• The Korea Journal of Herbology
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• v.34 no.4
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• pp.1-8
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• 2019

Objectives : Rosae Multiflorae Fructus is a traditional medicine derived from the fruit of Rosa multiflora Thunb. a member of the Rosaceae family. Even though it has a single origin, the possibility of adulterants has always existed. In fact, we had discovered suspicious commercial samples of Rosae Multiflorae Fructus, imported from China. Methods : To define the taxonomic origin of Rosae Multiflorae Fructus and its adulterants, DNA barcode analysis of the internal transcribed spacer, trnL-F intergenic spacer, and psbA-trnH sequences was carried out. These DNA barcode sequences from the correct origin of Rosae Multiflorae Fructus were analyzed and compared with those of other samples from genus Rosa used as medicinal herbs. Results : The analyses of the three DNA barcode sequences efficiently distinguished Rosae Multiflorae Fructus from six other species in genus Rosa and also separated each species used in this study. According to the DNA barcoding results, none of the suspicious commercial samples were Rosae Multiflorae Fructus. RMF09 was identified as Rosa acicularis, whereas RMF10 and RMF11 were identified as Rosa davurica and Rosa rugosa, respectively. These results corroborated the existence of adulterants of Rosae Multiflorae Fructus. Conclusions : Our research provides useful information that could be used as a criterion for distinguishing between Rosae Multiflorae Fructus and its adulterants. These results will help in the prevention of adulteration and also suggest effective methods for verifying the origin of commercial herbal medicines derived from genus Rosa.

• Journal of Species Research
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• v.10 no.2
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• pp.154-158
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• 2021

An endemic endangered species, Aster altaicus var. uchiyamae (Danyang aster) B2015-0044, is cultivated at the Shingu Botanical Garden, which serves as the ex situ conservation institution for this species. In this work, we sequenced the chloroplast genome of A. altaicus var. uchiyamae B2015-0044. We found that the chloroplast (cp) genome of B2015-0044 was 152,457 base pairs(bps) in size: 84,247 bps of large single copy regions(LSC), 25,007 bps of inverted repeats(IRs), and 18,196 bps of small single copy regions. The B2015-0044 cp genome contains 79 protein-coding genes (PCGs), 4 RNA genes, 29 tRNA genes, and 3 pseudogenes. These results were identical to a previously reported cp genome (Park et al., 2017), except for two sites in introns and three in intergenic spacer (IGS) regions. For the intronic differences, we found that clpP.i1 had a 1-bp small simple repeat (SSR) (T) and petD.i had a 3-bp SSR (ATT). We found 1-bp SSRs in the IGSs of trnT_ggu~psbD and psbZ~trnG_gcc, C and A, respectively. The IGS of(ndhF)~rpl32 had a SNP. Based on our results, the cp genome of the A. altaicus var. uchiyamae can be classified into two genotypes, [C]¹-[A]¹²-[T]¹²-[ATT]⁴-C and [C]²-[A]¹¹-[T]¹¹-[ATT]²-A.

• Journal of Food Hygiene and Safety
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• v.27 no.4
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• pp.466-472
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• 2012

In this study, ribulose bisphosphate carboxylase (rbcL), RNApolymeraseC (rpoC1), intergenic spacer (psbA-trnH), and second internal transcribed spacer (ITS2) as identification markers for discrimination of P. mirifica in foods were selected. To be primer design, we obtained 719 bp, 520 bp, 348 bp, and 507 bp amplicon using universal primers from selected regions of P. mirifica. The regions of rbcL, rpoC1, and psbA-trnH were not proper for design primers because of high homology about P. mirifica, P. lobata, and B. superba. But, we had designed 4 pairs of oligonucleotide primers from ITS2 gene. Predicted amplicon from P. mirifica were obtained 137 bp and 216 bp using finally designed primers SFI12-miri-6F/SFI12-miri-7R and SFI12-miri-6F/SFI12-miri-8R, respectively. The species-specific primers distinguished P. mirifica from related species were able to apply food materials and processed foods. The developed PCR method would be applicable to food safety management for illegally distributed products in markets and internet shopping malls.

• Journal of Microbiology and Biotechnology
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• v.25 no.3
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• pp.321-327
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• 2015

Nocardia salmonicida is one of the main pathogens of fish nocardiosis. The purpose of this study was to build a loop-mediated isothermal amplification (LAMP) method for the rapid and sensitive detection of N. salmonicida. A set of four primers were designed from the 16S-23S rRNA intergenic spacer region of N. salmonicida, and conditions for LAMP were optimized as incubating all the reagents for 60 min at 64℃. LAMP products were judged with agar gel electrophoresis as well as with the naked eye after the addition of SYBR Green I. Results showed the sensitivity of the LAMP assay was 1.68 × 10³ CFU/ml (16.8 CFU per reaction) and 10-fold higher than that of PCR. The LAMP method was also effectively applied to detect N. salmonicida in diseased fish samples, and it may potentially facilitate the surveillance and early diagnosis of fish nocardiosis.

• Journal of Microbiology and Biotechnology
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• v.20 no.7
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• pp.1140-1151
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• 2010

Differences in DNA banding patterns, obtained by ribosomal intergenic spacer analysis (RISA), and nitrification were followed in a moving-bed biofilm reactor (MBBR) receiving municipal landfill leachate. Complete nitrification (>99%) to nitrate was obtained in the two-stage MBBR system with an ammonium load of 1.09 g N-$NH_4/m^2{\cdot}d$. Increasing the ammonium load to 2.03 g N-$NH_4/m^2{\cdot}d$or more caused a decline in process efficiency to 70-86%. Moreover, at the highest ammonium load (3.76 g N-$NH_4/m^2{\cdot}d$), nitrite was the predominant product of nitrification. Community succession was evident in both compartments in response to changes in ammonium load. Nonmetric multidimensional scaling (NMDS) supported by similarity analysis (ANOSIM) showed that microbial biofilm communities differed between compartments. The microbial biofilm was composed mainly of ammonia-oxidizing bacteria (AOB), with Nitrosomonas europeae and N. eutropha being most abundant. These results suggest that high ammonium concentrations suit particular AOB strains.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.10a
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• pp.66-66
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• 2018

국화과(Compositae) 다년생 초본인 산국속(Dendranthema)은 국내 약 13여종이 자생하는 것으로 알려져 있으며, 이 중 감국(D. indicum (L.) Des Moul.)과 산국(D. boreale (Makino) Ling ex Kitam.), 구절초(D. zawadskii var. latilobum (Maxim.) Kitam.)가 주로 차 또는 한약재 등의 원료로 이용되고 있다. 차로 이용되는 꽃은 산국이 감국에 비해 상대적으로 작아서 구분이 가능하지만 시중에는 건조된 형태로 가공 유통되므로 육안으로 구분이 쉽지 않고, 산국 유래 제품들은 국내에서 감국 또는 국화로 혼용해서 표기되어 유통되고 있어 그 기원을 명확히 정립할 필요가 있다. 이에 본 연구는 감국과 산국의 분자유전학적 판별을 위해 DNA 바코드 후보 유전자를 활용하여 염기서열분석으로 확보된 SNP 및 InDel 정보를 바탕으로 CAPS 마커를 개발하고자 수행되었다. 감국과 산국 모두 trnL-trnF intergenic spacer 구간에서 약 1kb의 PCR 산물이 확인되었고, 이들 염기서열에서 분석한 2 SNP 및 3 InDel을 대상으로 CAPS 마커 개발을 위한 제한효소 사이트를 탐색하였다. Gap을 포함한 774bp (감국/산국=A/G) 위치의 SNP에서 BstUI(GC^GC)처리로 CAPS 마커로 전환 가능함이 확인되었고, 이에 감국과 산국의 PCR 산물에 제한효소를 처리한 결과, 제한효소 인식 사이트가 존재하는 산국에서 두 개의 DNA 단편이 확인되었다. 위 결과는 다양한 형태로 가공 유통되는 감국과 산국의 판별을 위한 마커로 활용될 수 있으며, 본 연구에 활용된 기술은 추후 건강기능식품 개발을 위한 원료표준화 확립 연구에 유용할 것으로 판단된다.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.1
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• pp.124-130
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• 2008

Streptococcus (Strep.) agalactiae is one of the major pathogens of bovine mastitis and is the main cause of subclinical infection. This study attempted to develop a rapid PCR diagnosis procedure using milk samples collected on filter paper disks. Chromatographic filter paper was employed as the preservation media and kept at room temperature for one to four weeks. The revival rate of Strep. agalactiae kept on dried filter paper disks was affected by the pretreatment preservation time. The revival test suggested that not all the bacteria in artificially contaminated milk samples on the filter paper disks could be recovered. After that, a PCR based on the 16-23S intergenic spacer region of Strep agalactiae was performed. The results distinguished the strep. agalactiae from major pathogens of bovine mastitis at a $2{\times}10^2$ colony forming units (CFU)/ml level, which showed similar sensitivity to the results from liquid milk samples. The results also showed that milk samples collected on filter paper disks could be kept at room temperature for one to four weeks with little negative effect on sensitivity and specificity. The field test showed that the diagnostic sensitivity and specificity was 96.15% and 98.60%, respectively. In conclusion, the protocol will provide a rapid and economic procedure for the detection of bovine mastitis.

• Korean Journal of Veterinary Service
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• v.34 no.3
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• pp.201-208
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• 2011

Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) is a pathogen of a variety of infections in horse. We studied biochemical and molecular characteristics of S. zooepidemicus isolated from the genital tract of Thoroughbred mares in Korea. Seventy-nine isolates were identified as S. zooepidemicus by biochemical and PCR method from 374 horses. The biochemical characteristics of S. zooepidemicus isolates were positive reaction of lactose and sorbitol. However, S. zooepidemicus isoltes were negative reaction of inulin, mannitol, raffinose, trehalose, aesculin hydrolysis, growth in 6.5% NaCl and variable reaction of maltose. Epidemiological investigations of S. zooepidemicus isolates were performed by fragment analysis of SzP (S. zooepidemicus protective protein) gene, CNE (collagen binding protein) gene and ISR (16s rRNA intergenic spacer region) gene using ABI Prism $3,130{\times}1$ Genetic Analyzer System. All isolates were shown single amplification size of 906 bp in CNE gene, but SzP and ISR gene were shown variable patterns of fragment size. The characteristics of S. zooepidemicus investigated in this study will be very useful for the prevention of infection and the studies of epidemiologic characteristics of S. zooepidemicus, causing the severe economic losses due to reproductive failures.

• Journal of Environmental Science International
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• v.14 no.7
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• pp.649-655
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• 2005

Microcystis aeruginosa is common form of cyanobacteria (blue-green algae) capable of producing toxic heptapeptide (microcystin) that cause illness or death. The comparison of molecular genetic method with the morphological characteristics of cyanobacteria was conducted. We have designed PCR primers (JJM98F, JJM1141R) for cyanobacterial 16S rRNA and phycocyanin intergenic spacer (PC-IGS) gene domain. To confirm the production of microcystins, PCR primers for the N-methyltransferase (NMT) domain of microcystin synthetase gene mcyA were designed using 21 cyanobacteria strains Most of isolated strains from the Nakdong River was classified as Microcystis aeruginosa and the similarities were $99\%$ with M. aeruginosa AF 139292. $38.1\%$ of isolated strains contained microcystin synthesis gene. NMT (N-methyltransferase) were not detected in isolated strain in several strains, which means non-toxic. However, the NMTs of the strains were detected during the cultivation.

• Journal of fish pathology
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• v.19 no.2
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• pp.127-139
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• 2006

From 2002 to 2004, various vibrios were isolated from the olive flounder, Paralichthys olivaceus of culturing size with disease signs. During this survey, it was known that the high proportion of Vibrio ichthyoenteri was occupied among the isolated vibrios. Generally, V. ichthyoenteri is well known as the pathogen of bacterial enteritis of olive flounder larvae. The aim of the present study was the compare the characteristics of two groups of V. ichthyoenteri, culturing sized olive flounder, and larvae of olive flounder showing the intestinal necrosis. The research was focused on the physiology, biochemistry, genetics in the two bacterial groups. The physiological and biochemical characteristics of the tested strains were very similar. The intergenic spacer (IGS) region between the 16S and 23S rRNA genes of 21 isolated strains and 3 reference strains, V. ichthyoenteri, were investigated by PCR fragment length typing and DNA sequencing. After the isolated strains were identified as V. ichthyoenteri, not only phenotypic characteristics of the isolated and reference strains but also homology of 16S-23S IGS of all isolated strains and reference strains as 99.1~100%. The V. ichthyoenteri showed 4 specific 16S-23S patterns and contained no-tRNA, tRNAGlu(TTC) , tRNAIle(GAT) tRNAAla(TGC) type .