• Title/Summary/Keyword: Random amplified polymorphic DNA (RAPD)

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Characteristics and breeding of a new cultivar Pleurotus eryngii, Seolsong (큰느타리버섯 신품종 '설송'의 육성 및 그 특성)

  • Shin, Pyung-Gyun;Yoo, Young-Bok;Kong, Won-Sik;Jang, Kab-Yeul;Oh, Youn-Lee;Cheong, Jong-Chun
    • Journal of Mushroom
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    • v.11 no.2
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    • pp.77-81
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    • 2013
  • 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 strain ASI 2824-21 derived from ASI 2824(Keunneutari No. 2) and dikaryotic strain ASI 2887(Aeryni 3). The Pe21-53(G09-21-10 x ASI 2844-9) was shown the best cultural characteristics, selected to be a new cultivar and designated as 'Seolsong'. The 'Seolsong' was distinctly formed incompatibility line in the confrontation growth of parental strains Keunneutari No. 2, Aeryni 3 and ASI 2844. Analysis of the genetic characteristics of the new cultivar 'Seolsong' 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) primer URP4 was used. The optimum temperature and pH arrange for mycelial growth were $25{\sim}30^{\circ}C$ and pH 5~8, respectively. This new cultivar 'Seolsong' of fruiting body production per bottle was about $131.4{{\pm}}43.1$ g which is about 102% quantity compared to that of other cultivar Keunneutari No. 2. And also the stipe is thick and long, but the number of available stipe is few. Particularly, it was tolerant of high moisture above 90% during the growth period after primodia formation. We therefore expect that this new strain will save of labor and cost of cultivation by without culling work.

Characteristics and breeding of a new oyster mushroom (Pleurotus ostreatus) variety 'Chunhwashim' by using cytoplasmic hybrid (세포질전환 기법에 의한 신품종 느타리 '천화심'의 육성 및 자실체 특성)

  • Shin, Pyung-Gyun;Yoo, Young-Bok;Kong, Won-Sik;Oh, Youn-Lee
    • Journal of Mushroom
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    • v.13 no.2
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    • pp.129-134
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    • 2015
  • Hybrid strains was selected by crossing between dikaryotic strain ASI 0628 (Dagul) bred from ASI 2596 (Suhan No.3) and ASI 2782(Black pileus mutant) and monokaryotic strain ASI 2344-84. The strain 84 that shown the best cultural characteristics was selected to be a new cytoplasmic hybrid variety and named as 'Chunhwashim'. The 'Chunhwashim' shown incompatibility among them forming the confrontation growth line against parental strains ASI 0628 (Dagul) and ASI 2344 (chunchu 2ho). Fruiting body produced about $161.4{\pm}4.8g$ per bottle. And also the individual generation of fruit body is multiple than chunchu2ho as 40.9. The 'Chunhwashim' was cytoplasmic hybrid included hybrid DNA bands of parental strains ASI 0628 (Dagul) and ASI 2344 (chunchu 2ho) by Random Amplified Polymorphic DNA (RAPD) primer, and mitochondria DNA band of monokaryotic ASI 2344 (chunchu 2ho) using mitochondria microsatellite DNA marker. This new cytoplasmic hybrid variety 'Chunhwashim' of oyster mushroom is characterized by multiple of individual generation and deeply grey color of pileus.

Fruiting body development and genetic analysis of somatic hybrids by protoplast fusion in edible fungi (식용버섯의 원형질체 융합체의 자실체 발생 및 유전분석)

  • Yoo, Young Bok;Kong, Won Sik;Oh, Se Jong;Jhune, Chang Sung;Shin, Pyung Gyun;Kim, Beom Gi;Kim, Gyu Hyun;Park, Minsun;Min, Byung Re
    • Journal of Mushroom
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    • v.2 no.3
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    • pp.115-126
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    • 2004
  • Somatic hybrids of inter-compatible and inter-incompatible strains were obtained by protoplast fusion. The fusion products between compatible strains, Pleurotus ostreatus and P. florida, formed heterokaryons, while fusants between incompatible strains such as P. cornucopiae + P. florida, P. ostreatus + Ganoderma applanatum, P. florida + Ganoderma lucidum, and P. ostreatus + Flammulina velutipes formed synkaryons that retained genes from both parents. The heterokaryons showed the same level of basidioma development. In contrast, the synkaryons showed unique characteristics including clamp connection formation at mitosis, either partner basidioma development, and abnormal segregation and recombination compared with inter-compatible strains. Synkaryons can be classified into homokaryoyic and heterokaryotic type. A comparison of somatic hybrids with compatible and incompatible strains was made using random amplified polymorphic DNA (RAPD) analysis. The heterokaryons between compatible species showed the same level of variability and contained both parental RAPD bands. In contrast, most of the synkaryons between incompatible species showed similarity to those of either parental bands and non-parental RAPD bands. Synkaryons can be classified into microgenome insertion type and macrogenome insertion type. A tetrapolar mating system was found among monospore isolates in somatic hybrids and wild type P. ostreatus. Homokaryons from each somatic hybrid combination were paired with tester homokaryons of the initial wild type of P. ostreatus. The changed mating types were identified in progenies. The pattern of mating type switching in somatic hybrids depends on compatibility of fusion partner. There are several factors related to the mechanism of clamp connection formation and fruiting body development of synkaryons. Of these,the major factor may be associated with self-fertility and mating type switching such as homokaryotic fruiting of wild type P. ostreatus. This review will discuss these aspects.

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Reproductive Ecology and Genetic Variations in the Sea Urchins Anthocidaris crassispina and Hemicentrotus pulcherrimus in the Cheju Coast (제주연안에 서식하는 보라성게 Anthocidaris crassispina와 말똥성게 Hemicentrotus pulcherrimus의 지역별 번식생태학적 특성과 유전적 변이의 비교)

  • 이정재;김범규;강상균;정상철;이기완;최광식
    • Journal of Aquaculture
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    • v.13 no.2
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    • pp.129-135
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    • 2000
  • Genetic variabilities of local populations of Anthocidaris crassispina and Hemicentrotus pulcherrimus were analysed by random amplified polymorphic DNA (RAPD) technique with phenotypic variabilities in timing of gonadal maturation, reproductive output and size of individuals in Cheju. H, pulcherrimus, collected from 4 locations during March 1997, indicated that Daepo individuals were significantly smaller than that at Hamdok, Wimi site A and Wimi site B (ANOVA, p>0.001). Gonodal-somatic index (GSI) of the Wimi site B population was significantly higher than that of three other locations (ANOVA, P>0.0001). An ANOVA test conducted on test size of A. crassispina, harvested from six different locations of Cheju during June 1997, indicated that the size of individuals from Pophwan was significantly smaller (P>0.0001) than that from five other locations. GSIs of urchin m Wimi and Hanlim were significantly higher than that from Pohwan and Oedo (ANOVA, p>0.0001). Genetic similarity, calculated from k13 primer analysis of total DNA, among the six different populations of A. crassispina varied from 0.67 to 0.92, and the values for H. pulcherrimus from 0.60 to 0.73; thus there was no genetic variation among different populations of the same species. Therefore, the populations are genetically homologous and the observed phenotypic variabilities were possibly associated with water temperature and food.

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Genetic Characteristics and Virulence Factors of Pandemic Vibrio parahaemolyticus Isolated in South Korea and Japan (한국과 일본에서 유행하는 장염비브리오의 병원성 인자와 유전자의 특성)

  • Hong, Seuk-Won;Moon, Ji-Young;Lee, Bok-Kwon;Kim, Yung-Bu
    • Journal of Life Science
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    • v.17 no.3 s.83
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    • pp.386-395
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    • 2007
  • A total of 27 strains of Vibrio parahaemolyticus (18 strains isolated from Korea and 9 strains from Japan) were serotyped and examined for biochemical characteristics, antimicrobial susceptibility patterns, cytotoxicity assay, thermostable direct hemolysin (TDH) production and molecular epidemiology. Using polymerase chain reaction (PCR) method and DNA probe hybridization method, the strains were tested for toxR, tdh, trh and ORF 8 genes. The V. parahaemolyticus isolated from patients were belonged to 8 different serotypes : O3:K6, O1:K38, O3:K57, O4:K9, O4:Kl2, O4:K68, O5:Kl5 and O6:K46. Urease-positive strain possessed the trh gene, and conversely, urease-negative strains lacked the gene, indicating that urease production by V. parahemolyticus strains strongly correlates with the possession of the trh gene. Most strains showed multiple resistant to more than three antibiotics and the antibiogram could be classified into 6 group (I to VI). All of the O3:K6 strains isolated in South Korea and Japan producted TDH at high levels. The TDH titers ranged between 256 and 2.048, and the average titer was 1009. To distinguish the new and increasingly common V. parahaemolyticus strains from clinical isolates, ORF 8 is a useful genetic marker. After Southern hybridization, the HindIII restriction fragment patterns of the tdh gene were grouped one type, respectively. One type showed two bands one of which was 4.3kb and the other was 11.5kb in size. Variation between the O3:K6 serotype are minor when compared to the differences seen with the non O3:K6 strains. The migration patterns of Not I -digested of the total DNA of the O3:K6 strains were similar, and only slight variations were observed between the serotypes. By contrast, the O3:K6 strains and non O3:K6 had markedly different profiles. In conclusion, Random amplified polymorphic DNA (RAPD) profile using appropriate primers was an effective epidemiological marker.

Characterization of a new variety Auricularia polytricha (Mont.) Sacc.'Geoni' (털목이 신품종 '건이')

  • Kim, Kil-Ja;Lee, Sook-Jae;Kim, Myeong-Seok;An, Ho-Sub;Kim, Dong-Gwan;Kwon, Oh-Do;Park, Heung-Gyu
    • Journal of Mushroom
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    • v.14 no.4
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    • pp.211-214
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    • 2016
  • Geoni, a new variety of Auricularia polytricha, was bred in JARES(Jeollanamdo Agricultural Research and Extension Services) in 2012. Geoni was selected through the monospore hybridization with JNM21008 and JNM21014 in 2010. Based on a performance test conducted from 2011 to 2012, the Geoni was selected from a line showing an excellent light brown pileus and strong pest resistance. Geoni has a favorable chewiness, light brown and smooth pileus. In addition, Geoni was rich in dietary fiber. MCM(mushroom complete medium), Malt and PDA(Potato Dextrose Agar) mediums were suitable for cultivating the Geoni. The number of effective stipes was 39 ea/0.9 kg and minor axis and major axis of pileus were 6.9 cm and 8.7 cm respectively. The yield of Geoni was 291 g/0.9 kg in plastic bag. Geoni was required 40~54 days for culture at $20^{\circ}C$ and 24days for the primordia and growth period, which is longer than that of the control(Pung-un). Somatic incompatibility was formed between parental strains and Geoni. Analysis of the genetic diversity of the new variety "Geoni"revealed a different profile from the parental strains when RAPD(random Amplified Polymorphic DNA) primers were used.

Breeding of Garden Chrysanthemum Cultivar 'Nuri Ball' (Dendranthema grandiflorum Ramat.) with White Color Petals and Semi-Decorative Type Characteristics (흰색 반겹꽃의 화단국화 '누리볼' 육성)

  • Kim, Dong Chan;Choi, Hyun Gu;Pak, Ha Seung;Lee, Young Hye;Won, Mikyung;Jung, Yun Kyung;Lee, Jung-Soo
    • Horticultural Science & Technology
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    • v.33 no.5
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    • pp.789-795
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    • 2015
  • The new garden chrysanthemum (Dendranthema grandiflorum Ramat.) cultivar 'Nuri Ball' was developed at Yesan Chrysanthemum Experiment Station of Chungcheongnam-do Agricultural Research and Extension Services in 2011. 'Nuri Ball' was bred through a cross between the '02-145-01' line as the female parent with yellow flower color and '02-04-32' as the male plant with white flower color in 2004. Three years of adaptation trials were conducted from 2006 to 2009 under natural conditions. This study compared the external shape type with that of 'White Miri' and conducted ploidy and RAPD (Random amplified polymorphic DNA) marker analyses. These tests showed that 'Nuri Ball' cultivar has its own characteristics compared with the control 'White Miri'. 'Nuri Ball' was a shrub type variety with semi-double flowers of 4.0 cm in width with white petals. It could produce 1025.2 flowers per plant in autumn. Compared with the control 'White Miri', 'Nuri Ball' was similar in terms of shape and color of flowers, but was different in flower size and number. The natural flowering time of 'Nuri Ball' was late September. It had very vigorous growth and an early budding plant. 'Nuri Ball' was demonstrated to be a new cultivar based on ploidy test and RAPD analysis. 'Nuri Ball' is intended for use as a bed chrysanthemum and expected to contribute to farm incomes in landscaping.

High frequency somatic embryogenesis and plant regeneration of interspecific ginseng hybrid between Panax ginseng and Panax quinquefolius

  • Kim, Jong Youn;Adhikari, Prakash Babu;Ahn, Chang Ho;Kim, Dong Hwi;Kim, Young Chang;Han, Jung Yeon;Kondeti, Subramanyam;Choi, Yong Eui
    • Journal of Ginseng Research
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    • v.43 no.1
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    • pp.38-48
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    • 2019
  • Background: Interspecific ginseng hybrid, Panax ginseng ${\times}$ Panax quenquifolius (Pgq) has vigorous growth and produces larger roots than its parents. However, F1 progenies are complete male sterile. Plant tissue culture technology can circumvent the issue and propagate the hybrid. Methods: Murashige and Skoog (MS) medium with different concentrations (0, 2, 4, and 6 mg/L) of 2,4-dichlorophenoxyacetic acid (2,4-D) was used for callus induction and somatic embryogenesis (SE). The embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 Schenk and Hildebrandt (SH) medium. The developed taproots with dormant buds were treated with $GA_3$ to break the bud dormancy, and transferred to soil. Hybrid Pgq plants were verified by random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analyses and by LC-IT-TOF-MS. Results: We conducted a comparative study of somatic embryogenesis (SE) in Pgq and its parents, and attempted to establish the soil transfer of in vitro propagated Pgq tap roots. The Pgq explants showed higher rate of embryogenesis (~56% at 2 mg/L 2,4-D concentration) as well as higher number of embryos per explants (~7 at the same 2,4-D concentration) compared to its either parents. The germinated embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 SH medium to support the continued growth and kept until nutrient depletion induced senescence (NuDIS) of leaf defoliation occurred (4 months). By that time, thickened tap roots with well-developed lateral roots and dormant buds were obtained. All Pgq tap roots pretreated with 20 mg/L $GA_3$ for at least a week produced new shoots after soil transfer. We selected the discriminatory RAPD and ISSR markers to find the interspecific ginseng hybrid among its parents. The $F_1$ hybrid (Pgq) contained species specific 2 ginsenosides (ginsenoside Rf in P. ginseng and pseudoginsenosides $F_{11}$ in P. quinquefolius), and higher amount of other ginsenosides than its parents. Conclusion: Micropropagation of interspecific hybrid ginseng can give an opportunity for continuous production of plants.

Genotypic Variation of Helicobacter pylori Isolated from Gastric Antrum and Body in Korean Patients

  • Park, Seon-Mee;Kwon, Soon-Kil;Son, Bo-Ra;Shin, Kyeong-Seob;Woo, Chan-Won;Kim, Eung-Gook;Kim, Seok-Yong
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.1
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    • pp.19-29
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    • 2000
  • Although most persons infected with Helicobacter pylori harbor a single strain of the organism, multiple strain colonization in the same patient is also occasionally reported in developed countries. The aims of this study were to determine the prevalence of multiple strain colonization in Korean patients and to detect the cagA, iceA1, and babA status of H. pylori isolated from the antrum and body of the stomach. H. pylori was obtained from 35 patients from the antrum and body of the stomach. The genomic diversity of H. pylori was determined by random amplified polymorphic DNA analysis. The status of cagA, iceA1, and babA genes of H. pylori was assessed by polymerase chain reaction with appropriate primers. Clearly different diversity patterns were identified among the isolates from 35 individual patients. Eighteen (51.4%) patients had a single strain of H. pylori. Eight (22.9%) and nine (25.7%) patients had subtypically (one or two bands difference) and typically (clearly different pattern) different strains of H. pylori in the antrum and body, respectively. Among the 70 isolates of H. pylori from 35 patients, the positive rates of 349-bp and 208-bp cagA gene fragments and the iceA1 gene were 68/70 (97.1%), 68/70 (97.1%), and 58/70 (82.9%), respectively. However, the babA gene was found in 22/66 cases (31.4%). In five out of 18 patients with a single strain, the genetic status of cagA, iceA1, and babA varied between the isolates from the antrum and the body. In 8/17 patients with sub typically or typically different strains, the gene status differed between antrum and body isolates. The prevalence of co-colonization with typically or subtypically different strains is high in Korea, and sub-clones with different pathogenic gene status exist within strains of identical RAPD patterns.

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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.