The Experts below are selected from a list of 360 Experts worldwide ranked by ideXlab platform
Liguo Che - One of the best experts on this subject based on the ideXlab platform.
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comparative transcriptomic analysis identified differentially expressed genes and pathways involved in the interaction between Tremella fuciformis and annulohypoxylon stygium
Antonie Van Leeuwenhoek International Journal of General and Molecular Microbiology, 2019Co-Authors: Dongmei Liu, Liesheng Zheng, Yuanyua Wang, Dwi Pujiana, Zhaosong Zhang, Liguo CheAbstract:Tremella fuciformis is an edible and medicinal white jelly mushroom. It has a life cycle that interacts with its companion fungus Annulohypoxylon stygium, both in natural conditions and artificial cultivation. RNA sequencing (RNA-Seq) was used to study the interaction between T. fuciformis and A. stygium by constructing 5 libraries, including the individual T. fuciformis mycelium (1), the T. fuciformis mycelium after interaction with A. stygium (2), the dual mycelia after interaction (3), the A. stygium mycelium after interaction with T. fuciformis (4), and the individual A. stygium mycelium (5). 33.4 G data and 46,871 Unigenes were generated from de novo splicing. For identification of differentially expressed genes (DEGs) related to interaction, we analyzed the expression data of DEGs1-vs-2 ∩ DEGs1-vs-3, and DEGs5-vs-4 ∩ DEGs5-vs-3. DEGs1-vs-2 ∩ DEGs1-vs-3, and DEGs5-vs-4 ∩ DEGs5-vs-3 data showed 614 DEGs and 1537 DEGs, respectively. The 614 DEGs for T. fuciformis and 1537 DEGs for A. stygium were analyzed by GO annotation and were assigned to biology process, cell composition, and molecular functions. The DEGs were used to match the KEGG database. In T. fuciformis, the pathways are primarily enriched various amino acids metabolism, pentose and glucuronate interconversions. In A. stygium, the pathways are primarily enriched in the biosynthesis of secondary metabolites, biosynthesis of antibiotics, starch and sucrose metabolism. The expression patterns of DEGs determined by qRT-PCR were consistent with those obtained by RNA-Seq, thus validating the reliability of our RNA-Seq data. Future studies of the functions of these interesting genes will be helpful to understand the mechanisms by which T. fuciformis interacts with A. stygium. This will also provide a reference for other research on interacting microorganisms.
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an enzymolysis assisted agrobacterium tumefaciens mediated transformation method for the yeast like cells of Tremella fuciformis
Mycobiology, 2019Co-Authors: Yuanyua Wang, Lisheng Zheng, Liguo CheAbstract:Agrobacterium tumefaciens-mediated transformation (ATMT), as a simple and versatile method, achieves successful transformation in the yeast-like cells (YLCs) of Tremella fuciformis with lower effic...
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An Enzymolysis-Assisted Agrobacterium tumefaciens-Mediated Transformation Method for the Yeast-Like Cells of Tremella fuciformis
Taylor & Francis Group, 2019Co-Authors: Yuanyua Wang, Lisheng Zheng, Liguo CheAbstract:Agrobacterium tumefaciens-mediated transformation (ATMT), as a simple and versatile method, achieves successful transformation in the yeast-like cells (YLCs) of Tremella fuciformis with lower efficiency. Establishment of a more efficient transformation system of YLCs is important for functional genomics research and biotechnological application. In this study, an enzymolysis-assisted ATMT method was developed. The degradation degree of YLCs depends on the concentration and digestion time of Lywallzyme. Lower concentration (≤0.1%) of Lywallzyme was capable of formation of limited wounds on the surface of YLCs and has less influence on their growth. In addition, there is no significant difference of YLCs growth among groups treated with 0.1% Lywallzyme for different time. The binary vector pGEH under the control of T. fuciformis glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter was utilized to transform the enzymolytic wounded YLCs with different concentrations and digestion time. The results of PCR, Southern blot, quantitative real-time PCR (qRT-PCR) and fluorescence microscopy revealed that the T-DNA was integrated into the YLCs genome, suggesting an efficient enzymolysis-assisted ATMT method of YLCs was established. The highest transformation frequency reached 1200 transformants per 106 YLCs by 0.05% (w/v) Lywallzyme digestion for 15 min, and the transformants were genetically stable. Compared with the mechanical wounding methods, enzymolytic wounding is thought to be a tender, safer and more effective method
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Cloning and heterologous expression of a hydrophobin gene Ltr.hyd from the tiger milk mushroom Lentinus tuber-regium in yeast-like cells of Tremella fuciformis
Elsevier, 2018Co-Authors: Dongmei Liu, Liesheng Zheng, Hanyu Zhu, Yue Che, Liguo CheAbstract:Background: Hydrophobins are small proteins secreted by filamentous fungi, which show a highly surface activity. Because of the signally self-assembling abilities and surface activities, hydrophobins were considered as candidates in many aspects, for example, stabilizing foams and emulsions in food products. Lentinus tuber-regium, known as tiger milk mushroom, is both an edible and medicinal sclerotium-producing mushroom. Up to now, the hydrophobins of L. tuber-regium have not been identified. Results: In this paper, a Class I hydrophobin gene, Ltr.hyd, was cloned from L. tuber-regium and expressed in the yeast-like cells of Tremella fuciformis mediated by Agrobacterium tumefaciens. The expression vector pGEH-GH was under the control of T. fuciformis glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter. The integration of Ltr.hyd into the genome of T. fuciformis was confirmed by PCR, Southern blot, fluorescence observation and quantitative real-time PCR (qRT-PCR). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that recombinant hydrophobin rLtr.HYD with an expected molecular mass of 13 kDa was extracted. The yield of rLtr.HYD was 0.66 mg/g dry weight. The emulsifying activity of rLtr.HYD was better than the typical food emulsifiers sodium caseinate and Tween 20. Conclusions: We evaluated the emulsifying property of hydrophobin Ltr.HYD, which can be potentially used as a food emulsifier. Keywords: Agrobacterium tumefaciens, Emulsifier, Expression vector, Filamentous fungi, Gel electrophoresis, Glyceraldehyde-3-phosphate dehydrogenase, Heterogenous expression, Hydrophobin, Quantitative real-time PCR, Southern blot, Surface activit
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an improved total rna extraction method for white jelly mushroom Tremella fuciformis rich in polysaccharides
Mycobiology, 2017Co-Authors: Hanyu Zhu, Liesheng Zheng, Dongmei Liu, Liguo CheAbstract:An improved method for extracting high quality and quantity RNA from a jelly mushroom and a dimorphic fungus-Tremella fuciformis which is especially rich in polysaccharides, is described. RNA was extracted from T. fuciformis mycelium M1332 and its parental monokaryotic yeast-like cells Y13 and Y32. The A260/280 and A260/230 ratios were both approximately 2, and the RNA integrity number was larger than 8.9. The yields of RNA were between 108 and 213 µg/g fresh wt. Downstream molecular applications including reverse transcriptional PCR and quantitative real-time PCR were also performed. This protocol is reliable and may be widely applicable for total RNA extraction from other jelly mushrooms or filamentous fungi rich in polysaccharides.
Aogui Xie - One of the best experts on this subject based on the ideXlab platform.
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intra specific comparison of mitochondrial genomes reveals host gene fragment exchange via intron mobility in Tremella fuciformis
BMC Genomics, 2020Co-Authors: Aogui Xie, Youji Deng, Xunxiao Zhang, Tom Hsiang, Xingta Zhang, Wenjing Miao, Ray MingAbstract:Mitochondrial genomic sequences are known to be variable. Comparative analyses of mitochondrial genomes can reveal the nature and extent of their variation. Draft mitochondrial genomes of 16 Tremella fuciformis isolates (TF01-TF16) were assembled from Illumina and PacBio sequencing data. Mitochondrial DNA contigs were extracted and assembled into complete circular molecules, ranging from 35,104 bp to 49,044 bp in size. All mtDNAs contained the same set of 41 conserved genes with identical gene order. Comparative analyses revealed that introns and intergenic regions were variable, whereas genic regions (including coding sequences, tRNA, and rRNA genes) were conserved. Among 24 introns detected, 11 were in protein-coding genes, 3 in tRNA genes, and the other 10 in rRNA genes. In addition, two mobile fragments were found in intergenic regions. Interestingly, six introns containing N-terminal duplication of the host genes were found in five conserved protein-coding gene sequences. Comparison of genes with and without these introns gave rise to the following proposed model: gene fragment exchange with other species can occur via gain or loss of introns with N-terminal duplication of the host genes. Our findings suggest a novel mechanism of fungal mitochondrial gene evolution: partial foreign gene replacement though intron mobility.
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first report of lecanicillium aphanocladii causing rot of Tremella fuciformis in china
Plant Disease, 2018Co-Authors: J Liu, Aogui Xie, Ingzhi Che, X Qiu, Lingda Lia, Zhipeng Huang, Y JiangAbstract:The white jelly-leaf mushroom (Tremella fuciformis Berk.) is a widely consumed edible fungus in Asia, particularly in China (Stamets 2000). In 2013, fresh T. fuciformis yield reached 385,000 t from all over China. Fujian Province is the main producing area of T. fuciformis, with 358,000 t in 2013. However, in October 2015, a serious rot disease with more than 30% incidence was found on T. fuciformis at a mushroom factory in Sanming, Fujian, China (26.30°N, 118.42°E). The symptoms mainly occurred on the bottom surface of T. fuciformis. Infected fruiting bodies were covered by white mycelia that later turned red or fuchsia. In the final stages of infection, the T. fuciformis fruiting bodies turned dark red or brown before rotting. The aerial mycelium of the pathogen was isolated from surface tissues of symptomatic fruiting bodies. On potato dextrose agar (PDA), the pathogen colony appeared red, reddish, then white to cream-color, successively. Aerial mycelia were white and grew vigorously to a height of 1 t...
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morphological and molecular analysis identifies the associated fungus xianghui of the medicinal white jelly mushroom Tremella fuciformis as annulohypoxylon stygium
International Journal of Medicinal Mushrooms, 2016Co-Authors: Youji Deng, Lingda Lia, Yuji Jiang, Arend F Van Pee, Qingfu Wang, Aogui XieAbstract:White jelly mushroom, Tremella fuciformis, is a popular edible mushroom with interesting medicinal properties (e.g., immunostimulating, antidiabetic). The formation of T. fuciformis basidiomes is highly dependent on the presence of a specific host fungus, both in nature and for industrial production. This host has traditionally been indicated as "Xianghui" in China, yet which or how many fungal species Xianghui comprises is unclear, with various authorities claiming different species. At present, Annulohypoxilon archeri is generally assumed to be the main Xianghui species, but this has not yet been confirmed experimentally. The implementation of older, premolecular-based research data (i.e., morphological) with present, sequence-based data to solve the identity remains confusing and studies addressing both identification methods in combination are lacking. The unclear identity of Xianghui is a major obstacle for further understanding of the important relationship between the host(s) and T. fuciformis. In this study, we collected a wild specimen of T. fuciformis together with several nearby stroma of Xianghui, cocultivated T. fuciformis with the Xianghui isolates, and observed basidiome formation. Internal transcribed spacer (ITS) sequence analysis showed that all Xianghui spore isolates belonged to the same species and both morphological analysis of sexual stages and ITS β-tubulin and actin gene sequences of the Xianghui specified it as Annulohypoxylon stygium. The ITS sequences of the newly identified Xianghui further closely matched those of the Xianghui strains used in the mushroom industry, showing that wild and culture spawn Xianghui in China consist of A. stygium. In contrast with previous conclusions, A. stygium, and not A. archeris, seems to be the preferred host of T. fuciformis.
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isolation of gpd promoter from Tremella fuciformis and driving expression of egfp gene
DNA and Cell Biology, 2009Co-Authors: Dongxing Che, Aogui Xie, Jingui ZhengAbstract:Glyceraldehyde-3-phosphate dehydrogenase (GPD) cDNA was cloned by RT-PCR using total RNA from Tremella fuciformis as template with a pair of degenerate primers. Then, a 500-bp 5′-upstream promoter region of the gene encoding GPD from T. fuciformis genomic DNA was isolated by thermal asymmetric interlaced PCR. The cloned promoter was fused to 5′-upstream of enhanced green fluorescent protein gene to construct T. fuciformis expression vector pCB-TEGFP with hygromycin gene as a selectable marker. Electroporation was performed to transfer plasmid DNA of pCB-TEGFP into yeast-like conidia from T. fuciformis. Molecular evidence, including PCR analysis, fluorescence detection, fluorescence spectra assay, and SDS-PAGE, indicated that the EGFP gene had been integrated into the genome of transgenic T. fuciformis strains and was expressed successfully. The results also showed that this promoter could be used to carry out regulated expression of heterologous gene products in T. fuciformis.
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isolation and optimization of regeneration condition of Tremella fuciformis protoplasts
Mycosystema, 2003Co-Authors: Aogui Xie, Hu ZhuAbstract:Orthogonal design was used for research the high yield protoplast of Tremella fuciformis. The results showed that the material (conidia, mycelia and fruit badies) was the major factor affecting protoplast yield. Using conidia as material, the yield of protoplasts could reach 2.75×10~(7)/ml, while using mycelia and fruit body material, the yield of protoplasts was only 2.5×10~(6)/ml and 1.0×10~(6)/ml respectively. The optimal digesting temperature was 35℃. The lywallzyme concentration ranging from 1% to 3%, did not affect the yield of protoplasts significantly. There was also no significant difference in the yield of protoplasts from different strains. The effect of osmotic stabilizer (KCl) concentration no regeneration of protoplasts was also studied. The results showed that there was no significant difference in the rate of protoplsts regeneration in different concentration. The highest protoplast regeneration rate was 32.3% in KCl concentration ranging from 0.5mol/L to 0.7mol/L.
Liesheng Zheng - One of the best experts on this subject based on the ideXlab platform.
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comparative transcriptomic analysis identified differentially expressed genes and pathways involved in the interaction between Tremella fuciformis and annulohypoxylon stygium
Antonie Van Leeuwenhoek International Journal of General and Molecular Microbiology, 2019Co-Authors: Dongmei Liu, Liesheng Zheng, Yuanyua Wang, Dwi Pujiana, Zhaosong Zhang, Liguo CheAbstract:Tremella fuciformis is an edible and medicinal white jelly mushroom. It has a life cycle that interacts with its companion fungus Annulohypoxylon stygium, both in natural conditions and artificial cultivation. RNA sequencing (RNA-Seq) was used to study the interaction between T. fuciformis and A. stygium by constructing 5 libraries, including the individual T. fuciformis mycelium (1), the T. fuciformis mycelium after interaction with A. stygium (2), the dual mycelia after interaction (3), the A. stygium mycelium after interaction with T. fuciformis (4), and the individual A. stygium mycelium (5). 33.4 G data and 46,871 Unigenes were generated from de novo splicing. For identification of differentially expressed genes (DEGs) related to interaction, we analyzed the expression data of DEGs1-vs-2 ∩ DEGs1-vs-3, and DEGs5-vs-4 ∩ DEGs5-vs-3. DEGs1-vs-2 ∩ DEGs1-vs-3, and DEGs5-vs-4 ∩ DEGs5-vs-3 data showed 614 DEGs and 1537 DEGs, respectively. The 614 DEGs for T. fuciformis and 1537 DEGs for A. stygium were analyzed by GO annotation and were assigned to biology process, cell composition, and molecular functions. The DEGs were used to match the KEGG database. In T. fuciformis, the pathways are primarily enriched various amino acids metabolism, pentose and glucuronate interconversions. In A. stygium, the pathways are primarily enriched in the biosynthesis of secondary metabolites, biosynthesis of antibiotics, starch and sucrose metabolism. The expression patterns of DEGs determined by qRT-PCR were consistent with those obtained by RNA-Seq, thus validating the reliability of our RNA-Seq data. Future studies of the functions of these interesting genes will be helpful to understand the mechanisms by which T. fuciformis interacts with A. stygium. This will also provide a reference for other research on interacting microorganisms.
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Cloning and heterologous expression of a hydrophobin gene Ltr.hyd from the tiger milk mushroom Lentinus tuber-regium in yeast-like cells of Tremella fuciformis
Electronic Journal of Biotechnology, 2018Co-Authors: Yue Chen, Liesheng Zheng, Liguo Chen, Aimin MaAbstract:Background: Hydrophobins are small proteins secreted by filamentous fungi, which show a highly surface activity. Because of the signally self-assembling abilities and surface activities, hydrophobins were considered as candidates in many aspects, for example, stabilizing foams and emulsions in food products. Lentinus tuber-regium, known as tiger milk mushroom, is both an edible and medicinal sclerotium-producing mushroom. Up to now, the hydrophobins of L. tuber-regium have not been identified. Results: In this paper, a Class I hydrophobin gene, Ltr.hyd, was cloned from L. tuber-regium and expressed in the yeast-like cells of Tremella fuciformis mediated by Agrobacterium tumefaciens. The expression vector pGEH-GH was under the control of T. fuciformis glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter. The integration of Ltr.hyd into the genome of T. fuciformis was confirmed by PCR, Southern blot, fluorescence observation and quantitative real-time PCR (qRT-PCR). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that recombinant hydrophobin rLtr.HYD with an expected molecular mass of 13 kDa was extracted. The yield of rLtr.HYD was 0.66 mg/g dry weight. The emulsifying activity of rLtr.HYD was better than the typical food emulsifiers sodium caseinate and Tween 20. Conclusions: We evaluated the emulsifying property of hydrophobin Ltr.HYD, which can be potentially used as a food emulsifier.
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Cloning and heterologous expression of a hydrophobin gene Ltr.hyd from the tiger milk mushroom Lentinus tuber-regium in yeast-like cells of Tremella fuciformis
Elsevier, 2018Co-Authors: Dongmei Liu, Liesheng Zheng, Hanyu Zhu, Yue Che, Liguo CheAbstract:Background: Hydrophobins are small proteins secreted by filamentous fungi, which show a highly surface activity. Because of the signally self-assembling abilities and surface activities, hydrophobins were considered as candidates in many aspects, for example, stabilizing foams and emulsions in food products. Lentinus tuber-regium, known as tiger milk mushroom, is both an edible and medicinal sclerotium-producing mushroom. Up to now, the hydrophobins of L. tuber-regium have not been identified. Results: In this paper, a Class I hydrophobin gene, Ltr.hyd, was cloned from L. tuber-regium and expressed in the yeast-like cells of Tremella fuciformis mediated by Agrobacterium tumefaciens. The expression vector pGEH-GH was under the control of T. fuciformis glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter. The integration of Ltr.hyd into the genome of T. fuciformis was confirmed by PCR, Southern blot, fluorescence observation and quantitative real-time PCR (qRT-PCR). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that recombinant hydrophobin rLtr.HYD with an expected molecular mass of 13 kDa was extracted. The yield of rLtr.HYD was 0.66 mg/g dry weight. The emulsifying activity of rLtr.HYD was better than the typical food emulsifiers sodium caseinate and Tween 20. Conclusions: We evaluated the emulsifying property of hydrophobin Ltr.HYD, which can be potentially used as a food emulsifier. Keywords: Agrobacterium tumefaciens, Emulsifier, Expression vector, Filamentous fungi, Gel electrophoresis, Glyceraldehyde-3-phosphate dehydrogenase, Heterogenous expression, Hydrophobin, Quantitative real-time PCR, Southern blot, Surface activit
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an improved total rna extraction method for white jelly mushroom Tremella fuciformis rich in polysaccharides
Mycobiology, 2017Co-Authors: Hanyu Zhu, Liesheng Zheng, Dongmei Liu, Liguo CheAbstract:An improved method for extracting high quality and quantity RNA from a jelly mushroom and a dimorphic fungus-Tremella fuciformis which is especially rich in polysaccharides, is described. RNA was extracted from T. fuciformis mycelium M1332 and its parental monokaryotic yeast-like cells Y13 and Y32. The A260/280 and A260/230 ratios were both approximately 2, and the RNA integrity number was larger than 8.9. The yields of RNA were between 108 and 213 µg/g fresh wt. Downstream molecular applications including reverse transcriptional PCR and quantitative real-time PCR were also performed. This protocol is reliable and may be widely applicable for total RNA extraction from other jelly mushrooms or filamentous fungi rich in polysaccharides.
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use of the yeast like cells of Tremella fuciformis as a cell factory to produce a pleurotus ostreatus hydrophobin
Biotechnology Letters, 2017Co-Authors: Hanyu Zhu, Liesheng Zheng, Yuanyua Wang, Dongmei Liu, Liguo CheAbstract:Objectives To obtain hydrophobin, a Class I hydrophobin gene, Po.hyd from Pleurotus ostreatus, was transformed into the yeast-like cells of Tremella fuciformis using Agrobacterium tumefaciens.
Hanyu Zhu - One of the best experts on this subject based on the ideXlab platform.
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Cloning and heterologous expression of a hydrophobin gene Ltr.hyd from the tiger milk mushroom Lentinus tuber-regium in yeast-like cells of Tremella fuciformis
Elsevier, 2018Co-Authors: Dongmei Liu, Liesheng Zheng, Hanyu Zhu, Yue Che, Liguo CheAbstract:Background: Hydrophobins are small proteins secreted by filamentous fungi, which show a highly surface activity. Because of the signally self-assembling abilities and surface activities, hydrophobins were considered as candidates in many aspects, for example, stabilizing foams and emulsions in food products. Lentinus tuber-regium, known as tiger milk mushroom, is both an edible and medicinal sclerotium-producing mushroom. Up to now, the hydrophobins of L. tuber-regium have not been identified. Results: In this paper, a Class I hydrophobin gene, Ltr.hyd, was cloned from L. tuber-regium and expressed in the yeast-like cells of Tremella fuciformis mediated by Agrobacterium tumefaciens. The expression vector pGEH-GH was under the control of T. fuciformis glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter. The integration of Ltr.hyd into the genome of T. fuciformis was confirmed by PCR, Southern blot, fluorescence observation and quantitative real-time PCR (qRT-PCR). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that recombinant hydrophobin rLtr.HYD with an expected molecular mass of 13 kDa was extracted. The yield of rLtr.HYD was 0.66 mg/g dry weight. The emulsifying activity of rLtr.HYD was better than the typical food emulsifiers sodium caseinate and Tween 20. Conclusions: We evaluated the emulsifying property of hydrophobin Ltr.HYD, which can be potentially used as a food emulsifier. Keywords: Agrobacterium tumefaciens, Emulsifier, Expression vector, Filamentous fungi, Gel electrophoresis, Glyceraldehyde-3-phosphate dehydrogenase, Heterogenous expression, Hydrophobin, Quantitative real-time PCR, Southern blot, Surface activit
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an improved total rna extraction method for white jelly mushroom Tremella fuciformis rich in polysaccharides
Mycobiology, 2017Co-Authors: Hanyu Zhu, Liesheng Zheng, Dongmei Liu, Liguo CheAbstract:An improved method for extracting high quality and quantity RNA from a jelly mushroom and a dimorphic fungus-Tremella fuciformis which is especially rich in polysaccharides, is described. RNA was extracted from T. fuciformis mycelium M1332 and its parental monokaryotic yeast-like cells Y13 and Y32. The A260/280 and A260/230 ratios were both approximately 2, and the RNA integrity number was larger than 8.9. The yields of RNA were between 108 and 213 µg/g fresh wt. Downstream molecular applications including reverse transcriptional PCR and quantitative real-time PCR were also performed. This protocol is reliable and may be widely applicable for total RNA extraction from other jelly mushrooms or filamentous fungi rich in polysaccharides.
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use of the yeast like cells of Tremella fuciformis as a cell factory to produce a pleurotus ostreatus hydrophobin
Biotechnology Letters, 2017Co-Authors: Hanyu Zhu, Liesheng Zheng, Yuanyua Wang, Dongmei Liu, Liguo CheAbstract:Objectives To obtain hydrophobin, a Class I hydrophobin gene, Po.hyd from Pleurotus ostreatus, was transformed into the yeast-like cells of Tremella fuciformis using Agrobacterium tumefaciens.
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comparing the sugar profiles and primary structures of alkali extracted water soluble polysaccharides in cell wall between the yeast and mycelial phases from Tremella fuciformis
Journal of Microbiology, 2016Co-Authors: Hanyu Zhu, Liesheng Zheng, Jua Liu, Yua Yua, Liguo CheAbstract:To gain insights into dimorphism, cell wall polysaccharides from Tremella fuciformis strains were obtained from alkali-extracted water-soluble fractions PTF-M38 (from the mycelial form), PTF-Y3 and PTF-Y8 (from the yeast form) of T. fuciformis strains were used to gain some insights into dimorphism study. Their chemical properties and structural features were investigated using gel permeation chromatography, gas chromatography, UV and IR spectrophotometry and Congo red binding reactions. The results indicated that the backbones of PTF-M38, PTF-Y3 and PTF-Y8 were configured with α-linkages with average molecular weights of 1.24, 1.08, and 1.19 kDa, respectively. PTF-M38 was mainly composed of xylose, mannose, glucose, and galactose in a ratio of 1:1.47:0.48:0.34, while PTF-Y3 and PTF-Y8 were mainly composed of xylose, mannose and glucose in a ratio of 1:1.65:4.06 and 1:1.21:0.44, respectively. The sugar profiles of PTF-M38, PTF-Y3 and PTF-Y8 were also established for further comparison. These profiles showed that all three polysaccharides contained the same sugars but in different ratios, and the carbon sources (xylose, mannose, glucose, and galactose) affected the sugar ratios within the polysaccharides.
Hu Zhu - One of the best experts on this subject based on the ideXlab platform.
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a three stage culture process for improved exopolysaccharide production by Tremella fuciformis
Bioresource Technology, 2012Co-Authors: Hu Zhu, Wei Liu, Ya Zhang, Aozhe Tia, Shuaishuai Zhang, Chunxu Cao, Weisheng ZouAbstract:Tremella fuciformis produces several bioactive secondary metabolites including exopolysaccharides. Cultivation of the fungus was carried out in a three-stage process consisting of a 1.5-day cultivation with orbital shaking at 200 rpm, a 1.5-day cultivation with reciprocal shaking at 200 strokes, and a 1.5-day cultivation with orbital shaking at 200 rpm. Exopolysaccharide production and specific production rate reached 5.80 g L(-1) and 0.15 d(-1), respectively, which is an increase of 260% and 200% compared with the corresponding values for fermentations with orbital shaking only, and of 243% and 150% compared with the corresponding values for fermentations with reciprocal shaking only. The three-stage culture method is time-saving and easy to operate.
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ph control modes in a 5 l stirred tank bioreactor for cell biomass and exopolysaccharide production by Tremella fuciformis spore
Bioresource Technology, 2011Co-Authors: Hu Zhu, Ya Zhang, Shuaishuai Zhang, Chunxu Cao, Weisheng ZouAbstract:Abstract The effect of pH-control modes on cell growth and exopolysaccharide production by Tremella fuciformis was evaluated in a 5-L bioreactor. The results show that the maximal dry cell weight (DCW) and exopolysaccharide production were 23.57 and 4.48 g L−1 in pH-stat fermentation, where the maximal specific growth rate (μmax) and specific production rate of exopolysaccharide (PP/X) were 1.03 and 0.24 d−1, respectively; under pH-shift cultivation, the maximal DCW and exopolysaccharide production were 30.57 and 3.90 g L−1, where the μmax and PP/X were 1.21 and 0.06 d−1. Unlike batch fermentation, maximal DCW and exopolysaccharide production merely reached 15.04 and 2.0 g L−1, where the μmax and PP/X were 0.86 and 0.05 d−1, respectively. These results suggest that a pH-stat strategy is a more efficient way of performing the fermentation process to increase exopolysaccharide production. Furthermore, this research has also proved that the three-stage pH-control mode is effective for cell growth.
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chromosomal integration of the vitreoscilla hemoglobin gene and its physiological actions in Tremella fuciformis
Applied Microbiology and Biotechnology, 2006Co-Authors: Hu Zhu, Tianwe Wang, Yaling She, Dongzhi WeiAbstract:The Vitreoscilla hemoglobin (VHb) gene was expressed in yeast-like conidia (YLCs) of Tremella fuciformis (T. fuciformis) to increase cell density in submerged fermentation by enhancing oxygen uptake. With the intention of doing this, an integrated expression vector containing the VHb gene and the hygromycin B phosphotransferase (hph) gene derived from Escherichia coli (E. coli) as the selectable marker was constructed, and then transformed into protoplasts of YLCs from T. fuciformis with restriction enzyme-mediated DNA integration (REMI). Hygromycin-resistant transformants had been generated during the transformation. Molecular evidences including PCR assay, Southern blotting, and Western blot analysis indicated the VHb gene had been integrated into the genome of transgenic T. fuciformis strains and was expressed successfully. Shake-flask fermentation and bioreactor cultivation results showed that the expression of VHb in this fungus could enhance growth of YLCs. The final cell density was higher in the culture of VHb-expressing strain than that of the wild-type strain. Moreover, these results also suggested that CaMV35S promoter was capable of driving the expression of heterologous genes in T. fuciformis.
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isolation and optimization of regeneration condition of Tremella fuciformis protoplasts
Mycosystema, 2003Co-Authors: Aogui Xie, Hu ZhuAbstract:Orthogonal design was used for research the high yield protoplast of Tremella fuciformis. The results showed that the material (conidia, mycelia and fruit badies) was the major factor affecting protoplast yield. Using conidia as material, the yield of protoplasts could reach 2.75×10~(7)/ml, while using mycelia and fruit body material, the yield of protoplasts was only 2.5×10~(6)/ml and 1.0×10~(6)/ml respectively. The optimal digesting temperature was 35℃. The lywallzyme concentration ranging from 1% to 3%, did not affect the yield of protoplasts significantly. There was also no significant difference in the yield of protoplasts from different strains. The effect of osmotic stabilizer (KCl) concentration no regeneration of protoplasts was also studied. The results showed that there was no significant difference in the rate of protoplsts regeneration in different concentration. The highest protoplast regeneration rate was 32.3% in KCl concentration ranging from 0.5mol/L to 0.7mol/L.
