• Title/Summary/Keyword: RAPD method

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Analysis of Microbial Community Structure in Soil and Crop Root System II. Analysis of soil microbial community structure in different soil Environmental conditions by MIDI and DNA analyses (토양과 작물근계의 미생물군집 구조 해석 II. MIDI 및 DNA 분석에 의한 토양환경별 미생물 군집 해석)

  • Ryu, Jin-Chang;Kwon, Soon-Wo;Kim, Jong-Shik;Suh, Jang-Sun;Jung, Beung-Gan;Choi, Sun-Shik
    • Korean Journal of Soil Science and Fertilizer
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    • v.35 no.2
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    • pp.118-126
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    • 2002
  • To evaluate the correlations of microbial populations with soil healthiness and crop production and establish the criteria for microbial population of soil types. We analyzed the microbial community structure of 13 soils which were different in physical and chemical properties and cultivation methods. According to the analysis of microbial population suing the dilution plate method, the large differences of the microbial population structures among soil types were shown: aerobic bacteria $2-27{\times}10^6$, fluorescent Pseudomonas $1-1,364{\times}10^5$, Gram negative bacteria $1-126{\times}10^4$, and mesophilic Bacillus $1-110{\times}10^5$. The density of Gram negative bacteria was highest on red pepper cultivating soils (sample no. 4 and 6) of Umsung and Gesan, Chungbuk, and the density of the fluorescent Pseudomonas was highest on greenhouse soil (sample no. 7) of Jinju, Kyungnam. The crop productivity of three soils was high as compared with those of other soils. It was supposed that the density of fluorescent Pseudomonas and mesophilic Bacillus were correlated with the incresed crop production. By MIDI analysis, 579 strains isolated from 13 soils composed of a variety of microbes including 102 isolates of Agrobacterium, 112 isolates of Bacillus, 32 isolates of Pseudomonas, 44 isolates of Kocuria, and 34 isolates of Pseudomonas. Among the 624 isolates of Gram negative bacteria, Pseudomonas including P. putida and p. fluorescens occupied the highest density (51%), and Stenotrophomonas maltophilia and Burkholderia cepacia also appeared at high density. From RAPD analysis, the fluorescent Pseudomonas strains isolated from 13 soil types showed a high level of strain diversities and were grouped into 2 - 14 patterns according to soil types. Many of unknown bacteria were recovered from the paddy soil, and needed to be further characterized on the molecular basis.

Characterization and breeding of a new cultivar Pleurotus ostreatus 'Heuksol' (느타리버섯 신품종 '흑솔'의 육성 및 특성)

  • Oh, Min-Ji;Im, Ji-Hoon;Shin, Pyung-Gyun;Oh, Youn-Lee;Jang, Kab-Yeul;Kon, Won-Sik
    • Journal of Mushroom
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    • v.15 no.3
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    • pp.129-133
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    • 2017
  • Oyster mushroom is a type of mushroom that is commonly cultivated and consumed in Korea. P. ostreatus 'Suhan' is a preferred cultivar for many mushroom farmers because it has a dark pileus and thick stipe. However, as it is very sensitive to environmental conditions, farmers consistently demand an alternative cultivar. To develop a new cultivar, the parental strains KMCC01680 ('Suhan') and KMCC00478 ('Gosol') were selected from various collected P. ostreatus strains by cultivating genetic resources. P. ostreatus 'Heuksol' was developed by the method of Mon-Mon crossing between monokaryotic strains derived from 'Suhan' and 'Gosol'. Thirty strains of 174 crossed strains were initially selected by cultivation experiments. After bulk cultivation tests, 'Heuksol' was selected. The nuclear DNA profile of 'Heuksol' was similar to those of the parental strains, 'Suhan' and 'Gosol', when RAPD (random amplified polymorphic DNA) primers and UPF (Universal PCR Fingerprinting) 2, 3, and 4 were used. The optimum temperature for mycelial growth was $30^{\circ}C$ for 'Heuksol', but medium-high temperatures were also appropriate, especially $13-20^{\circ}C$. The fruiting body production per bottle (1,100 mL) was approximately 140.8 g. When compared to the control strain 'Suhan', the thickness of the stipe of 'Heuksol' was greater than that of 'Suhan' (13.5 mm vs 9.4 mm). The pileus diameter of 'Heuksol' was similar to that of 'Suhan' and the pileus thickness of 'Heuksol' and 'Suhan' was 19.7 mm and 21.8 mm, respectively. 'Heuksol' had more a productive stipe number than 'Suhan' and the pileus of 'Heuksol' was dark gray, even at high temperatures. Therefore, it was suggested that this new cultivar, 'Heuksol', could provide an alternative to 'Suhan' and contribute to the profit of oyster mushroom farms.

Characteristics and breeding of a new cultivar Pleurotus eryngii, Song-A (큰느타리버섯 신품종 '송아'의 육성 및 그 특성)

  • Shin, Pyung-Gyun;Park, Yun-Jung;Yoo, Young-Bok;Kong, Won-Sik;Jang, Kab-Yeul;Cheong, Jong-Cheon;Oh, Se-Jong;Lee, Keum-Hee
    • Journal of Mushroom
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    • v.9 no.2
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    • pp.59-62
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    • 2011
  • To develop a new cultivar of King oyster mushroom(Pleurotus eryngii), G09-21 as parental strain was selected by the method of Di-mon crossing between monokaryotic strains derived from ASI 2824(Keunneutari No.2) and dikaryotic strain ASI 2887(Aeryni 3). The Pe21-51($G09-21-10{\times}2844-11$) was shown the best cultural characteristics, selected to be a new cultivar and named as 'Song-A'. The 'Song-A' was formed incompatibility line distinctly in the confrontation growth of parental strains Keunneutari No.2, Aeryni 3 and ASI 2844. The optimum temperature for mycelial growth, fruiting body development and pH arrange were $25{\sim}30^{\circ}C$, $14{\sim}16^{\circ}C$ and pH5~8, respectively. Fruiting body production per bottle was about $94.7{\pm}29.5$ g which is almost 106% quantity compared to that of other cultuvar Keunneutari No.2. And also the stip is thick and long but the number of available stipe is few. Analysis of the genetic characteristics of the new cultivar 'Song-A' showed a different DNA profile as that of the control strains, Keunneutari No.2, Aeryni 3 and ASI 2844, when RAPD(Random Amplified Polymorphic DNA) primers URP4 and 7 were used. This new cultivar 'Song-A' of Pleurotus eryngii is characterized by a small number of primordia formation and the stip is thick and long. Therefore, we expect that this new strain will save of labor and cost by without culling work.

Characteristics and breeding of a new multi-generation oyster mushroom (Pleurotus ostreatus) variety 'Dagul' (다발성 신품종 느타리 '다굴'의 육성 및 자실체 특성)

  • Shin, Pyung-Gyun;Kim, Hee-Jung;Choi, Chan-Sik;Yoo, Young-Bok;Kong, Won-Sik;Jang, Kab-Yeul;Oh, Youn-Lee;Cheong, Jong-Chun;Suh, Jang-Sun;Oh, Se Jong;Lee, Keum-Hee
    • Journal of Mushroom
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    • v.11 no.3
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    • pp.154-158
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    • 2013
  • To develop a new variety of oyster mushroom(Pleurotus ostreatus), parental strains was selected by the method of Mon-Mon crossing between monokaryotic strains derived from ASI 2596(Suhan No.3) and ASI 2782(Black pileus mutant). The SB-73(ASI 2596-11 x 2782-8) was shown the best cultural characteristics, selected to be a new variety and named as 'Dagul'. The 'Dagul' was formed incompatibility line distinctly in the confrontation growth of parental strains Suhan No.3 and ASI 2782. The optimum temperature for mycelial growth, fruiting body development and pH arrange were $25{\sim}30^{\circ}C$, $14{\sim}17^{\circ}C$ and pH5~8, respectively. Fruiting body production per bottle was about $68.0{\pm}24.1$ g which is almost 115% quantity compared to that of other variety Suhan No.3. And also the stipe is long and individual generation is multiple. Analysis of the genetic characteristics of the new variety 'Dagul' showed different DNA bands as that of the control strains, Suhan No.3 and ASI 2782, when RAPD(Random Amplified Polymorphic DNA) primers URP7 and Rcb1 were used. This new variety 'Dagul' of oyster mushroom is characterized by multiple of individual generation and the stipe is long. We therefore expect that this new strain will increase of the income by cultivation of field.

Development of artificial cultivation conditions on Tricholoma gigantium (왕송이버섯(Tricholoma gigantium)의 인공재배를 위한 환경조건 구명)

  • Jang, Kab-Yeul;Park, Jeong-Sik;Cheong, Jong-Chun;Kong, Won-Sik;Yoo, Young-Bok;Jhune, Chang-Sung;Sung, Jae-Mo
    • Journal of Mushroom
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    • v.5 no.3_4
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    • pp.98-102
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    • 2007
  • Tricholoma gigantium, an edible mushroom, belongs to Tricholomataceae of Tricholoma and is distributed at Jeju-Do in Korea. It is also well-known as the medicinal mushroom in Taiwan. The cultivation method using the compost was developed in Korea in 1995. To develop a mass cultivation method and a superior strain, four strains were collected and tested. To establish the optimal cultivation conditions, various examinations were accomplished. Bag cultivation was more effective than box cultivation and the optimal relative humidity was more than 80%. Although the mycelial growth was tested in the substrate supplemented with different additives, such as rice bran and wheat bran, there's no significant difference between them. It suggested that the environmental conditions were more important than the substrate additives for cultivation.

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Detection of DNA from Dermatophytes by Polymerase Chain Reaction (Polymerase chain reaction에 의한 동물 유래 피부사상균 DNA의 검출)

  • Kim, Young-Wook;Yeo, Sang-Geon;Choi, Woo-Pil
    • Korean Journal of Veterinary Research
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    • v.42 no.3
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    • pp.363-370
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    • 2002
  • For the development of diagnostic polymerase chain reaction (PCR) to fungal infection by dermatophytes Trichophyton and Microsporum, detection of the fungal DNA by PCR and analysis of the DNA pattern were undertaken in the present study. A total of 15 strains were tested and those consisted of 3 reference strains and 12 isolates such as: reference strains of T mentagrophytes (downy type, ATCC 9533), T rubrum (IFO 6204) and M gypseum (ATCC 9083), and each isolate of T mentogrophytes (powdery type), T mentagrophytes (granular type), T mentogrophytes (purple-red type), T rubrum, T raubitschekii, T tonsurans, T equinum, T ajelloi, T verrucosum, M cookei, M nanum and M gypseum. The DNA were purely isolated from all strains of Trichophyton spp. and Microsporum spp. by a simple method partly consisted of disruption of fungal cells by lyophilization and grinding and extraction of fungal DNA without phenol treatment which is a routine procedure in DNA isolation. For the detection of fungal DNAs, optimal condition of PCR was determined as preheating once at $94^{\circ}C$ for 5 min, 35 cycles of denaturation at $94^{\circ}C$ for 1 min, annealing at $38^{\circ}C$ for 1 min and polymerization at $72^{\circ}C$ for 2 min, and 1 cycle of final extension at $72^{\circ}C$ for 5 min. In PCR using arbitrary primers AP-1 (5' ACCCGACCTG3') and AP-2 (5' ACGGGCCAGT3'), DNAs in various numbers and sizes were detected from different species of Trichophyton and Microsporum, while DNAs in similar size were also detected in all strains of Trichophyton spp. and Microsporum spp. There were unique DNAs observed from certain dermatophytes by AP-1 such as 1,900 bases in T rubrum, 950 and 1,100 bases in T raubitscheldi, 2,100 bases in T equinum, 400 bases in T verrucosum and 1,150 bases in M gypseum. The unique DNAs were also observed by AP-2 such as 1,200 bases in T ajelloi, 250 bases in T verrucosum, 1,150 bases in M cookei and 2,000 bases in M nanum. The results indicated that PCR can detect a specific DNA from certain Trychophyton and Microsporum spp, which can be the information for further development of diagoomc PCR to dennatophytes.