Entomopathogenic fungus Cordyceps militaris is famous for its medicinal efficacies. It has been reported to have various pharmacological activities such as anti-tumour, insecticidal, antibacterial, immunomodulatory and antioxidant. In this study, we investigated the effect of the extract of C. militaris (MPUN8501), which was identified by the analysis of the nucleotide sequences of 5.8S ribosomal RNA, on the function of liver. C. militaris powder was extracted using hot water extracts method as time, volume and temperature and using method as differential polarity of organic solvent. Each fraction was tested for the improvement of hepatic enzyme alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activity. The BuOH extracts (CME) had highest activity which was used for the test of toxicity and efficacy of C. militaris. The enhancing effect of CME on the activity of ADH and ALDH was much more than medicine, drink, natural tea etc. Thus CME promoted the resolution of alcohol and acetaldehyde in rats, inducing recovery to normal condition rapidly. Furthermore, oral administration of CME effectively protected the carbon tetrachloride-induced acute hepatic injury as revealed by the hematological parameters (levels of sGOT and sGPT) and histological observation. CME was ascertained to be safe by regulatory toxicity studies of single dose toxicity and genotoxicity. These results suggest that CME would be useful for the maintaining normal hepatic activity as a functional health food.
Four species of nematodes attacking mushroom beds were found in samples taken from 35 mushroom farms throughout Korea. These were Rhabditis sp., Aphelenchoides sp., Ditylenchus sp. and Aphelenchus sp. ,Rhabditis sp. was found from compost and casing from all mushroom farms and the frequencies of Aphelenchoides sp. was 31.4% in the both compost and casing. Both Ditylenchus sp. and Aphelenchus sp. showed 2.7% of frequencies in the compost, none in casing. Temperature and moisture content of compost affected pathogenicity of Aphelenchoides sp. on mushroom mycelia grown in compost. The higher temperature and moisture content the sooner the damage became apparent, and the more rapid was subsequent destruction of mycelia. There was no mycelial destruction at the lowest temperature of $10^{\circ}C$. Rhabditis sp. completely disintegrated mycelia grown in the compost, in the early stage, the numbers of Rhabditis sp. rose gradually and then increased suddenly to reach a peak but soon declined. At first, the pH of Rhabditis-infested spawned compost declined but then rose gradually as mycelia was disintegrated by nematodes. The trend in pH of infested unspawned compost was similar to those of uninfested, unspawned compost. Cultures inoculated with surface-disinfected dead Rhabditis sp. and with tap water used in the nematode extraction procedures showed no mycelial injury associated microorganisms containing within or outside the nematodes even though added by artificial wounding of the mycelia. Cultures artificially wounded showed no injury away from the wounds without the presence of living Rhabditis sp., such wounded mycelia slowly regenerated. On the other hand, artificial wounding accelerated the breakdown of mycelia in the presence of living Rhabditis sp.
Lee, Mun Haeng;Lee, Hee Kyoung;Kim, Sung Eun;Lee, Hwan Gu;Lee, Sun Gye;Yu, Seung Hun;Kim, Young Shik;Kim, Sang Woo;Lee, Youn Su
The Korean Journal of Mycology
/
v.41
no.3
/
pp.172-180
/
2013
Grey mold infection rate in tomato was investigated with the inoculation of dead flowers on Botrytis selective media. The grey mold infection rate of flower after fruiting were higher in the order of after 45 days, after 25 days, and fruiting day with 100%, 87% and 65%, respectively. The number of infected flowers were increased with time increase after the flowering before fruiting. BSM (Botrytis selective medium) was used to check grey mold infection rate depending on the flowering stage and cultivar. Grey mold infection rate depending on the flowering stage was similar in all the beef-tomato cultivar as 1.5~5% at preflowering, 1.5~45% at flowering and 75~90% at fruiting. On the other hand, cherry tomato cultivar "KoKo" had lower infection rates of 0~3.5% at pre-flowering, 10~30% at flowering and 20~50% at fruiting. These resulted from the fact that beaf-tomato cultivar have much bigger flowers and larger amount of pollens compared to those of cherry tomato cultivar. The amounts of falling pollens of Botrytis spp. were checked for beaf-tomato cultivar and cherry tomato cultivar using BSTM. The amounts of falling pollens were increased as growth period was extended, and the amount of spores increased rapidly during the outbreak of grey mold. Twelve field trials in Buyeo and Iksan areas showed that Fluazinam, and Diethofencarb+Carbendazim were effective fungicides to control tomato grey mold, and these results were similar to those of field trials with BSTM. This is the first report of Fluazinam as a effective fungicide for the control of grey mold of tomato even though it has not been registered yet for the control of gray mold in tomato.
From soil samples, 380 antagonistic microorgnisms were isolated. Among the isolates, 42 strains had mycelia growing inhibition ability against Fusariun solani, ginseng root rot causing pathogen. Isolates CHA 1 and S-PFHR 6 were proposed as antagonists for this study and they were identified as Promicromonospora sp. and Pseudomonas pseudoalcaligenes respectively. As an antagonism against hyphae of F. solani in dual culture test, CHA 1 and S-PFHR 6 inhibited linear growing, caused abnormal branching, and the membrane projection which formed by cell wall destruction. The secondary metabolites contained in the culture filtrates which prepared from PD broth and Nutrient broth inhibited the spore germination to 14.3%. The culture filtrate of S-PFHR 6 which prepared by a little amount of soil extract addition to nutrient rich medium had more strongly. inhibited the spore germination and spore germination decreased to less than 4.0% in it. The soil used in this study had fungistasis and the germination rate of macroconidia and chlamydospore of F.solani was 19.4% and 17.7% respectively. The steam sterilized soil lost fungistasis and germination rate of conidia increased to more than 97.9%. The soils amended with the propagule of CHA 1 and S-PFHR 6 increased fungistasis and the germination rate of macroconidia decreased to 14.7% and 11.7% respectively in each treatments. But the soil ammended with glucose and asparagine annulled fungistatic ability and the germination rate of macroconidia increased to more than 48.0%. As an antagonistic activity of the secondary metabolites of two antagonistic isolates in soil, the germination rate of macroconidia of F. solani was 9.3% in the soil amended with the culture filtrate of CHA 1 but the culture filtrate of S-PFHR 6 had no such activity. In the soil which treated with antagonist propagule or culture filtrate, the chlamydospore germination rate was lower than that in natural soil. The addition of glucose and asparagine to antagonist propagule treated soil did not enhanced the chlamydospore germination.
Kim, Jung-Mi;Hong, Sung-Kee;Kim, Wan-Gyu;Lee, Young-Kee;Yu, Seung-Hun;Choi, Hyo-Won
The Korean Journal of Mycology
/
v.38
no.1
/
pp.75-79
/
2010
A total of 25 isolates of Fusarium fujikuroi were obtained from diseased rice plants in Korea from 2006 to 2007 to assess their resistance against fungicides prochloraz and benomyl + thiram. Minimal inhibitory concentration (MIC) values of F. fujikuroi isolates were examined by agar dilution method. Most of the isolates were sensitive to the fungicides. Out of 25 isolates, six were resistant to prochloraz and three to benomyl + thiram. In addition, the isolates CF245, CF249 and CF337 showed resistant to both fungicides. The progenies ($F_1$ isolates) obtained through two different crosses between sensitive parental isolates(CF202, CF232 and CF179) and resistant parental isolate (CF337) were evaluated for their mycelial growth at different temperatures and resistance against fungicides. Mycelial growth rate of $F_1$ isolates originated from CF202 $\times$ CF232 was similar to the parental isolates. However mycelial growth rate of $F_1$ isolates originated from CF179 $\times$ CF337 was faster than their parent isolates. In case of prochloraz, distribution ratio of sensitivity(S) to resistance(R) against to the fungicide of $F_1$ isolates originated from CF202 $\times$ CF232 and CF179 $\times$ CF337 was 86 : 14 and 78 : 22, respectively. In case of benomyl+thiram, all the $F_1$ isolates originated from CF202 $\times$ CF232 were sensitive to the fungicide, however ratio of sensitivity(S) to resistance(R) against to the fungicide of $F_1$ isolates originated from CF179 $\times$ CF337 was 35 : 65.
Ha, Hyo-Cheol;Park, Shin;Park, Kyung-Sook;Lee, Chun-Woo;Jung, In-Chang;Kim, Seon-Hee;Kwon, Yong-Il;Lee, Jae-Sung
The Korean Journal of Mycology
/
v.23
no.2
s.73
/
pp.121-128
/
1995
The characteristics of protein-bound polysaccharides (PBP) which were isolated and purified from the sawdust mycelia of Agrocybe cylindracea were investigated. The yield of crude protein-bound polysaccharides (Fr.CB) extracted with boiling water and precipitated with 95% ethanol, was 0.74% based on the original sawdust mycelia. The Fr.CB was purified by the membrane filtration, ion exchange chromatography and gel filtration. The Fr.B fraction of which the molecular weight is over 300 KDa, was isolated from the Fr.CB using membrane filtration, and the yield was 38.6% based on the Fr.CB. This result indicates that high molecular protein-bound polysaccharides are the dominent components of the Fr.CB. Two fractions (Fr.B-1, Fr.B-2) were also isolated from the Fr.B using ion exchange chromatography, and the yields were 17.3% (Fr.B-1) and 10.3% (Fr. B-2), respectively. The Fr.B-1 was concentrated and gel-filtrated, and the single peak, thought to be nearly pure protein-bound polysaccharides, was obtained. The yield of final fraction $(Fr.B-1-{\beta})$ was 42.5% based on the Fr.B-1. The molecular weight of $Fr.B-1-{\beta}$ was nearly 710 KDa. The monosaccharides' composition of $Fr.B-1-{\beta}$ was analized by HPLC, and glucose was the dominent component, and fucose and galactose were also detected. The result of amino acid analysis was that glutamic acid and analysis were detected to a significant level, and cysteine was not detected.
The studies were carried out to obtain the basic data for maximizing the protoplast yields from the mycelia of P. ostreatus and P. sajor-caju. Some factors affecting the regeneration of the protoplast of both species and the productivity of their reversion were also examined. The maximum yields of protoplasts were obtained from four days cultured mycelia of both species on cellophan membrane placed on the surface of PSA or MCM media in a petri dish. The optimal concentration of lytic enzyme Novozym 234 for protoplast releasing was 5 mg per ml of 0.5 M phosphate buffer solution with 0.6 M sucrose or 0.6 M $MgSO_4$ at pH 6.0. The greatest number of protoplasts was released 3 hours after incubation of the mycelia of P. ostreatus and after 4 hours for the P. sajor-caju in the lytic enzyme solution. Among the osmotic stabilizer solutions tested 0.6 M sucrose and 0.6 M KCl showed the best regeneration rates of the protoplasts of both species. When 0.75 % agar solution was over-layed on the regeneration media immediately after inoculation of the protoplast the regeneration rates were greatly enhanced. The ampicillin added to the agar solution prevented bacteria from infection. The reverted isolates produced the sporophores and basidial spores just like their parents without any mutations when they were cultivated in a broad mouth bottle with sawdust substrates.
This study was conducted to investigate the morphological characteristics and cultural conditions for artificial fruiting body(synnemata) production of Paecilomyces japonica. In the morphological characteristics of P. japonica, the size of it's conidia was ranged from $5.0{\sim}1.5\;to\;7.9{\sim}2.4\;{\mu}m$. The artificial fruiting body showed yellow in color, shape was confirmed ellipsoidal or obovoid type, and the length was $50.6{\sim}104.5\;mm$. The mycelial growth on the PDA medium treated with pH7, at $25^{\circ}C$ was superior to that of other treatments. The formation period of an artificial fruiting body of P. japonica treated with polypropylene and glass bottle culture was 30 days and 50 days, respectively. The length and number of fruiting body was longer and higher in the polypropylene bottle culture than those of the glass bottle culture. As the results, the artificial fruiting body production in the polypropylene bottle increased 1.2g per bottle compared to that of the glass bottle. It also increased in $100{\sim}400\;lx$ illumination, whereas the elongation of synnemata, pinheading and fruiting body growth were inhibited by continuous use of 900 lx illumination. The results of these experiment indicated that fruiting body formation seemed to be lower as the light intensity increased. The fruiting body formation was also dependent on the light color. There was a higher incidence in red color light and fluorescent light treatment than that of incandescent and blue color light. The fruiting body of the naked barley medium had so much better growth compared to other media that it would be able to use for it's production. The growth of fruiting body was affected by $CO_2$ concentration. It increased after putting the lid on the bottle.
Park, Ji-Hyun;Choi, Gyung-Ja;Lee, Seon-Woo;Jang, Kyoung-Soo;Choi, Yong-Ho;Chung, Young-Ryun;Cho, Kwang-Yun;Kim, Jin-Cheol
The Korean Journal of Mycology
/
v.32
no.1
/
pp.31-38
/
2004
An endophytic bacterial strain EB215 that was isolated from cucumber (Cucumis sativus) roots displayed a potent in vivo antifungal activity against Colletotrichum species. The strain was identified as Burkholderia cepacia based on its physiological and biochemical characteristics, and 16S rDNA gene sequence. Optimal medium and incubation period for the production of antifungal substances by B. cepacia EB215 were nutrient broth (NB) and 3 days, respectively. An antifungal substance was isolated from the NB cultures of B. cepacia EB215 strain by centrifugation, n-hexane partitioning, silica gel column chromatography, preparative TLC, and in vitro bioassay. Its chemical structure was determined to be pyrrolnitrin by mass and NMR spectral analyses. Pyrrolnitrin showed potent disease control efficacy of more than 90% against pepper anthracnose (Colletotrichum coccodes), cucumber anthracnose (Colletotrichum orbiculare), rice blast (Magnaporthe grisea) and rice sheath blight (Corticium sasaki) even at a low concentration of $11.1\;{\mu}g/ml$. In addition, it effectively controlled the development of tomato gray mold (Botrytis cinerea) and wheat leaf rust (Puccinia recondita) at concentrations over $33.3\;{\mu}g/ml$. However, it had no antifungal activity against Phytophthora infestans on tomato plants. Further studies on the development of microbial fungicide using B. cepacia EB215 are in progress.
Seo, Geon-Sik;Kim, Byung-Ryun;Park, Myeung-Soo;Kim, Min-Kyung;Yu, Seung-Hun
The Korean Journal of Mycology
/
v.30
no.2
/
pp.86-94
/
2002
Recently a serious outbreak of weed mould caused by a species of Hypocrea occurred in oyster mushroom (Pleurotus ostreatus) substrates in Korea. The disease was characterized by a rapid infestation of the oyster mushroom substrates by Hypocrea sp. and subsequent inhibition of fructification of the mushroom. In spite of it's serious losses to the oyster mushroom industry in Korea, etiology and ecology of the disease have not been studied. Morphological characteristics of the fungus were examined and molecular characteristics of the fungus were compared with those of the green moulds (Trichoderma spp.) isolated from oyster mushroom bed. Stromata formed superficially on suface of the substrates were pulvinate to effuse or irreguler, initially white but becoming yellowish brown, measuring $6.0{\sim}13.0{\times}3.0{\sim}11.0mm$. Perithecia were globose to subglobose, immersed in stroma, $223{\sim}263\;(Ave.239.9){\times}167.3{\sim}231\;(Ave.204.1){\mu}m$ in size. Asci were unitunicate, cylindrical, nonamyloid, $82.7{\sim}124.8\;(Ave.103.3){\times}4.1{\sim}5.1\;(Ave.4.9){\mu}m$ in size, 16 part-spored. Ascospores were bullet-shaped or somewhat oblong, hyaline, bicellular, roughened or warted, $5.4{\sim}7.4\;(Ave.6.5){\times}3.6{\sim}5.5\;(Ave.4.7){\mu}m$ in size. This fungus readily form the stroma on PDA. Mycelia on PDA nearly invisible and without cottony aerial mycelium. Optimum temperature for mycelial growth of this fungus was $25^{\circ}C$ on PDA and its growth rate was 15 mm per day. This species did not grow at below 10 and above $35^{\circ}C$. Phialides in culture enlarged in the middle and aggregated to penicillate type. They were very variable, shorted ampulliform and occasionally curved when matured, but cylinderical when young, measuring $11.9{\sim}24.3\;(Ave.\;14.7){\times}2.9{\sim}3.9\;(Ave.\;3.4){\mu}m$ when matured and $7.2{\sim}14.0\;(Ave.\;10.8){\times}2.8{\sim}4.9\;(Ave.\;3.5){\mu}m$ when young. Phialosopres were ovoid to ellipsoid, smooth, measuring $3.5{\sim}7.2\;(Ave.\;4.5){\times}2.6{\sim}3.3\;(Ave.\;2.9){\mu}m$. Nineteen isolates of Hypocrea sp. were analyzed on the basis of molecular characteristics and classified into phenotypic groups. On the basis of RAPD, URP-PCR, the fungus was confirm to monoclonal, and was classified as a different taxon from reported species of Hypocrea and Trichoderma and supposed to be a new species not previously reported in literature.
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