• Title/Summary/Keyword: fungal metabolism

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Botrytis cinerea hypovirulent strain △BcSpd1 induced Panax ginseng defense

  • Shuhan Zhang;Junyou Han;Ning Liu;Jingyuan Sun;Huchen Chen;Jinglin Xia;Huiyan Ju;Shouan Liu
    • Journal of Ginseng Research
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    • v.47 no.6
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    • pp.773-783
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    • 2023
  • Background: Gray mold, caused by Botrytis cinerea, is one of the major fungal diseases in agriculture. Biological methods are preferred over chemical fungicides to control gray mold since they are less toxic to the environment and could induce the resistance to pathogens in plants. In this work, we try to understand if ginseng defense to B. cinerea could be induced by fungal hypovirulent strain △BcSpd1. BcSpd1 encodes Zn(II)2Cys6 transcription factor which regulates fungal pathogenicity and we recently reported △BcSpd1 mutants reduced fungal virulence. Methods: We performed transcriptomic analysis of the host to investigate the induced defense response of ginseng treated by B. cinerea △BcSpd1. The metabolites in ginseng flavonoids pathway were determined by UPLC-ESI-MS/MS and the antifungal activates were then performed. Results: We found that △BcSpd1 enhanced the ginseng defense response when applied to healthy ginseng leaves and further changed the metabolism of flavonoids. Compared with untreated plants, the application of △BcSpd1 on ginseng leaves significantly increased the accumulation of p-coumaric acid and myricetin, which could inhibit the fungal growth. Conclusion: B. cinerea △BcSpd1 could effectively induce the medicinal plant defense and is referred to as the biological control agent in ginseng disease management.

Induction of Fungal Secondary Metabolites by Co-Culture with Actinomycete Producing HDAC Inhibitor Trichostatins

  • Gwi Ja Hwang;Jongtae Roh;Sangkeun Son;Byeongsan Lee;Jun-Pil Jang;Jae-Seoun Hur;Young-Soo Hong;Jong Seog Ahn;Sung-Kyun Ko;Jae-Hyuk Jang
    • Journal of Microbiology and Biotechnology
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    • v.33 no.11
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    • pp.1437-1447
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    • 2023
  • A recently bioinformatic analysis of genomic sequences of fungi indicated that fungi are able to produce more secondary metabolites than expected. Despite their potency, many biosynthetic pathways are silent in the absence of specific culture conditions or chemical cues. To access cryptic metabolism, 108 fungal strains isolated from various sites were cultured with or without Streptomyces sp. 13F051 which mainly produces trichostatin analogues, followed by comparison of metabolic profiles using LC-MS. Among the 108 fungal strains, 14 produced secondary metabolites that were not recognized or were scarcely produced in mono-cultivation. Of these two fungal strains, Myrmecridium schulzeri 15F098 and Scleroconidioma sphagnicola 15S058 produced four new compounds (1-4) along with a known compound (5), demonstrating that all four compounds were produced by physical interaction with Streptomyces sp. 13F051. Bioactivity evaluation indicated that compounds 3-5 impede migration of MDA-MB-231 breast cancer cells.

Alteration of Gas Exchange in Rice Leaves Infected with Magnaporthe grisea

  • Yun, Sung-Chul;Kim, Pan-Gi;Park, Eun-Woo
    • The Plant Pathology Journal
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    • v.16 no.5
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    • pp.257-263
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    • 2000
  • Infection with rice blast fungus (Magnaporthe grisea) significantly reduced foliar net photosynthesis (A) of rice cultivars: Ilpoom, Hwasung, and Choochung in greenhouse experiments. By measuring the amount of diseased leaf area with a computer image analysis system, the relation between disease severity (DS) and net photosynthetic rate was curvilinearly correlated (r=0.679). Diseased leaves with 35% blast symptom can be predicted to have a 50% reduction of photosynthesis. The disease severity was linearly correlated (r=0.478) with total chlorophyll (chlorophyll a and chlorophyll b) per unit leaf area(TC). Light use efficiency was reduced by the fungal infection according to the light response curves. However, dark respiration (Rd) did not change after the fungal infection (p=0.526). Since the percent of reduction in photosynthesis greatly exceeded the percent of leaf area covered by blast lesions, loss of photosynthetic tissue on an area basis could not by itself account for the reduced photosynthesis. Quantitative photosynthetic reduction can be partially explained by decreasing TC, but cannot be explained by decreasing Rd. By photosynthesis (A)-internal CO$_2$ concentration (C$_i$ curve analysis, it was suggested that the fungal infection reduced ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity, ribulose-1,5-bisphosphate (RuBP) regeneration, and inorganic phosphate regeneration. Thus, the reduction of photosynthesis by blast infection was associated with decreased TC and biochemical capacity, which comprises all carbon metabolism after CO$_2$ enters through the stomata.

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Catabolic Pathway of Lignin Derived-Aromatic Compounds by Whole Cell of Phanerochaete chrysosporium (ATCC 20696) With Reducing Agent

  • Hong, Chang-Young;Kim, Seon-Hong;Park, Se-Yeong;Choi, June-Ho;Cho, Seong-Min;Kim, Myungkil;Choi, In-Gyu
    • Journal of the Korean Wood Science and Technology
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    • v.45 no.2
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    • pp.168-181
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    • 2017
  • Whole cell of Phanerochaete chrysosporium with reducing agent was applied to verify the degradation mechanism of aromatic compounds derived from lignin precisely. Unlike the free-reducing agent experiment, various degraded products of aromatic compounds were detected under the fungal treatment. Our results suggested that demethoxylation, $C_{\alpha}$ oxidation and ring cleavage of aromatic compounds occurred under the catabolic system of P. chrysosporium. After that, degraded products stimulated the primary metabolism of fungus, so succinic acid was ultimately main degradation product of lignin derived-aromatic compounds. Especially, hydroquinone was detected as final intermediate in the degradation of aromatics and production of succinic acid. In conclusions, P. chrysosporium has an unique catabolic metabolism related to the production of succinic acid from lignin derived-aromatic compounds, which was meaningful in terms of lignin valorization.

Metabolic Engineering of Saccharomyces cerevisiae for Redox Balance of Xylose Fermentation

  • Kim, Soo Rin;Jin, Yong-Su
    • Current Research on Agriculture and Life Sciences
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    • v.32 no.4
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    • pp.199-202
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    • 2014
  • The bioconversion of cellulosic biomass hydrolyzates consisting mainly of glucose and xylose requires the use of engineered Saccharomyces cerevisiae expressing a heterologous xylose pathway. However, there is concern that a fungal xylose pathway consisting of NADPH-specific xylose reductase (XR) and $NAD^+$-specific xylitol dehydrogenase (XDH) may result in a cellular redox imbalance. However, the glycerol biosynthesis and glycerol degradation pathways of S. cerevisiae, termed here as the glycerol cycle, has the potential to balance the cofactor requirements for xylose metabolism, as it produces NADPH by consuming NADH at the expense of one mole of ATP. Therefore, this study tested if the glycerol cycle could improve the xylose metabolism of engineered S. cerevisiae by cofactor balancing, as predicted by an in-silico analysis using elementary flux mode (EFM). When the GPD1 gene, the first step of the glycerol cycle, was overexpressed in the XR/XDH-expressing S. cerevisiae, the glycerol production significantly increased, while the xylitol and ethanol yields became negligible. The reduced xylitol yield suggests that enough $NAD^+$ was supplied for XDH by the glycerol cycle. However, the GPD1 overexpression completely shifted the carbon flux from ethanol to glycerol. Thus, moderate expression of GPD1 may be necessary to achieve improved ethanol production through the cofactor balancing.

Involvement of Mrs3/4 in Mitochondrial Iron Transport and Metabolism in Cryptococcus neoformans

  • Choi, Yoojeong;Do, Eunsoo;Hu, Guanggan;Caza, Melissa;Horianopoulos, Linda C.;Kronstad, James W.;Jung, Won Hee
    • Journal of Microbiology and Biotechnology
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    • v.30 no.8
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    • pp.1142-1148
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    • 2020
  • Mitochondria play a vital role in iron uptake and metabolism in pathogenic fungi, and also influence virulence and drug tolerance. However, the regulation of iron transport within the mitochondria of Cryptococcus neoformans, a causative agent of fungal meningoencephalitis in immunocompromised individuals, remains largely uncharacterized. In this study, we identified and functionally characterized Mrs3/4, a homolog of the Saccharomyces cerevisiae mitochondrial iron transporter, in C. neoformans var. grubii. A strain expressing an Mrs3/4-GFP fusion protein was generated, and the mitochondrial localization of the fusion protein was confirmed. Moreover, a mutant lacking the MRS3/4 gene was constructed; this mutant displayed significantly reduced mitochondrial iron and cellular heme accumulation. In addition, impaired mitochondrial iron-sulfur cluster metabolism and altered expression of genes required for iron uptake at the plasma membrane were observed in the mrs3/4 mutant, suggesting that Mrs3/4 is involved in iron import and metabolism in the mitochondria of C. neoformans. Using a murine model of cryptococcosis, we demonstrated that an mrs3/4 mutant is defective in survival and virulence. Taken together, our study suggests that Mrs3/4 is responsible for iron import in mitochondria and reveals a link between mitochondrial iron metabolism and the virulence of C. neoformans.

FUNGAL EXTRACELLULAR POLYSACCHARIDES INVOLVED IN RECYCLING OF METABOLITES AND OSMOTOLERANCE OF PENICILLIUM FELLUTANUM : APPLICATION OF $^{13}$ C-NMR SPECTROSCOPY FOR THE STUDY ON FUNGAL PHYSIOLOGY AND METABOLISM

  • Park, Yong-Il;Gander, John.-E.
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.04a
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    • pp.208-213
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    • 2000
  • Penicillium fellutanum produces a phosphorylated, choline-containing extracellular peptido-polysaccharide, peptidophosphogalactomannan (pPxGM) (8). The $\^$13/C-methyl labeled pPxGM ([methyl-$\^$13/C]pPxGM) was prepared from the cultures supplemented with L-[methyl-$\^$13/C]methionine or [2-$\^$13/C]glycine and was used as a probe to monitor the fate of phosphocholine in this polymer. Addition of purified [methyl-$\^$l3/C]pPxGM to growing cultures in low phosphate medium resulted in the disappearance of [methyl-$\^$13/C]phosphocholine and -N,N'-dimethyl-phosphoethanolamine from the added [methyl-$\^$13/C]pPxGM. Two $\^$l3/C-methyl-enriched cytoplasmic solutes, choline-O-sulfate and glycine betaine, were found in mycelial extracts, suggesting that phosphocholine-containing extracellular pPxGM of P.fellutanum is a precursor of intracellular choline-O-sulfate and glycine betaine and thus of phosphatydilcholine (l0). $\^$13/C-Methyl-labeled cells grown in 3 M NaCl-containing medium showed 2.6- and 22-fold more accumulation of $\^$13/C-methyl labeled choline-O-sulfate and glycine betaine, respectively, originated from the extracellular [$\^$13/C-methyl]pPxGM than those grown without added NaCl. The results suggest that, in addition to glycerol and erythritol, glycine betaine and choline-O-sulfate and thus choline are also osmoprotectants and hence that pPxGM is involved in osmotolerance of this fungus (11). Taken collectively, the $\^$l3/C- and $\^$31/P-NMR analyses of cytosolic solute pools and structural modulation of extracellular pPxGM corresponding to environmental stimuli in P. fellutanum, provided evidence that pPxGM is involved in cellular choline metabolism, osmotolerance, and recycling of metabolites.

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Profiles of Bacillus spp. Isolated from the Rhizosphere of Suaeda glauca and Their Potential to Promote Plant Growth and Suppress Fungal Phytopathogens

  • Lu, Ping;Jiang, Ke;Hao, Ya-Qiao;Chu, Wan-Ying;Xu, Yu-Dong;Yang, Jia-Yao;Chen, Jia-Le;Zeng, Guo-Hong;Gu, Zhou-Hang;Zhao, Hong-Xin
    • Journal of Microbiology and Biotechnology
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    • v.31 no.9
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    • pp.1231-1240
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    • 2021
  • Members of the genus Bacillus are known to play an important role in promoting plant growth and protecting plants against phytopathogenic microorganisms. In this study, 21 isolates of Bacillus spp. were obtained from the root micro-ecosystem of Suaeda glauca. Analysis of the 16S rRNA genes indicated that the isolates belong to the species Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus subtilis, Bacillus pumilus, Bacillus aryabhattai and Brevibacterium frigoritolerans. One of the interesting findings of this study is that the four strains B1, B5, B16 and B21 are dominant in rhizosphere soil. Based on gyrA, gyrB, and rpoB gene analyses, B1, B5, and B21 were identified as B. amyloliquefaciens and B16 was identified as B. velezensis. Estimation of antifungal activity showed that the isolate B1 had a significant inhibitory effect on Fusarium verticillioides, B5 and B16 on Colletotrichum capsici (syd.) Butl, and B21 on Rhizoctonia cerealis van der Hoeven. The four strains grew well in medium with 1-10% NaCl, a pH value of 5-8, and promoted the growth of Arabidopsis thaliana. Our results indicate that these strains may be promising agents for the biocontrol and promotion of plant growth and further study of the relevant bacteria will provide a useful reference for the development of microbial resources.

Identification of Genes Associated with Fumonisin Biosynthesis in Fusarium verticillioides via Proteomics and Quantitative Real-Time PCR

  • Choi, Yoon-E.;Shim, Won-Bo
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.648-657
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    • 2008
  • In this study, we used functional genomic strategies, proteomics and quantitative real-time (qRT)-PCR, to advance our understanding of genes associated with fumonisin production in the fungus Fusarium verticillioides. Earlier studies have demonstrated that deletion of the FCC1 gene, which encodes a C-type cyclin, leads to a drastic reduction in fumonisin production and conidiation in the mutant strain (FT536). The premise of our research was that comparative analysis of F. verticillioides wild-type and FT536 proteomes will reveal putative proteins, and ultimately corresponding genes, that are important for fumonisin biosynthesis. We isolated proteins that were significantly upregulated in either the wild type or FT536 via two-dimensional polyacrylamide gel electrophoresis, and subsequently obtained sequences by mass spectrometry. Homologs of identified proteins, e.g., carboxypeptidase, laccase, and nitrogen metabolite repression protein, are known to have functions involved in fungal secondary metabolism and development. We also identified gene sequences corresponding to the selected proteins and investigated their transcriptional profiles via quantitative real-time (qRT)-PCR in order to identify genes that show concomitant expression patterns during fumonisin biosynthesis. These genes can be selected as targets for functional analysis to further verify their roles in $FB_1$ biosynthesis.

The Effect of Estragole Identified and Extracts from Agastache rugosa O. Kuntze on the Fungal Growth and Metabolism (진균류의 증식과 대사에 미치는 방아(Agastache rugosa)추출물과 Estragole의 효과)

  • 박재림;박송희;김정옥;김수원;이수영
    • Journal of Food Hygiene and Safety
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    • v.12 no.1
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    • pp.63-70
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    • 1997
  • The extracts from Agastache rugosa O. Kuntze, their chloroform and hexane fractions, and estragole identified from hexane fraction were tested to investigate the effects on the growth and metabolic activities of several true fungi. The fungi used were: Aspergillus oryzae KFCC 890, Aspergillus niger KCCM 11240, Saccharomyces cerevisiae IAM 4597, Saccharomyces ellipsoideus PNU 2215. The growth of S. Cerevisiae by treatment of water extract(1%), hexane fraction (0.05%), and estragole (0.05%) were inhibited 93%, 50%, and 33% respectively, and S. ellipsoideus was also inhibited markedly with delaying the alg phase maximum 12 hrs. The growth of A. oryzae was inhibited by treatment of extracts and fractions. The echanol production by S. cerevisiae was increased more than two times in the highest value around 42 hrs incubation by water extract, but chloroform fraction inhibited its production. The glucoamylase actibities by A. niger were strongly inhibited by hexane and chloroform fractions (0.05%). The invertase activity by S. cerevisiae using estragole (0.05%) reached to 57.5% of control group. S. cerevisiae treated with the estragole was damaged the cell wall and cell membrane, leaked the protoplasm, and observed broken pieces of cell.

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