrinin content. Other mycotoxins whose production in pure culture has been reported were simultaneously detected with high-resolution liquid chromatography–mass spectrometry (LC-MS) analysis with the positive ion mode of electrospray ionization. Along with patulin and citrinin, expansolides A and B were identified based on the HPLC and LC-MS spectral data and detected in 88% (35) of the extracts. The results indicate that P. expansum is a consistent producer of expansolides A and B in rotten areas of apple fruits. The findings raise the possibility that products from decayed apples might contain expansolides A and B in addition to patulin and citrinin.

scholarly journals Međuovisnost različitih indikatora vitaliteta stabala i njihov odziv na klimatske uvjete na plohi obične bukve (Fagus sylvatica L.)

2020 ◽  
Vol 144 (7-8) ◽  
pp. 351-365
Author(s):  
Mladen Ognjenović ◽  
Ivan Seletković ◽  
Krunoslav Indir ◽  
Damir Ugarković ◽  
Nenad Potočić ◽  
...  
Keyword(s):  
High Temperatures ◽  
Radial Growth ◽  
Negative Influence ◽  
Climatic Conditions ◽  
Common Beech ◽  
Wide Range ◽  
Crown Defoliation ◽  
Beech Stand ◽  
Negative Effect ◽  
Low Precipitation

Interrelations of various common beech vitality indicators (crown defoliation, foliar chemistry, radial growth) as well as their possible dependencies on climatic conditions were investigated over the course of 12 years in a mature and healthy beech stand. Our results confirm the importance of temperature variables for defoliation, as high temperatures during spring and summer months induce the increase of defoliation. The same negative influence was observed with high maximum temperatures and low precipitation during previous year summer months. Phosphorus, calcium and magnesium nutrition of beech trees suffers from high temperatures during current year summer and benefits from more precipitation. High temperatures in current year May positively influence beech radial growth, while a wide range of minimum temperatures during March and June has a negative effect. In summary, high summer temperatures and low precipitation were shown to have a negative effect on all vitality indicators, and for defoliation and nutrition this effect can last into the following year.

scholarly journals Cross talk between increased intracellular zinc (Zn2+) and accumulation of reactive oxygen species in chemical ischemia

2017 ◽  
Vol 313 (4) ◽  
pp. C448-C459 ◽  
Author(s):  
Kira G. Slepchenko ◽  
Qiping Lu ◽  
Yang V. Li
Keyword(s):  
Reactive Oxygen Species ◽  
Nadph Oxidase ◽  
Cross Talk ◽  
Reactive Oxygen ◽  
Glucose Deprivation ◽  
Ros Generation ◽  
Oxygen Species ◽  
Mitochondrial Ros ◽  
Ros Accumulation ◽  
Ischemic Stress

Both zinc (Zn2+) and reactive oxygen species (ROS) have been shown to accumulate during hypoxic-ischemic stress and play important roles in pathological processes. To understand the cross talk between the two of them, here we studied Zn2+ and ROS accumulation by employing fluorescent probes in HeLa cells to further the understanding of the cause and effect relationship of these two important cellular signaling systems during chemical-ischemia, stimulated by oxygen and glucose deprivation (OGD). We observed two Zn2+ rises that were divided into four phases in the course of 30 min of OGD. The first Zn2+ rise was a transient, which was followed by a latent phase during which Zn2+ levels recovered; however, levels remained above a basal level in most cells. The final phase was the second Zn2+ rise, which reached a sustained plateau called Zn2+ overload. Zn2+ rises were not observed when Zn2+ was removed by TPEN (a Zn2+ chelator) or thapsigargin (depleting Zn2+ from intracellular stores) treatment, indicating that Zn2+ was from intracellular storage. Damaging mitochondria with FCCP significantly reduced the second Zn2+ rise, indicating that the mitochondrial Zn2+ accumulation contributes to Zn2+ overload. We also detected two OGD-induced ROS rises. Two Zn2+ rises preceded two ROS rises. Removal of Zn2+ reduced or delayed OGD- and FCCP-induced ROS generation, indicating that Zn2+ contributes to mitochondrial ROS generation. There was a Zn2+-induced increase in the functional component of NADPH oxidase, p47phox, thus suggesting that NADPH oxidase may mediate Zn2+-induced ROS accumulation. We suggest a new mechanism of cross talk between Zn2+ and mitochondrial ROS through positive feedback processes that eventually causes excessive free Zn2+ and ROS accumulations during the course of ischemic stress.

scholarly journals Protection of Transplanted Hematopoietic Stem Cells from Inflammatory Recipient Marrow Environment through Specific TNF-α Signal Blockade

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 1867-1867
Author(s):  
Takashi Ishida ◽  
Sachie Suzuki ◽  
Chen-Yi Lai ◽  
Masaaki Higashihara ◽  
Hiromitsu Nakauchi ◽  
...  
Keyword(s):  
Stem Cell ◽  
Nadph Oxidase ◽  
Conflicts Of Interest ◽  
Dependent Manner ◽  
Ros Production ◽  
Cell Protection ◽  
Hematopoietic Stem ◽  
Control Peptide ◽  
Ros Accumulation ◽  
Tnf Α

Abstract Introduction Hematopoietic stem cell (HSC) transplantation (HSCT) for hematological malignancy generally requires aggressive chemotherapy and irradiation, which exceed dose-limiting toxicity. These treatments are necessary to eradicate malignant cells and to create a niche for graft HSCs. However, as the intense preconditioning may also induce inflammation in recipient bone marrow (BM), transplanted HSCs thus should be inevitably exposed to this devastating environment. Whilst triggered inflammation within BM is hypothesized to deteriorate transplanted HSCs, how this BM environmental change affects graft HSCs remains largely unknown. We therefore sought to clarify how the pre-conditioned BM environment might affect donor HSCs, focusing especially on inflammatory effects and on developing protective measures against them. Methods & Results Our experiments revealed that total body irradiation (TBI) could induce local inflammation peaking around 2-3 days within marrow environment. In vivo exposure of HSCs to irradiated BM environment 2-3 days after TBI exhibited negative effects on HSC function. Then, we tested how the TBI-conditioned BM environment could affect donor HSCs. We first conducted comprehensive gene expression analysis on BM-resident stromal cells considered to constitute the HSC niche. We found that expression of 3 major inflammatory cytokines, IFN-γ, IL-1β, and TNF-α was induced after TBI treatments, thus we compared the effects of these cytokines by an in vitro HSC colony forming assay. Only TNF-α inhibited colony formation of HSCs in a dose dependent manner. We then sought to elucidate mechanisms by which TNF-α impaired HSCs' reconstitution abilities. Based on previous reports, TNF-α is a major stimulus of reactive oxygen species (ROS) through activating Nicotinamide Adenine Dinucleotidemono Phosphate Hydride (NADPH) oxidase. We therefore determined whether TNF-α stimulation produced excessive levels of ROS in highly purified murine HSPCs by culturing them with or without TNF-α for up to 48 hours and staining them with dichlorodihydrofluorescein (DCF) to quantify the accumulation of ROS. The addition of TNF-α was found to induce ROS production in HSPCs in a dose- and time-dependent manner. We next examined if overproduction of TNF-α-mediated ROS impaired reconstitution ability of HSCs; we compared reconstitution abilities of HSCs between groups, one exhibiting high levels of ROS and the other with low/medium levels of ROS. Transplantation experiments using flow-cytometry-sorted populations revealed the negative effect of high levels of ROS on HSCs, suggesting a causal role of elevated ROS levels in TNF-α-mediated impairment of stem cell ability. Accordingly we examined the hypothesis that specific inhibition of the TNF-α-ROS signaling could preserve graft HSCs' functions. The TNF receptor 1 (TNFR1) blocking peptide (PepTNFR1) that specifically blocks NADPH oxidase-mediated ROS production in cells after stimulation with TNF-α was evaluated for this purpose. Pre-incubation of murine HSPCs with PepTNFR1, but not with a scrambled control peptide, inhibited TNF-α-mediated ROS accumulation. Eventually, to determine if reduction of TNF-α-mediated ROS accumulation in graft-HSCs by PepTNFR1 could improve transplantation outcomes, we compared reconstitution kinetics of highly purified HSCs pre-incubated for 2 hours either with PepTNFR1 or a scrambled control peptide. Pre-treatment with PepTNFR1 successfully protected transplanted HSCs from an inflammatory BM environment, showing higher donor-cell chimerism in recipients of "protected" HSCs than in those of non-protected HSCs. Conclusion We here provide a proof of concept that stem cell protection measures through specific TNF-α signal blockade in the context of HSCT will eventually lead to better engraftment and reconstitution kinetics in transplantation, thereby ameliorating outcomes of HSCT. Disclosures No relevant conflicts of interest to declare.

Sign in / Sign up

Export Citation Format

Share Document