• Horticultural Science & Technology
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• 제28권4호
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• pp.638-647
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• 2010

The genetic relationships in 5 species of $Ligularia$ were investigated using RAPD (Randomly Amplified Polymorphic DNA) and ITS (Internal Transcribed Spacer) sequences analyses. In RAPD analysis, sixty three of 196 arbitrary primers showed polymorphism. The amplified fragments ranged from 0.2 to 1.6 kb in size. The dendrogram was constructed by the UPGMA clustering algorithm based on genetic similarity of RAPD markers. A total of 16 accessions were classified into 5 major groups corresponding each species at the similarity coefficient value of 0.77. In the ITS sequence analysis, the size of ITS 1 was varied from 248 to 256 bp, while ITS 2 was varied from 220 to 222 bp. The 5.8S coding region was 164 bp in lengths. Forty nine sites (10.2%) of the 478 nucleotides were variable, and the G+C content of ITS region ranged from 49.4 to 53.5%. In the ITS tree, five species of $Ligularia$ were monophyletic, and $L.$ $taquetii$ was the first branching within the clade. $Ligularia$ $intermedia$ formed a clade with $L.$ $fischeri$ var. $spiciformis$ (BS=79), and $L.$ $stenocephala$ and $L.$ $fischeri$ were also claded. Two data sets were congruent, except of the position of $L.$ $fischeri$ var. $spiciformis$.

• Korean Journal of Veterinary Service
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• 제31권1호
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• pp.1-16
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• 2008

Salmonella infections cause the disease in pigs but also some zoonotic Salmonella serotypes can be transmitted to human through swine products, resulting in food poisoning. The objective of this study was to investigate the bacteriological prevalence and detection of invA gene using Salmonella specific polymerase chain reaction (PCR), the epidemiological characteristics related to Salmonella strains cultured from pig samples in Gangwon areas using serotyping, random amplified polymorphic DNA (RAPD) and pulsed field gel electrophoresis (PFGE) methods. During the period of November 2001 through April 2002, 1,174 ileocecal lymph node were collected from the slaughtered pigs raised in 38 farms located in Gangwon province. The samples were submerged in boiling water and macerated in saline and lymph node homogenates were inoculated into Tetrathionate broth with iodine (TTB, Difco, 0.5% iodine was added) for enrichment growth. Then additional tests were performed using several mediums, and suspects were identified by API 20E kit (BioMerieux) and PCR. Of total 1,174 samples from 38 farms, 44 (3.7%) were isolated as Salmonella spp from 13 farms (34.2%). Of 44 isolates, 31 were in Yangyang region, followed by 9 in Goseong, 2 in both Gangneung and Sokcho. However, there was no difference in regional isolation frequency. All isolates have a 521bp amplified product in Salmonella specific PCR with primer invA which encodes in proteins for invasion of epithelial cells. Of 44 recovered serotypes, 23 (52.3%) were S Eingedi, 10 (22.7%) S Schwarzengrund, 9 (20.5%) S Typhimurium, and 2 (4.5%) S Mbandaka. In RAPD analysis, there appeared to be unique bands distinguishing each serotype, although similarities exist between the different serotypes. Four serotypes of 44 Salmonella isolates appeared to fall into 14 different RAPD types. In PFGE analysis, 9 S Typhimurium were tested with XbaI enzyme and SpeI enzyme. The combination of results obtained with two enzymes subdivided the 9 S Typhimurium into 4 PFGE types.

• Journal of Life Science
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• 제31권1호
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• pp.52-58
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• 2021

Pseudomonas syringae pv. actinidiae, which causes bacterial canker in kiwifruit, is divided into five biovars (1, 2, 3, 5, 6) on the basis of genetic characteristics and toxin productivity. Among them, biovar 3 is responsible for the current global outbreak, and has been isolated in Korea since 2011. Biovar 3 strains isolated from Korea are subdivided into six genetically different lineages (subgroup I, IV, V, VI, VII, and VIII) based on random amplified polymorphic DNA (RAPD) analysis. In this work, the subgroup-specific sequence characterized amplified region (SCAR) primers were developed from sequenced differential RAPD bands. Distribution of the subgroups of the biovar 3 strains collected in Korea from 2011-2017 were examined using these subgroup-specific primer sets. Among the 54 strains tested, 35 strains (64.8%) belonged to subgroup V, 9 strains (16.7%) belonged to subgroup IV, 4 strains (7.4%) belonged to subgroup VI, 3 strains (5.6%) belonged to subgroup VII, 2 strains (3.7%) belonged to subgroup VIII, and 1 (1.9%) strain belonged to subgroup I. Strains belonging to subgroups IV, V, and VI were shown to be related to strains isolated from China, New Zealand, and Chile, respectively. The study revealed that the biovar 3 strains in Korea are genetically diverse and are estimated to have been introduced through pollen sourced from foreign countries.

• Horticultural Science & Technology
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• 제28권5호
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• pp.828-835
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• 2010

Conventional methods for identification of apple cultivars are based on the evaluation of sets of morphological characteristics, however, closely related cultivars often cannot be distinguished by morphological traits. This study was conducted to develop DNA markers for discrimination of the apple cultivars bred in Korea. Thirty random primers generated eighty-three random amplified polymorphic DNA (RAPD) markers from thirty-one Korean bred and introduced apple cultivars. Fifty-two RAPD fragments were cloned and sequenced for conversion into sequence characterized amplified region (SCAR) markers. Among them only seventeen SCAR markers resulted in the amplification of single major bands the same size as the RAPD fragment cloned. Several combinations of six (AN11_433, AN08_566, A408_592, AK17_653, AO04_711, AO04_779 or AW15_368, AN11_433, A408_592, AK17_653, AO04_711, AO04_779, or AL1_427, AN11_433, AN08_566, A408_592, AK17_653, AO04_779) to seven (AL1_427, AN11_433, AN08_566, A408_592, AK17_653, AM16_708, AO04_779 or A330_424, AN11_433, AG14_502, AN08_566, A408_592, AK17_653, AO04_779 or A330_424, AN11_433, AK14_564, A408_592, AK17_653, AM16_708, AT14_789) SCAR markers provided enough polymorphism to identify sixteen Korean apple cultivars among thirty-one tested cultivars. Therefore, application of the seventeen SCAR markers was sufficient to identify the thirty-one tested apple cultivars. These markers could be utilized as a reliable tool for cultivar discrimination of Korean apples.

• Molecules and Cells
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• 제26권2호
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• pp.146-151
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• 2008

The brown planthopper (BPH) is a major insect pest in rice, and damages these plants by sucking phloem-sap and transmitting viral diseases. Many BPH resistance genes have been identified in indica varieties and wild rice accessions, but none has yet been cloned. In the present study we report fine mapping of the region containing the Bph1 locus, which enabled us to perform marker-aided selection (MAS). We used 273 F8 recombinant inbred lines (RILs) derived from a cross between Cheongcheongbyeo, an indica type variety harboring Bph1 from Mudgo, and Hwayeongbyeo, a BPH susceptible japonica variety. By random amplification of polymorphic DNA (RAPD) analysis using 656 random 10-mer primers, three RAPD markers (OPH09, OPA10 and OPA15) linked to Bph1 were identified and converted to SCAR (sequence characterized amplified region) markers. These markers were found to be contained in two BAC clones derived from chromosome 12: OPH09 on OSJNBa0011B18, and both OPA10 and OPA15 on OSJNBa0040E10. By sequence analysis of ten additional BAC clones evenly distributed between OSJNBa0011B18 and OSJNBa0040E10, we developed 15 STS markers. Of these, pBPH4 and pBPH14 flanked Bph1 at distances of 0.2 cM and 0.8 cM, respectively. The STS markers pBPH9, pBPH19, pBPH20, and pBPH21 co-segregated with Bph1. These markers were shown to be very useful for marker-assisted selection (MAS) in breeding populations of 32 F6 RILs from a cross between Andabyeo and IR71190, and 32 F5 RILs from a cross between Andabyeo and Suwon452.

• Microbiology and Biotechnology Letters
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• 제43권3호
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• pp.195-203
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• 2015

Two bacterial strains, Bacillus licheniformis YB-1413 and YB-1414, producing extracellular α-galactosidase, were obtained from homemade Doenjang. On the basis of their biochemical properties, 16S rRNA sequences and random amplified polymorphic DNA patterns by polymerase chain reaction, they were found to be somewhat different from one another. α-Galactosidase productivities of the two isolates were increased by wheat bran, but drastically decreased by melibiose, raffinose and sucrose which were used as carbon sources. The enzyme productivities were increased by yeast extract as a nitrogen source with maximum levels of 1.87 U/ml for YB-1413 and 1.69 U/ml for YB-1414, respectively. The enzymes of both isolates exhibited maximum activity for hydrolysis of para-nitrophenyl-α-D-galactopyranoside (pNP-αGal) under reaction conditions of pH 6.0 and 45℃. Their hydrolyzing activities for pNP-αGal were drastically decreased by the addition of low concentrations of ribose and galactose. They were capable of hydrolyzing completely α-1,6 linked galactosyl residue in melibiose, raffinose and stachyose, which are known to be anti-nutritional factors in products of soybean and legume. In relation to the latter, the isolates YB-1413 and YB-1414 have potential applicability in improving soybean-fermented foods and the nutritional value of soybean feed.

• The Korean Journal of Mycology
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• 제39권1호
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• pp.39-43
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• 2011

A new cultivar "Dan Bi" of Pleurotus eryngii was developed by the method of mono-mono crossing between monokaryotic strains derived from KNR2312 and KNR2596. The parental strains, KNR2312 and KNR2596, are characterized by the property of high quality and a small number of primordia formation, respectively. The optimum temperature of mycelial growth was 25 and that of fruiting body development was $15{\sim}16^{\circ}C$. The period of harvesting including primordia formation was 0.7~1.3 days later than that of control strain Knneutari No. 3 in the culling cultivation. The color of pileus and stipe surface was neutral-brown and pure white, respectively. The shape of pileus was dome and has a scale like as cobweb. The yield was $93{\pm}9.7$ g per 850 cc of plastic bottle. Analysis of the genetic characteristics of the new commercial variety "Dan Bi" showed a different profile as that of the control strain, Knneutari No. 3, when RAPD (Random Amplified Polymorphic DNA) primer #8005 was used. This new variety "Dan Bi" of Pleurotus eryngii is characterized by a small number of primordia formation after scratching.

• Journal of Mushroom
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• 제11권2호
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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.

• Journal of Mushroom
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• 제11권2호
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• pp.63-68
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• 2013

The fruiting body of honey mushroom, Armillaria gallica, was collected from Gastrodia elata cultivated fields. Pure cultures were isolated from fruiting body tissue of the mushrooms, and cultured on MCM (mushroom complete medium) or PDA (potato dextrose agar) medium. Then, 12 different types of mycelial growth characteristics such as growth rate, colony morphology and rhizomorph formation were obtained. The vitality of the mycelial growth and rhizomorph formation of the fruiting body culture isolates were better on MCM than PDA, suggesting that the optimal culture medium for A. gallica mycelia was MCM. To observe the feature of colony morphology, the subculture of isolates were incubated on MCM. Consequently, we could find the segregated or differentiated colony morphology from isolate type 11 that was similar morphology to isolate type 12. For phylogenetic analysis of the 12 isolates, RAPD (Random Amplified Polymorphic DNA) were performed. The isolate type 12 was not only shown different band patterns of RAPD variation in other 11 isolates, but also commercial strain known as Chunmagyun No. 1. Among the tissue culture isolates of fruiting, strains with better mycelial growth characteristics than Chunmagyun No. 1 were selected. We expect that the new strain can be substituted to commercial strain Chunmagyun No. 1.

• Korean Journal of Food Science and Technology
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• 제40권1호
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• pp.101-105
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• 2008

Enterobacter sakazakii is implicated in severe forms of neonatal infections such as meningitis and sepsis. This organism has been isolated from a wide range of foods, including cheese, vegetables, grains, herbs, and spices, but its primary environment is still unknown. Generally, dried infant milk formula has been epidemiologically identified as the source of E. sakazakii. Sunsik (a powdered mixture of roasted grains and other foodstuffs) is widely consumed in Korea as a side dish or energy supplement. Sunsik is consumed without heat treatment; thus, lacking an additional opportunity to inactivate foodborne pathogens. Therefore, its microbiological safety should be guaranteed. In this study, the prevalence of E. sakazakii was monitored in 23 different sunsik component flours, using FDA recommended methods; but E. sakazakii medium (Neogen) and Chromogenic E. sakazakii medium (Oxoid) were used as the selective media. In total, presumptive E. sakazakii strains were isolated from 8 different sunsik powders. Subsequently, an API 20E test was conducted, and 15 strains from 5 different sunsik flours (sea tangle, brown rice, non-glutinous rice, cheonggukjang, dried anchovy) were confirmed as E. sakazakii. Fifteen strains were again confirmed by PCR amplification, using three different primer sets (tDNA sequence, ITS sequence, 16S rRNA sequence), and compared to ATCC strains (12868, 29004, 29544, 51329). They were once again confirmed by their enzyme production profiles using an API ZYM kit. Finally, RAPD (random amplified polymorphic DNA)-genotyping was carried out as a monitoring tool to determine the contamination route of E. sakazakii during processing.