• Title/Summary/Keyword: URP-PCR

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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.

Genetic Variation in Geographic Crayfish (Cambaroides similis) Populations

  • Yoon, Jong-Man;Kim, Yong-Ho;Kim, Sol
    • Journal of fish pathology
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    • v.19 no.2
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    • pp.141-153
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    • 2006
  • Genomic DNA samples isolated from two geographical crayfish (Cambaroides similis) populations in the inland of the Korean Peninsula, at Jeonju (Jeonju crayfish; JJC) and Jeongup (Jeongup crayfish; JUe), were PCR-amplified repeatedly. The six arbitrarily selected primers OPC-03, OPC-06, OPC-09, URP-02, URP07 and URP-09 generated the common, specific, and polymorphic fragments. The sizes of DNA fragments also varied widely, from 100 bp - 2,600 bp. Here, 521 fragments were identified in the JJC population, and 354 in the JUC population: 6 primers generated 60 specific fragments (60/521 fragment, 11.5%) in the JJC population, and 90 (90/354 fragments, 25.4%) in the JUC population. These primers produced 42 polymorphic fragments (8.1%) in the DC population, and 18 (5.1%) in the mc population. Especially these results demonstrate that the primers detected numerous specific fragments. Especially, the decamer primer OPC-06 generated inter-population-common DNA fragments, approximately 400 and 800 bp, respectively, in both the JJC and JUC populations. The universal primer URP-02 also generated inter-population-identical DNA fragments, approximately 350 bp and 600 bp, between the two geographical crayfish populations. Based on the average bandsharing values of all samples, the bandsharing value of individuals within the JJC population was much higher than in the JUC population. The bandsharing value between individuals no. 10 and no. 15 was 0.683, which was the highest between the two geographical populations. The dendrogram obtained by the six primers indicates two genetic clusters: cluster I (CRAYFISH 01 - CRAYFISH II), and cluster 2 (CRAYFISH 12 - CRAYFISH 22). The genetic distance between the two geographical populations ranged from 0.053 to 0.605. Ultimately, the longest genetic distance displaying significant molecular differences was found to exist between individuals in the two crayfish populations, between individuals CRAYFISH no. 02 of Jeonju and CRAYFTSH no. 15 of Jeongup (genetic distance = 0.605).

Characteristics of a new Pleurotus eryngii cultivar, Aeryni 6 (큰느타리버섯 신품종 '애린이6'의 생육특성)

  • Lee, Song Hee;Kim, Min-Keun;Ryu, Jae-San
    • Journal of Mushroom
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    • v.16 no.1
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    • pp.16-21
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    • 2018
  • In order to breed a new Pleurotus eryngii cultivar exhibiting intermediate size and rapid growth, single crosses between $24{\times}46$ and KNR2539 were performed. The resulting new cultivar, $17{\times}15$, was chosen based on the required days to harvest (15.4 days), quality (7.5), and yield (81.5 g/850 cc bottle) obtained in a preliminary test. The strain was named Aeryni 6 and cultivated at mushroom farms on a large scale for comparison with the reference cultivar Kenneutari No. 2. The yield of Aeryni 6 (76.0 g) was 113.0% compared with the reference cultivar, Kenneutari No. 2, and its quality was 6.8, while that of the reference strain was 5.7. An independent t-test revealed significant differences between the days for harvest, yield, and quality of the two cultivars. The new and reference cultivars were discriminated by PCR amplification of URP2 and URP11 and a confrontation cultivation.

Phylogenic Relationship of Allium Species in Subgenus Rhizirideum by PCR DNA Fingerprint

  • Kim, Haeng-Hoon;Kang, Hee-Wan;Park, Yong-Jin;Baek, Hyung-Jin;Gwag, Jae-Kyun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.4
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    • pp.328-333
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    • 2001
  • Allium is one of the largest genera, which has more than 700 species. PCR by URP (universal rice primer) primers was carried out to get phylogenetic information on 26 species, 62 accessions of subgenus Rhizirideum. The accessions were divided into seven groups at 0.76 similarity level. A. tuberosum (Chinese chives) and A. ramosum represented high similarity of 0.91. A. montanum, A. nutans, A. senescens, A. libani, A. odorum, A. austrosibiricum, and A. narcissiflorium grouped at 0.80 similarity. Some of the wild species, such as A. prostratum, A. polyrhizum, A. odorum, and A. mongolicum, showed different band patterns according to polyploidy, occurrence of B-chromosome, collection site, and origin.

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DNA Fingerprinting Analysis of the Genus Phytophthora in Korea

  • Park, Dong-Suk;Kang, Hee-Wan;Lee, Mi-Hee;Park, Young-Jin;Lee, Byoung-Moo;Hahn, Jang-Ho;Go, Seung-Joo
    • Mycobiology
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    • v.31 no.4
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    • pp.235-247
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    • 2003
  • In order to investigate biodiversity and establish identification system for Phytophthora spp. in Korea, a variety of band pattern was produced by using the URP(universal rice primer). The fingerprint patterns of Phytophthora spp. showed many common and variable fragments according to their isolates in distinct genotypes. In particular, P. drechsleri was classified into four distinct types(I to IV). P. drechsleri(KACC 40498 and KACC 40499) and P. cryptogea(KACC 40413) appeared to have almost equal bands despite their being different species. Ninety isolates of Phytophthora spp. were clustered into 13 groups based on UPGMA(unweighted pair group method with arithmetic means) analysis. These DNA fingerprinting data would be helpful for inter- and intra-species identification of Phytophthora species.

Mutagenesis of of Hypsizygus marmoreus by Gamma Ray Irradiation (감마방사선 조사에 의한 느티만가닥버섯의 변이)

  • Kim, Jong-Kun;Moon, Deok-Hun;Seo, Geon-Sik;Kang, Hee-Wan
    • The Korean Journal of Mycology
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    • v.39 no.3
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    • pp.231-234
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    • 2011
  • This study was performed to develop new Hypsizygus marmoreus cultivars that have enhanced functional materials and improved physiological characteristics with mutagenesis by gamma ray irradiation. Protoplasts of H. marmoreus brown strain HYM-056 were irradiated by gamma ray for mutagenesis, and then 2,000 clones of mutants were randomly selected and the fruiting bodies were induced by bottle culture. Among them, 157 isolates with fast-growing, heavy and many fruiting body-producing were selected. The isolates were cultured in plastic bottle containing rice bran, barley hulls and fir sawdust to form the fruiting bodies. About 100 days after inoculation, characteristic of fruiting bodies were investigated. The isolates were divided into 6 groups based on color, shape and size of pileus, and length, diameter, number and weight of stipe. In addition, the genetic variation of the isolates was analyzed by URP-PCR fingerprinting.

Cultural characteristics of a new spore-less cultivar 'chungheung 1ho' for Lentinula edodes sawdust cultivation (톱밥 재배용 무포자 신품종 '청흥1호'의 생육특성)

  • jeong, Gu-Min;Joung, Eui-Young;Jung, Hea-Young;Jang, Myoung-Jun
    • Journal of Mushroom
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    • v.19 no.3
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    • pp.251-255
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    • 2021
  • The competitiveness of the mushroom industry can be increased by diversifying the domestic Lentinula edodes cultivar. Therefore, 'Chungheung 1ho' was cultivated by hybrid breeding method using strain 'E140025,' which was collected by the Chungheung Mushroom Farming Association. Regarding the morphological characteristics of the fruit, the cap of 'Chungheung 1ho' was smaller than that of the control cultivar, whereas, its stem was longer and thicker than that of the control cultivar. In addition, the weight of the individual fruits of the new cultivar was heavier than that of the control cultivar. The period for primordia formation was 24 days for the new cultivar, which was 16-20 days later than that of the control cultivar. Furthermore, it was found that the patterns of the PCR amplification band using six URP primers were different for 'Chungheung 1ho' than those of the mating strains.

Characteristics of a newly bred Pleurotus eryngii cultivar, Gat_Aeryni (큰느타리(새송이)버섯 신품종 '갓애린이'의 생육특성)

  • Lee, Song Hee;Kim, Min-Keun;Jung, HwaJin;Ryu, Jae-San
    • Journal of Mushroom
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    • v.16 no.3
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    • pp.186-191
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    • 2018
  • In order to breed a new Pleurotus eryngii cultivar with a large pileus and convex shape, which are favorite traits of customers from Europe and North America, single crosses between monokaryotic mycelia derived from basidiospores of KNR2555 were performed to yield the new cultivar $2{\times}12$ chosen by cap shape (convex), diameter of pileus (60.7 mm), and quality (4.9) in a preliminary cultivation. The strain was named Gat_aeryni and was cultivated on a large scale for comparison with Kenneutari 2 ho at the GNARES and at mushroom farms. The yield of Gat_aeryni (71.7 g) was not significantly different from that of Kenneutari 2 ho (71.4 g), and the quality of the new cultivar was 6.8, which was not significantly different from that of a reference cultivar (6.5). Days for harvest, length, and diameter of the pileus in the two cultivars were statistically different by an independent t-test (P < 0.001, P < 0.05, P < 0.001 and P < 0.05 respectively). The new and reference cultivars were discriminated by PCR reactions with a primer set (URP1 and URP10) and simultaneous cultivation.

Differentiation Among Commercial Strains of Pleurotus spp. Based on DNA Fingerprinting Using Universal Rice Primer (URP) (Universal Rice Primer(URP)에 의한 DNA 핵산지문법을 이용한 느타리의 유통 품종간 구분)

  • Seo, Kyoung-In;Jang, Kab-Yeul;Yoo, Young-Bok;Park, Soon-Young;Kim, Kwang-Ho;Kong, Won-Sik
    • The Korean Journal of Mycology
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    • v.36 no.2
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    • pp.130-137
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    • 2008
  • To distinct the commercial strains in Pleurotus spp., 81 strains in eight Pleurotus species were used. DNA fingerprinting using URP-PCR was conducted to determine the phylogenetic relationships among Pleurotus strains. DNA profiles of Pleurotus species obtained by twelve URP primers were analyzed for genetic similarity by NTSYS program. We could divide strains into ten clusters, in which three of them belong to P. ostreatus and the others to the different species, respectively. At the 76% similarity level, 70 P. ostreatus strains were distinguished into three clusters. Cluster I contained 35 strains and some of them showed almost 100% similarity, one strain closely related to Weonhyeong and six strains closely related to Wangheukpyeong. In cluster II, twenty-one out of 23 strains showed 100% to Suhan. Cluster III contained twelve strains, including six strains closely related to Chunchu-2. The results suggested that there are many same strains with different names in mushroom spawn market.