• Journal of Mushroom
• /
• v.12 no.3
• /
• pp.193-200
• /
• 2014

Flammulina velutipes fruit body, Flammulina velutipes mycelium and Cordyceps militaris mycelium were analyzed for their proximate composition, protein-bound polysaccharide, nucleic acid and amino acids. The content of ash and crude fiber in F. velutipes fruit body were higher than F. mycelium and C. militaris mycelium. C. militaris mycelium showed the highest crude fat content while F. velutipes fruit body had lowest. Nitrogen free extract content of the samples varied from 56.8% in F. velutipes fruit body to 61.9% in F. velutipes mycelium. The compositions of total protein and total free sugars of protein-bound polysaccharide were found to be significant differences for all samples. Nucleic acid related compounds were identified the 5'-GMP, 5'-XMP, 5'-IMP in all samples. The content of total nucleic acids were high in the orders of F. velutipes myclial (286.71 mg%), F. velutipes fruit body(187.36 mg%) and C. militaris mycelial(76.85 mg%). The highest content of 5'-GMP was found in F. velutipes fruit body. The most nucleic acid of F. velutipes mycelial and C. militaris mycelial were the 5'-XMP. As for the analysis of total amino acids, seventeen amino acids were identified by HPLC and the major amino acid was glutamic acid in all samples. The content of total amino acids were high in the orders of F. velutipes fruit body(19,919 mg%), F. velutipes mycelium(19,018 mg%) abd C. militaris mycelium(18,965 mg%). We determined the developing new food product such as amino acid drink and amino acid containing food using extracts of Flammulina velutipes fruit body, Flammulina velutipes mycelium and Cordyceps militaris mycelium.

• Mycobiology
• /
• v.43 no.3
• /
• pp.327-332
• /
• 2015

Phenylalanine ammonia-lyase (PAL) gene is known to be expressed in plants, and is involved in the differentiation, growth and synthesis of secondary metabolites. However, its expression in fungi remains to be explored. To understand its expression in mushroom fungi, the PAL gene of the edible mushroom Flammulina velutipes (Fvpal) was cloned and characterized. The cloned Fvpal consists of 2,175 bp, coding for a polypeptide containing 724 amino acids and having 11 introns. The translated amino acid sequence of Fvpal shares a high identity (66%) with that of ectomycorrhizal fungus Tricholoma matsutake. Distinctively, the Fvpal expression in the mycelium was higher in minimal medium supplemented with L-tyrosine than with other aromatic amino acids. During cultivation of the mushroom on sawdust medium, Fvpal expression in the fruit body correspondingly increased as the mushroom grew. In the fruiting body, Fvpal was expressed more in the stipe than in the pileus. These results suggest that F. velutipes PAL activity differs in the different organs of the mushroom. Overall, this is first report to show that the PAL gene expression is associated with mushroom growth in fungi.

• Journal of Microbiology and Biotechnology
• /
• v.19 no.8
• /
• pp.774-780
• /
• 2009

Flammulina velutipes is a popular edible basidiomycete mushroom found in East Asia and is commonly known as winter mushroom. Mushroom development showing dramatic morphological changes by different environmental factors is scientifically and commercially interesting. To create a genetic database and isolate genes regulated during mushroom development, cDNA libraries were constructed from three developmental stages of mycelium, primordium, and fruit body in F. velutipes. We generated a total of 5,431 expressed sequence tags (ESTs) from randomly selected clones from the three cDNA libraries. Of these, 3,332 different unique genes (unigenes) were consistent with 2,442 (73%) singlets and 890 (27%) contigs. This corresponds to a redundancy of 39%. Using a homology search in the gene ontology database, the EST unigenes were classified into the three categories of molecular function (28%), biological process (29%), and cellular component (6%). Comparative analysis found great variations in the unigene expression pattern among the three different unigene sets generated from the cDNA libraries of mycelium, primordium, and fruit body. The 19-34% of total unigenes were unique to each unigene set and only 3% were shared among all three unigene sets. The unique and common representation in F. velutipes unigenes from the three different cDNA libraries suggests great differential gene expression profiles during the different developmental stages of F. velutipes mushroom.

• The Korean Journal of Mycology
• /
• v.16 no.2
• /
• pp.70-78
• /
• 1988

To obtain basic information for the genetic analysis and breeding of Flammulina velutipes, some factors affecting the release of protoplasts from the fungus were studied. Potato Dextrose peptone Agar medium was suitable for the growth of the mycelium and the protoplast formation of F. velutipes. The culture age for the high yields of protoplast was 5 days on PDPA. Few protoplasts were formed from the mycelium cultured on Mushroom minimum Media. The highest yield of protoplasts was obtained in enzyme solution containing Novozyme 234 plus cellulase CP at 10 mg $ml^{-1}$ concentration, while a half amount of protoplasts was obtained in enzyme solution containing Novozyme 234 only. The optimal reaction time of the mycelium in the Iytic enzyme mixtures was 3 hours. The best osmotic stabilizer for the protoplast formation of the mycelium was 0.6M sucrose without buffer at pH 6.2.

• The Korean Journal of Mycology
• /
• v.39 no.3
• /
• pp.235-238
• /
• 2011

Agrobacterium harboring vector pCHBs with hygromycin phosphotransferase(hph) and hepatitis B virus surface antigen(HBsAg)gene was transformed into the mycelial culture of Flammulina velutipes. In particular, mild NaOH solution was treated to the mycelia before Agrobacterium infection step. This was purposed to generate putative surface wounds in the mycelial cell walls. The results showed that hygromycin-resistant($hyg^r$) mycelia could be obtained only from NaOH-treated mycelia but not from intact mycelia. The integration of $hyg^r$ gene in fungal genome was confirmed by PCR. In addition, a single transgene integration and heterologous protein expression in F. velutipes could be verified by Southern blot hybridization and western blot analysis, respectively. This study demonstrated an efficient tool for the Agrobacterium-mediated transformation of F. velutipes mycelia.

• Journal of agriculture & life science
• /
• v.44 no.6
• /
• pp.141-146
• /
• 2010

In this study, Agrobacterium-mediated transformation was tested to the mycelium culture of Flamulina velutipes which is very popular as an edible mushroom in Korea. Particularly, aluminum oxide particles were used to generate wounds in F. velutipes mycelia via vigorous shaking prior to agro-infiltration. The result showed that transformants resistant to hygromycin could be obtained only from the mycelia with physical wounds. Gene transfer was verified by genomic DNA PCR. This study suggested a convenient tool to improve Agrobacterium-mediated transformation of F. velutipes.

• The Korean Journal of Mycology
• /
• v.23 no.1 s.72
• /
• pp.53-60
• /
• 1995

Nutritional requirements for the growth of Flammulina velutipes were studied. Mannitol, glutamic acid and ammonium nitrate were chosen for the maximum mycelial growth when various carbon and nitrogen sources tested. Optimum C : N ratio for the mycelial growth was 20 : 1. Potassium dihydrogen phosphate was selected among the phosphate sources. Magnesium sulphate and thiamine HCl stimulate mycelial growth. Final compositions of optimized chemically defined medium were 1.5% mannitol, 0.082% $NH_4NO_3$, 0.312% glutamic acid, 0.25% $KH_2PO_4$, 0.06% $MgSO_4{\cdot}7H_2O\;and\;0.3\;{\mu}g/l$ thiamin HCl. This medium not only support mycelial growth but also induce fruit body formation.

• Journal of Animal Science and Technology
• /
• v.54 no.5
• /
• pp.341-347
• /
• 2012

The purpose of this study was to evaluate the effects of dietary Flammulina velutipes mycelium (FVM) on broiler chick performance, pathogenic bacterial (E. coli, Salmonella) counts in caecal contents and amount of $NH_3$ in excreta. Ninety-six broiler chicks (HanHyup No. 3, Korea) were divided into four groups: 1) Control (basal diet), 2) T1 (supplemented with 1% FVM), 3) T2 (3% FVM), and T3 (5% FVM), and rose for 7 weeks. In results, there were no significant differences among treatments in weight gain, feed intake, feed efficiency and carcass yield. However, giblets were significantly increased in FVM treatments compared with control group (p<0.05). The number of Salmonella in caecum was significantly decreased in FVM treatments compared with control group (p<0.05). The number of E. coli was decreased in T2 but increased in T1 and T3 compared with control group. The emission of fecal $NH_3$ gas was significantly decreased in accordance with increasing the feeding level of FVM (p<0.05). In conclusion, our data indicated that the supplementation of high level of FVM could inhibit the Salmonella in caecum and reduce the emission of fecal $NH_3$ gas. Therefore, FVM at 5% level could be added in the diet of broiler chicks.

• Mycobiology
• /
• v.47 no.4
• /
• pp.441-448
• /
• 2019

Two new SAM-dependent methyltransferase encoding genes (fvsmt1 and fvsmt2) were identified from the genome of Flammulina velutipes. In order to make a comprehensive characterization of both genes, we performed in silico analysis of both genes and used qRT-PCR to reveal their expression patterns during the development of F. velutipes. There are 4 and 6 exons with total length of 693 and 978 bp in fvsmt2 and fvsmt1, respectively. The deduced proteins, i.e., FVSMT1 and FVSMT2 contained 325 and 230 amino acids with molecular weight 36297 and 24894 Da, respectively. Both proteins contained a SAM-dependent catalytic domain with signature motifs (I, p-I, II, and III) defining the SAM fold. SAM-dependent catalytic domain is located either in the middle or at the N-terminal of FVSMT2 and FVSMT1, respectively. Alignment and phylogenic analysis showed that FVSMT1 is a homolog to a protein-arginine omega-N-methyltransferase, while FVSMT2 is of cinnamoyl CoA O-methyltransferase type and predicted subcellular locations of these proteins are mitochondria and cytoplasm, respectively. qRT-PCR showed that fvsmt1 and fvsmt2 expression was regulated in different developmental stages. The maximum expression levels of fvsmt1 and fvsmt2 were observed in stipe elongation, while no difference was found in mycelium and pileus. These results positively demonstrate that both the methyltransferase encoding genes are involved in the stipe elongation of F. velutipes.

• Journal of Mushroom
• /
• v.10 no.1
• /
• pp.3-8
• /
• 2012

In this study, to produce Flammulina velutipes mushroom liquid spawn efficiently and effectively the effects of explosive aeration (supplying air with tiny bubbles) of the liquid culture medium on carbon dioxide concentration and residual sugar content in the medium on carbon dioxide concentration and residual sugar contentin the medium were measured. Carbon dioxide concentrations were measured at the outlet of the incubator. On the third day the explosive aeration greatly increased mycelial growth of the liquid spawn, and carbon dioxide concentration also greatly increased but decreased after 5 days. Free sugar contents in the liquid culture consistantly decreased up to 7 days and thereafter was not detected. The weight of the mycelia were maintained similar levels after 3 days. Total nitrogen content in the liquid medium constantly decreased during the 11days of explosive aeration. The content of free sugars in 7 days of culture was the lowest level, thus the inoculum incubated for 6~7 days was thought to be the most effective. Carbon dioxide concentration measurement at the outlet of the container during the liquid spawn incubation required low cost but was efficient to estimate the degree of mycelial growth to be used as a simple indicator.