• Title/Summary/Keyword: stromata

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Fruiting Body Formation of Cordyceps militaris from Multi-Ascospore Isolates and Their Single Ascospore Progeny Strains

  • Shrestha, Bhushan;Han, Sang-Kuk;Sung, Jae-Mo;Sung, Gi-Ho
    • Mycobiology
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    • v.40 no.2
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    • pp.100-106
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    • 2012
  • Interest in commercial cultivation and product development of Cordyceps species has shown a recent increase. Due to its biochemical and pharmacological effects, Cordyceps militaris, commonly known as orange caterpillar fungus, is being investigated with great interest. Cultivation of C. militaris has been practiced on a large scale in order to fulfill a demand for scientific investigation and product development. Isolates of C. militaris can be easily established from both spores and tissue. For isolation of spores, ascospores released from mature stromata are trapped in sterile medium. Multi-ascospore isolates, as well as combinations of single ascospore strains, are used for production of fruiting bodies. Progeny ascospore strains can be isolated from artificial fruiting bodies, thus, the cycle of fruiting body production can be continued for a long period of time. In this study, we examined fruiting body production from multi-ascospore isolates and their progeny strains for three generations. $F_1$ progeny strains generally produced a larger number of fruiting bodies, compared with their mother multi-ascospore isolates; however, $F_2$ and $F_3$ progeny strains produced fewer fruiting bodies. Optimum preservation conditions could help to increase the vitality of the progeny strains. In order to retain the fruiting ability of the strains, further testing of various methods of preservation and different methods for isolation should be performed.

Scratching Stimuli of Mycelia Influence Fruiting Body Production and ROS-Scavenging Gene Expression of Cordyceps militaris

  • Liu, Gui-Qing;Qiu, Xue-Hong;Cao, Li;Han, Ri-Chou
    • Mycobiology
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    • v.46 no.4
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    • pp.382-387
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    • 2018
  • The entomopathogenic fungus Cordyceps militaris is a valuable medicinal ascomycete, which degenerates frequently during subsequent culture. To avoid economic losses during industrialized production, scratching stimuli of mycelia was introduced to improve the fruiting body production. The present results indicated that higher yields and biological efficiency were obtained from two degenerate strains (YN1-14 and YN2-7) but not from g38 (an insertional mutant in Rhf1 gene with higher yields and shorter growth periods). Furthermore, the growth periods of the fruiting bodies were at least 5 days earlier when the mycelia were scratched before stromata differentiation. Three ROS-scavenging genes including Cu/Zn superoxide dismutase (CmSod1), Glutathione peroxidase (CmGpx), and Catalase A (CmCat A) were isolated and their expression profiles against scratching were determined in degenerate strain YN1-14 and mutant strain g38. At day 5 after scratching, the expression level of CmGpx significantly decreased for strain g38, but that of CmSod1 significantly increased for YN1-14. These results indicated that scratching is an effective way to promote fruiting body production of degenerate strain, which may be related at least with Rhf1 and active oxygen scavenging genes.

Varietal characteristics of new Cordyceps militaris 'Dowonhongcho 2ho' improved by mating type molecular markers (교배형 분자마커를 이용한 신품종 밀리타리스 동충하초 '도원홍초 2호'의 품종 특성)

  • Lee, Byung-joo;Lee, Mi-Ae;Kim, Yong-Gyun;Lee, Sun-Gye;Choi, Young-sang;Lee, Byung-eui
    • Journal of Mushroom
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    • v.15 no.3
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    • pp.111-117
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    • 2017
  • The mushroom species Cordyceps militaris has been studied and cultivated as a medicinal mushroom due to its multiple valuable biological and pharmaceutical activities. For breeding new strains of C. militaris, multiplex PCR assays were performed using primers specific for its mating type genes, MAT1-1 and MAT1-2. Mating types and mating status were confirmed, as evidenced by DNA bands at 233-bp and 191-bp for MAT1-1 and MAT1-2 respectively. The novel 'Dowonhongcho 2ho' was developed through mating; they were found to possess high-quality fruiting bodies when grown in artificial media. The stromata of the new strain were club-shaped, with a bright orange-red color, and measured 7.1 cm in length. They had an average cordycepin content of 0.33%. Compared to 'Dowonhongcho,' the new strain had a 7% higher yield, as well as firm fruiting bodies. The optimum temperature for mycelial growth was $20{\sim}25^{\circ}C$, and the optimum temperature for stroma development was $18{\sim}22^{\circ}C$. The fruiting bodies developed after 49.1 days from inoculation. The use of mating type molecular markers improved the breeding efficiency of the new strain 'Dowonhongcho 2ho.' Thus, they may be valuable for artificial cultivation and industrial-scale production of C. militaris with excellent characteristics.

Varietal characteristics of cross-bred Cordyceps militaris 'Dowonhongcho' (교배육종에 의한 밀리타리스 동충하초 '도원홍초'의 품종특성)

  • Lee, Byung-joo;Lee, Mi-Ae;Kim, Yong-Gyun;Lee, Kwang-won;Choi, Young-sang;Lee, Byung-eui
    • Journal of Mushroom
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    • v.13 no.3
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    • pp.151-156
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    • 2015
  • Cordyceps militaris is being studied and cultivated as a medicinal mushroom having many valuable biological and pharmaceutical activities. In the breeding of new C. militaris mushroom, single ascospores were isolated and examined their mycelial growth, mycelial density, and production of stroma and perithecia. Among them selected isolates were crossed and hybrids were produced showing high quality fruiting bodies on artificial media. Mycelial growth rate of new strain 'Dowonhongcho' was higher than that of better on 'Yedang 3' on SDAY at $10-25^{\circ}C$. The stromata of new strain were club-shaped and bright orange-red. Its height was 6.1 cm and the cordycepin content was 0.34% on average. The new strain showed 9% higher yield than 'Yedang 3' with producing firmer fruit bodies. The optimum temperature for mycelial growth was $22{\sim}25^{\circ}C$ and the optimum temperature for stroma development was $18{\sim}22^{\circ}C$. Fruiting bodies were began to produce 45 days later after inoculation. This new cultivar may serve as a valuable one for artificial cultivation and industrial-scale production of C. militaris.

Biological Activities and Cultural Characteristics of an Entomogenous fungus, Paecilomyces tenuipes (Peck) Samson (눈꽃동충하초의 배양적 특성 및 생리활성)

  • Ha, Nam-Gyu;Kim, Seung-Yul;Kang, Jin-Ho;Kang, Pil-Don;Sung, Gyoo-Byung;Hong, In-Pyo
    • Journal of Sericultural and Entomological Science
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    • v.47 no.1
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    • pp.12-17
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    • 2005
  • To develop technique for the production of P. tenuipes stromata on a large scale, the infection of P. tenuipes and the growth of stroma were investigated by silkworm (Bombyx mori) variety. Also, studied about biological activites of fruiting body formed on silkworm. Infection rate of the 5th instar larvae of the silkworm with P. tenuipes was the highest in Yangwonjam, followed by Hachojam, Baegokjam and Chilbojam in that order. Also, as the inoculation times was increased, infection rate tended to be raised. The rate of fruiting body formation of the silkworm pupae infected with P. tenuipes was the highest in Baegokjam, followed by Yangwonjam and Chilbojam in the order. But, actually the fruiting body formation of the 5th instar larvae of the silkworm tested was good in Chilbojam, followed by Yangwonjam and Baegokjam in that order in 3 times spraying inoculation. The fruiting bodies of Yangwonjam and Chilbojam infected with P. tenuipes had high amount of Mannitol, but Baegokjam and Hachojam had high concentration of Glucose on a dry weight basis. The mean content of total amino acid in the fruiting bodies of P. tenuipes was 1.03 ${\mu}mole/g$. The distribution rate of amino acid components decreased in the order of Arginine (12.2%)>Glycine (10.5%)> Proline (9.6)>Tyrosine (8.9%)>Serine>Leucine>Threonine. The most abundant amino acid in the fruiting bodies of the Baegokjam, Chilbojam and Hachojam infected with P. tenuipes was arginine, while Yangwonjam was Glycine. The most abundant fatty acid in P.tenuipes was Oleic acid on a dry weight basis. The unsaturated fatty acids such as Oleic acid, Linoleic acid and Linolenic acid accounted for more than 78% of the total fatty acids.

Morphological Characteristics and URP-PCR Analysis of Hypocrea sp., a Weed Mould of Oyster Mushroom Cultivation (느타리버섯 유해균 Hypocrea sp.의 형태 및 URP-PCR 분석)

  • Seo, Geon-Sik;Kim, Byung-Ryun;Park, Myeung-Soo;Kim, Min-Kyung;Yu, Seung-Hun
    • The Korean Journal of Mycology
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    • v.30 no.2
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    • pp.86-94
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    • 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.

Biological Activities and Artificial Cultivation of Cordyceps pruinosa Petch (붉은자루동충하초의 자실체 증식 특성)

  • Hong, In-Pyo;Nam, Sung-Hee;Jung, I-Yeon;Sung, Gyoo-Byung;Kim, Hyun-Bok;Jung, In-Mo;Cho, Soo-Muk;Lee, Min-Woong
    • Journal of Mushroom
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    • v.3 no.4
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    • pp.133-139
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    • 2005
  • Fruiting bodies of Cordyceps have been regarded as popular folk and effective medicines to treat human diseases such as asthma, bronchial and lung inflammation, and kidney disease. Cordyceps pruinosa (Clavicipitaceae; Hypocreales; Ascomycotina) has received special attention for medicinal purpose due to its various physiological activites. The nucleoside derivative N6-(2-hydroxyethyl) adenosine (HEA) isolated from it showed a $Ca^{2+}$ antagonistic effect and negative inotropic response. The artificial production of fruiting body of C. pruinosa has not been tried successfully yet by using living silkworm substrate. To develop techniques for the production of C. pruinosa stromata on a large scale, the infection of Bombyx mori with C. pruinosa and the growth characteristics of stroma of C. pruinosa were investigated. Also, studied about biological activities of fruiting body formed on silkworm. Infection rate of the silkworm pupae with C. pruinosa was the highest in injection inoculation. The formation of the fruiting body of C. pruinosa was quite good in the room controlled at $21{\sim}25^{\circ}C$, over 91% of relative humidity and over 1500 lx. Glucose concentration was high in the fruiting bodies of the silkworm pupae infected with C. pruinosa on a dry weight basis. The most abundant amino acid in the fruiting bodies was arginine and phenylalanine. The fruiting bodies of silkworm pupae infected with C. pruinosa was rich in oleic acid. The high amount of citric acid was found in the fruiting bodies of silkworm pupae infected with C. pruinosa.

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