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22 June 2019, Volume 38 Issue 6
    

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  • MYCOSYSTEMA. 2019, 38(6): 730-761.
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  • Research papers
  • GUO Mei,ZHOU Ya-Nan,XIAO Qian,LI Bao-Du,LIANG Chen
    MYCOSYSTEMA. 2019, 38(6): 761-767. https://doi.org/10.13346/j.mycosystema.180329
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    As a new disease, Chinese cabbage root rot was found in Qingdao, Shandong Province. The lower part of taproot becomes rot, and the root vascular bundle turns brown and black. The overground plant grows slowly, dwarfs, and even dies. The pure isolate HMQAU170164 of the pathogen was obtained by tissue isolation method and single mycelium isolation method. Pathogenicity test by Koch?s postulate, observation of morphological and biological characteristics, and sequence analyses of ITS and coxⅠ, proved that the pathogen was Pythium brassicum. It is the first report of P. brassicum as the pathogen of Chinese cabbage root rot.

  • LIU Si-Rui,SONG Li-Sha,REN Jing,LI Zhong
    MYCOSYSTEMA. 2019, 38(6): 768-777. https://doi.org/10.13346/j.mycosystema.180333
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    The pathogen of a kind of leaf spot disease on Polygonatum sibiricum in Shibing County, Guizhou Province was isolated, and the pathogenicity and biological characteristics of the pathogen was investigated. Morphological characteristics and the gene analysis of rDNA-ITS, β-tubulin and tef1 sequence indicated that the pathogen was Pestalotiopsis trachicarpicola. The mycelium could grow in 15-30°C, and 28°C was the most suitable temperature. The optimum pH value was 5. Glucose was the best carbon source and yeast extract was the best nitrogen source for mycelial growth. PDA was the best culture medium. Light had less effect on mycelial growth. Mycelium lethal temperature was 45°C.

  • ZENG Chun-Han,DENG Jing-Jing,WANG Chuan-Hua
    MYCOSYSTEMA. 2019, 38(6): 778-791. https://doi.org/10.13346/j.mycosystema.190010
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    Some species of Armillaria are essential to the growth and development of Gastrodia elata. We proposed that there should be potential interspecific interactions between Armillaria rhiozomorph when different Armillaria species were cultivated together. Eight biological species of Armillaria including A. sinapina (CBS A), A. gallica (CBS B), A. stoyae (CBS D), A. borealis (CBS M), and four unnamed species (CBS F, CBS H, CBS J and CBS N) were selected as material for revealing the interspecific rhizomorph interactions. The whole rhizomorph growth rate and unilateral rhizomorph growth rate, and the number of rhizomorph growing tips per unit length (cm) under mixed culture of different species of Armillaria were investigated. The results showed that there were four interaction types among experimented species pairs. Pair of CBS D-CBS F was antagonistic to each other. CBS A showed unaffected or even favorable in mixed culture of CBS A-CBS H. That was the case with CBS J in CBS F-CBS J, CBS A in CBS A-CBS F, CBS A in CBS A-CBS N, CBS M in CBS F-CBS M, and CBS B in CBS B-CBS J. Some pairs demonstrated a synergistic growth to each other, e.g. pair of CBS A-CBS M. In some pairs, only one species grew favorably in the intermixed regions of mixed culture, e.g. CBS B in CBS A-CBS B, CBS J in CBS A-CBS J, CBS F in CBS B-CBS F, CBS H in CBS D-CBS H, CBS N in CBS D-CBS N and CBS F in CBS F-CBS M. This study provides reference for the identification of Armillaria species and the selection of Armillaria strains for cultivation of Gastrodia elata.

  • FENG Wan-Yan,HU Dan,FENG Jing-Wei,CHEN Yu-Qian,GUO Qi-Qiang,SUN Xue-Guang
    MYCOSYSTEMA. 2019, 38(6): 792-801. https://doi.org/10.13346/j.mycosystema.190031
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    Edible mycorrhizal fungi (EMF) are valuable forest resources, in terms of their ecological functions as symbiont of forest trees and the economic value of their edible basidiomata. They are supposed to play important roles in sustaining forest tree and ecosystem health, and contributing to economic development. In this study, basidiomata and mycorrhizas of two Suillus species associated with Pinus massoniana were morphologically observed, and a three-year field survey was conducted to clarify the phenology of the formation of basidiomata. It was proved that S. bovinus and S. luteus coexisted in forest of P. massoniana in Guizhou Province. The ectomycorrhizas of the two fungi formed in P. massoniana roots are usually coralloid in shape with dichotomous branches. The mycorrhizas formed by S. bovinus had smooth surface without emanating hyphae but with rhizomorph, while those formed by S. luteus had densely emanating hyphae but had no rhizomorph. The basidiomata formed by the two fungi have long fruiting season, considerable production, and staggered occurrence of fruiting peak. Compared with the precipitation, the temperature seemed to be more important in determining basidioma production.

  • SU Zhen-Zhu,FENG Xiao-Xiao,WANG Yan-Xin,LIU Xiao-Hong,ZHANG Chu-Long,LIN Fu-Cheng
    MYCOSYSTEMA. 2019, 38(6): 802-810. https://doi.org/10.13346/j.mycosystema.180308
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    The symbiosis of dark septate endophyte Falciphora oryzae isolated from Oryza granulata and rice was reestablished. The colonization pattern was monitored via inoculation of DsRED fluorescent protein-expressing F. oryzae on rice roots. To investigate the resistance induced by F. oryzae, the leaves of control and F. oryzae-colonized plants were inoculated with pathogenic Xanthomonas oryzae pv. oryzae (Xoo). The results showed that foliar infection of control plants with Xoo resulted in serious blight lesions. The disease ranged at rank 7 and 9 which accounted for 43.33% and 34.67%, respectively. The disease index was up to 79.26. Upon infestation with F. oryzae, leaves were protected from Xoo infection as evidenced by reductions in lesion size and severity, only with a few hypersensitive spots. It was clear that 8% of F. oryzae- colonized plants was not infected by Xoo. A large proportion of F. oryzae-infected plants ranged at rank 1, accounting for 72%, while its disease index was reduced to 15.26. The biocontrol efficiency of F. oryzae against Xoo was up to 80.75%. F. oryzae-inoculated roots showed 5.26-fold greater superoxide dismutase (SOD) activity in comparison with controls. Meanwhile, catalase (CAT) and peroxidase (POD) activities were elevated by 12.08- and 10.53-fold after infestation with F. oryzae, respectively. Due to the colonization of F. oryzae, the expressions of PR1a and PR1b were significantly up-regulated by 8.71- and 3.37-fold, while the expressions of AOS, OsSAUR2 and EL5 were significantly down-regulated by 0.28-, 0.57- and 0.65-fold, respectively. It was first reported to apply endophytic fungi isolated from wild rice against rice bacterial blight. It provided a novel measure for managing plant disease.

  • XU Ling-Ling,ZHAO Ming-Yang,LI Jing,LI Shu-Qi,GAO Li-Jun,ZHANG Yan,XU Jing
    MYCOSYSTEMA. 2019, 38(6): 811-821. https://doi.org/10.13346/j.mycosystema.180257
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    The roots of three Cypripedium species (C. tibeticum, C. flavum and C. bardolphianum) growing at different altitude (3 170-3 340m) in Huanglong valley, Sichuan Province of China, were sampled. Culture-independent method was used to obtain orchid mycorrhizal fungal (OrMF) ITS sequences from the roots. The correlation between vertical distribution of three Cyprepedium species and mycorrhizal fungal community composition was investigated. A total of 18 different operational taxonomic units (OTU) were detected. Among them, 14 OTUs were assigned to family Tulasnellaceae composing dominant taxa (99.6%), two OUTs were affiliated in the family Sebacinaceae, and the others in Thelephoraceae. With the incease of altitude, the OrMF diversity of C. tibeticum decreased, while that of the other two Cypripedium species was unchanged. There were no significant differences in OrMF community composition between the three Cypripedium species at different altitude. Different Cypripedium species growing in the same habitat showed significant differences in OrMF community composition, and the dominant mycorrhizal OTUs were also different.

  • SHAO Qiu-Yu,DONG Chun-Bo,ZHANG Zhi-Yuan,ZHENG Huan,ZOU Xiao,HAN Yan-Feng,LIANG Zong-Qi
    MYCOSYSTEMA. 2019, 38(6): 822-830. https://doi.org/10.13346/j.mycosystema.190023
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    Tree hole is one special habitat of the terrestrial ecosystems, being insufficiently attracted by investigators. Based on high-throughput sequencing, the fungal community composition in some soil samples in tree holes of Robinia pseudoacacia grown with different plants in Guizhou urban green field was investigated. The plants growing in the tree hole soil include Solanum cathayanum, Ligustrum lucidum, Chenopodium serotinum, Hydrangea chinensis, Cichorium endivia and Achyranthes bidentata. The dominant genus of fungi is Metarhizium, Trechispora, an unknown fungus, an unclassified genus in Boliniaceae, Guehomyces and an unclassified genus in Nectriaceae, with relative abundance of 57.67%, 42.96%, 30.60%, 86.45%, 33.15% and 12.26%, respectively. Some potential pathogens of insects and human beings and other unique fungi need further identification.

  • YAN Lian-Lian,XU Rui-Ping,DAI Sheng-Hong,WANG Gang-Zheng,BIAN Yin-Bing,GONG Yu-Hua,ZHOU Yan
    MYCOSYSTEMA. 2019, 38(6): 831-840. https://doi.org/10.13346/j.mycosystema.180309
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    Fungal laccase is an important polyphenol oxidase which plays important roles in fungal development. In the present study, the shiitake mushroom Lentinula edodes strain W1 was used as acceptor, and a laccase gene Lelcc1 was overexpressed under the control of actin promoter. The vector was transferred into L. edodes mycelia via Agrobacterium tumefaciens-mediated transformation method. Seven transformants were single-copy inserted, and qRT-PCR analysis showed that the expression of Lelcc1 in these seven transformants was 1.5-8 times higher than that in wild type strain W1. The genetic stability of these seven transformants were further analyzed, and three stable transformants were selected for phenotypic analysis including growth rate in different media, browning degree of substitution cultivation bags, and the ultrastructure of mycelial cells observed by transmission electron microscopy. The results indicated that the mycelium growth rate of three overexpression transformants growing in different medium was not significantly different from that of wild type strain. In sawdust-based cultivation, the transformants browned faster and the surface color was darker than W1. Significant differences were detected in the cell wall thickness and cell membrane morphology between the two overexpressed transformants and W1. These results indicated that Lelcc1 gene of L. edodes might be involved in the synthesis or accumulation of pigments during the process of brown film formation, and might also be related to the formation of cell wall.

  • WANG Pei,LIU Wen,ZHOU Yan,SHEN Xiang-Ling
    MYCOSYSTEMA. 2019, 38(6): 841-850. https://doi.org/10.13346/j.mycosystema.190024
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    Agaricus bisporus is an edible mushroom possessing very high economic value. Recent years, a high-efficiency functional genomic research method, Agrobacterium-mediated genetic transformation system, has been widely used for Agaricus bisporus, but the problems of low efficiency need to be improved. In this study, the method for improvement of transformation efficiency and finding the optimal genetic transformation conditions was proposed by detection of resistance of Agaricus bisporus to hygromycin. The results showed that hygromycin at concentration of 20mg/L could weaken the growth of mycelia, and at concentration of 50mg/L completely inhibit mycelia growth. The orthogonal test of L16(4 5) was carried out to analyse the effects of different strain of Agrobacterium, concentration of Agrobacterium, concentration of acetosyringone, culture duration of mycelia and infection duration of agrobacteria on optimizing the conditions of Agrobacterium-mediated genetic transformation in Agaricus bisporus. The results indicated that the culture duration of mycelia and selected strain of Agrobacterium were important factors influencing transformation efficiency. The highest genetic transformation efficiency was obtained by using Agrobacterium tumefaciens LBA4404 at concentration of OD600 0.8, under the acetosyringone concentration of 100mmol/L, mycelial culture duration of 40 days and infection duration of 10 minutes.

  • YE Li-Yun,MENG Guo-Liang,SONG Yan-Jiao,HAO Jin-Bin,LIU Xin-Rui,WU Xiao-Ping
    MYCOSYSTEMA. 2019, 38(6): 851-862. https://doi.org/10.13346/j.mycosystema.180325
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    Isonuclear alloplasmic strains N1M1 and N1M2 and isocytoplasmic allonuclear strains N1M1 and N2M1 of Pleurotus pulmonarius were obtained by hybridization technique. In order to analyze the effects of mitochondrial genes on mycelium and search into the interaction between mitochondrial genes and nuclear genes, mycelium growth rate, nutrient composition and the expression of several common nuclear and mitochondrial genes of N1M1, N1M2 and N2M1 were compared. Observation of mycelium growth showed that N1M1 and N1M2 had similar mycelial morphology and no significant difference in growth rate, however mycelial morphology and growth rate of strains N1M1 and N2M1 had significant difference, indicating that nuclear genes played a greater role than mitochondrial genes in mycelial morphology and growth rate. Further determination of the main nutrient components showed that the content of essential amino acids and total hydrolyzed amino acids in different strains was significantly different. The protein content of strain N1M2 was significantly higher than that of strain N1M1. The vitamin C content of strain N1M1 was 1.67 times higher than that of strain N1M2. The content of polysaccharide and protein of strain N2M1 was significantly higher than that of strain N1M1, while the content of iron and vitamin C was significantly lower than that of strain N1M1. Facts had proved that both nuclear genes and mitochondrial genes could affect the nutrient content of Pleurotus pulmonarius. The expression of seven common genes in nuclei of isonuclear alloplasmic strains N1M1 and N1M2 was detected. As a result, six of the genes in N1M2 was significantly higher in genes expression as compared with those in N1M1. It indicated that the different mitochondrial genes of Pleurotus pulmonarius could affect the expression of nuclear genes. There were significant differences in the expression of 14 common mitochondrial coding protein genes in isocytoplasmic allonuclear strains N1M1 and N2M1, indicating that the expression of mitochondrial genes significantly differs in accordance with different nuclear genes. In conclusion, mitochondrial and nuclear genes of Pleurotus pulmonarius can interact each other and act on life activities jointly.

  • XU Ying-Yin,WANG Yong,PENG Wei-Hong,TANG Jie,CHEN Ying,LIU Li-Xu,GAN Bing-Cheng
    MYCOSYSTEMA. 2019, 38(6): 863-874. https://doi.org/10.13346/j.mycosystema.180314
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    Single factor experiment, Plackett-Burman experiment and response surface Box-Behnkeh experiment were implemented for optimization of massive industrialized production of smaller-pelletized Morchella sextelata liquid spawn. The optimized medium contained 2.5% corn starch, 6% bran, 0.4% yeast powder, 2.5% glucose, 0.25% peptone, 0.1% KH2PO4, and 0.05% MgSO4. Under the conditions of inoculation volume of 10%, pH value of 5.5, rotation speed of 200-500r/min, and incubation temperature of 25°C, the biomass of Morchella sextelata reached 16mg/mL in 3 days. The diameter of mycelium pellet reduced to 1.5mm and the density reached 2 967/mL. The culture period from the stock culture to the applicable spawn was 13-19 days. Our research offered reference for Morchella sextelata liquid spawn industrialized production.

  • YAN Jun,BAO Hai-Ying,YANG Ming,ZHANG Jun,BAU Tolgor,LI Yu
    MYCOSYSTEMA. 2019, 38(6): 875-885. https://doi.org/10.13346/j.mycosystema.180332
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    The correlation between the content of nutrients in raw and unprocessed soybean and soybean meal and the yield and effective substance content of the fermentation product was analyzed. The results showed that calcium and daidzin in raw and unprocessed soybean were positively correlated with adenosine content in fermentation products; calcium, daidzin, daidzein and genistein in soybean meal were positively correlated with adenosine content in fermentation products; calcium, magnesium, daidzin, daidzein, genistein and total isoflavones in soybean meal were negatively correlated with adenine content in fermentation products; iron in soybean meal was positively correlated with dry weight of mycelium. It was concluded that nutritional component content of the soybean meal prepared from different soybean varieties had significant effects on the quality of fermentation products of Paecilomyces hepiali.

  • GAO Kun,Feng Jie,Yan Meng-Qiu,Tang Qing-jiu,Zhang Jin-song,Zhou Shuai,Yang Yan,Feng Na,Liu Yan-fang
    MYCOSYSTEMA. 2019, 38(6): 886-894. https://doi.org/10.13346/j.mycosystema.190015
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    Ten strains of Ganoderma lingzhi were cultured by liquid fermentation to compare the mycelial biomass, the yields and content of extracellular polysaccharides, and monosaccharide composition characteristics. Results indicated that the mycelial biomass increased with extending culture period, however, different strains showed a difference in the content of intracellular polysaccharide and the yields of extracellular polysaccharide. Extracellular crude polysaccharide yields of two strains, G0023 and G0160, reached 3.02 and 3.14g/L in seven days, and polysaccharide content was 84.11% and 91.03%, respectively. G0023 and G0160 could be regarded as excellent strains for producing extracellular polysaccharides. Analysis of ethanol fractional precipitated extracellular polysaccharides showed that the yields and polysaccharide content of 20E fractions (precipitated with 20% ethanol) were higher than those of 50E fractions (precipitated with 50% ethanol), indicating that the fermentation broth mainly contained high molecular weight polysaccharide fractions. 20E fractions were composed principally of glucose with small amount of xylose and mannose. 50E fractions mainly contained glucose and mannose together with small amount of xylose and galactose. Dependence of the apparent viscosity on the shear rate analysis for culture broth of different strains showed that culture broth exhibited non-Newtonian fluid and shear-thinning behavior. The values of apparent viscosity were positively correlated to the yields and polysaccharide content of 20E.

  • HOU Xiao-Xiao,LIN Fan,CHANG Ming-Chang,LIU Jing-Yu,MENG Jun-Long,WEI Yin
    MYCOSYSTEMA. 2019, 38(6): 895-906. https://doi.org/10.13346/j.mycosystema.190038
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    The effects of two key factors, ultrasound and deproteinization, on the quality of extracting polysaccharides (HEFPs) from Hericium erinaceus fruit-body were investigated using water extraction and alcohol precipitation method. The extracted samples were sequentially eluted by DEAE-52 ion exchange column and Sephacryl-S400 gel column. The result was that the extraction rate of HEFPs could be improved by ultrasound-assisted treatment, showing significant effect on the purity of HEFPs and increasing small peak areas of elution curves of partial HEFP-α component distilled by water and total HEFP-β component by 0.1mol/L NaCl, and presenting a shift in absorbance at the main peak eluted by Sephacryl-S400 gel column. Among three deproteinization methods, the method of combining ferrocyanide-zinc acetate with Sevage achieved the highest rate of protein removal, however, the polysaccharide retention rate was the lowest. The potassium ferrocyanide-zinc acetate method was simple and effective in protein removal and polysaccharide retention, but it could reduce the content and recovery rate of partial polysaccharide HEFP-β-2 component. The Sevage method had the lowest deproteinization efficiency and comparatively higher retention rate of HEFPs, showing highest main peak under elution of Sephacryl-S400 gel column, and it was suitable for protein removal when the bioactive components of polysaccharides were unknown. The data provide reference for extraction of macromolecular bioactive substances and further studies in exploring the related products of fruiting bodies of macrofungi.

  • ZHANG Meng-Li,CHAI Yi-Qiu,FANG Ming,ZHANG Si-Si,HAN Bao-Yu
    MYCOSYSTEMA. 2019, 38(6): 907-916. https://doi.org/10.13346/j.mycosystema.180299
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    Antihypertensive effect and intravascular transport mechanism of N 6-(2-hydroxyethyl) adenosine (HEA) isolated from Ophiocordyceps sobolifera, and interaction between HEA and human serum albumin (HSA) were studied. HEA with purity over 95% was obtained by using water-sinking extraction. Intraperitoneal injection of HEA at low (2.5mg/kg), medium (5mg/kg), and high (7.5mg/kg) concentration showed significant antihypertensive effect on the rats suffered from hypertension, and medium concentration particularly showed more effectiveness. The antihypertensive mechanism of HEA might be activation of adenosine A1 receptor. Van der Waals force, hydrogen bonding and hydrophobic force were the main forces in the binding of HEA to human serum albumin. HEA bound to human serum albumin at site I. The structure and microenvironment of the human serum albumin was slightly changed during the binding process. Molecular docking results indicated that Ser-192, Lys-195, Arg-257, Ser-287 and Arg-291 were important amino acid residues for the interaction between HEA and human serum albumin.

  • HAO Meng-Meng,WANG Jin-Yan,FENG Na,ZHANG Jing-Song,TANG Qing-Jiu,HAN Wei
    MYCOSYSTEMA. 2019, 38(6): 917-925. https://doi.org/10.13346/j.mycosystema.190028
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    To investigate the activities of triterpenoids with different polarity from the fruiting body of Ganoderma lingzhi, the total triterpenoid extract was separated into different polarity extracts by macroporous resin. Fingerprints and the content of triterpenoids were analyzed by HPLC. Anti-tumor and anti-inflammatory activities of different polarity extracts were compared. The results showed that the total triterpenoids can be separated by D-101 macroporous resin effectively into two polar parts, medium polar triterpenoid and low polar triterpenoid, with the content of (279.00±2.90)mg/g and (94.52±2.03)mg/g respectively. The anti-tumor activity of low polar triterpenoid is much stronger than that of medium polar triterpenoid. The IC50 of inhibition to K562 cells, SW620 cells, and L1210 cells are (74.12±1.94)μg/mL, (121.45±2.13)μg/mL and (13.52±1.13)μg/mL, respectively. The suppression effect of low polar triterpenoid on PMA-stimulated respiratory burst in RAW264.7 macrophages is also obviously stronger than that of medium polar triterpenoid. The comparison of the suppression effect between different triterpene monomers on PMA-stimulated respiratory burst in RAW264.7 macrophages shows that ganoderenic acid F, ganoderic acid DM and ganoderol B, with low polarity, have stronger anti-inflammatory activity than ganoderic acid C2 and ganoderic acid A with medium polarity. It is proved that the antitumor and anti-inflammatory activities in vitro of triterpenoids with different polarity from the fruiting bodies of Ganoderma lingzhi are different significantly. This study provides reference for improving quality standards of the Ganoderma lingzhi products.

  • YAN Jun,BAO Hai-Ying,YAN Fang,YANG Ming,BAU Tolgor,LI Yu
    MYCOSYSTEMA. 2019, 38(6): 926-937. https://doi.org/10.13346/j.mycosystema.190029
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    A mouse model of kidney and testis injury was established using adenine to explore the recovery effect of fermented powder products of Paecilomyces hepiali Cs-4 on adenine-induced kidney and testis damage. The mice were randomly divided into control group, model group, five treatment groups orally treated with different doses of fermented powder products of Paecilomyces hepiali Cs-4 (350mg/kg, 550mg/kg, 750mg/kg, 1 500mg/kg, 3 000mg/kg). The mice were treated daily for consecutive 21 days. The histopathological observation of the kidney and testis of each group of mice was carried out by using HE and Masson staining. The content of BUN, Cr, FSH and LH in blood serum were detected. In addition, the content of lactate dehydrogenase (LDH), acid phosphatase (ACP) and succinate dehydrogenase (SDH) in testis homogenate were detected. The results showed that the body weight of mice gradually recovered and increased in the experimental group treated with fermented powder products of Paecilomyces hepiali Cs-4, manifesting a recovery effect on damaged kidney and testis structure, and normalization of BUN, Cr, FSH, LH, ACP, SDH and LDH levels.

  • WANG Yi-Fei,YU Xiao-Dan,TIAN Xue-Mei,WANG Xue-Wei,WEI Yu-Lian,JIANG Ji-Hang,ZHOU Li-Wei
    MYCOSYSTEMA. 2019, 38(6): 938-950. https://doi.org/10.13346/j.mycosystema.190034
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    Sanghuangporus is a genus of important medicinal wood-inhabiting fungi; however, only a few species have been artificially cultivated in a small scale. The medicinal functions of most species are still imperfectly known. In this study, a recently newly described species S. quercicola and an unattractive species S. lonicericola are used as materials for evaluation of antioxidant activity of the fungi. Mycelial biomass and 12 indicative data (polysaccharide content, polyphenol content, flavonoid content, ascorbic acid content, DPPH radical scavenging activity, ABTS radical scavenging activity, total antioxidant capacity, superoxide dismutase activity, hydroxyl radical scavenging activity, superoxide anion scavenging activity, ferric ion reducing antioxidant power and ferrous ion chelating ability) of fermentation broths were tested under different duration of liquid cultivation. The result was that fermentation broth of the two species both showed strong antioxidant activities. S. quercicola produced higher content of polysaccharides, ascorbic acid and superoxide dismutase, while the content of polyphenols and flavonoids from S. lonicericola were higher. These results provide new data for medicinal value of Sanghuangporus.

  • DU Ping,He Shuang-hui,Qian Xi,Shen Di,Xu Jing-Han,Wu Yi,Si Jing
    MYCOSYSTEMA. 2019, 38(6): 951-969. https://doi.org/10.13346/j.mycosystema.190013
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    Liquid medium of Porodaedalea laricis was optimized by response surface methodology for increasing exopolysaccharide yield, and in vitro antioxidant activities of the preliminarily purified exopolysaccharide were evaluated. Plackett-Burman design, the steepest ascent path and Box-Behnken design were employed to establish mathematical model using exopolysaccharide yield as response value. The optimal medium for high production of P. laricis exopolysaccharide was composed of peeled potato 415.70g/L, peptone 10.80g/L, glucose 20.0g/L, Tween 80 7.27mL/L, KH2PO4 1.0g/L, MgSO4?7H2O 0.5g/L, CaCO3 0.57g/L, FeSO4?7H2O 0.3g/L, ZnSO4?7H2O 0.3g/L, vitamin B1 0.01g/L and rutin 6.09g/L. By use of the optimized medium, the exopolysaccharide yield was 3.07g/L, being closely consistent with the theoretical value of 3.06g/L predicted by the developed model, and this yield was 2.87-fold higher than that obtained in unoptimized medium. The result indicated that the response surface methodology is available for optimizing the medium composition with the purpose of increasing exopolysaccharide yield of P. laricis. The exopolysaccharide preliminarily purified by ethanol precipitation, dialysis and deproteination exhibited effective scavenging activities against hydroxyl, superoxide, DPPH and ABTS radicals.

  • HUA Ji,YAN Jun-Qing,HUO Guang-Hua,ZHANG Cheng,HU Dian-Ming,LIU Ying-Jie,WU Xin
    MYCOSYSTEMA. 2019, 38(6): 970-981. https://doi.org/10.13346/j.mycosystema.190042
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    The secondary metabolite of Gymnopus sp. 0612-9 has strong anti-Penicillium activity and can be used as microbial preservative for Citrus preservation, however, long fermentation cycle, higher energy consumption and lower efficiency are a bottleneck in production of the antifungal product. In this study, the medium and shake flask culture condition of Gymnopus sp. 0612-9 liquid inoculum were optimized, the optimum age and inoculation quantity of the optimized liquid inoculum were investigated, and the amplification culture of the inoculum culture was carried out by using 5L fermenter. The number of mycelial pellet and the dry weight of filtrated mycelium was determined. The accumulation of active substance Ⅱ was monitored by HPLC, and the activity of anti-Penicillium was evaluated by the Oxford Cup method. The results showed that the best carbon source was corn meal and maltose, and the best nitrogen source was peptone. The best culture medium for obtaining liquid fermented product consisted of corn meal 30g/L, maltose 10g/L, peptone 15g/L, KH2PO4 2g/L, and MgSO4·7H2O 1g/L. The best culture conditions included initial pH 5, inoculum (mycelial cake) 3×Ф7mm, liquid volume 100mL/250mL triangle bottle, temperature 28°C, and rotation speed 160r/min. The number of mycelial pellet before optimization is 46 pieces per 10mL, and the dry weight of mycelial pellet is 0.28g per 100mL. After optimization, the number of mycelial pellet is 985 pieces per 10mL, and the dry weight of mycelial pellet is 0.69g per 100mL, increasing by 21.4 times and 2.43 times respectively over pre-optimization. Subsequent fermentation was carried out using optimized liquid inoculum aged 9d and inoculative quantity of 7.5%. The optimized fermentation period in the fermenter was shortened from 10d to 5d, and the yield increased 8.28% over pre-optimization.

  • YU Dong-Sheng,WU Ying-Ying,FENG Ting,ZHANG Jing-Song,ZHANG He-Nan,WU Di,YANG Yan,LIU Xiao-Xiao,LIU Zhen-Dong,JIA Wei,WANG Wen-Han
    MYCOSYSTEMA. 2019, 38(6): 982-992. https://doi.org/10.13346/j.mycosystema.190022
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    The homogeneous polysaccharides FVPB2 from the fruiting body of Flammulina filiformis were separated and purified for the first time. The purified polysaccharide was a heteropolysaccharide consisting of D-glucose, D-galactose, L-fucose, and D-mannose, with an average molecular weight of about 1.5×10 4Da. The immunoactivity of FVPB2 in vitro was evaluated on spleen lymphocyte and bone marrow macrophage in C57BL/6 mice. In vitro immunomodulatory studies showed FVPB2 induced T lymphocyte activation, and stimulated TNF and IFN-γ secretion of T lymphocyte. FVPB2 also could induce NO production and IL-1β, IL-6 and TNF-α secretion of bone marrow macrophage. It was proved that the homogenous polysaccharide FVPB2 could enhance non‐specific immune response and specific immune responses in vitro.

  • LV Jun,YU Cun
    MYCOSYSTEMA. 2019, 38(6): 993-1002. https://doi.org/10.13346/j.mycosystema.190027
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    Irpex lacteus, a white rot fungus isolated from fallen wood, which can secrete laccase and manganese peroxidase, was tested for its ability to decolorize reactive black, reactive red, crystal violet, alizarin red and malachite green under solid conditions. The decolorizing conditions of alizarin red by I. lacteus were optimized by single factor and orthogonal test, and the toxicity changes of alizarin red before and after decolorization were measured with germination rate of three crops as indicators. The results showed that I. lacteus could decolorize all of the five direct dyes, especially alizarin red. The optimized conditions for alizarin red decolorization included medium pH value of 7.0, 20.0g/L of glucose, 0.66g/L of ammonium sulfate, and two pieces of inoculum cake (diameter 8.0mm) in 20.0mL alizarin red solution. Under such optimized conditions, the decolorization rate of alizarin red was 88.26% in 10d, 60.50% increase as compared with the decolorization rate before optimization. The toxicity of alizarin red dye before and after decolorization decreased progressively in order of dye stock solution > decolorized solution > solution treated with PDB medium. Alizarin red dye has certain toxicity, but I. lacteus can reduce its toxicity during decolorization. This study provides reference for application of I. lacteus in decolorization of dye wastewater such as alizarin red and reduction of toxicity of dye wastewater.

  • YANG Dan,WANG Zuo-Huan,JIANG Xiao-Wu,FANG Wei-Huan,SONG Hou-Hui,ZHANG Xian
    MYCOSYSTEMA. 2019, 38(6): 1003-1013. https://doi.org/10.13346/j.mycosystema.180330
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    Ochratoxin A (OTA) produced by Aspergillus and Penicillium species, is one of the most widespread toxic members of ochratoxins and has been shown to be nephrotoxic, teratogenic, carcinogenic and immunotoxic to several animal species. Rapid detection of OTA is important to prevent contaminated food from entering into the food chain. A rapid and sensitive fluorescent immunochromatographic assay (FICGA) based on the use of quantum dots (QDs) was prototyped and applied for detecting OTA in cereal samples. The detection signal was enhanced which in turn improved the sensitivity of the assay. The limit of detection of FICGA was 0.04ng/mL while the IC50 was 0.18ng/mL. The linear working range was 0.05-0.59ng/mL. The cross-reactivities to the OTA analogues, ochratoxin B and ochratoxin C, were 7.3% and 11.9% respectively. No cross-reactivity (<0.01%) was observed to other co-occurring mycotoxins (AFB1, ZEN, FB1 and DON). The recovery rates in spiked corn, wheat and soybean samples were 83.2%-117.8%, and the intra-day and inter-day relative standard deviations were <10%. Simultaneous analysis of commercial samples (corn, wheat, and feedstuff) showed a good correlation between FICGA and liquid chromatography tandem mass spectrometry (LC/MS-MS). Hence, this developed quantum dots-based FICGA shows great potential for rapid, sensitive, simple and cost-effective quantitative detection of OTA in food safety control.