Ten species of Cordyceps species were collected throughout Kangwon province including Chuncheon Dongsanmyun KNU forest experiment from June to September, 1993. Collected Cordyceps species were identified as Cordyceps militaris, C. roseostromata, C. kyushuensis, C. scarabaeicola, Phytocordyceps ninchukiospora, C. nutans, Paecilomyces tenuipes, C. sphecocephala, Hymenostilbe odonatae, Torrubiella sp.. C. militaris, type species of Cordyceps species, was mainly formed on pupae of Lepidoptera and found after the rainy season around July. Fruiting body of C. roseostromata was morphologically similar to those of C. militaris, but relatively small in size and they were also found on lawn or pupa of Lepidoptera. Fruiting body of C. scarabaeicola was found on adult Scarabaeidae specifically and collect fruiting bodies of C. kyushuensis were on larva of moth. C. nutans and C. sphecocephala had host specificity on Hemiptera and Hymenoptera, respectively. Each species formed elliptical fertile part attach to the slim and carneous stalk and they were collected the most in specimen number through whole season of the summer. Ascospore of Phytocordyceps ninchukiospora on seed was characterized by two viable, multiseptate, fusiform units linked end-to-end by a long, filiform connective. Paecilomyces tenuipes, imperfect stage of the genus Cordyceps is multi-infective fungi that attack all stages of all groups of insects. Hymenostilbe odonatae attacks only adult Odonata and Torrubiella sp. formed on spider was difficult to collect because it was found the back side of leaf. As results of cultural test PDA medium showed the best mycelial growth. In the experiment of effect of the acidity inside of the media, C. militaris was good on pH 5, C. nutans and Phytocordyceps ninchukiospora were good on pH 6 and Paecilomyces tenuipes was on pH 7 and C. scarabaeicola was on pH 9. All isolates tested showed the best mycelial growth at $20^{\circ}C$. Morphologically similar isolates were used to analyze protein banding pattern among and within species. As a result, C. militaris, C. roseostromata and C. kyushuensis were clustered into close species and C. scarabaeicola and Phytocordyceps ninchukiospora were relatively distant from those species.
This study was carried out to identify the phylogenetic relationship among Phellinus species by comparing the DNA sequences of the 5.8S ribosomal DNA (rDNA) and the internal transcribed spacers (ITSs), ITS1 and ITS2 regions. Two primers from the 3' end of 18S rDNA and the 5' end of 28S rDNA sequences were chosen to amplify the specific ITS regions of Phellinus spp. Phellinus strains used in the study were divided into four clusters by the phylogenetic tree based on the amplified regions of ITS and 5.8S rDNA sequences. The first cluster consist of Phellinus hartigii IMSNU 32041 and Phellinus robustus IMSNU 32068, and the second cluster consists of Phellinus linteus strains and Phellinus weirianus IMSNU 32021. Phellinus laevigatus KCTC 6229, KCTC 6230 and Phellinus igniarius KCTC 6227, KCTC 6228 belong to the third cluster. Finally, Phellinus chrysoloma KCTC 6225 and Phellinus chrysoloma KCTC 6226 are the fourth cluster. In the second cluster the differentiation between Phellinus linteus strains and Phellinus weirianus species were not possible by the comparison of the ITS sequences. These results revealed that Phellinus linteus and Phellinus weirianus cannot be established the concept of species level only by the ITS sequences. Therefore, both physiological and molecular biological methods as well as the sequences of type strains are necessary to classify the strains of these two species accurately. The comparison of the ITS sequences of four Phellinus species indicated that the sequences of the ITS1 generally are more divergent than those of the ITS2. Although the ITS sequences are varied in some species, the conserved regions in both ITS1 and ITS2 are useful tool to differentiate the species. Phellinus linteus and related species have their specific sequences in the ITS1 compared to the other species.
Taxonomic and genetic analysis of Phytophthora species belonging to six different morphological groups (GI, GII, GIII, GIV, GV, GVI) was conducted using RAPD method. Amplified fragments ranged $0.3{\sim}3.2$ kb in their molecular weights. Among total of 145 bands, there were 109 polymorphic bands. Seven isolates of P. infestans showed high similarities of $0.92{\sim}0.99$, and P. infestans isolate 3 from potato showed similarities of $0.93{\sim}0.95$ compared with other P. infestans. Among isolates of P. capsici, similarities of $0.77{\sim}0.86$ were observed and they were grouped in 80% level. P. cinnamomi and P. cryptogea isolates which belonging to group GVI showed very similar RAPD fingerprinting pattern. Primers OPA-04, OPA-17, OPA-18, OPA-19, and OPB-12 showed high level of differences among the tested isolates in major bands and molecular weights. The similarity between the isolates was 0.67. P. megasperma and P. sojae in group GV showed similarity of 0.65. These two isolates showed big differences in single major band in reactions with primers OPA-08, OPA-17, and OPA-19. Phytophthora-specific and P. infestans-specific molecular markers were also selected with one of the random primers tested. In reaction with primer OPA-20, all the genus Phytophthora showed common band at 600 bp, and all the P. infestans isolates showed specific band at 680 bp. These markers can be useful for identification of Phytophthora speices or P. infestans. As a result, P. infestans isolated from tomato and/or potato can easily be differentiated from other Phytophthora species with this primer.
Kim, Han-Kyoung;Cheong, Jong-Chun;Seok, Soon-Ja;Kim, Gwang-Po;Cha, Dong-Yeul;Moon, Byung-Ju
The Korean Journal of Mycology
/
v.25
no.4
s.83
/
pp.311-319
/
1997
These studies were carried out to develop an artificial cultivation method. The diameter and thickness of pileus ranged $1.5{\sim}7.0\;cm$ and $0.8{\sim}3.0\;cm$, respectively. The diameters of stipe were $1.2{\sim}2.5\;cm$ and the lengthes were $4.5{\sim}9.0\;cm$. The spore fingerprint was white. The sizes were spore $10.8{\sim}12.2{\times}4.35{\sim}5.65\;{\mu}m$, basidia $50.0{\sim}59.2{\times}7.4{\sim}7.8\;{\mu}m$, nalsistidia $21.75{\sim}28.7{\times}4.8{\sim}6.1\;{\mu}m$, pileus hymenium cell $50.6{\sim}66.0{\times}4.4{\sim}6.7\;{\mu}m$, and stipe hymenium cell $28.6{\sim}33.0{\times}5.5{\sim}6.6\;{\mu}m$. The thirty percent mixture of rice and wheat bran into sawdust gives the high density of mycelia and the good development of fruiting structure. The optimum water contents of sawdust substrates were $60{\sim}65%$ in which condition the mycelium grows well and gives high density. In PP bottle cultivation, the first fruiting period was $6{\sim}8$ days earlier in nonscratching samples than scratching ones, but the quantity of fruiting body was higher in scratching samples than nonscratching ones. In the case of PP bag cultivation, the first fruiting was 10 days faster, and the quantity of fruiting bodies was 30% higher in samples with 30% wheat bran than those with rice bran. The fleshiness of stipe was $2{\sim}3$ times harder than that of pileus.
Morphological and cultural characteristics of fungi causing rice sclerotial diseases were examined. Hyphal widths of R. solani and R. oryzae were same and ranged $6.0-12.0\;{\mu}m$ with average $9.0\;{\mu}m$, the widest among those of the sclerotial fungi examined. Hyphal width of R. oryzae sativae ranged $6.0-9.0{\mu}m$ with average $7.4{\mu}m$. Hyphal width of R. cerealis was the narrowest among those of Rhizoctonia species examined, and the same was hyphal width of S. oryzae among those of Sclerotium species. Nuclear staining by HCL-Giemsa method showed that R. solani and R. oryzae had many nuclei within one hyphal cell, S. oryzae one nucleus, and the other sclerotial fungi mostly two nuclei. The nuclear number of R. solani was the largest, which ranged 2-17 with average 6.3. Average size of sclerotia of the sclerotial fungi except S. hydrophilum and S. oryzae produced in lesions ranged 1.0-2.0mm. Average size of sclerotia of S. hydrophilum and S. oryzae was 0.5mm and 0.24mm, respectively. Sclerotia of R. solani and R. oryzae produced in culture were more variable in size and larger than those produced in lesions. However, the sclerotial sizes of the other sclerotial fungi produced in culture were almost the same as those produced in lesions. Sclerotial colors of sclerotial fungi produced in lesions were similar to those produced in culture, but sclerotial shapes of some sclerotial fungi exhibited somewhat difference between the sclerotia produced in lesions and in culture. Optimum temperature for mycelial growth of R. cerealis was $23^{\circ}C$, and that of the other sclerotial fungi ranged from $27\;to\;33^{\circ}C$. Maximum temperature for mycelial growth of some sclerotial fungi was as high as $41^{\circ}C$, while that of R. cerealis was as low as $31^{\circ}C$. Minimum temperature for mycelial growth of R. cerealis was $2^{\circ}C$, and that of the other sclerotial fungi ranged from $6\;to\;10^{\circ}C$.
To investigate nutritive values of a feed fermented with basidiomycetes, among the isolated strains, Lyophyllum decastes (Fr.) Sing. was found with the greatest enzyme productivity and rapid mycelial growth in rice straw medium. Optimum temperature, pH and moisture content for mycelial growth and enzyme production of the strain were $25{\sim}30^{\circ}C,\;pH\;4.0{\sim}7.0\;and\;70{\sim}75\;%$, respectively. Fifteen days of culture were required for the highest enzyme productivity. Among the sub-materials added, $30{\sim}40\;%$ of rice bran and $10{\sim}20\;%$ of defatted perilla seeds were effective for the enzyme production, but caused a reduced mycelial growth. The greatest effect of an addition of inorganic salts was obtained with $0.36{\sim}0.72\;%\;of\;(NH_4)_2HPO_4$. When 40 mesh or smaller rice straw and steam treatment at $0.5\;kg/cm^2$ were used, the mycelial growth decreased, whereas the enzyme production increased. The mycelial growth and enzyme production increased when $Ca(OH)_2$ was used as the alkali treatment, but decreased with increasing concentration of NaOH. As the fermentation proceeded, the amounts of ash, reducing sugar and total nitrogen increased, but cellulose, lignin and pentosan decreased. When the rice straw was treated with alkali, the amounts of ash, total nitrogen and lignin decreased, but reducing sugar and cellulose increased. At higher NaOH concentration, the variation become greater. The in vitro dry matter digestibility of the products increased from 55.03 % at the beginning of the fermentation to 62.72 % at 45 days after fermentation. The most effective alkali treatment on the digestibility of rice straw was KOH followed by NaOH. However, the digestibility increased with increasing concentration of NaOH. The digestibility of pretreated with alkali increased after fermentation as well.
As a preliminary study in order to develop new varieties of Hericium species, this study was carried out to investigate the optimal temperature for mycelial growth, to figure out the applicability to sawdust cultivation on Quercus mongolica substrate, and to analyze the antioxidant capacity of ergothioneine and polyphenols in Hericium strains preserved in Korea Forest Research Institute (KFRI). In the results of optimal temperature for mycelial growth of eight Hericium erinaceus, it was $20^{\circ}C$ in a strain (KFRI 842), $25^{\circ}C$ in five strains (KFRI 507, 508, 509, 843, 845), and $30^{\circ}C$ in two strains (KFRI 582, 844). Optimal temperature for mycelial growth of H. coralloides (KFRI 713) was $25^{\circ}C$. Four strains (KFRI 508, 843, 844, 713) out of the total nine Hericium strains showed full mycelium growth within 20 days at the optimal temperature on PDA medium in petri-dish (85 mm in diameter). The other strains have need of more time for full mycelium growth. Mushroom production of H. erinaceus ranged from 215 to 384 g of fresh weight and its dry weight was 7 to 9% of it, whereas that of H. coralloides was 299 g of fresh weight and its dry weight was 10% of it. The contents of ergothioneine and polyphenols of H. erinaceus strains were different by strains and those were in the range of $1.6{\sim}3.7$ mg/g dw. and $5.9{\sim}7.8$ mg/g dw., respectively. On the other hand, those of H. coraloides were in the range of 1.7 mg/g dw. and 3.9 mg/g dw., respectively. From the results of correlation ($R^2$ = 0.1) between ergothioneine and polyphenols in the strains, it was found that the total contents of them differ by strains but the ratio of the two compounds was not very different in the strains.
The survival or colonization of beneficial organsisms and suppression of root rot of ginseng (Panax ginseng) by two distinct bacteria, Pseudomonas cepacia, Bacillus cereus and three mycorrhiza in pot soil were investigated and compared with uninoculated root. In separate inoculation, colonization of roots by P. cepacia was maintained at 6.25 (log cfu/g root) during growth for 10 days under pot culture conditions comparing to $5.62{\sim}6.19$ by mixed treatment with other organisms. Colonizations of P. cepacia were gradually decreased from 6.25 (log cfu/g root) in 10 days growth to 3.01 (log cfu/g root) in 270 days incubation period. This reduction was also investgated in combination treatments by B. cereus or F. solani. The numbers of Fusarium spp. were colonized high number in rhizosphere soil from 3.33 to 3.67 (log cfu/g root) in control within $10{\sim}60$days after treatment of pathogen F. solani, but it's numbers were markedly decreased in 270 days cultivation of plant from 3.33 to 1.02 (log cfu/g root) after treatment. In treatment of beneficial strains of P. cepacia and B. cereus, P. cepacia significantly suppressed the development of root rot from 4.3 in control to 1.2 in treatment, whereas B. cereus alone had no effect on the rate of disease suppression. The disease index $(1.8{\sim}2.3)$ in combination of two bacteria was reduced in plants inoculated with both P. cepacia and B. cereus comparing to the index (4.3) of control. As an effect of inoculation with mycorrhiza on disease suppression, suppression of root rot by F. solani was reduced to $1.2{\sim}1.6$ in disease index in treatment of Glomus albidum and Acaulospora longular comparing to 4.3 of control. In the treatment of bacterial strain P. cepacia and mycorrhizal fungus Glomus albidum, the disease suppression was apparent to 1.2 and 1.2 comparing to 4.3 of control in disease index respectively.
A hot water extract was prepared from the artificially grown Gastrodia elata to investigate its chemical composition and various in vitro biological activities as an effort to develop G. elata as health/functional food materials. The contents of crude protein, ash, fat, fiber, moisture and total sugar were 5.4, 2.6, 3.6, 3.3, 8.1 and 77% (w/w), respectively. The extract of G. elata had greater amount of potassium (1,150 mg/100 g) than phosphorus (300 mg/100 g). Dose-dependence against human carcinoma (Hep3B, MCF-7, A549 and AGS) were observed from 0.2 mg/ml to 1.0 mg/ml. Especially, the treatment of 1.0 mg/ml extracts showed the highest cytotoxicity with 83% against gastric carcinoma (AGS). The extracts showed weak antimicrobial activities against Bacillus subtilis and Pseudomonas aeruginosa, but practically no antimicrobial activity against the other microorganisms tested. The effect of ${\alpha}$-glucosidase inhibition was 64% at the concentration of 1.0 mg/ml. The inhibitory effect of angiotensin converting enzyme (ACE) of the extract in the range of $0.2{\sim}1.0mg/ml$ showed $63{\sim}89%$, and the highest ACE inhibition was 89% at the concentration of 1.0 mg/ml of extracts. The highest activity of glutathion S-transferase (GST) was 221 % at the concentration of 1.0 mg/ml of the G. elata extracts. These results suggest that G. elata may be used as health/functional food materials.
Min-Jun Kim;Yeun Sug Jeong;Eunjin Kim;Yeongseon Jang;Kang-Hyeon Ka
The Korean Journal of Mycology
/
v.51
no.2
/
pp.81-90
/
2023
The cultivation method of shiitake is divided into two methods: log cultivation and sawdust cultivation. Recently, the importance of sawdust cultivation has been highlighted due to problems such as environmental problems, rising labor costs, and a reduced labor force. Among the 24 Sawdust-cultivated Lentinula edodes cultivars in Korea stored by the National Institute of Forest Science, this study was conducted to select excellent cultivars under the sawdust block cultivation method. After inoculation, the sawdust mediums were cultured for 100 days (60 days in dark and then 40 days in light), and only Sanjo 712ho sprouted the primordia on the 10th day of light culture (total 70 days of cultivation). As a result of cultivation, the average total yield of the 24 cultivars were 1,816 g and Sanjo 712ho was 2,267 g. The fruiting body yield was the highest in Sanjo 713ho with 3,443 g followed by Sanjo 710ho (3,355 g). Sanlim 10ho, Sansanhyang, and Sanjo 716ho showed low production with 174, 238, and 214 g, respectively. As a result of investigating the morphological characteristics of the fruit bodies, the fresh weight of Sansanhyang and Bambithyang was about twice as heavy as the overall average, and Sulbaekhyang was about twice as light. Bambithyang was the largest and longest in diameter and length of the stipe, and the thickness of the stem was the thickest in Sanjo 716ho. As for the hardness of pileus, Bambithyang showed the highest value at 1,276 g/5 mm, and Sanjo 711ho showed the lowest value at 542 g/5 mm. In summary, Bambithyang showed the best fruiting body characteristics and Sanjo 713ho showed the highest yield in the sawdust block cultivation method. This study, the cultivar with shortened cultivation periods and cultivars with excellent morphological characteristics and high yields.
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