1.Isolation and determination of anamorph of Shiraia bambusicola.
Du WEN ; Yanfeng HAN ; Jiandong LIANG ; Zongqi LIANG
China Journal of Chinese Materia Medica 2009;34(13):1640-1642
OBJECTIVETo isolate and identify the anamorph of Shiraia bambusicola.
METHODFungus strains were isolated from mature ascospores and stroma. They were identified by means of morphological identification including colony and microscope characteristic, the molecular identification was done by 5.8S-ITS rDNA.
RESULTThe strains GZDXIFR-171 and GZDXIFR-181 were isolated and obtained with the separation of different methods, which had the same colony morphology. With the universal primers of the ITS1-5.8S rDNA-ITS2, the 5.8S-ITS rDNA sequence of GZDXIFR-171 and GZDXIFR-181 were obtained by the PCR amplification and sequencing. Compared with the published nucleotide sequence of 5.8S-ITS rDNA in NCBI (National Center for Biotechnology Information), GZDXIFR-171 and GZDXIFR-181 were highly identical with S. bambusicola.
CONCLUSIONThe isolated strains GZDXIFR-171 and GZDXIFR-181 were confirmed to be the true anamorph of S. bambusicola.
Ascomycota ; classification ; genetics ; isolation & purification ; DNA, Fungal ; genetics ; DNA, Ribosomal Spacer ; genetics ; Molecular Sequence Data ; Mycological Typing Techniques ; Phylogeny ; RNA, Ribosomal, 5.8S ; genetics
2.Isolation and identification of Sclerotinia stem rot causal pathogen in Arabidopsis thaliana.
Ai-Rong WANG ; Wen-Wei LIN ; Xiao-Ting CHEN ; Guo-Dong LU ; Jie ZHOU ; Zong-Hua WANG
Journal of Zhejiang University. Science. B 2008;9(10):818-822
A new stem rot disease is found to occur naturally on Arabidopsis plants in greenhouses of Fuzhou, China. In order to identify its pathogen, we conducted a series of fungal isolation and purification, plant reinoculation, and ascus and ascospore induction from the sclerotia. The isolate caused typical water-soaked lesions after reinoculation and produced sclerotia both on Arabidopsis plants and culture medium plates, and the sclerotia could be induced to produce discal apothecia and 8 binucleate ascospores per ascus. These disease symptom and fungal morphology data revealed that the fungus Sclerotinia sclerotiorum (Lib.) de Bary was the pathogen for Arabidopsis stem rot. To confirm this, we further amplified its large subunit ribosomal DNA (LSU rDNA) by polymerase chain reaction (PCR), and compared the sequence with the known LSU rDNA sequences in GenBank. The results show that the sequence shares the highest identities with the LSU rDNAs of different S. sclerotiorum strains. Taking all these data together, we concluded that the fungus that caused the Arabidopsis stem rot is S. sclerotiorum (Lib.) de Bary. This is the first report that Arabidopsis is naturally infected by S. sclerotiorum.
Arabidopsis
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microbiology
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Ascomycota
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classification
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genetics
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isolation & purification
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pathogenicity
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Base Sequence
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China
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DNA, Fungal
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genetics
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DNA, Ribosomal
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genetics
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Phylogeny
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Plant Diseases
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microbiology
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Plant Stems
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microbiology
3.Studies on purification and properties of antagonistic protein from bacteria SS02 of Paenibacillus daejeonensis.
Kai ZHU ; Xiao-Yu ZHANG ; Zhi REN ; Ding-Sheng FENG ; Yi-Ding WANG
Chinese Journal of Biotechnology 2007;23(4):681-685
The antifungal, anti-bacterical, anti-brine shrimp activities of SD22 isolated from Paenibacillus daejeonensis Bacteria SS02 were studied. The separation steps included ultracentrifugation, ultrafiltration and (NH4)2SO4 fractional precipitation, further purification was performed by SephadexG-75 and DEAE-32 chromatography. Its molecular weight determined by SDS-PAGE was 56.0 kD and its isoelectfic point was 6.4. SD22 was thermostable to some extent and stable to ultraviolet, but sensitive to some of the enzyme. SD22 could kill most pathogens from propagation, such as Rhizoctonia cerealis, Sclerotinia sclerotiorum Physalospora piricala, Trichodema viride, Gliocladium viride, Curvularia leaf spot, Fusarium sp, Fusarium head blight, Beauveria Bassiana, Escherichia coli, Staphylococcus aureus, Bacillus subtilis , Candidal vaginitis, Fusarium oxysporum Schl. emend. Sayder & Hansem et al. The results will be helpful to find out a novel antifungal protein.
Antifungal Agents
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isolation & purification
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pharmacology
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Ascomycota
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drug effects
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Bacillus
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chemistry
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genetics
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Bacterial Proteins
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isolation & purification
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pharmacology
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Gliocladium
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drug effects
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Plants
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microbiology
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Rhizoctonia
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drug effects
4.A new lactone derivative from plant endophytic fungus Periconia sp. F-31.
De-wu ZHANG ; Ji-mei LIU ; Ri-dao CHEN ; Min ZHANG ; Li-yan YU ; Jun WU ; Jun-gui DAI
China Journal of Chinese Materia Medica 2015;40(12):2349-2351
To investigate the secondary metabolites of endophytic fungi Pericinia sp. F-31. Column chromatography on silica gel, Sephadex LH-20 and semi-preparative HPLC were used to separate and purify the compounds. Two compounds were isolated from the fermentation broth of Periconia sp. Their structures were identified as 5-(1-hydroxyhexyl) -6-methyl-2H-pyran-2-one (1) and 2-(3-hydroxy-4-methylphenyl) -propanoic acid (2). Compound 1 was a new lactone compound, compound 2 was new natural product, and the NMR data of compound 2 was reported for the first time.
Annona
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microbiology
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Ascomycota
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chemistry
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genetics
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isolation & purification
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metabolism
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Drugs, Chinese Herbal
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chemistry
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isolation & purification
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metabolism
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Endophytes
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chemistry
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genetics
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isolation & purification
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metabolism
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Lactones
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chemistry
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isolation & purification
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metabolism
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Mass Spectrometry
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Molecular Structure
5.Isolation, idetification and anti-HIV-1 integrase activity of culturable endophytic fungi from Tibetan medicinal plant Phlomis younghusbandii Mukerjee.
Da-Wei ZHANG ; Ming-Ming ZHAO ; Juan CHEN ; Chao LI ; Shun-Xing GUO
Acta Pharmaceutica Sinica 2013;48(5):780-789
A total of 52 endophytic fungi were isolated from roots and stems of Tibetan medicinal plant Phlomis younghusbandii Mukerjee. These fungal isolates were molecularly identified based on ITS sequnces and 28S sequences distributed to 12 genera, including Phoma, Chaetosphaeronema, Fusarium and Leptosphaeria, etc. Among them, the dominant genus was Phoma. Extracts of all strains were evaluated for anti-HIV-1 integrase activity by using soluable integrase expressed in E. coli BL21 (DE3). The results showed that seven samples from five fungal endophytes PHY-24, PHY-38, PHY-40, PHY-51, PHY-53, which belonged to genus Chaetosphaeronema, inhibited strand transfer reaction catalyzed by HIV-1 integrase with IC50 values, of 6.60, 5.20, 2.86, 7.86, 4.47, 4.56 and 3.23 microg x mL(-1) respectively. In conclusion, the endophytic fungi of Phlomis younghusbandii Mukerjee are valuable for further screening anti-HIV-1 integrase agents.
Ascomycota
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enzymology
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isolation & purification
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Chaetomium
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enzymology
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isolation & purification
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Endophytes
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enzymology
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isolation & purification
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Escherichia coli
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enzymology
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HIV Integrase
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genetics
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metabolism
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HIV Integrase Inhibitors
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pharmacology
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Phlomis
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microbiology
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Phylogeny
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Plant Roots
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microbiology
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Plant Stems
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microbiology
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Plants, Medicinal
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microbiology
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Plasmids
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Recombinant Proteins
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genetics
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metabolism
6.Subcutaneous Phaeohyphomycosis Caused by Phaeoacremonium Species in a Kidney Transplant Patient: The First Case in Korea.
Jonghyeon CHOI ; Yangsoon LEE ; Hae Sun CHUNG ; Ja Seung KOO ; Dongeun YONG ; Yu Sun KIM ; Kyungwon LEE ; Yunsop CHONG
The Korean Journal of Laboratory Medicine 2011;31(3):201-204
Phaeohyphomycosis is a subcutaneous infection caused by dark pigmented fungi, including fungi of the species Phaeoacremonium, Alternaria, Exophiala, and Pyrenochaeta. In August 2005, a 54-yr-old man who had received a renal transplant 5 yr ago was admitted to our hospital with a subcutaneous mass on the third finger of the right hand; the mass had been present for several months. He had been receiving immunosuppressive agents for several years. He underwent excision of the mass, which was followed by aspiration of the wound for bacterial and fungal cultures. Many fungal hyphae were observed on the histology slide treated with periodic acid-Schiff stain. A few white waxy colonies with a woolly texture grew on the Sabouraud dextrose agar at 30degrees C and changed to dark brown in color. Nucleotide sequencing of internal transcribed spacer regions revealed 100% homology to the Phaeoacremonium aleophilum anamorph and Togninia minima teleomorph (514 bp/514 bp). The patient completely recovered after wide surgical excision. Here, we report the first case of phaeohyphomycosis caused by Phaeoacremonium species in a kidney transplant patient in Korea.
Antifungal Agents/therapeutic use
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Ascomycota/genetics/*isolation & purification
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Dermatomycoses/drug therapy/etiology/*microbiology
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Fingers/surgery
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Humans
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Immunosuppressive Agents/adverse effects
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*Kidney Transplantation
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Male
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Middle Aged
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Republic of Korea
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Sequence Analysis, DNA
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Subcutaneous Tissue/microbiology