1.Study of production of sesquiterpenes of Aquilaria senensis stimulated by Lasiodiplodia theobromae.
Xiao-Min HAN ; Liang LIANG ; Zheng ZHANG ; Xiu-Jin LI ; Yun YANG ; Hui MENG ; Zhi-Hui GAO ; Yan-Hong XU
China Journal of Chinese Materia Medica 2014;39(2):192-196
To investigate the mechanism of agarwood formation in Aquilaria sinensis induced by Lasiodiplodia theobromae, the fermentation liquor of L. theobromae was analyzed qualitatively and quantitatively by gas chromatography-mass spectrometry (GC-MS). JAs were detected in the fermentation liquor. The effect of the fermentation liquor on the abundance of sesquiterpenes in the callus of A. sinensis was analyzed by solid phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS). And the fermentation liquor stimulated alpha-guaiene, alpha-humulene and delta-guaiene biosynthesis in calli. It was inferred that L. theobromae produced JAs, which resulted in a significant increase of sesquiterpenes in A. sinensis.
Ascomycota
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physiology
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Fermentation
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Sesquiterpenes
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metabolism
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Thymelaeaceae
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metabolism
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microbiology
2.Agrobacterium tumefaciens-mediated transformation of Aureobasidium pullulans and high-efficient screening for polymalic acid producing strain.
Guangwei TU ; Yongkang WANG ; Jun FENG ; Xiaorong LI ; Meijin GUO ; Xiang ZOU
Chinese Journal of Biotechnology 2015;31(7):1063-1072
To develop a genetic transformation method of Aureobasidium pullulans and T-DNA insertion for high-efficient screening of polymalic acid (PMA) producing strain. Agrobacterium tumefaciens-AGL1, containing the selection genes encoding hygromycin B phosphotase or phosphinothricin acetyltranferase, was used to transform Aureobasidium pullulans CCTCC M2012223 and transformants were confirmed by colony PCR method. Transferred DNA (T-DNA) insertional mutants were cultured in microwell plate, and screened for high-titer PMA producing strain according to the pH response model. DNA walking was used to detect the insertion sites in the mutant. Results show that the selection markers could stably generated in the transformants, and 80 to 120 transformants could be found per 10(7) single cells. A high-titer PMA mutant H27 was obtained, giving a good PMA production caused by the disruption of phosphoglycerate mutase, that increased by 24.5% compared with the control. Agrobacterium tumefaciens-mediated transformation and high-efficient screening method were successfully developed, which will be helpful for genetic transformation of Aureobasidium pullulans and its functional genes discovery.
Agrobacterium tumefaciens
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Ascomycota
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genetics
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metabolism
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DNA, Bacterial
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Malates
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metabolism
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Polymerase Chain Reaction
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Polymers
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metabolism
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Transformation, Genetic
3.Protoplast isolation and transformation of Exserohilum turcicum.
Yong-Li ZHOU ; Chihiro TANAKA ; Satoshi MATSUSHIM ; Mitsuya TSUDA
Chinese Journal of Biotechnology 2003;19(3):364-367
Northern corn leaf blight, caused by the fungus Exserohirum turcicum Pass. (Leonard and Suggs), is one of the major diseases in most corn-growing areas of the world. Research on gene tagging of E. turcicum has been limited due to the lack of an efficient transformation system. Since E. turcicum produces and accumulates melamin in cell walls during vegetative growth, it is difficult to efficiently isolate its protoplast. To isolate the protoplast of this pathogen with a high frequency, the effects of cell wall degradation enzymes, including beta-1,3-glucanase (Fungase, Funcelase, Novozyme and Glucanex) and beta-glucuronidase (Driselase, Uskizyme and Kitalase), enzyme concentrations, combinations, strains and medium on the isolation frequency were tested. The isolation frequencies were high enough for transformation when the combinations of (Kitalase + Glucanex + Driselase), (Kitalase + Glucanex) or (Kitalase + Uskizyme) were used. Moreover, the isolation frequencies of protoplast were significantly affected by the cultural morphologies of strain and the growth stage of mycelia. Among the plasmids tested, only plasmid pAN71 is efficient for transformation of E. turcicum. This result will provide some useful information for gene tagging of E. turcicum and other species in Exserohirum.
Ascomycota
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cytology
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metabolism
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Cell Wall
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metabolism
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Fungal Proteins
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metabolism
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Glycoside Hydrolases
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metabolism
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Protoplasts
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cytology
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metabolism
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Transformation, Genetic
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genetics
4.Genetic algorithm for fermentation kinetics of submerged fermentation by Morchella.
Ying WANG ; Meizi PIAO ; Yonghai SUN
Chinese Journal of Biotechnology 2008;24(8):1454-1457
Fermentation kinetics is important for optimizing control and up-scaling fermentation process. We studied submerged fermentation kinetics of Morchella. Applying the genetic Algorithm in the Matlab software platform, we compared suitability of the Monod and Logistic models, both are commonly used in process of fungal growth, to describe Morchella growth kinetics. Meanwhile, we evaluated parameters involved in the models for Morchella growth, EPS production and substrate consumption. The results indicated that Logistic model fit better with the experimental data. The average error of this model was 5.8%. This kinetics model can be useful for optimizing and up-scaling fungal fermentation process.
Algorithms
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Ascomycota
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growth & development
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metabolism
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Culture Media
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Fermentation
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Kinetics
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Models, Biological
5.Secondary metabolites from Epicoccum nigrum 14one,an endophytic fungus isolated from plant Leptogium masiaticum.
Chao YUAN ; Yu-Hua GUO ; Ying-Bo ZHANG ; Xuan HU ; Dan WANG ; Fu-Lai YU ; Gang LI
China Journal of Chinese Materia Medica 2019;44(18):4021-4025
Phytochemical investigation of the culture of Epicoccum nigrum,an endolichenic fungus inhabiting Leptogium masiaticum,led to the isolation of 11 compounds. Based on NMR spectroscopy and HRESIMS data,their structures were determined as one alkaloid fusaricide( 1),and seven benzofuran derivatives including epicoccone( 2),4,6-dihydroxy-5-methoxy-7-methyl-1,3-dihydro isobenzofuran( 3),5-methyl-epicoccone B( 4),3,6,7-trihydroxy-5-methoxy-4-methylisobenzo furan-1( 3 H)-one( 5),3-methoxyepicoccone B( 6),2,3,4-trihydroxy-6-( hydroxymethyl)-5-methylbenzyl-alcohol( 7),and isoochracinic acid( 8),together with three epicoccolide analogs epicocconigrones A( 9),epicoccolide B( 10),and epicocconigrones B( 11). Compounds 1,9 and 10 showed potent microorganism inhibitory effects. These results indicated the potential perspective of this endophytic fungus as an eco-friendly biocide.
Ascomycota/chemistry*
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Endophytes/chemistry*
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Magnetic Resonance Spectroscopy
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Microbial Sensitivity Tests
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Secondary Metabolism
6.Study on breeding up high-yield strain of taxol by protoplast mutagensis.
Kai ZHAO ; Dong-Po ZHOU ; Wen-Xiang PING ; Jun LIU ; Xi MA ; Tao JIN
Chinese Journal of Biotechnology 2005;21(5):848-851
In order to obtain resistant mutants to nystatin, ultraviolet radiation and LiCl were used to mutagenize the protoplasts of taxol-producing fungi NCEU-1, and four positive mutants with high yield of taxol were screened out on nystatin flat. After further screening experiments on fermentation, a mutant strain--UL04-5 which was able to produce taxol with high yield and could be stably passed on in genetics was eventually found, it's ability to produce taxol was improved from 314.07 microg/L (strain NCEU-1) to 418.24 microg/L (strain U04-5).
Ascomycota
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drug effects
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genetics
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growth & development
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metabolism
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Fermentation
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Genetic Variation
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Mutagenesis
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Nystatin
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pharmacology
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Paclitaxel
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biosynthesis
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Protoplasts
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metabolism
7.Cluster analysis on hypocrellin A content in anamorphs of Shiraia bambusicola.
Wen DU ; Chunlong SUN ; Yanfeng HAN ; Jiandong LIANG ; Zongqi LIANG
China Journal of Chinese Materia Medica 2010;35(8):960-963
OBJECTIVEBy examining the colony growth rate, colony texture, substrate mycelium color and separation method of 40 anamorph strains of Shiraia bambusicola to investigate the relationship between these indexes and content of hypocrellin A.
METHODIn UV-spectrophotometry and microscopic observation were applied in the experiments. The results were analyzed by the clustering and variance analysis.
RESULTSPSS clustering analysis showed that the colony characters of S. bambusicola anamorph strains had significant differences with hypocrellin A content.
CONCLUSIONIn the tested conditions, substrate mycelium color of S. bambusicola anamophs has the most distinct correlation with the yield of hypocrellin A.
Ascomycota ; growth & development ; isolation & purification ; metabolism ; Cluster Analysis ; Mycelium ; growth & development ; isolation & purification ; metabolism ; Perylene ; analogs & derivatives ; metabolism ; Pigmentation ; Quinones ; metabolism ; Spectrophotometry, Ultraviolet
8.Two Cases of Mushroom Poisoning by Podostroma Cornu-Damae.
Jin Young AHN ; Soon Ja SEOK ; Je Eun SONG ; Jung Ho CHOI ; Sang Hoon HAN ; Jun Yong CHOI ; Chang Oh KIM ; Young Goo SONG ; June Myung KIM
Yonsei Medical Journal 2013;54(1):265-268
Podostroma cornu-damae is a rare fungus that houses a fatal toxin in its fruit body. In this case report, two patients collected and boiled the wild fungus in water, which they drank for one month. One patient died, presenting with desquamation of the palms and soles, pancytopenia, severe sepsis and multiple organ failure. The other patient recovered after one month of conservative care after admission. We found a piece of Podostroma cornu-damae in the remaining clusters of mushrooms. Mushroom poisoning by Podostroma cornu-damae has never been previously reported in Korea.
Agaricales/metabolism
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Anti-Bacterial Agents/therapeutic use
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Ascomycota/*metabolism
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Fatal Outcome
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Female
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Fever
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Hospitalization
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Humans
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Male
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Middle Aged
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Mushroom Poisoning/*diagnosis
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Pancytopenia/chemically induced
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Republic of Korea
9.Isolation and identification of endophytic fungi from Huperzia serrata and their metabolites' inhibitory activities against acetylcholinesterase and anti-inflammatory activities.
Bo-Wen QI ; Ting MO ; Xin ZHANG ; Ya-Ru YAN ; Xi-Ping XU ; Hong-Yun YANG ; Xiao-Hui WANG ; Jun LI ; She-Po SHI ; Xiao LIU
China Journal of Chinese Materia Medica 2019;44(15):3213-3220
A total of 27 endophytic fungal strains were isolated from Huperzia serrata,which were richly distributed in the stems and leaves while less distributed in roots. The 27 strains were identified by Internal Transcribed Spacer( ITS) r DNA molecular method and one of the strains belongs to Basidiomycota phylum,and other 26 stains belong to 26 species,9 general,6 families,5 orders,3 classes of Ascomycota Phylum. The dominant strains were Colletotrichum genus,belonging to Glomerellaceae family,Glomerellales order,Sordariomycetes class,Ascomycota Phylum,with the percentage of 48. 15%. The inhibitory activities of the crude extracts of 27 endophytic fungal strains against acetylcholinesterase( ACh E) and nitric oxide( NO) production were evaluated by Ellman's method and Griess method,respectively. Crude extracts of four fungi exhibited inhibitory activities against ACh E with an IC50 value of 42. 5-62. 4 mg·L~(-1),and some fungi's crude extracts were found to inhibit nitric oxide( NO) production in lipopolysaccharide( LPS)-activated RAW264. 7 macrophage cells with an IC50 value of 2. 2-51. 3 mg·L~(-1),which indicated that these fungi had potential anti-inflammatory activities.The chemical composition of the Et OAc extract of endophytic fungus HS21 was also analyzed by LCMS-IT-TOF. Seventeen compounds including six polyketides,four diphenyl ether derivatives and seven meroterpenoids were putatively identified.
Acetylcholinesterase
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Animals
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Anti-Inflammatory Agents
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isolation & purification
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pharmacology
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Ascomycota
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chemistry
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classification
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isolation & purification
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Cholinesterase Inhibitors
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isolation & purification
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metabolism
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Endophytes
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classification
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isolation & purification
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Huperzia
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microbiology
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Mice
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RAW 264.7 Cells
10.Study on difference of biological characteristics and resistance to powdery mildew of different Astragalus populations.
Jian-Jun CAO ; Zong-Suo LIANG ; Wei-Ling WANG ; Qi-Mei DUAN
China Journal of Chinese Materia Medica 2008;33(9):992-996
OBJECTIVETo study difference among populations which belong to Astragalus membranaceus and A. membranaceus var. mongholicus on morphology, habit, characteristics of physiology and resistance to powdery mildew, and classify them in order to provide theoretical basis for breeding and improving varieties.
METHODMorphology, habits, isozyme and soluble protein electrophoretograms were compared among the populations. They were categorized by cluster analysis based on those electrophoretograms. Different ability of resistance to powdery mildew was also studied through comparing disease indices among six populations.
RESULTThe results showed A. membranaceus var. mongholicus was distinctly different from A. membranaceus. There was a special type in colonies of A. membranaceus, which showed hairy upper epidermis of leaflets and later florescence.
CONCLUSIONAstragalus for medicine could be categorized in three types A. membranaceus var. mongholicus, A. membranaceus early florescence type and A. membranaceus late florescence type. Among them A. membranaceus var. mongholicus is most resistant to powdery mildew, while A. membranaceus is easily infected, and the early florescence type is even more easily infected.
Ascomycota ; pathogenicity ; Astragalus Plant ; classification ; growth & development ; metabolism ; microbiology ; Astragalus membranaceus ; growth & development ; metabolism ; microbiology ; Peroxidase ; metabolism ; Phylogeny ; Plant Diseases ; microbiology ; Plant Proteins ; metabolism ; Plants, Medicinal ; classification ; growth & development ; metabolism ; microbiology