• Journal of Ecology and Environment
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• 제29권2호
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• pp.143-149
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• 2006

This study was conducted to identify native ectomycorrhizal (ECM) fungi colonizing Pinus densiflora for revegetation of abandoned coal mines in Korea. Seedlings of P. densiflora growing on coal mining spoils of a study site in Samcheok were collected. ECM roots were observed under stereomicroscope and their DNA were extracted from each root tip for a seedling for molecular identification. A PCR primer pair specific to fungi, ITS1F and ITS4, was used to amplify fungal DNA. Restriction enzymes, Alul and Hinfl were used for restriction fragment length polymorphism (RFLP). Combined with RFLP profiles and sequence analysis, total twenty one taxa were identified from the ECM root tips. Basidiomycetous fungi including Thelephoraceae, Pezizales, Laccaria, Pisolithus and Ascomycetous fungi including ericoid mycorrhizal fungi were identified from this study. Results showed that the most frequently found in the study sites was a species in Thelephoraceae. A possible use of ECM fungi identified in this study for the revegetation of abandoned coal mines with P. densiflora was discussed.

• 대한의생명과학회지
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• 제19권3호
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• pp.213-223
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• 2013

The objectives of this study are to develop a molecular diagnosis to differentiate serotypes and mating-types of C. neoformans/C. gattii complex isolates from pigeon droppings in Korea and to elucidate molecular epidemiology of the isolates. Phenotypes and genotypes of C. neoformans/C. gattii complex isolates were identified by biochemical properties and PCR using specific CNLAC1 gene, respectively. To classify serotypes and mating-types of C. neoformans/C. gattii complex isolates, the five reference strains and thirty-three isolates in Korea were investigated by restriction fragment length polymorphism (RFLP) analysis using CNLAC1 gene for varieties, by random amplified polymorphic DNA (RAPD) for serotyping, and by PCR using specific primer sets for mating typing. All isolates in Korea were belonged to C. neoformans var. grubii (serotype A) by RFLP and RAPD patterns which showed high sensitivity and specificity. Therefore, RFLP and RFLP were available to differentiate varieties and serotypes of C. neoformans. Amplification patterns of the five reference strains by specific PCR for mating typing were differentiable, and all isolates were classified into $MAT{\alpha}$. All C. neoformans environmental isolates in Korea were Cr. neoformans serotype A and $MAT{\alpha}$ which is a more virulent pathogen. This study suggests that RFLP and RAPD are rapid and correct molecular diagnosis tools for epidemiology of C. neoformans/C. gattii complex isolates.

• 대한수의학회지
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• 제36권3호
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• pp.627-633
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• 1996

To analyze the genomic diversity of transmissible gastroenteritis virus (TGEV), the N-terminal half of the spike (S) glycoprotein gene and nonstructural protein gene (open reading frames 3 and 3-1) were amplified by reverse transcriptase reaction and polymerase chain reaction (RT-PCR), and analyzed using restriction fragment length polymorphism (RFLP) patterns of the amplified DNA. In this study, TGEV Miller (M6) and Purdue (P115) strains were used as reference strains, and two vaccine strains (MSV and STC3) and four Korea isolates (P44, VRI-WP, VRI-41, and VRI-48) were analyzed. All TGEV strains were amplified with three TGEV primer pairs. Although there was some exception in RFLP analysis, this method differentiated TGEV strains into following groups : Miller group (M6 and MSV), Purdue group (PUS, STC3, P44, VRI-WP, VRI-41, and VRI-48). Using Sau3AI and SspI, VRI-48 was differentiated from the Miller and Purdue type viruses. The RT/PCR in conjuction with RFLP analysis was a rapid and valuable tool for differentiating several strains of TGEV. This study revealed the occurences of distinct difference in genome of TGEV strains.

• Molecules and Cells
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• 제21권3호
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• pp.411-417
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• 2006

We have developed a polymerase chain reactionrestriction fragment length polymorphism (PCR-RFLP) marker that can distinguish male-fertile (N) and male-sterile (S) cytoplasm in onions. The PCR-RFLP marker was located in a chloroplast psbA gene amplicon. Digesting the amplicons from different cytoplasm-containing varieties with the restriction enzyme MspI revealed that N-cytoplasm plants have a functional MspI site (CCGG), whereas the S-cytoplasm plants has a substitution in that site (CTGG), and thus no MspI target. The results obtained using this PCR-RFLP marker to distinguish between cytoplasmic male sterile factors in 35 onion varieties corresponded with those using a CMS-specific sequence-characterized amplified region (SCAR) marker. Moreover, the PCR-RFLP marker can identify N- ot S-cytoplasms in DNA sample mixtures in which they are in up to a 10-fold minority, indicating that use of the marker has high diagnostic precision. We also demonstrated the usefulness of the SNP detected in the psbA gene for high-throughput discrimination of CMS factors using Real-time PCR and a TaqMan probe assay.

• The Plant Pathology Journal
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• 제38권3호
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• pp.254-260
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• 2022

Fusarium incarnatum-equiseti species complex (FIESC) contain over 40 members. The primer pair Smibo1FM/Semi1RM on the RPB2 partial gene has been reported to be able to identify Fusarium semitectum. The F. fujikuroi species complex (FFSC) contains more than 50 members. The F. verticillioides as a member of this complex can be identified by using VER1/VER2 primer pair on the CaM partial gene. In this research, the Smibo1FM/Semi1RM can amplify F. sulawesiense, F. hainanense, F. bubalinum, and F. tanahbumbuense, members of FIESC at 424 bp. The VER1/VER2 can amplify F. verticillioides, F. andiyazi, and F. pseudocircinatum, members of FFSC at 578 bp. Polymerase chain reaction-restriction fragment length polymorphism by using the combination of three restriction enzymes EcoRV, MspI, and HpyAV can differentiate each species of FIESC used. The two restriction enzymes HpaII and NspI can distinguish each species of FFSC used. The proper identification process is required for pathogen control in the field in order to reduce crop yield loss.

• Animal cells and systems
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• 제13권2호
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• pp.179-186
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• 2009

Discriminating between eel species of the genus Anguilla using morphological characteristics can be problematic, particularly in the glass eel and elver stages. In this study, sequence-characterized amplified region (SCAR) and polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) markers were developed for the identification of Anguilla japoniea, Anguilla btcoior bicaor. Anguilla rostrata, and Anguilla anguilla. Random amplified polymorphic DNA (RAPD) fragments from A. japoniea (362 bp), A. bicolor bicctor (375 bp), A. rostrata (375 bp), and A. anguilla (375 bp) were isolated, sequenced, and converted to SCAR markers. The principal difference between the SCARs of A. japoniea and the three other species is the absence of a 13 bp deletion in the A. japoniea SCAR. Specific PCR primers amplified a 290 bp fragment for A. japoniea and 303 bp fragments for A. bicolor bicoior. A. rostrata, and A. anguilla. Restriction enzyme digestion with Taql, Mael, and Tru9l yielded PCR-RFLP patterns with differences that, when analyzed together, are sufficient for distinguishing each of the four eel species. In addition, RAPD fragments for A. japoniea (577 bp), A. bicoior bicoor (540 bp), A. rostrata (540 bp), and A. anguilla (509 bp) were also isolated and sequenced. The A. japoniea, A. bicoior blcoior. A. rostrata, and A. anguilla PCR products contain ten, nine, nine, and eight tandem repeats, respectively, of a 37 bp sequence. These results suggest that SCAR and PCR-RFLP markers and repeat numbers for specific loci will be useful for the identification of these four Anguilla eel species.

• 한국응용곤충학회지
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• 제48권4호
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• pp.547-551
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• 2009

팥나방(Matsumuraeses phaseoli)과 어리팥나방(M. falcana)은 유사한 종으로 작물에 상이한 피해를 주고 있다. 그러나 형태적 특징만으로 이 두 동소적 유사종을 쉽게 구분하기 어렵다. 본 연구는 이 두 종을 뚜렷하게 판별할 수 분자마커를 개발했다. 두 종의 시토크롬 옥시다아제-I의 부분(약 500 bp) 염기서열이 밝혀졌다. 이를 바탕으로 두 종을 구분하는데 이용될 수 있는 판별 제한효소인 Rsa I이 선발되었고 PCR-RFLP를 통해 입증되었다.

• The Plant Pathology Journal
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• 제18권1호
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• pp.1-5
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• 2002

The availability of nucleotide sequences of the coat protein gene of Potato leafroll virus (PLRV) permitted the construction of DNA primers that were utilized for cDNA synthesis. Polymerase chain reaction (PCR) products of a 487 bp. and approximately 500 bp DNA fragments were amplified from nucleic acid extracts of PLRV-infected tissue and strawberry mild yellow-edge (SMYE) diseased strawberry tissue, respectively. The amplified DNA fragments were further differentiated by hybridization analysis with a CDNA probe for the coat protein gene of PLRV and restriction fragment length polymorphism (RFLP) analysis. These results suggest that a luteovirus is associated with the SMYE disease.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 추계학술대회
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• pp.126-126
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• 2018

Many of researches have revealed that human intestinal microbiota is related to health. Several diseases like obesity, diabetes, and hypertension are affected by the microbiota directly and indirectly. So, interventions with food and drug have been tried to change a composition of the microbiota to better condition. However, few natural medicines have elucidated to date. To understand an influence on microbiota by plant materials including Glycyrrhizae Radix, the extract of medicines were administered to mice and the feces were collected before and after the administration. The feces were analyzed by terminal restriction fragment length polymorphism (T-RFLP). The changes in composition of mice gut microbiota were detected and analyzed. The data could be utilized to further study about biological activities of the plant medicines.

• 식물병연구
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• 제17권1호
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• pp.99-101
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• 2011

자낭균인 Taphrina wiesneri는 한국의 공원과 가로수에 주로 식재되는 왕벚나무에 빗자루병을 일으키는 병원균이다. 한국과 일본에서 분리한 13개의 병원균에 대해 18S rDNA 염기서열 분석을 통한 계통학적 분석과 rDNA-IGS 영역에 대한 RFLP 분석을 실시하였다. 18S rDNA 염기서열 분석을 통한 계통도 분석결과 병원균은 2그룹으로 분류되었다. Hha I 제한효소를 이용한 rDNA-IGS 영역에 대한 RFLP 분석결과 B, C, D, G 4개의 패턴으로 나타났으며, 그중 G 패턴은 새로운 패턴이었다.