• Proceedings of the Korean Aquaculture Society Conference
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• 2003.10a
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• pp.92-92
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• 2003

본 연구는 부착규조류에 따른 부착성 요각류 Tigriopus japonicus의 nauplius 생산력을 알아보기 위하여 실시하였다. 실험에 사용된 T. japonicus는 부산 동백섬 부경대학교 수산과학연구소 부근에 있는 tidal pool에서 동물성부유생물망으로 채집하였다. 먹이생물로는 부경대학교 한국해양 미세조류 은행에서 보관중인 부착규조류 중 중형종인 Caloneis schroder를 대표종으로 이와 크기가(14.8~27.5$\mu\textrm{m}$) 유사한 종들을 대상으로 형태를 현미경 하에서 관찰하고, 지역과 분리일자 등을 고려하여 유사종을 선별한 후 최종적으로 종의 유전적 유사성을 밝히기 위하여 RAPD-PCR (random amplified polymorphic DNA polymerase chain reaction)을 실시하였다. 이 중 Caloneis schroder와 유전적으로 유사성이 낮은 Navicula spp. (KMCC B-245, 393, 394, 581) 4종을 선별하여 T. japonicus의 먹이로 사용하여 포란한 암컷의 nauplius 생산력을 3반복 조사하였다. Genomic DNA는 대부분의 종에서 성공적으로 검출되었으며, 종에 따라 PCR 증폭산물이 나타나지 않은 경우도 있었으므로, PCR 산물이 나타난 종에 대해서만 분석하였고, 증폭된 DNA band는 대부분 크기 0.5~2.0kb 범위에서 나타났다. 실험에 사용된 부착규조류 간의 유사성을 알아보기 위하여 similarity matrix를 분석한 결과 F값의 범위는 0.00에서 1.00까지 였으며, Caloneis schroder와 유사성이 낮은 종들에 비하여 유사성이 높은 종들이 더 많이 나타났다. 이들을 먹이로 하여 포란한 T. japonicus의 실험구별 nauplius 평균 개체수를 살펴보면, KMCC B-394가 255.7마리로 가장 높았던 반면 KMCC B-581가 29.7마리로 가장 낮았다. 그 외 KMCC B-245가 120.0마리, KMCC B-393가 76.0마리, Caloneis schroder가 32.3마리 각각 나타났다. 이와같은 결과를 볼 때 T. japonicus의 nauplius 생산력은 규조 종에 따라 큰 차이가 있는 것으로 판단된다.

• Molecules and Cells
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• v.25 no.1
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• pp.20-29
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• 2008

Cytoplasmic male sterility (CMS), one of the most important traits in crop breeding, has been used for commercial seed production by $F_1$ hybrid cultivars of pepper (Capsicum annuum L.). To develop reliable molecular markers for allelic selection of the Restorer-of-fertility (Rf) gene, which is known to be a major determinant of pollen fertility restoration in peppers, a sequence of approximately 10 kb flanking an RAPD fragment closely linked to the Rf locus was obtained by genome walking. A homology search revealed that this sequence contained an LTR retrotransposon and a non-LTR LINE-like retrotransposon. Sequencing of this Rf-linked region to search for polymorphisms between a dominant and recessive allele revealed 98% nucleotide sequence identity between them. A third polymorphic haplotype of the Rf-linked sequence, which has 94-96% nucleotide sequence identity with the two previously isolated haplotypes, was identified among a large number of breeding lines. Utilizing polymorphic sequences in the haplotypes, PCR markers were developed for selection of particular haplotypes and used to examine the distribution of the haplotypes in diverse breeding lines, cultivars, and C. annuum germplasms. Surprisingly, the third haplotype was the predominant type in C. annuum germplasms, while its frequency in $F_1$ hybrid cultivars was relatively low. Meanwhile, analysis of breeding lines whose Rf allele genotypes and male-sterility phenotypes were already known revealed that the third haplotype was mainly present in exotic breeding lines that cause unstable male-sterility when combined with sterile cytoplasms.

• The Plant Pathology Journal
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• v.22 no.1
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• pp.7-10
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• 2006

Anthracnose severely occurred on begonia (Begonia semperflorens Link.) nurseries in Gyeongju, Gyeongbuk in July, 2004. More than $80\%$ of begonia seedlings were diseased in the greenhouse surveyed and diseased leaves per plant were $12.1\%$ in average. Yellowish spots occurred on the leaves of begonia as initial symptoms, and they coalesced irregularly to form large brown pleomorphic lesions. Severely infected leaves were defoliated, resulting in abnormal growth of the entire plant. Colletotrichum sp. was repeatedly isolated from the diseased plants and was identified as Colletotrichum acutatum on the basis of the mycological characteristics on potato dextrose agar and RAPD analysis. Pathogenicity of the fungus was also confirmed by artificial inoculation on healthy plants. The optimum temperature for mycelial growth of C. acutatum was around $25^{\circ}C$. The fungus was sensitive to azoxystrobin, bitertanol, diethofencarb-carbendazim, difenoconazole and tebuconazole. This is the first report on the anthracnose of begonia caused by C. acutatum in Korea.

• Journal of Plant Biotechnology
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• v.3 no.3
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• pp.145-149
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• 2001

This study was conducted to analyze the morphological characteristics such as flower color, flower type, flower diameter and flower stalk, and the main annual production yield, and genetic similarity of twenty four Gerbera species growing in Korea. Most of flower colors were pink. The numerical order of flower color was pink, orange, red, double-colored, and milk-white. Majority of flower types were sin81e or semidouble flowers. A few species were double flowers. flower diameters were from 7 ㎝ to 12 ㎝, showed significant differences compared to other characteristics. Flower stalks were ranged from 55 ㎝ to 65 ㎝. Only one species was the shortest as 55 ㎝. The others were similar size as about 65 ㎝. Main annual production yields were between 190 and 400 blossoms. Fifty seven reproducible polymorphic bands from eighty primers were used for analyses of genetic similarity. The genetic similarity of 24 collected Gerberas was largely classified into five groups. The average similarity coefficient was 0.72 ranged from 0.50 to 0.90. The highest similarity coefficient was shown between 'Sardana' with red/white flower color and double flower type, and 'Tamara' with orange flower color and double flower type as 0.90.

• Development and Reproduction
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• v.13 no.2
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• pp.97-103
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• 2009

Genomic DNA samples isolated from four geographical freshwater crab (Eriocheir sinensis) populations collected in the inland of the Korean Peninsula (Gunsan, Paju, and Nampo) and a Chinese site, were used for PCR amplification. Seven decamer primers generated 19 specific loci (19/243 loci, 7.81%) in the Gunsan population, 32 (32/215 loci, 14.88%) in the Paju population, 19 (19/231 loci, 8.23%) in the Nampo population and 62 (62/340 loci, 18.24%) in a Chinese population. The average 8.9 specific loci exhibited inter-individual-specific characteristics, thus revealing DNA polymorphisms in the Chinese population. The number of unique shared loci to each population and number of shared loci by the four populations were generated by molecular analysis using seven primers in four populations. 35 unique shared loci to each population, with an average of 5.0 per primer, were observed in the Gunsan population, and 50 loci, with an average of 7.1 per primer, were observed in the Chinese population. The hierarchical dendrogram indicates three main branches: cluster 1 (GUNSAN 01$\sim$GUNSAN 05, PAJU 06$\sim$PAJU 10 and NAMPO 11$\sim$NAMPO 15) and cluster 2 (CHINESE 16, 17, 18, 19 and 20). Conclusively individual no. 20 of the PAJU 10 freshwater crab was most distantly related to CHINESE no. 20 (genetic distance = 0.667). Taken together, these results demonstrate the potential of RAPD analysis to identify diagnostic markers for the identification of four freshwater crab populations.

• Microbiology and Biotechnology Letters
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• v.46 no.1
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• pp.1-8
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• 2018

Gul (oyster) jeotgals (GJs) were prepared using different types of salt (23%, w/v): purified salt, solar salt aged for 3 years, and bamboo salt crystalized 3 times. One set of GJs was fermented with Bacillus subtilis JS2 ($10^6CFU/g$), while the other GJ set was fermented without starter. During fermentation for 24 weeks at $15^{\circ}C$, the starter GJs showed 10-fold higher bacilli counts than the no-starter GJs, where the maximum bacilli count was $8{\times}10^3CFU/g$. All 28 bacilli strains isolated from the 6-week GJs were identified as B. subtilis by using a RAPD-PCR, indicating that some of the B. subtilis JS2 cells remained viable. Lactic acid bacteria (LAB) and yeasts were present at low levels, $10^1-10^2CFU/g$. LAB with protease activities isolated from 10-week samples were identified as Enterococcus species. The isolates obtained at 16 weeks were all Staphylococcus species. The GJs with bamboo salt showed higher pH and lower titratable acidity (TA) values than the other GJs due to the strong alkalinity of bamboo salt. The amino-type nitrogen in the GJs increased slowly during the fermentation. At 24 weeks, the GJs with purified salt showed the highest amino-type nitrogen (412-430 mg%), followed by the GJs with solar salt (397-406 mg%) and GJs with bamboo salt (264-276 mg%). Meanwhile, the GJs with bamboo salt showed the highest ammonia-type N (63.67 mg%), followed by the GJs with purified salt (49 mg%) and solar salt (48 mg%).

• Microbiology and Biotechnology Letters
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• v.47 no.4
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• pp.498-508
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• 2019

Bacillus species were isolated from iru, a traditional fermented condiment in Nigeria. Polyphasic approach was used to evaluate the phylogenetic relationship and strain sub-type of the isolated species. Additionally, the phylogenetic profiles of the species isolated from iru were compared with those of bacilli isolated from different continents. The phylogenetic diversity analysis was performed using the combination of 16S rRNA gene sequencing, ITS-PCR, ITS-PCR-RFLP, and M13 RAPD-PCR. The analysis revealed that Bacillus subtilis U170B and B. subtilis U146A isolated from iru were the closest relatives of strains belonging to the phylogeny of B. subtilis sensu stricto and were related to other bacilli isolated from different continents that had functional benefits. The two isolated species exhibited resistance to acidic pH (pH 2.0). The survival rates of B. subtilis U170B, B. subtilis U146A, and B. clausii UBBC-07 (commercial probiotic strain) cultured at pH 2.0 for 3 h were 33.45, 12.44, and 9.53%, respectively. The strains were highly tolerant to bile salts [0.3% (w/v)]. B. subtilis U170B exhibited the highest cell viability (43.45%) when cultured for 3 h in the presence of bile salts, followed by B. subtilis U146A (25%) and B. clausii UBBC-07 (18.94%). B. subtilis U170B and B. subtilis U146A did not exhibit haemolytic activity and were susceptible to different antibiotics. Additionally, these two strains exhibited weak antagonistic activity against B. cereus. The diverse wild strains of B. subtilis can be used as a safe multifunctional starter culture for the industrial production of condiments with health benefits.

• Journal of Microbiology and Biotechnology
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• v.30 no.6
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• pp.856-867
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• 2020

Vibrio parahaemolyticus is a major gastroenteritis-causing pathogen in many Asian countries. Antimicrobial resistance in V. parahaemolyticus has been recognized as a critical threat to food safety. In this study, we determined the prevalence and incidence of antimicrobial resistance in V. parahaemolyticus in the southern Fujian coast, China. A total of 62 isolates were confirmed in retail aquatic products from June to October of 2018. The serotype O3:K6 strains, the virulence genes tdh and trh, antibiotic susceptibility and molecular typing were investigated. Then plasmid profiling analysis and curing experiment were performed for multidrug-resistant strains. The results showed that the total occurrence of V. parahaemolyticus was 31% out of 200 samples. Five strains (8.1%) out of 62 isolates were identified as the V. parahaemolyticus O3:K6 pandemic clone. A large majority of isolates exhibited higher resistance to penicillin (77.4%), oxacillin (71%), ampicillin (66.1%) and vancomycin (59.7%). Seventy-one percent (44/62) of the isolates exhibited multiple antimicrobial resistance. All 62 isolates were grouped into 7 clusters by randomly amplified polymorphic DNA, and most of the isolates (80.6%) were distributed within cluster A. Plasmids were detected in approximately 75% of the isolates, and seven different profiles were observed. Seventy-six percent (25/33) of the isolates carrying the plasmids were eliminated by 0.006% SDS incubated at 42℃, a sublethal condition. The occurrence of multidrug-resistant strains could be an indication of the excessive use of antibiotics in aquaculture farming. The rational use of antimicrobial agents and the surveillance of antibiotic administration may reduce the acquisition of resistance by microorganisms in aquatic ecosystems.

• Journal of Microbiology and Biotechnology
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• v.12 no.1
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• pp.166-171
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• 2002

The genus Leuconostoc is generally recognized as a favorable microorganism associated with a good taste of Kimchi and Lactobacillus plantarum is responsible for the overripening and acidification of Kimchi. A rapid and reliable PCR-based method to monitor the change of these lactic acid bacterial populations during Kimchi fermentation was attempted. A Leuconostoc-specific primer set was chosen from the conserved sequences of 16S rRNA genes among Leuconostoc species. The Lb. plantarum-specific primer set was the internal segments of a Lb. plantarum-specific probe which was isolated after randomly amplified polymorphic DNA (RAPD) analysis and tested for identification. The specificity of this protocol was examined in DNA samples isolated from a single strain. In agarose gel, as little as 10 pg of template DNA could be used to visualize the PCR products, and quantitative determination was possible at the levels of 10 pg to 100 ng template DNA. For the semi-quantitative determination of microbial changes during Kimchi fermentation, total DNAs from the 2 h-cultured microflora of Kimchi were extracted for 16 days and equal amounts of DNA templates were used for PCR. The intensities of DNA bands obtained from PCR using Leuconostoc-specific and Lb. plantarum-specific primer sets marked a dramatic contrast at the 1 ng and 100 ng template DNA levels during Kimchi fermentation, respectively. As the fermentation proceeded, the intensity of the band for Leuconostoc species increased sharply until the 5th day and the levels was maintained until the 11 th day. The sharp increase for Lb. plantarum occurred after 11 days with the decrease of Leuconostoc species. The results of this study indicate that Leuconostoc species were the major microorganisms at the beginning of Kimchi fermentation and reach their highest population during the optimum ripening period of Kimchi.

• Journal of Plant Biotechnology
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• v.36 no.2
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• pp.124-129
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• 2009

In general, the root color of Codonopsis lanceolata is white, but red or blue-colored root is found at a low frequency in nature. Red or blue-colored roots have scarcity value, thus farmers wish to produce colored roots. The factors for determining the color of roots are unclear whether the color is controlled by genetically or simply by environmentally such as soil environment. Using in vitro culture system which is advantageous for setting of the same culture condition, we analyzed the physiological and morphological characteristics and genetic differences among red-, blue- and white lines of C. lanceolata. In the red colored roots, stems of in vitro cultured plantlet were colored in dark red pigment. Histological analysis revealed that the red pigment was accumulated in the outer cortex layer of the stem and determined as anthocyanin. Chlorophyll contents in red root lines were higher than those in white- and blue root lines. Plantlets from red roots were smaller in both shoot length and total leaf area than those from white- and blue roots. Genetic differences among the three different colored C. lanceolata were determined by RAPD (Randomly Amplified Polymorphic DNA) analysis. Each line of colored roots had clear DNA polymorphism. These results indicate that the occurrence of red- and blue colored roots in nature was determined by genetic factors rather than soil enviromental conditions.