Journal of Microbiology

Journal Articles

Identification and characterization of a novel light-induced promoter for recombinant protein production in Pleurotus ostreatus: Chaomin Yin , Xiuzhi Fan , Kun Ma , Zheya Chen , Defang Shi , Fen Yao , Hong Gao , Aimin Ma; J. Microbiol. 2020;58(1):39-45. Published online November 4, 2019; DOI: https://doi.org/10.1007/s12275-020-9230-4

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A lectin gene (plectin) with a high level of expression was previously identified by comparative transcriptome analysis of Pleurotus ostreatus. In this study, we cloned a 733-bp DNA fragment from the start codon of the plectin gene. Sequence analysis showed that the plectin promoter (Plp) region contained several eukaryotic transcription factor binding motifs, such as the TATA-box, four possible CAAT-box, light responsiveness motifs and MeJA-responsiveness motifs. To determine whether the Plp promoter was a light-regulated promoter, we constructed an expression vector with the fused egfp-hph fragment under the control of the Plp promoter and transformed P. ostreatus mycelia via Agrobacterium tumefaciens. PCR and Southern blot analyses confirmed the Plpegfp- hph fragment was integrated into the chromosomal DNA of transformants. qRT-PCR, egfp visualization, and intracellular egfp determination experiments showed the Plp promoter could be a light-induced promoter that may be suitable for P. ostreatus genetic engineering. This study lays the foundation for gene homologous expression in P. ostreatus.

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The GATA transcription factor BcWCL2 regulates citric acid secretion to maintain redox homeostasis and full virulence in Botrytis cinerea
Weiheng Ren, Chen Qian, Dandan Ren, Yunfei Cai, Zhaohui Deng, Ning Zhang, Congcong Wang, Yiwen Wang, Pinkuan Zhu, Ling Xu, Regine Kahmann
mBio.2024;[Epub] CrossRef

Promising cellulolytic fungi isolates for rice straw degradation: Diana Catalina Pedraza-Zapata , Andrea Melissa Sánchez-Garibello , Balkys Quevedo-Hidalgo , Nubia Moreno-Sarmiento , Ivonne Gutiérrez-Rojas; J. Microbiol. 2017;55(9):711-719. Published online September 2, 2017; DOI: https://doi.org/10.1007/s12275-017-6282-1

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Abstract

The objective of this study was to evaluate the potential of eight fungal isolates obtained from soils in rice crops for straw degradation in situ. From the initial eight isolates, Pleurotus ostreatus T1.1 and Penicillium sp. HC1 were selected for further characterization based on qualitative cellulolytic enzyme production and capacity to use rice straw as a sole carbon source. Subsequently, cellulolytic, xylanolytic, and lignolytic (Pleurotus ostreatus) activity on carboxymethyl cellulose, oat xylan, and rice straw with different nitrogen sources was evaluated. From the results obtained it was concluded both isolates are capable to produce enzymes necessary for rice straw degradation. However, their production is dependent upon carbon and nitrogen source. Last, it was established that Pleurotus ostreatus T1.1 and Penicillium sp. HC1 capability to colonize and mineralize rice straw, in mono-and co-culture, without affecting nitrogen soil content.

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Unravelling of cellulolytic fungal consortium from humus soil for efficient lignocellulosic waste degradation
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Sustainability in residue management: a review with special reference to Indian agriculture
Meenakshi Verma, Pooja Singh, Manikprabhu Dhanorkar
Paddy and Water Environment.2024; 22(1): 1. CrossRef
Crop residue heterogeneity: Decomposition by potential indigenous ligno-cellulolytic microbes and enzymatic profiling
Sandeep Sharma, Kailash Chand Kumawat, Paawan Kaur, Sukhjinder Kaur, Nihar Gupta
Current Research in Microbial Sciences.2024; 6: 100227. CrossRef
Recent developments in microbial degradation of crop residues: a comprehensive review
K. S. Sruthy, S. Puranik, V. Kumar, A. Kaushik, K. V. Vikram, M. Manoj, L. Shukla, S. K. Singh, A. Kumar
International Journal of Environmental Science and Technology.2024;[Epub] CrossRef
Fungal Saprotrophic Promotion and Plant Pathogenic Suppression under Ditch-Buried Straw Return with Appropriate Burial Amount and Depth
Jie Zhou, Yanling Li, Jiawen Lou, Yuekai Wang, Zhengrong Kan, Reinhard W. Neugschwandtner, Fengmin Li, Jian Liu, Ke Dong, Yaguang Xue, Haishui Yang, Lingling Shi
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Rice crop residue management by the microbial consortium for rapid decomposition of straw
Kunvar Gyanendra Kumar, Raja Husain, Anurag Mishra, Nitin Vikram, Devendra Kumar Dwivedi, Saurabh Pandey, Ashutosh Singh
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Shir Nee Ong, Chin Mei Lee
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Yao Jiang, Xinyue Du, Qianqian Xu, Chunhua Yin, Haiyang Zhang, Yang Liu, Xiaolu Liu, Hai Yan
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Research Support, Non-U.S. Gov'ts

Effect of Light and Reductones on Differentiation of Pleurotus ostreatus: Seung-Rock Lee , Woo-Jeong Joo , Yong-Un Baek , Inyoung Kim , Kee-Oh Chay , Seung-Hyun Cho , Seung-Jae Lee , Sa-Ouk Kang; J. Microbiol. 2011;49(1):71-77. Published online March 3, 2011; DOI: https://doi.org/10.1007/s12275-011-0507-5

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Abstract: Vegetative mycelia of Pleurotus ostreatus were differentiated into primordia and subsequently into fruit bodies in synthetic sucrose-asparagine medium when exposed to light at low temperature. During photomorphogenesis, L-ascorbic acid-like substances called reductones were produced. L-Ascorbic acid, D-erythroascorbic acid, 5-O-(α-D-glucopyranosyl)-D-erythroascorbic acid, 5-O-(α-D-xylopyranosyl)-D-erythroascorbic acid, 5-methyl-5-O-(α-D-glucopyranosyl)-D-erythroascorbic acid and 5-methyl-5-O-(α-D-xylopyranosyl)-D-erythroascorbic acid were accumulated initially in the illuminated mycelia before the initiation of fruiting. The content of glycosides of erythroascorbic acid and their methylated compounds increased again in the primordia and the fruit bodies. Exogenous L-ascorbic acid induced the formation of primordia from the mycelia in the dark in a dose-dependent manner. Thus, this suggests that these reductones might play a role in mediating the light stimulus in photomorphogenesis.

Intracellular Substrates of a Heme-Containing Ascorbate Oxidase in Pleurotus ostreatus: Seung-Rock Lee , Woo-Jeong Joo , Yong-Un Baek , Youn-Kyong Lee , Seong-Woon Yu , Yeon-Ran Kim , Kee-Oh Chay , Seung-Hyun Cho , Sa-Ouk Kang; J. Microbiol. 2009;47(2):178-186. Published online May 2, 2009; DOI: https://doi.org/10.1007/s12275-008-0307-8

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Abstract: A novel heme-containing ascorbate oxidase isolated from oyster mushroom, Pleurotus ostreatus, catalyzes oxidation of ascorbic acid (Kim et al., 1996). In this report, we describe the identification of intracellular substrates of the enzyme in the mushroom. Six compounds, which can serve as substrate of the heme-containing ascorbate oxidase, were identified as L-ascorbic acid, D-erythroascorbic acid, 5-O-(α-D-glucopyranosyl)-D-erythroascorbic acid, 5-O-(α-D-xylopyranosyl)-D-erythroascorbic acid, 5-methyl-5-O-(α-D-glucopyranosyl)-D-erythroascorbic acid, and 5-methyl-5-O-(α-D-xylopyranosyl)-D-erythroascorbic acid. All of the compounds were oxidized at a significant rate by the heme-containing ascorbate oxidase. Oxidation of the compounds produced equimolar amounts of hydrogen peroxide per mole of substrate.

A Bacterium Belonging to the Burkholderia cepacia Complex Associated with Pleurotus ostreatus: Ricardo Yara , Walter Maccheroni Junior , Jorge Horii , Joao Lucio Azevedo; J. Microbiol. 2006;44(3):263-268.; DOI: https://doi.org/2387 [pii]

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Abstract: Pleurotus ostreatus is a widely cultivated white-rot fungus. Owing to its considerable enzymatic versatility P. ostreatus has become the focus of increasing attention for its possible utility in biobleaching and bioremediation applications. Interactions between microorganisms can be an important factor in those processes. In this study, we describe the presence of a bacterial species associated with P. ostreatus strain G2. This bacterial species grew slowly (approximately 30 days) in theliquid and semi-solid media tested. When P. ostreatus was inoculated in solid media containing Tween 80 or Tween 20, bacterial microcolonies were detected proximal to the fungal colonies, and the relevant bacterium was identified via the analysis of a partial 16S rDNA sequence; it was determined to belong to the Burkholderia cepacia complex, but was not closely related to other fungus-isolated Burkholderiaceae. New specific primers were designed, and confirmed the presence of in vitro P. ostreatus cultures. This is the first time that a bacterial species belonging to the B. cepacia complex has been found associated with P. ostreatus.

Journal Article

Laccase Production Using Pleurotus ostreatus 1804 Immobilized on PUF Cubes in Batch and Packed Bed Reactors: Influence of Culture Conditions: K. Krishna Prasad , S. Venkata Mohan , Y. Vijaya Bhaskar , S. V. Ramanaiah , V. Lalit Babu , B. R. Pati , P. N. Sarma; J. Microbiol. 2005;43(3):301-307.; DOI: https://doi.org/2209 [pii]

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Abstract: The feasibility of laccase production by immobilization of Pleurotus ostreatus 1804 on polyurethane foam (PUF) cubes with respect to media composition was studied in both batch and reactor systems. Enhanced laccase yield was evidenced due to immobilization. A relatively high maximum laccase activity of 312.6 U was observed with immobilized mycelia in shake flasks compared to the maximum laccase activity of free mycelia (272.2 U). It is evident from this study that the culture conditions studied, i.e. biomass level, pH, substrate concentration, yeast extract concentration, Cu^2^+ concentration, and alcohol nature, showed significant influence on the laccase yield. Gel electrophoretic analysis showed the molecular weight of the laccase produced by immobilized P. ostreatus to be 66 kDa. The laccase yield was significantly higher and more rapid in the packed bed reactor than in the shake flask experiments. A maximum laccase yield of 392.9 U was observed within 144 h of the fermentation period with complete glucose depletion.

Infectious RNA viruses in the edible mushroom pleurotus spp: Park, Jeong Soo , Kim, Young Ho; J. Microbiol. 1996;34(1):61-67.

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Abstract: Double-stranded RNA (dsRNA) viruses and single-stranded RNA(ssRNA) viruses were detected in a strain of Pleurotus mushroom cultivated in a farm. Those fungal viruses were purified in the pH 6.0 or pH 7.2 using CsCI or Cs₂SO₄buoyant density centrifugation. Each viral particles were not completely separated at any trials. However, mushroom bacili-form virus contains a single major nucleic acid with 0.7 Kb ssRNA, which might code for 20 Kd viral capsid protein. The dsRNAs are encapsidatred into spherical-form viruses, whereas ssRNA viral genomes are encapsidated into two different sizes of bacili-form particles. A healthy-looking mushroom also contained some spherical-form viruses with dsRNAs. Laboratory strains of Pleurotus ostreatus and a cultivated strain of P. sajor-caju did not show any viral particles. Mushrooms with specific disease symptoms. however, contained at least four different sizes of spherical-form viruses. Thus, we concluded that a bacilli-form virus case a severe disease symptoms of abnormal on mushroom development.

Effect of culture parameters on the decolorization of remazol brilliant blue R by pleurotus ostreatus: Kim, Bok Sun , Ryu, Seong Joo , Shin, Kwang Soo; J. Microbiol. 1996;34(1):101-104.

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Abstract: The influences of culture parameters on the decolorization of anthron-type dye, Remazol brilliant blue R(RBBR) by Pleurotus ostreatus were studied in defined media. In the decolorization, 1-10 mM nutrient nitrogen and 40 mM glucose were effective whereas agitation and Tween 80 were not suitable. The decolorization occurred and the activity of extracellular peroxidase was detected during the stationary phase.

The Genetic Organization of the Linear Mitochondrial Plasmid mlp1 from Pleurotus ostreatus NFFA2: Kim, Eun Kyung , Youn, Hye Sook , Koo, Yong Bom , Roem Jung Hye; J. Microbiol. 1997;35(4):264-270.

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Abstract: The structure of plasmid mlp1, a linear 10.2kb mitochondrial plasmid of Pleurotus ostreatus NFF A2 was determined by restriction enzyme mapping and partial sequencing. The plasmid encodes at least two proteins; a putative RNA polymerase showing homology to yeast mitochondrial RNA polymerase and to viral-encoded RNA polymerases, and a putative DNA polymerase showing significant homology to the family B thpe DNA polymerases. It also contains terminal inverted repeat sequences at both ends which are longer than 274 bp. A 1.6 kb EcoRI restriction fragment of m1p1 containing the putative RNA polymerase gene did not hybridize to the nuclear or motochondrial genomes from P. ostreatus, suggesting that it may encode plasmidspecific RNA polymerase. The gene fragment also did not hybridize with the RNA polymerase gene (RPO41) from Saccaromyces cerevisiae. The relationship between genes in m1p1 and those in another linear plasmid pC1K1 of Claviceps purpurea was examined by DNA hybridization. The result indicates that the genes for DNA and RNA polymerases are not closely related with those in C. purpurea.

Purification and Characterization of Dehydroascorbate Reductase from Pleurotus ostreatus: Kim, Yeon Ran , Kang, Sa Ouk; J. Microbiol. 1998;36(3):164-170.

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Abstract: Dehydroascorbate reductase was purified 93-fold relative to the crude cell extracts from the cytosolic fraction of the mycelia of Pleurotus ostreatus with an overall yield of 5%. The molecular mass of the native enzyme determined by gel filtration chromatography was 86 kDa. Sodium dodecylsulfate-polyacrylamide gel electrophoresis revealed that the enzyme is a single polypeptide. Dehydroascorbate kreductase from P. ostreatus contained relatively quite a lot of lysine and a relatively small amount of glutamate/glutamine. The enzyme was optimally active at pH 7.5 and at 45℃. Apparent K_m values of dehydroascorbate reductase were 2.5 mM and 0.7 mM for dehydroascorbate and glutathione. The enzyme was significantly unstable under acidic and highly alkaline conditions. The absorption spectrum of the purified enzyme showed an unusual flavin peak, the result of which suggests that the enzyme might form flavin adduct.

Function of mORF1 Protein as a Terminal Recognition Factor for the Linear Mitochondrial Plasmid pMLP1 from Pleurotus ostreatus: Eun-Kyoung Kim , Jung-Hye Roe; J. Microbiol. 1999;37(4):229-233.

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Abstract: The mitochondrial plasmid pMLP1 from a white-rot fungus, Pleurotus ostreatus, is a double-stranded DNA containing 381 bp terminal inverted repeat (TIR) whose 5'-ends are covalently bound by terminal proteins. The plasmid contains two major open reading frames (ORFs), encoding putative DNA and RNA polymerases, and a minor ORF encoding a small, highly basic protein. To identify the DNA binding activity that recognizes the TIR region of pMLP1, gel retardation assays were performed with mitochondrial extracts. A specific protein binding to a region between 123 and 248 nt within TIR was observed. We examined whether the gene product of mORF1 bindes to this region specifically. E. coli cell extract which contains an overproduced mORF1 protein formed a complex specific to the region between 123 and 248 nt. Inclusion of mORF1 protein in the specific complex formed between P. ostreatus mitochondrial extract and TIR was confirmed by a supershift assay using polyclonal antibodies against the mORF1 protein. Our result suggest that the product of mORF1 may function as a terminal region recognition factor (TRF), recognizing an internal region in TIR.

Mutation Spectrum of Manganese (II) Peroxidase Gene in the Pleurotusostreatus Mutants Induced by Gamma Radiation: Hwa-Hyoung Chang^ , Young-Keun Lee^ , Jae-Sung Kim^ , Ki-Sung Lee^ , Kyu Seong Cho^; J. Microbiol. 2003;41(1):52-57.

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Abstract: The mutational spectra in the manganese (II) peroxidase gene (mnp) of the Pleurotus ostreatus mutants induced by gamma radiation (Co^60) give evidence to prove the effect of gamma radiation on the gene. mnp of each mutant was cloned, sequenced and analyzed. Among the 1941 base pairs of the sequenced region of the mnp genes of 4 mutants (PO-5, -6, -15 and -16), nine mutational hotspots on which the same base was mutated simultaneously were found, additionally 6 mutations were also found at different positions in the mnp gene. These mutation-spectra were predominantly A:T_G:C transitions (50.1%). By the analysis of putative amino acid sequences, PO-5 and PO-16 mutants have 3 and 1 mutated residues, respectively. Since the mutational spectra reported herein are specific to the mnp gene, we propose that the mutational hotspots for the gamma radiation could be in the gene(s) within cells.

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