In this study,Tremella fuciformis residues as raw material,dietary fibers from tremella were prepared by multiple enzymes.The structure of dietary fibers from tremella was studied by Fourier transform infrared(FTIR),X...In this study,Tremella fuciformis residues as raw material,dietary fibers from tremella were prepared by multiple enzymes.The structure of dietary fibers from tremella was studied by Fourier transform infrared(FTIR),X-ray diffraction analysis(XRD)and scanning electron microscopy(SEM).We analyzed their lipidlowering properties in vitro(water holding,oil holding swelling cholesterol and sodium cholate binding capacitises)and the hypolipidemic effects in mice.The results showed that tremella dietary fibers presented the infrared absorption spectrum characteristics of polysaccharides and the characteristic diffraction peaks of cellulose type I.SEM results indicated that the surface of insoluble dietary fiber(IDF)was porous,while the soluble dietary fiber(SDF)was relatively compact and spongy.IDF exhibited significantly higher water holding,oil holding,and swelling binding capacities than the corresponding SDF.However,SDF exhibited significantly higher viscosity than IDF.The results showed tremella dietary fibers were significant in swelling,water holding and oil holding,cholesterol and bile acids.In vivo experiment results in mice indicated that SDF has the best effect on hyperlipidemia mice than IDF and total dietary fiber(TDF).SDF showed that the total cholesterol(TC),triglyceride(TG)and low density lipoprotein cholesterol(LDL-C)contents dropped by 28.33%,18.65%,and 48.97%,respectively,while high density lipoprotein cholesterol(HDL-C)content increased by 43.80%.Compared with the high-fat control(HCM)group,the arteriosclerosis index(AI)and liver index(LI)of the SDF group mice showed significant differences,indicating that SDF has a good auxiliary effect of lowering blood lipids.The administration of tremella fibers improved the lipid metabolism disorderly situation of hyperlipidemia mice.These results provide a reference for further research and rational development of T.fuciformis.展开更多
Tremella fuciformis polysaccharide(TFP) is an important bioactive substance in T. fuciformis, which contributes to its application as medicine and food. In the present work, TFP was extracted with enzymatic combined w...Tremella fuciformis polysaccharide(TFP) is an important bioactive substance in T. fuciformis, which contributes to its application as medicine and food. In the present work, TFP was extracted with enzymatic combined with wet beating and its physicochemical as well as biological activities were investigated in the study. The result showed that the purity of TFP1 reached 92.14%(m/m). The TFP1, purified by DEAE column, had a molecular weight(Mw) of 5.8 × 10^(3) k Da and consisted mainly of mannose, xylose, fucose, glucose and glucuronic acid at a molar ratio of 1.91 : 0.1 : 2.49 : 6.23 : 0.95. The structure of TFP1 was preliminarily investigated by infrared spectroscopy, nuclear magnetic resonance and atomic force microscope(AFM) analysis. Moreover, the antioxidant assays showed that TFP1 could scavenge DPPH, ABTS+ and hydroxyl radicals. The excellent water holding capacity of TFP1 implied its application in the food, pharmaceutical andcosmetic industries.展开更多
The effect of Tremella fuciformis polysaccharide(TFP)on the retrogradation property and aroma profile changes of tteok during storage was investigated.Results indicated that addition of TFP significantly increased the...The effect of Tremella fuciformis polysaccharide(TFP)on the retrogradation property and aroma profile changes of tteok during storage was investigated.Results indicated that addition of TFP significantly increased the stability to thermal and mechanical shearing of starch,decreased short-term retrogradation,the hardness and the retrogradation enthalpy(ΔH)of tteok during storage,but had no significant effect on the amylopectin chain length of tteok.The possible mechanism for the retarding staling effect of TFP is related to the interaction between TFP and starch chains that interferes with the alignment of starch chains.Electronic nose and g as chromatography-mass spectrum(GC-MS)analysis results showed that the difference in the concentrations of volatile compounds and fatty acids of tteoks at different storage time gradually increased with the advancement of storage period.The addition of TFP to tteok inhibited the development of unpleasant volatile compounds,probably by delaying the oxidation and decomposition of lipids and preserving the antioxidant phenolic compounds in tteok,thus slowing down the flavor deterioration of tteok and contributing to flavor maintainace.Overall,this study could help food manufacturers to choose a high-effective and natural polysaccharide to control the retrogradation rate and flavor loss of tteok.展开更多
TSP could markedly enhance the proliferative response of the murine splenocyte to LPS and induce the mitogenesis of the spleen cells.Furthermore,it was able to augment the activity of natural killer cell and ADGG;at a...TSP could markedly enhance the proliferative response of the murine splenocyte to LPS and induce the mitogenesis of the spleen cells.Furthermore,it was able to augment the activity of natural killer cell and ADGG;at a dosage of 25-250μg/ml,the ability of splenocytes to produce IL-2 induced by onA had been improved; at the concentration of 250μg／ml or more,TSP could inhibit the proliferative response of the murine lymphocyte to GonA and the ~3-HTdR spontaneous incorporation rate of thymocytes,and the inhibitory action ran in paralell with the increase in concentration of TSP.展开更多
Tremella fuciformis polysaccharides have rich bioactivities,while low molecular weight polysaccharides have higher bioavailability and nutritional value.In this study,it was investigated that the structural characteri...Tremella fuciformis polysaccharides have rich bioactivities,while low molecular weight polysaccharides have higher bioavailability and nutritional value.In this study,it was investigated that the structural characteristics of low molecular weight polysaccharides from Tremella fuciformis(TFLP)prepared by acid hydrolysis method,its impact on antioxidant capacity and stress resistance of Caenorhabditis elegans,and potential mechanisms.TFLP was an acidic polysaccharide mainly composed of mannose and xylose with a molecular weight of 106 kDa.TFLP could prolong the median lifespan of C.elegans under heat stress and acute oxidative stress conditions through significantly reducing the malondialdehyde(MDA)level and increasing the activity of antioxidant enzymes total-superoxide dismutase(T-SOD),glutathione peroxidase(GSH-Px)and catalase(CAT).The intervention of high-dose TFLP significantly prolonged the median lifespan of C.elegans under heat stress and acute oxidative stress conditions by 24.47%and 7.84%,respectively.At the same time,the MDA level significantly decreased by 69.59%,and the levels of T-SOD,GSH-Px and CAT increased significantly by 2.22,1.28 and 0.53 times,respectively.Meanwhile,the transcription levels of daf-16,fat-5,fat-7,and hsf-1 mRNA in C.elegans treated with TFLP were significantly increased,while the transcription levels of akt-1 and daf-2 mRNA were significantly reduced.In addition,propanoate metabolism,valine,leucine and isoleucine degradation metabolism and vitamin B6 metabolism were key metabolic pathways for TFLP to enhance antioxidant capacity and stress resistance in C.elegans.These results indicated that TFLP could enhance in vivo antioxidant capacity and stress resistance by improving the levels mRNA transcription and metabolites,and it provided new evidence for TFLP to exert in vivo antioxidant activity.展开更多
Tremella fuciformis is one of higher basidiomycetes.Its basidiospore can reproduce yeast-like conidia,also called the blastospore by budding.The yeast-like conidia of T.fuciformis is monokaryotic and easy to culture b...Tremella fuciformis is one of higher basidiomycetes.Its basidiospore can reproduce yeast-like conidia,also called the blastospore by budding.The yeast-like conidia of T.fuciformis is monokaryotic and easy to culture by submerged fermentation similar to yeast.So it is a good recipient cell for exogenous gene expression.In this study,two expression vectors pGlg-gfp containing gpd-Gl promoter and gfp gene and pGlg-hph containing gpd-Gl promoter and hph gene were constructed.The lowest sensitive concentration of hygromycin for the blastospore was determined on three types of media.Our ex-periments showed that the lowest sensitive concentration of hygromycin for the blastospore was 5μg/mL on MA medium.The intact blastospores were transformed with the expression vector pGlg-hph by electroporation.The putative transformants were obtained by the MA selective medium.Experi-mental results showed that the most effective parameters for the electroporation of intact blastospores were obtained by using STM buffer,1.0×108 cells/mL of blastospores,200μL in transformation volume,6μg plasmid,2.0 kV/cm of electric pulse voltage,stillness culturing on MB liquid medium for 48 h after electroporation.In these transformation conditions,the efficiency reached 277 colonies/μg DNA.Co-transformation of plasmid pGlg-gfp and pGlg-hph with ratio of 1:1 was performed by electroporation with the optimal parameters.The putative co-transformants were obtained by the MA selective medium.Eight randomly selected colonies from the vast putative co-transformants were analyzed by PCR de-tection and Southern blotting.The experiments showed that the gfp was integrated into the genomes of three transformants.The co-transformation efficiency was 37.5%.Green fluorescence was observed under laser scanning confocal microscope in these gfp positive transformants.This indicates that the exogenous gfp can be expressed effectively in the yeast-like conidia of T.fuciformis.展开更多
This study evaluated and compared the fermentative and flavor properties of Hericium erinaceus and Tremella fuciformis fermented beverage(HTFB)produced by four commercial lactic acid bacteria(LAB),i.e.,L.plantarum,L.r...This study evaluated and compared the fermentative and flavor properties of Hericium erinaceus and Tremella fuciformis fermented beverage(HTFB)produced by four commercial lactic acid bacteria(LAB),i.e.,L.plantarum,L.rhamnosus,L.paracasei or S.thermophilu s.All four strains showed the probiotic potential of utilizing Hericium erinaceus and Tremella fuciformis as fermentation substrates.Among four LAB,L.plantarum produced significantly highest concentrations of total acids(76.16 g/L),while the viable count of HTFB inoculated with L.paracasei was the highest(7.18 log CFU/mL).The contents of bioactive components such as polyphenols and triterpenoids significantly increased after fermentation regardless of the strains.The fermentation process enriched the volatile profile and reduced the unpleasant flavors such as nonanal and 1-octen-3-ol.According to the relative odor activity values and PLS-DA analysis,a total of 8 volatile organic compounds were identified as markers with odor contributions.Furthermore,the olfactory characteristics of the four beverages could be clearly distinguished by electronic nose.The electronic tongue results showed that the fermentation process increased the sourness and decreased the bitterness and astringency of the beverage,resulting in an improved taste property.The sensory evaluation finally confirmed that HTFB inoculated with L.paracasei was the most acceptable product to consumers with high overall flavor coordination.Overall,this work provided valuable information for the selection of premium fermentation strains to produce fermented edible fungi beverages with better nutritional and sensory qualities.展开更多
Tremella fuciformis Berk(TF)is an edible fungus with multiple nutritional values,renowned for its antiinflammatory and immunomodulatory properties.This research aimed to elucidate the mechanism by which TF regulates d...Tremella fuciformis Berk(TF)is an edible fungus with multiple nutritional values,renowned for its antiinflammatory and immunomodulatory properties.This research aimed to elucidate the mechanism by which TF regulates depressive symptoms in chronic unpredictable mild stress(CUMS)mice.CUMS modeling was conducted on 6-week-old C57BL/6 male mice,which were subsequently evaluated for depression-like behavior.Immunofluorescence(IF)staining,intestinal microbiota analysis,metabolomics analysis,and Western blot were utilized to investigate the underlying mechanisms.TF significantly mitigated depression-like behaviors and reversed the reduction of synaptophysin(SYN)and postsynaptic density-95(PSD-95)in the brain hippocampus.Furthermore,the expression level of brain-derived neurotrophic factor(BDNF)was significantly upregulated.The antidepressant effect of TF may be ascribed to its anti-inflammatory properties.Further investigation disclosed that TF significantly repressed microglia and astrocyte activation,as well as the phosphorylation of nuclear transcription factor-κB(NF-κB)and inhibitor of NF-κB(IκB)in the brain,while reducing the production of interleukin-6(IL-6),interleukin-1 beta(IL-1β)and tumor necrosis factor-alpha(TNF-α).TF may modulate neuroinflammation by inhibiting NF-κB signaling,thereby exerting an antidepressant effect,suggesting TF as a potential candidate for the prevention and treatment of depression.展开更多
Tremella fuciformis,one of the most approved mushrooms worldwide,possesses both edible and medicinal value.However,low transformation efficiency and high false positive rate,long verification period,and low expression...Tremella fuciformis,one of the most approved mushrooms worldwide,possesses both edible and medicinal value.However,low transformation efficiency and high false positive rate,long verification period,and low expression of heterologous proteins hinder further genetic research.In this study,a polyethylene glycol-mediated genetic transformation system was established by using the homologous resistance marker carboxin in T.fuciformis.By optimizing the protoplast quality and transformation conditions,the transformation efficiency reached 8.23 CFU/μg,and the positive rate reached 64.95%.Moreover,Driselase-Heating-Freezing screening system was developed for rapid transformants verification,which took only 35-45 min and made the colony PCR 100%accurate.Considering the rationality of tRNA use frequency in heterologous genes,the coordinated relative synonymous codon usage(RSCU)strategy was adopted to recode the heterologous gene sequence,which ensured the protein expression level.This study comprehensively enhances molecular biology tools in T.fuciformis,and lays a foundation for further development.展开更多
TSP could markedly enhance the proliferative response of the murine splenocyte to LPS and induce the mitogenesis of the spleen cells.Furthermore,it was able to augment the activity of natural killer cell and ADGG;at a...TSP could markedly enhance the proliferative response of the murine splenocyte to LPS and induce the mitogenesis of the spleen cells.Furthermore,it was able to augment the activity of natural killer cell and ADGG;at a dosage of 25-250μg/ml,the ability of splenocytes to produce IL-2 induced by onA had been improved; at the concentration of 250μg/ml or more,TSP could inhibit the proliferative response of the murine lymphocyte to GonA and the ￣3-HTdR spontaneous incorporation rate of thymocytes,and the inhibitory action ran in paralell with the increase in concentration of TSP.展开更多
基金financially supported by the Key Projects of the National Research and Development Program of China(2018YFD0400204)。
文摘In this study,Tremella fuciformis residues as raw material,dietary fibers from tremella were prepared by multiple enzymes.The structure of dietary fibers from tremella was studied by Fourier transform infrared(FTIR),X-ray diffraction analysis(XRD)and scanning electron microscopy(SEM).We analyzed their lipidlowering properties in vitro(water holding,oil holding swelling cholesterol and sodium cholate binding capacitises)and the hypolipidemic effects in mice.The results showed that tremella dietary fibers presented the infrared absorption spectrum characteristics of polysaccharides and the characteristic diffraction peaks of cellulose type I.SEM results indicated that the surface of insoluble dietary fiber(IDF)was porous,while the soluble dietary fiber(SDF)was relatively compact and spongy.IDF exhibited significantly higher water holding,oil holding,and swelling binding capacities than the corresponding SDF.However,SDF exhibited significantly higher viscosity than IDF.The results showed tremella dietary fibers were significant in swelling,water holding and oil holding,cholesterol and bile acids.In vivo experiment results in mice indicated that SDF has the best effect on hyperlipidemia mice than IDF and total dietary fiber(TDF).SDF showed that the total cholesterol(TC),triglyceride(TG)and low density lipoprotein cholesterol(LDL-C)contents dropped by 28.33%,18.65%,and 48.97%,respectively,while high density lipoprotein cholesterol(HDL-C)content increased by 43.80%.Compared with the high-fat control(HCM)group,the arteriosclerosis index(AI)and liver index(LI)of the SDF group mice showed significant differences,indicating that SDF has a good auxiliary effect of lowering blood lipids.The administration of tremella fibers improved the lipid metabolism disorderly situation of hyperlipidemia mice.These results provide a reference for further research and rational development of T.fuciformis.
基金the the Open Project Program of National R&D Center for Edible Fungus Processing Technology(20200110)。
文摘Tremella fuciformis polysaccharide(TFP) is an important bioactive substance in T. fuciformis, which contributes to its application as medicine and food. In the present work, TFP was extracted with enzymatic combined with wet beating and its physicochemical as well as biological activities were investigated in the study. The result showed that the purity of TFP1 reached 92.14%(m/m). The TFP1, purified by DEAE column, had a molecular weight(Mw) of 5.8 × 10^(3) k Da and consisted mainly of mannose, xylose, fucose, glucose and glucuronic acid at a molar ratio of 1.91 : 0.1 : 2.49 : 6.23 : 0.95. The structure of TFP1 was preliminarily investigated by infrared spectroscopy, nuclear magnetic resonance and atomic force microscope(AFM) analysis. Moreover, the antioxidant assays showed that TFP1 could scavenge DPPH, ABTS+ and hydroxyl radicals. The excellent water holding capacity of TFP1 implied its application in the food, pharmaceutical andcosmetic industries.
基金financially supported by the Key Projects of the National Research and Development Program of China (2018YFD0400103-6)
文摘The effect of Tremella fuciformis polysaccharide(TFP)on the retrogradation property and aroma profile changes of tteok during storage was investigated.Results indicated that addition of TFP significantly increased the stability to thermal and mechanical shearing of starch,decreased short-term retrogradation,the hardness and the retrogradation enthalpy(ΔH)of tteok during storage,but had no significant effect on the amylopectin chain length of tteok.The possible mechanism for the retarding staling effect of TFP is related to the interaction between TFP and starch chains that interferes with the alignment of starch chains.Electronic nose and g as chromatography-mass spectrum(GC-MS)analysis results showed that the difference in the concentrations of volatile compounds and fatty acids of tteoks at different storage time gradually increased with the advancement of storage period.The addition of TFP to tteok inhibited the development of unpleasant volatile compounds,probably by delaying the oxidation and decomposition of lipids and preserving the antioxidant phenolic compounds in tteok,thus slowing down the flavor deterioration of tteok and contributing to flavor maintainace.Overall,this study could help food manufacturers to choose a high-effective and natural polysaccharide to control the retrogradation rate and flavor loss of tteok.
文摘TSP could markedly enhance the proliferative response of the murine splenocyte to LPS and induce the mitogenesis of the spleen cells.Furthermore,it was able to augment the activity of natural killer cell and ADGG;at a dosage of 25-250μg/ml,the ability of splenocytes to produce IL-2 induced by onA had been improved; at the concentration of 250μg／ml or more,TSP could inhibit the proliferative response of the murine lymphocyte to GonA and the ~3-HTdR spontaneous incorporation rate of thymocytes,and the inhibitory action ran in paralell with the increase in concentration of TSP.
基金funded by Science and Technology Planning Project of Fujian Province(2021L3007).
文摘Tremella fuciformis polysaccharides have rich bioactivities,while low molecular weight polysaccharides have higher bioavailability and nutritional value.In this study,it was investigated that the structural characteristics of low molecular weight polysaccharides from Tremella fuciformis(TFLP)prepared by acid hydrolysis method,its impact on antioxidant capacity and stress resistance of Caenorhabditis elegans,and potential mechanisms.TFLP was an acidic polysaccharide mainly composed of mannose and xylose with a molecular weight of 106 kDa.TFLP could prolong the median lifespan of C.elegans under heat stress and acute oxidative stress conditions through significantly reducing the malondialdehyde(MDA)level and increasing the activity of antioxidant enzymes total-superoxide dismutase(T-SOD),glutathione peroxidase(GSH-Px)and catalase(CAT).The intervention of high-dose TFLP significantly prolonged the median lifespan of C.elegans under heat stress and acute oxidative stress conditions by 24.47%and 7.84%,respectively.At the same time,the MDA level significantly decreased by 69.59%,and the levels of T-SOD,GSH-Px and CAT increased significantly by 2.22,1.28 and 0.53 times,respectively.Meanwhile,the transcription levels of daf-16,fat-5,fat-7,and hsf-1 mRNA in C.elegans treated with TFLP were significantly increased,while the transcription levels of akt-1 and daf-2 mRNA were significantly reduced.In addition,propanoate metabolism,valine,leucine and isoleucine degradation metabolism and vitamin B6 metabolism were key metabolic pathways for TFLP to enhance antioxidant capacity and stress resistance in C.elegans.These results indicated that TFLP could enhance in vivo antioxidant capacity and stress resistance by improving the levels mRNA transcription and metabolites,and it provided new evidence for TFLP to exert in vivo antioxidant activity.
基金National High-tech Research and Development Program(863Program)of China(GrantNo.2006AA10Z301)the National Natural Science Foundation of China(GrantNos.30371000 and 30671457)the Natural Science Foundati on of Guangdong Province,China(GrantNo.032239)
文摘Tremella fuciformis is one of higher basidiomycetes.Its basidiospore can reproduce yeast-like conidia,also called the blastospore by budding.The yeast-like conidia of T.fuciformis is monokaryotic and easy to culture by submerged fermentation similar to yeast.So it is a good recipient cell for exogenous gene expression.In this study,two expression vectors pGlg-gfp containing gpd-Gl promoter and gfp gene and pGlg-hph containing gpd-Gl promoter and hph gene were constructed.The lowest sensitive concentration of hygromycin for the blastospore was determined on three types of media.Our ex-periments showed that the lowest sensitive concentration of hygromycin for the blastospore was 5μg/mL on MA medium.The intact blastospores were transformed with the expression vector pGlg-hph by electroporation.The putative transformants were obtained by the MA selective medium.Experi-mental results showed that the most effective parameters for the electroporation of intact blastospores were obtained by using STM buffer,1.0×108 cells/mL of blastospores,200μL in transformation volume,6μg plasmid,2.0 kV/cm of electric pulse voltage,stillness culturing on MB liquid medium for 48 h after electroporation.In these transformation conditions,the efficiency reached 277 colonies/μg DNA.Co-transformation of plasmid pGlg-gfp and pGlg-hph with ratio of 1:1 was performed by electroporation with the optimal parameters.The putative co-transformants were obtained by the MA selective medium.Eight randomly selected colonies from the vast putative co-transformants were analyzed by PCR de-tection and Southern blotting.The experiments showed that the gfp was integrated into the genomes of three transformants.The co-transformation efficiency was 37.5%.Green fluorescence was observed under laser scanning confocal microscope in these gfp positive transformants.This indicates that the exogenous gfp can be expressed effectively in the yeast-like conidia of T.fuciformis.
基金financially supported by Transformation Program of Scientific and Technological Achievements of Jiangsu Province(BA2021062).
文摘This study evaluated and compared the fermentative and flavor properties of Hericium erinaceus and Tremella fuciformis fermented beverage(HTFB)produced by four commercial lactic acid bacteria(LAB),i.e.,L.plantarum,L.rhamnosus,L.paracasei or S.thermophilu s.All four strains showed the probiotic potential of utilizing Hericium erinaceus and Tremella fuciformis as fermentation substrates.Among four LAB,L.plantarum produced significantly highest concentrations of total acids(76.16 g/L),while the viable count of HTFB inoculated with L.paracasei was the highest(7.18 log CFU/mL).The contents of bioactive components such as polyphenols and triterpenoids significantly increased after fermentation regardless of the strains.The fermentation process enriched the volatile profile and reduced the unpleasant flavors such as nonanal and 1-octen-3-ol.According to the relative odor activity values and PLS-DA analysis,a total of 8 volatile organic compounds were identified as markers with odor contributions.Furthermore,the olfactory characteristics of the four beverages could be clearly distinguished by electronic nose.The electronic tongue results showed that the fermentation process increased the sourness and decreased the bitterness and astringency of the beverage,resulting in an improved taste property.The sensory evaluation finally confirmed that HTFB inoculated with L.paracasei was the most acceptable product to consumers with high overall flavor coordination.Overall,this work provided valuable information for the selection of premium fermentation strains to produce fermented edible fungi beverages with better nutritional and sensory qualities.
基金funded by the earmarked fund for Edible Mushroom(CARS-20).
文摘Tremella fuciformis Berk(TF)is an edible fungus with multiple nutritional values,renowned for its antiinflammatory and immunomodulatory properties.This research aimed to elucidate the mechanism by which TF regulates depressive symptoms in chronic unpredictable mild stress(CUMS)mice.CUMS modeling was conducted on 6-week-old C57BL/6 male mice,which were subsequently evaluated for depression-like behavior.Immunofluorescence(IF)staining,intestinal microbiota analysis,metabolomics analysis,and Western blot were utilized to investigate the underlying mechanisms.TF significantly mitigated depression-like behaviors and reversed the reduction of synaptophysin(SYN)and postsynaptic density-95(PSD-95)in the brain hippocampus.Furthermore,the expression level of brain-derived neurotrophic factor(BDNF)was significantly upregulated.The antidepressant effect of TF may be ascribed to its anti-inflammatory properties.Further investigation disclosed that TF significantly repressed microglia and astrocyte activation,as well as the phosphorylation of nuclear transcription factor-κB(NF-κB)and inhibitor of NF-κB(IκB)in the brain,while reducing the production of interleukin-6(IL-6),interleukin-1 beta(IL-1β)and tumor necrosis factor-alpha(TNF-α).TF may modulate neuroinflammation by inhibiting NF-κB signaling,thereby exerting an antidepressant effect,suggesting TF as a potential candidate for the prevention and treatment of depression.
基金supported by the National Key Research and Development Program of China(No.2019YFA0905200)the National Natural Science Foundation of China(No.22178177)+1 种基金the Jiangsu Synergetic Innovation Center for Advanced Bio-Manufacture(No.XTD2201)the Jiangsu Funding Program for Excellent Postdoctoral Talent(2023ZB445).
文摘Tremella fuciformis,one of the most approved mushrooms worldwide,possesses both edible and medicinal value.However,low transformation efficiency and high false positive rate,long verification period,and low expression of heterologous proteins hinder further genetic research.In this study,a polyethylene glycol-mediated genetic transformation system was established by using the homologous resistance marker carboxin in T.fuciformis.By optimizing the protoplast quality and transformation conditions,the transformation efficiency reached 8.23 CFU/μg,and the positive rate reached 64.95%.Moreover,Driselase-Heating-Freezing screening system was developed for rapid transformants verification,which took only 35-45 min and made the colony PCR 100%accurate.Considering the rationality of tRNA use frequency in heterologous genes,the coordinated relative synonymous codon usage(RSCU)strategy was adopted to recode the heterologous gene sequence,which ensured the protein expression level.This study comprehensively enhances molecular biology tools in T.fuciformis,and lays a foundation for further development.
文摘TSP could markedly enhance the proliferative response of the murine splenocyte to LPS and induce the mitogenesis of the spleen cells.Furthermore,it was able to augment the activity of natural killer cell and ADGG;at a dosage of 25-250μg/ml,the ability of splenocytes to produce IL-2 induced by onA had been improved; at the concentration of 250μg/ml or more,TSP could inhibit the proliferative response of the murine lymphocyte to GonA and the ￣3-HTdR spontaneous incorporation rate of thymocytes,and the inhibitory action ran in paralell with the increase in concentration of TSP.
