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  • Technical Strategies in Cross Breeding of Edible Fungi(Ⅰ)
    BAO Dapeng
    2024, 31(06): 1-16. https://doi.org/10.16488/i.cnki.1005-9873.2024.06.001
    Abstract ( ) PDF ( ) Knowledge map Save
    Cross breeding is an important approach in edible fungi breeding, but the practice of cross breeding involves many uncertainties. The author raises the notion of establishing a rational cross breeding system for edible fungi, and analyzes the technical strategies in this system from the following nine aspects: breeding goals, crossing concepts, core parental strains, crossing volume, crossing generations, variety pedigrees, variety series, crossing paths, and germplasm resources. A clear breeding goal is the No.1 element of edible fungi breeding practice. Accurate understanding and correct application of cross breeding concepts are crucial for developing new varieties. The orderly inheritance of variety pedigrees is not only the result of extensive breeding practices but also the cornerstone of rational cross breeding. Core parental strains serve as the core material in cross breeding and thus are the key material element of rational cross breeding. Appropriate crossing volume and reasonable crossing combinations can increase the probability of obtaining superior hybrid offspring. Reasonable design of hybrid generations and the accurate judgment of crossing paths need breeder’s deep understanding and precise control of breeding goals and breeding materials, facilitating enhancement of superior agronomic traits through genetic approaches such as gene recombination and reduction of genetic instability of hybrid offspring. A rich and diverse collection of varieties is the embodiment of fruitful achievements in cross breeding of edible fungi, showing adaptation to diverse cultivation environments and broad market demands. Rich and diverse germplasm resources are important materials for cross breeding, and their genetic diversity should be explored with emphasis. A rational cross breeding system for edible fungi not only can increase the certainty of breeding practices and the predictability of breeding results, but also provides a crucial reference for evaluating the effectiveness, outcomes and quality of cross breeding practices.
  • Identification and Analysis of Lignin-Responsive Transcription Factors in Ganoderma lucidum
    WANG Lining, WANG Qingfu
    2024, 31(06): 17-28. https://doi.org/10.16488/i.cnki.1005-9873.2024.06.002
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    Lignin-responsive transcription factors in Ganoderma lucidum were identified through comparison of the transcriptome data of G. lucidum GL0102 mycelia cultured with glucose as the carbon source versus lignin as the carbon source. Differentially expressed transcription factors (TFs) were identified, validated, and then determined for expression levels under different agricultural and forestry wastes (bagasse, rice straw, wheat straw, oak sawdust, spruce sawdust, and pine sawdust). Three interference strains of the differential transcription factor chr8g0158731 were constructed, and then determined for mycelial growth rate under different carbon sources and lignin utilization. The results showed that there were 26 up-regulated and 40 down-regulated TFs in the transcriptome of mycelia cultured on lignin compared with that of mycelia cultured on glucose. When cultured on the agricultural and forestry wastes, seven of the up-regulated TFs showed a high expression level in mycelia, and six of the down-regulated TFs showed a low expression level. When cultured with glucose as the carbon source, the mycelial growth rates of the three interference strains were not significantly different from that of the wild-type strain. When cultured on lignin and the six agricultural and forestry wastes, the mycelial growth rates of the three interference strains were greater than that of the wild-type strain, and the expression level of chr8g0158731 in the the three interference strains was lower than that in the wild-type strain. When cultured with lignin as the carbon source, all three interference strains degraded lignin to a greater extent than the wild-type strain. The results provided candidate genes for subsequent lignin degradation studies.
  • Effect of Cultivation Temperature and Time on Flammulina filiformis Yield in Factory Production
    LIU Yalan, LI Hewen, CAI Weiming, FENG Zhan, SHEN Yingyue, YANG Ruiheng, QIU Ya, LI Na
    2024, 31(06): 29-33. https://doi.org/10.16488/i.cnki.1005-9873.2024.06.003
    Abstract ( ) PDF ( ) Knowledge map Save
    The effect of different cultivation temperatures, LT (15 ℃ for the first 6 d and then 13 ℃ for the rest time), MT (16 ℃ for the first 6 d and then 14 ℃ for the rest time) and HT (17 ℃ for the first 6 d and then 15 ℃ for the rest time), and cultivation time (20-27 d) on Flammulina filiformis yield was studied. The results showed that the temperature of culture substrate inside culture bottles gradually increased as the mycelium cultivation time extended, and the maximum temperature difference between the inside and outside of culture bottles reached 4 ℃. The average substrate temperature of HT was 3.16 ℃ higher than that of LT. Fruiting bodies started to emerge after mycelium cultivation for 22-27 d, and the yield of MT with mycelium cultivation for 22 d was the highest, reaching 472.13 g per bottle (1 200 mL). Based on the accumulated daily culture temperature of MT, the cultivation time of LT an HT was prolonged and shortened respectively to keep their accumulated daily temperature consistent with that of MT, and the yields of LT (23 d, 465.69 g per bottle) and HT (20 d, 393.94 g per bottle) were lower than that of MT (22 d, 472.13 g per bottle). In summary, cultivation under 16 ℃ during day 1 to day 6 and then 14 ℃ from day 7 to day 22 improved the yield of F. filiformis.
  • Fruiting Characteristics after Pairing of Single Mating Type Strains of Cordyceps militaris
    LI Xiu’e, QIU Wenxu, SA Fangping, SONG Guoyue, LI Weihuan, CHENG Xianhao
    2024, 31(06): 34-42. https://doi.org/10.16488/i.cnki.1005-9873.2024.06.004
    Abstract ( ) PDF ( ) Knowledge map Save
    Cordyceps militaris strains D1, D2, S1, S2 and S3 were cultivated and then determined for their mating type. Monoconidial strains were isolated from the five strains and determined for mating type. Paired crossing between different mating types of monoconidial strains isolated from S1, S2 and S3 were carried out. The MAT1-2 type monoconidial strains of S1, S2 and S3 were also crossed with monoconidial strains of D1 and D2. Superior hybrid strains were then screened out of the crossing progenies. The results showed that D1, D2, S1, S2 and S3 fruited stably. Fruiting bodies of D1 and D2 were smooth, lack of perithecium, and exclusively MAT1-1 mating type. On the other hand, fruiting bodies of S1, S2 and S3 had abundant perithecium on the surface, and contained either MAT1-1 or MAT1-2 mating type gene. All paired-crossing progenies developed perithecium on the surface of their fruiting bodies, and produced ascospores. After screening for agronomic traits, superior strains D1S1-38, D2S1-15, D2S1-43 (MAT1-2), D2S1-35 and D2S1-44 (MAT1-1) were selected. These findings provided a reference for genetic breeding of C. militaris.
  • Effects of Light Quality on Mycelial Growth and Extracellular Enzyme Activity of Pleurotus ostreatus
    SONG Zhibo, DUAN Yakui, HU Sujuan, YUAN Ruiqi, LU Guangfan, WANG Yan, KONG Weiwei
    2024, 31(06): 43-49. https://doi.org/10.16488/i.cnki.1005-9873.2024.06.005
    Abstract ( ) PDF ( ) Knowledge map Save
    Mycelia of Pleurotus ostreatus were cultivated under darkness, blue light, and red light, respectively. Then mycelia of different light quality groups were measured for growth parameters, activities of lignocellulose-degrading enzymes, i.e., laccase, cellulase, and xylanase, and relative expression of lignocellulose-degrading enzyme genes, including laccase gene (LAC), exoglucanase gene (EXO), endoglucanase gene (END), and glucosidase gene (GLU). The results showed that red light resulted in the highest mycelial growth rate and biomass. After cultivation for 3 d, darkness resulted in the highest activities of laccase, cellulase and xylanase, and the highest expression levels of LAC and END. After cultivation for 7 d, red light resulted in the highest activities of laccase and cellulase, and the highest expression levels of EXO and END, and mycelia cultivated under blue light showed the highest xylanase activity and GLU expression. After cultivation for 10 d, mycelia under darkness showed the highest laccase activity and the highest expression level of GLU; the activities of cellulase and xylanase under darkness and red light treatments were significantly higher than those under blue light treatment; red light resulted in the highest expression levels of LAC and END.
  • Ameliorating Effect of Sparassis latifolia Polysaccharide on Hippocampal Injury in Lead-Exposed Mice
    WU Rui, FENG Rui, NIU Xukai, XU Mengyan, HAN Xiaoyue, YUN Shaojun, FENG Cuiping, CAO Jinling
    2024, 31(06): 50-58. https://doi.org/10.16488/i.cnki.1005-9873.2024.06.006
    Abstract ( ) PDF ( ) Knowledge map Save
    Kunming mice were randomly divided into blank control (NC) group, lead exposure (PB) group, Sparassis latifolia polysaccharide (SLP) group, and lead exposure (Co-PB) group co-housed with SLP treatment (Co-SLP) group. Except NC group, mice in the rest groups were given 1 g·L-1 lead acetate solution as drinking water, and mice in SLP and Co-SLP groups were also administered 400 mg·kg-1 SLP by gavage. After 8 weeks, mice in different groups were measured for lead content in hippocampus by graphite furnace atomic absorption spectrometry, observed for hippocampal tissue structure by hematoxylin-eosin staining, determined for contents of malondialdehyde (MDA) and glutathione (GSH), and determined for activities of total superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) in hippocampus. Real-time quantitative PCR was used to detect relative expression of inflammation-related genes IL-4, IL-6, IL-10, IL-1β, TNF-α, TGF-β, and autophagy related genes Beclin-1, LC3-I, LC3-II, Lamp2, P62 and Atg5 in hippocampus. The results showed that the hippocampal lead contents of SLP, Co-SLP and Co-PB were significantly lower than that of PB. There were significant increases of CAT activity and GPx activity in SLP mice and Co-SLP mice compared with PB mice. The SOD activity and GSH content of SLP, Co-SLP and Co-PB groups were also increased significantly. On the other hand, the contents of MDA in SLP, Co-SLP and Co-PB groups decreased significantly compared with that in PB group. Compared with PB mice, mice treated with SLP showed significantly decreased expression of IL-1β, IL-6 TNF-α, LC3II, Lamp2, Beclin1, LC3-I and Atg5, and significantly increased expression of IL-4, TGF-β and IL-10. Compared with PB mice, Co-SLP mice showed significantly decreased expression of IL-6, TNF-α, LC3 II, Lamp2, Beclin1, LC3-I and Atg5, and significantly increased expression of IL-4, TGF-β, IL-10 and P62. Compared with PB mice, Co-PB mice showed significantly decreased expression of TNF-α, LC3-I and Atg5, and significantly increased expression of TGF-β and IL-10. The results provided a reference for ameliorating lead injury by S. latifolia polysaccharide.
  • Inhibition Mechanism of Ethyl Acetate Extract of Schizophyllum commune on Fusarium oxysporum
    LI Fengfu, PAN Meichen, HU Huiyue, WANG Liying, LI Changtian
    2024, 31(06): 59-68. https://doi.org/10.16488/i.cnki.1005-9873.2024.06.007
    Abstract ( ) PDF ( ) Knowledge map Save
    Fusarium oxysporum is a major pathogen causing root rot of Panax ginseng. To investigate the antifungal mechanism of ethyl acetate extract from Schizophyllum commune against Fusarium oxysporum, the pathogen was cultured with the extract, and then observed for hyphal growth, spore germination, and microscopic hyphal morphology; determined for cell membrane permeability by measuring electrical conductivity and nucleic acid content; and also determined for the degree of membrane lipid peroxidation by measuring malondialdehyde (MDA) content, hydrogen peroxide content, superoxide dismutase activity, peroxidase activity and catalase activity. The results showed that the S. commune ethyl acetate extract significantly inhibited the hyphal growth and spore germination of F. oxysporum, with an inhibition rate of 56.75% after cultivation at the minimum inhibitory mass concentration (5.00 mg·mL-1) for 7 d, and an inhibition rate of 100% after cultivation at the minimum bactericidal mass concentration (20.00 mg·mL-1) for 7 d. The ethyl acetate extract caused F. oxysporum hyphae to shrivel, and increased cell membrane permeability and leakage of cellular contents. Compared with the blank control, 5.00 mg·mL-1 (minimum inhibitory mass concentration) of the ethyl acetate extract significantly increased the conductivity and nucleic acid content of F. oxysporum at 10 h, and the contents of MDA and H2O2 in hyphae after 24 h. The activities of POD, CAT and SOD initially increased and then decreased, peaking at 12 h. In summary, the ethyl acetate extract of S. commune induced production of excessive reactive oxygen species and exacerbated membrane lipid peroxidation to cause membrane damage and loss of cellular contents, which destroyed the fungal defense system and reduced the content of protective enzymes, and thus significantly inhibited the hyphal growth and spore germination of F. oxysporum.
  • Effects of Solid-State Multienzyme Complex Hydrolysis and Nano Ball Milling on Physical and Nutritional Characteristics and In vitro Immune Activity of Hericium erinaceus Fruiting Body Powder
    DU Minjie, YANG Yan, LIU Peng, ZHANG Zhong, CHEN Wanchao, LI Wen, WU Di, JIN Yueling
    2024, 31(06): 69-80. https://doi.org/10.16488/i.cnki.1005-9873.2024.06.008
    Abstract ( ) PDF ( ) Knowledge map Save
    Using ultrafine pulverized Hericium erinaceus fruiting body powder (PS) as the control, the fruiting body powder was further processed by either solid-state multienzyme complex hydrolysis (PE) or nano ball milling (PN). The resultant fruiting body powders were observed for microstructure, measured for digestibility in vitro, analyzed for nutritional components, and evaluated for immune activity in vitro. The results showed that compared with PS, PN had a smaller particle size and altered fiber columnar structure, and PE had increases in surface porosity, surface area, and digestibility in vitro. Further processing with solid-state multienzyme complex hydrolysis or nano ball milling resulted in increases of polysaccharide, soluble solids, soluble protein and peptide in H. erinaceus fruiting body powders. The yield (13.42±0.64)% and polysaccharide content (67.05±0.18)% of the crude polysaccharide of PE (PLE) were higher than those of the crude polysaccharide of PS (PLS). The crude polysaccharide yield of PN (PLN) was also higher than that of PS, but the polysaccharide contents in PLN and PLS were statistically the same. Compared with PLS, the proportion of million molecular weight component (P1) in PLE decreased, whereas the proportion of P1 component in PLN increased. PLE significantly increased the release of NO, TNF-α and IL-1β in RAW264.7 macrophages, showing good immune activity in vitro. In summary, solid-state multienzyme complex hydrolysis improved the physical and nutritional characteristics and efficacy of H. erinaceus fruiting body powder, and thus provided a reference for the development of functional dietary supplements from H. erinaceus.
  • Ethyl Acetate Extract of Inonotus obliquus Reduced Uric Acid in Hyperuricemia Model Mice
    LIN Yuqi, MA Jisheng, BAO Haiying
    2024, 31(06): 81-91. https://doi.org/10.16488/i.cnki.1005-9873.2024.06.009
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    The uric acid lowering effect of ethyl acetate extract from Inonotus obliquus on hyperuricemic mice was studied. Mice were randomly divided into the following groups: normal group, hyperuricemia model group, allopurinol group (5 mg·kg-1, positive BA), benzbromarone group (7.8 mg·kg-1, positive BB), low-dose I. obliquus ethyl acetate extract (IO-E) group at 50 mg·kg-1, and high-dose I. obliquus ethyl acetate extract group at 100 mg·kg-1. Except the normal group, all the other groups of mice were administered potassium oxyzinate (300 mg·kg-1) intraperitoneally and hypoxanthine (500 mg·kg-1) by gavage for 14 consecutive days to establish a hyperuricemia model. After successful modeling, mice in different groups were administered corresponding interventions once a day for 28 consecutive days. One hour after the last administration, mice in different groups were measured for levels of uric acid, urea nitrogen, creatinine, xanthine oxidase, and adenosine deaminase in the blood by enzyme colorimetry, calculated for kidney and liver indices, observed for pathological changes in the kidneys and livers by hematoxylin-eosin staining, determined for URAT1 and GLUT9 protein contents in the kidneys by enzyme linked immunosorbent assay, observed for the localization of URAT1 and GLUT9 in the kidney tissues by immunohistochemical staining, and determined for the expression of URAT1 and GLUT9 by immunoblotting. Compared with the model group, mice in positive BA, positive BB and I. obliquus ethyl acetate extract groups showed ameliorated hyperuricemic symptoms, including decreased kidney and liver indices, decreased levels of blood uric acid, urea nitrogen and creatinine, decreased xanthine oxidase and adenosine deaminase activity, and ameliorated pathological changes in the kidney and liver tissues. The levels of URAT1 and GLUT9 in the kidneys of BA, BB and IO-E treated mice were significantly reduced, and the xanthine oxidase activity in the livers of these groups was also significantly reduced. Immunohistochemical staining of URAT1 and GLUT showed that URAT1 was primarily expressed in the cytoplasm of renal tubular epithelial cells, and GLUT9 was primarily expressed in the lateral membrane and cytoplasm of renal tubular epithelial cells. Compared with the model, BA, BB and IO-E resulted in significant decreases in average optical density (AOD) value and down-regulated expression of URAT1 and GLUT in kidney. Ethyl acetate extract of I. obliquus showed a uric acid lowering effect and effectively ameliorated kidney and liver damage caused by hyperuricemia.
  • Preparation of Auricularia auricula Polysaccharide-Ergosterol Complex and Its Cholesterol-Lowering Effect In vitro
    LI Liang, LI Jiahui, WU Jiaman, WANG Zheming, JIA Fengying, JIN Wen, YUN Shaojun, FENG Cuiping
    2024, 31(06): 92-98. https://doi.org/10.16488/i.cnki.1005-9873.2024.06.010
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    Polysaccharides in Auricularia auricula were obtained by water extraction and alcohol precipitation, and then used to prepare A. auricula polysaccharide-ergosterol (APE) complex by the antisolvent method. The complex was observed for microstructure, analyzed for structural characteristics and photothermal stability, and determined for its effect on cholesterol solubility in micelles. The results showed that ergosterol encapsulation efficiency reached the highest when the mass ratio of ergosterol to A. auricula polysaccharides was 1∶16. The APE prepared at this mass ratio had a rough and fibrous surface, and its average particle size was 1 265 nm. Compared with free ergosterol, APE showed better photothermal stability. APE significantly reduced the solubility of cholesterol in micelles. These results provided a theoretical basis for further development and utilization of A. auricula polysaccharides and ergosterol.
  • Extraction Process Optimization of Polysaccharides from Mycelium and Fruiting Body of Morchella importuna and Their Antioxidant and Immunomodulatory Activities
    DAI Zongyi, CHEN Hongyu, XU Aiguo, HAN Yucui, BAO Dapeng, WANG Weixia, LI Fuhou, TANG Lihua
    2024, 31(06): 99-111. https://doi.org/10.16488/i.cnki.1005-9873.2024.06.011
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    The effects of extraction time, temperature, and liquid-to-solid ratio on contents of polysaccharides from Morchella importuna mycelium (MIMP) and fruiting bodies (MIFP) were studied by single-factor experiment. Then the optimal extraction parameters were determined through response surface design. MIMP and MIFP were then analyzed for their structures, and compared for their antioxidant and immunomodulatory activities. The results showed that the optimal extraction conditions for MIMP were extraction time 166 min, extraction temperature 86 ℃, and a liquid-to-solid ratio of 31∶1 (mL∶g). Under the optimal conditions, the polysaccharide content reached (54.25±1.30)%, closely matching the theoretical predicted value of 55.14%. MIMP was primarily composed of glucose, mannose, and galactose in a molar ratio of 0.71∶0.11∶0.07, and its weight-average molecular weight (Mw) and number-average molecular weight (Mn) were 5.082×104 and 3.160×104 g·mol-1, respectively. For MIFP, the optimal extraction conditions were extraction time 161 min, extraction temperature 86 ℃, and a liquid-to-solid ratio of 31∶1 (mL∶g), under which the polysaccharide content was (39.85±1.55)% and consistent with the response surface prediction value of 40.14%. MIFP was mainly composed of glucose, mannose, and galactose in a molar ratio of 0.42∶0.31∶0.22, and its Mw and Mn were 1.263×104 and 8.345×103 g·mol-1, respectively. Both MIMP and MIFP showed scavenging activities against ABTS, DPPH, and hydroxyl radicals, with scavenging rates of 74% and 53% against ABTS, 61% and 46% against DPPH, and 78% and 51% against hydroxyl radicals, respectively. In the range of 0.1-1.0 mg·mL-1, the free radical scavenging rates of MIMP and MIFP gradually increased, with MIMP showing higher activity than MIFP at the same concentration. Both MIMP and MIFP stimulated NO release from RAW264.7 macrophages at 25-100 μg·mL-1, and there was no significant difference between them. This study provided a reference for the development of M. importuna polysaccharides in mycelia and fruiting bodies.
  • Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry for Detection of Ergosterol Peroxide in Edible Fungi and Their Processed Products
    E Hengchao, PENG Shuting, ZHAO Zhiyong, ZHANG Yanmei, LI Xiaobei, ZHOU Changyan, ZHAO Xiaoyan
    2024, 31(06): 112-118. https://doi.org/10.16488/i.cnki.1005-9873.2024.06.012
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    Using ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), a method was established for measuring ergosterol peroxide (EP) content in edible fungi and their processed products through optimizing chromatographic conditions, mass spectrometric parameters, extraction solvents, extraction time, and temperature. The method was validated through matrix effect assessment, spike-and-recovery testing, and evaluation of repeatability and reproducibility. The established method was used to measure EP content in 31 samples of edible fungi and processed products. The results showed that the chromatographic conditions were using a C18 chromatographic column and isocratic elution with 0.1% formic acid in methanol and 0.1% formic acid in water at a volumetric ratio of 92:8. Aacetonitrile was used as the extraction solvent for ultrasonic extraction at 30 ℃ for ٣٠ min. The quantitative ion transition was m/z 429.0/191.0. The optimal extraction conditions were There was good linearity in the range of 5 to 2 000 ng·mL-1 with minimal matrix effects. The EP detection limit for fresh and pickled edible fungi products was 0.020 mg·kg-1, and that of dried and ready-to-eat edible fungi products was 0.040 mg·kg-1. The spike recovery rates were 78.4%-111.5%, which comply with the national standard of 60%-120% in GB 2763-2021. The intraday variation coefficients and interday variation coefficients were 0.3%-4.8%, and 0.4%-2.0%, respectively. Fresh L. edodes had a relatively high EP content.
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