POSTHARVEST BIOLOGY AND TECHNOLOGY
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2022
·
IF 6.751
Okwong Oketch Reymick,Dazhao Liu,Yue Cheng,Qiuli Ouyang,Nengguo Tao
Cuminaldehyde (CA) has a strong antimicrobial activity against an array of pathogens. In this study, its growth inhibitory effect on a wild type strain (Pds01) and an imazalil-resistant strain (Pdw03) of Penicillium digitatum , the causative agent of postharvest green mold in citrus fruit was examined. CA strongly inhibited mycelial growth of both strains with minimum inhibitory concentration (MIC) of 0.3 mL L -1 and minimum fungicidal concentration (MFC) of 0.6 mL L -1 . In vivo assays revealed that wax + CA (WCA, 10 × MFC) treatment could significantly reduce green mold incidence in Ponkan fruit incubated with P. digitatum . After 5 d of incubation, the disease incidence of 10 × MFC WCA treated fruit was 65.0 ± 3.4 % for Pds01 and 50.0 ± 2.1 % for Pdw03 which were much lower than the corresponding values in the control samples. The antifungal activity was attributed to activation of massive oxidative stress , as evidenced by the accumulation of reactive oxygen species, increased lipid peroxidation , damage to plasma membrane permeability of hyphal cells, leakage of intracellular components and eventual cell death. The imazalil-resistant strain was more susceptible to the damaging effect of CA than the wild type strain. Collectively, our present results suggest that CA may potentially be a new antifungal drug molecule for the control of postharvest green mold in citrus fruit.
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高影响因子
Journal of Fungi
·
2021
·Vol. 7
·期 6
·432
·
IF 5.816
Qiuli OuYang,Yangmei Liu,Okwong Reymick Oketch,Miaoling Zhang,Xingfeng Shao,Nengguo Tao
Ergosterol (ERG) is a potential target for the development of antifungal agents againstPenicillium digitatum, the pathogen of green mold in citrus fruits. This study examined the mechanism by which citronellal, a typical terpenoid ofCymbopogon nardusessential oil, acts on ergosterol to exhibit its antifungal activity againstP. digitatum. We previously reported that citronellal inhibited the growth ofP. digitatumwith minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of 1.36 and 2.72 mg/mL, respectively. In citronellal-treated cells, the membrane integrity and ergosterol contents significantly decreased, whereas lanosterol, which serves as a precursor for ergosterol biosynthesis, massively accumulated. Addition of 150 mg/L of exogenous ergosterol decreased the inhibitory rate of citronellal, restoring the ergosterol content and hence the membrane structure to normal levels, and triggered expression of nearly all ERG genes. Based on our findings, we deduce that citronellal damages the cell membrane integrity ofP. digitatumby down-regulating the ERG genes responsible for conversion of lanosterol to ergosterol, the key downregulated gene beingERG3, due to the observed accumulation of ergosta-7,22-dienol.Keywords:citrus;pathogen;ergosterol;ERG genes;cell membrane integrity
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PubMed
高影响因子
LWT-FOOD SCIENCE AND TECHNOLOGY
·
2022
·
IF 6.056
Ruopeng Yang,Jinyu Miao,Zuyun Zhang,Chunpeng Wan,Liqiang Zou,Chuying Chen,Jinyin Chen
Eugenol (EUG), cinnamaldehyde (CA), and carvacrol (CAR) have been used to prevent fungal invasion due to their potent inhibitory activity. However, nanoemulsion can overcome these limitations because essential oils are hydrophobic, volatile, and have limited bioavailability. The previous study has shown that cytomembrane lipids play an essential role in fungal infection and pathogenesis. Changes in lipid domains have been shown to disrupt the orientation of virulence determinants and influence fungus pathogenicity. Understanding P. digitatum's lipid metabolic process and control are critical for understanding its pathogenesis. Untargeted lipidomics was used in this study to evaluate lipid change in P. digitatum exposed to compound EUG, CAR, and CA nanoemulsions. There were 511 lipid species and 28 classes discovered, with the substantially changed lipids belonging to eight classes can be employed as biomarkers for antifungal nanoemulsion exploitation. Our findings shed new light on nanoemulsion-induced lipid homeostasis disruption in P. digitatum .
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高影响因子
Applied Sciences-Basel
·
2019
·Vol. 9
·期 11
·2240
·
IF 2.217
Chunpeng Wan,Yuting Shen,Muhammad Farrukh Nisar,Wenwen Qi,Chuying Chen,Jinyin Chen
Carvacrol (5-Isopropyl-2-methylphenol), a volatile oil constituent, mainly exists in Labiaceae family plants. Carvacrol has long been studied for its natural antifungal potential and food preservative potential. However, its exact mode of action, especially againstPenicillium digitatum(P. digitatum), remains unexplored. Herein, a1H-NMR-based metabolomic technique was used to investigate the antifungal mechanism of carvacrol againstP. digitatum. The metabolomic profiling data showed that alanine, aspartate, glutamate, and glutathione metabolism were imbalanced in the fungal hyphae. A strong positive correlation was seen between aspartate, glutamate, alanine, and glutamine, with a negative correlation among glutathione and lactate. These metabolic changes revealed that carvacrol-induced oxidative stress had disturbed the energy production and amino acid metabolism ofP. digitatum. The current study will improve the understanding of the metabolic changes posed by plant-based fungicides in order to control citrus fruit green mold caused byP. digitatum. Moreover, the study will provide a certain experimental and theoretical basis for the development of novel citrus fruit preservatives.
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高影响因子
PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
·
2023
·
IF 4.7
Yonghua Zhang,Yuanzhen Tan,Qiuli OuYang,Bin Duan,Ziting Wang,Kuixian Meng,Xiaoli Tan,Nengguo Tao
The volatility of essential oils greatly limits their industrial applications. Here, we successfully prepared γ -cyclodextrin ( γ -CD) inclusion compounds ( γ -CDTL) containing thymol (TL) for the control of green mold caused by Penicillium digitatum ( P. digitatum ) in citrus fruit. In vitro experiment showed that the minimum fungicidal concentration (MFC) of γ -CDTL against the hyphae growth of P. digitatum was 2.0 g/L, and 8 × MFC treatment significantly reduced the occurrence of green mold in citrus fruit and had no adverse effect on fruit quality in vivo test compared to prochloraz. Scanning electron microscopy (SEM), x-ray diffraction (XRD), fourier transform-infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), physical properties and sustained release properties were also performed, results indicated that the hydrogen bonds between TL and γ -CD were the basis for the formation of γ -CDTL. We further investigated the inhibition mechanism of γ -CDTL. SEM and TEM experiments showed that γ -CDTL treatment caused severe damage to the hyphal morphology and cells in 30 min and disrupted the permeability of P. digitatum mycelial cell walls by increasing the chitinase activity, thus accelerating the leakage of intracellular lysates. However, the integrity of the cell membrane was obviously damaged only after 60 min of treatment. In conclusion, we prepared a novel inclusion complex γ -CDTL with obvious antifungal effects and preliminarily elucidated its inclusion mechanism and antifungal mechanism. γ -CDTL might be a potent alternative to chemical fungicides for controlling the postharvest decay of citrus.
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PubMed
高影响因子
Journal of Fungi
·
2022
·Vol. 8
·期 11
·1199
·
IF 5.724
Yonghua Zhang,Yuanzhen Tan,Okwong Oketch Reymick,Qiuli Ouyang,Nengguo Tao
In this study, aγ-cyclodextrin-cinnamaldehyde inclusion compound (γ-CDCL) was prepared to control green mold caused byPenicillium digitatum(P. digitatum) in citrus. The results showed that the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) ofγ-CDCL against the mycelial growth ofP. digitatumwere 2.0 g L−1and 4.0 g L−1, respectively. Simultaneously, eight × MFCγ-CDCL could effectively reduce the incidence of green mold in citrus fruit without impairment of the fruit qualities, meanwhile, eight × MFCγ-CDCL was comparable to Prochloraz in controlling fruit under natural storage conditions. The structure ofγ-CDCL was characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transform-infrared spectroscopy (FT-IR), and nuclear magnetic resonance (NMR) analyses. Results showed that the successful preparation ofγ-CDCL was due to the spatial interaction between H-4,8 of cinnamaldehyde and H-5′ ofγ-cyclodextrin. Meanwhile, the cell membrane permeability ofP. digitatumwas impaired byγ-CDCL through massive accumulation of reactive oxygen species, whereas the cell wall integrity was barely affected. These results indicated thatγ-CDCL might inhibit the growth ofP. digitatumthrough a membrane damage mechanism and it is a promising alternative to chemical fungicides in controlling the post-harvest citrus decay.Keywords:inclusion compounds;inhibitory activity;green mold;action mechanism;membrane permeability
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高影响因子
POSTHARVEST BIOLOGY AND TECHNOLOGY
·
2020
·
IF 4.303
Zehua Zhang,Shuhua Li,Dandan Sun,Ying Yang,Zhanhai Wei,Changlu Wang,Laifeng Lu
Penicillium digitatum acidifies the extracellular environments of fruit during decay development, this pH fluctuation decreased the efficacy of biological approaches under commercial conditions. Here, we explored the interaction between P. digitatum and an antagonistic yeast Rhodosporidium paludigenum to investigate potential mechanism behind the battle. P. digitatum was found in acidified citrus fruit tissue at pH 4.25 to 3.98 as mold infection progressed. Metabonomic data showed that gluconic, glucaric, and malic acids markedly increased upon infection at pathogen-fruit interaction sites. At concentrations > 1 × 10 6 cells mL −1 in liquid culture containing glucose or fructose instead of lactose, R. paludigenum could effectively control green mold. High concentrations of six different acids suppressed yeast growth even in the presence of sufficient glucose. Conversely, at ≤200 mmol L −1 , malic and gluconic acids promoted yeast growth in reverse. Cultivation of R. paludigenum with gluconic acid enhanced its ability to control green mold. This study provides new insights into the mechanisms by which extracellular acidification caused by acids produced by P. digitatum discriminates against competing microbes, thereby enhancing antagonistic yeast biocontrol of green mold in citrus.
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高影响因子
Frontiers in Nutrition
·
2021
·
IF 6.576
Yang Ruopeng,Chen Xiu,Huang Qiang,Chen Chuying,Rengasamy Kannan R. R.,Chen Jinyin,Wan Chunpeng (Craig)
Penicillium digitatum is the most severe pathogen that infects citrus fruits during storage. It can cause fruit rot and bring significant economic losses. The continuous use of fungicides has resulted in the emergence of drug-resistant strains. Consequently, there is a need to develop naturally and efficiently antifungal fungicides. Natural antimicrobial agents such as clove oil, cinnamon oil, and thyme oil can be extracted from different plant parts. They exhibited broad-spectrum antimicrobial properties and have great potential in the food industry. Here, we exploit a novel cinnamaldehyde (CA), eugenol (EUG), or carvacrol (CAR) combination antifungal therapy and formulate it into nanoemulsion form to overcome lower solubility and instability of essential oil. In this study, the antifungal activity evaluation and transcriptional profile of Penicillium digitatum exposed to compound nanoemulsion were evaluated. Results showed that compound nanoemulsion had a striking inhibitory effect on P. digitatum in a dose-dependent manner. According to RNA-seq analysis, there were 2,169 differentially expressed genes (DEGs) between control and nanoemulsion-treated samples, including 1,028 downregulated and 1,141 upregulated genes. Gene Ontology (GO) analysis indicated that the DEGs were mainly involved in intracellular organelle parts of cell component: cellular respiration, proton transmembrane transport of biological process, and guanyl nucleotide-binding molecular function. KEGG analysis revealed that metabolic pathway, biosynthesis of secondary metabolites, and glyoxylate and dicarboxylate metabolism were the most highly enriched pathways for these DEGs. Taken together, we can conclude the promising antifungal activity of nanoemulsion with multiple action sites against P. digitatum. These outcomes would deepen our knowledge of the inhibitory mechanism from molecular aspects and exploit naturally, efficiently, and harmlessly antifungal agents in the citrus postharvest industry.
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高影响因子
Frontiers in Microbiology
·
2020
·
IF 4.236
Zhang Lu-Lu,Huang Wen,Zhang Ying-Ying,Fan Gang,He Jin,Ren Jing-Nan,Li Zhi,Li Xiao,Pan Si-Yi
α-Terpineol has been widely used in daily chemical, pharmaceutical, food, and flavor industries due to its pleasant odor with high economic value and pharmacological action. Our previous study showed that Penicillium digitatum DSM 62840 was an efficient biocatalyst for the transformation of limonene to α-terpineol. Thus, it was meaningful to explore the genome features and the gene expression differences of strain DSM 62840 during limonene biotransformation, and the detailed bioconversion pathways. In this study, the functional genes related to limonene bioconversion were investigated using genome and transcriptome sequences analysis. The results showed that the P. digitatum DSM 62840 genome was estimated to be 29.09 Mb and it encoded 9,086 protein-encoding genes. The most annotated genes were associated to some protein metabolism and energy metabolism functions. When the threshold for differentially expressed genes (DEGs) was set at twofold ratio, a total of 4,128, and 4,148 DEGs were identified in P_L_12h (limonene-treated condition) compared with P_0h (blank) and P_12h (limonene-untreated blank), respectively. Among them, the expression levels of genes involved in the biosynthesis of secondary metabolites, energy metabolism and ATP-binding cassette (ABC) transporters were significantly altered during the biotransformation. And the reliability of these results was further confirmed by quantitative real-time polymerase chain reaction (RT-qPCR). Moreover, we found that the enzyme participated in limonene biotransformation was inducible. This enzyme was located in the microsome, and it was inhibited by cytochrome P450 inhibitors. This indicated that the cytochrome P450 may be responsible for the limonene bioconversion. Several differentially expressed cytochrome P450 genes were further identified, such as PDIDSM_85260 and PDIDSM_67430, which were significantly up-regulated with limonene treatment. These genes may be responsible for converting limonene to α-terpineol. Totally, the genomic and transcriptomic data could provide valuable information in the discovery of related-genes which was involved in limonene biotransformation, pathogenicity of fungi, and investigation of metabolites and biological pathways of strain DSM 62840.
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高影响因子
PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
·
2025
·
IF 1.9
Zehao Li,Zixuan Wang,Lulu Zhang,Feng Yingjie,Jinchu Yang,Haoliang Li,Weimin Song,Qiuling Wang,Yongfeng Yang,Zhenzhen Huang,Yongming Xu,Yufeng Fu,Meizhou Ding,Gang Fan,Jingnan Ren,Aiqun Yu,Jian Feng,Guijie Li,Linhua Huang,Yujiao Cheng
α-Terpineol, an abundant oxygenated monoterpene compound, shows a varied range of beneficial bioactivities. This study focused on the efficient extraction and purification of α-terpineol from the biotransformation of limonene using Penicillium digitatum DSM 62840 mutant (PdTP1-overexpressed OE2 strain). The α-terpineol was primarily distributed in the supernatant, with minimal association with the biomass. And the optimal extraction was achieved using ethyl acetate as the extractant, which can directly obtain the highest recovery of α-terpineol without sequential extractions. The equilibrium of mass transfer was quickly reached (≤20 s in vortex). Additionally, the purification method of α-terpineol using column chromatography was described, further improving the purity of product to 96.86% in gas chromatography. Structural identification of purified α-terpineol was confirmed using gas chromatography-mass spectrometry, Fourier-transform infrared spectroscopy, and nuclear magnetic resonance. This simplified and efficient strategy for the extraction and purification of α-terpineol not only provides a solid theoretical basis for the final step in the relevant study of microbial synthesis of α-terpineol but also is of great significance for its industrial application.
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高影响因子
Journal of Fungi
·
2025
·Vol. 11
·期 9
·612
·
IF 4
Mingchen Yang,Yonghua Zhang,Xiaoli Tan,Lu Li,Qiuli OuYang,Nengguo Tao
Green mold formed byPenicillium digitatumis a major disease that limits the yield and overall value of postharvest citrus fruits. The antifungal activity of sodium cuminate (SC) againstP. digitatumand the corresponding mechanism were explored in this research. The minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) of SC againstP. digitatumwere 0.4 and 0.8 g L−1, respectively. SC (8× MFC) reduced the incidence of disease in Ponkan fruits without compromising their quality. The results of CFW staining and extracellular alkaline phosphatase assays revealed that 1/2MIC SC for 30 min had no impact on the cell wall integrity ofP. digitatum. In contrast, 1/2MIC SC apparently destroyed cell membrane integrity, as shown by the increase in the content of reactive oxygen species (ROS), malondialdehyde, and H2O2. The addition of exogenous cysteine (Cys) or diphenyleneiodonium chloride (DPI) significantly mitigated the cytotoxic effects of SC. At the same time, mitochondrial membrane potential was significantly decreased by 1/2MIC SC, and the addition of exogenous Cys or DPI restored it to normal levels. In summary, the antifungal capacity of SC might be attributable to membrane damage inP. digitatumcaused by oxidative stress.
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PubMed
高影响因子
FOOD CONTROL
·
2025
·
IF 6.3
Shuqi Liu,Yuxin Jiang,Jiancheng Sun,Fan Yang,Yuqing Wang,Yongqing Lu,Weiqiang Lai,Chao-an Long
Perillaldehyde (PAE) is a green and safe active ingredient of natural plants. The inhibitory activity of PAE against the citrus postharvest pathogen Penicillium digitatum and its ability to control green mold were evaluated by in vitro and in vivo experiments. The results revealed that the minimum inhibitory concentration (MIC) and the minimum fungicidal concentration (MFC) of PAE against P. digitatum were 0.5 and 1 μL/mL, respectively. The results of conductivity, cell activity staining, and calcium fluoride white staining revealed that PAE damaged the membrane and cell wall integrity of P. digitatum . PAE promoted ROS accumulation in P. digitatum , as shown by 2',7'-Dichlorofluorescein diacetate (DCHF-DA) staining, which may induce cell death. To further investigate the molecular regulatory network of PAE in P. digitatum , we performed an RNA-sequencing analysis. The results displayed that differentially expressed genes were significantly enriched in the biogenetic pathway of ribosomes, and all of these genes were downregulated, suggesting that PAE treatment interfered with ribosome biogenesis in P. digitatum and inhibited the normal life processes of cells. Furthermore, PAE treatment reduced the incidence and severity of citrus green mold, and the overall control effect of the PAE treatment group with 5 × MFC was greater than that with 10 × MFC. Therefore, we determined the activity of enzymes related to fruit disease resistance. The results revealed that PAE treatment significantly increased the activities of CAT, POD, and PAL in citrus. These findings demonstrate that PAE prevents green mold disease by increasing the activity of citrus disease-resistant enzymes.
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高影响因子
危险品化学品经营许可证（带存储）