• Mycobiology
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• 제37권4호
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• pp.258-266
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• 2009

Pleurotus eryngii, known as king oyster mushroom has been widely used for nutritional and medicinal purposes. This study was initiated to screen the suitable conditions for mycelial growth and to determine the phylogenetic relationship of the selected strains. Optimal mycelial growth was observed at $30{^{\circ}C}$ and minimum mycelial growth observed at $10{^{\circ}C}$. This mushroom tolerates a broad pH range for mycelial growth, with most favorable growth observed at pH 6. Results also indicated that glucose peptone, yeast malt extract and mushroom complete media were favorable growth media, while Hennerberg and Hoppkins media were unfavorable. Dextrin was the best and xylose the least effective carbon sources. Results revealed that inorganic nitrogen sources were less effective than organic sources for the mycelial growth of P. eryngii. Investigation of genetic diversity is necessary to identify the strains. The ITS region of rDNA were amplified using PCR. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 214 to 222 bp and 145 to 236 bp, respectively. The sequence of ITS2 was more variable than that of ITS1, and the 5.8S sequences were identical. A phylogenetic tree based on the ITS region sequences indicated that selected strains could be classified into six clusters. Fourteen IUM and ATCC- 90212 strains were also analyzed by RAPD with 20 arbitrary primers. Fourteen of these primers were efficiently amplified the genomic DNA. The number of amplified bands varied with the primers and strains, with polymorphic fragments in the range from 0.2 to 2.3 kb.

• 미생물학회지
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• 제44권1호
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• pp.63-68
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• 2008

참외 경작지 토양으로부터 순수 분리된 선충포식성 곰팡이 Kan-23 균주는 배양 30일 이후에 분생포자가 형성되는 증식이 매우 더딘 특성을 나타내었다. 주사전자현미경(SEM)을 이용하여 Kan-23 균주의 미세형태 구조를 관찰한 결과 분생포자병의 각 가지마다 $5{\sim}10$개의 분생포자를 형성하였으며, 분생포자는 3개의 격막을 형성하고 타원형을 나타내었고, 수축성 고리형태(constricting ring)의 포식 구조체를 형성하는 특징을 나타내었다. 다양한 배지를 이용하여 포식 구조체의 형태학적 특성을 비교 검토한 결과 GPA 배지에서 2배 이상 크기의 대형 수축성 고리 형태의 포식 구조체를 형성하는 특징을 나타내었다. Kan-23 균주의 rDNA의 ITS 영역의 염기서열을 분석한 결과, Drechslerella 속의 계통군에 속하였으며, 특히 Drechslerella brochopaga (U51950)와 99%의 높은 염기서열 유사도를 나타내어 Drechslerella brochopaga로 동정되어 국내 미기록종임이 확인되었다.

• Fisheries and Aquatic Sciences
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• 제13권4호
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• pp.263-271
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• 2010

G protein-coupled receptors (GPCR) constitute the largest superfamily of cell membrane receptors, mediating diverse signal-transduction pathways. The melanocortin 4 receptor (MC4R) has been of interest for its physiological role and size, one of the smallest among the GPCRs, which makes it a good model system for the structural study of GPCRs. To study the molecular structure and tissue-specific expression of MC4R in olive flounder (Paralichthys olivaceus), the full-length MC4R gene was obtained using PCR amplification of genomic DNA as well as cDNA synthesis. Sequence analysis of the gene indicates that 978 bp of the MC4R gene encodes 325 amino acids without introns. Sequence alignment with the MC4Rs from other fish shows the highest degree of identity (96%) between Paralichthys olivaceous and Verasper moseri, followed by Takifugu rubripes and Tetraodon nigroviridis (89%). RNA was isolated from various tissues to examine the tissue distribution of MC4R by using RT-PCR. The results showed major expression of MC4R in the liver, brain, and eye, which is consistent with the expression pattern in other fish belonging to the order Pleuronectiformes.

• 한국산림과학회지
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• 제81권4호
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• pp.320-324
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• 1992

들메나무(Fraxinus mandshurica Rupr.) 는 우리나라에서 두가지의 서로 다른 형태(形態)가 자생(自生)하고 있는 것으로 알려져 있다. 최근(最近)에 개발(開發)된 PCR기법(技法)을 이용(利用)하여 이 두 형태(形態)의 들메나무 DNA의 변이(變異)를 조사(調査)하였다. DNA 합성(合成) 효소(酵素)와 인공합성(人工合成)된 primer를 이용(利用)하여 이 수종(樹種)의 DNA를 증폭(增幅)시켜 비교(比較)한 결과(結果)이 두 형태(形態)는 DNA 비례(排列)에서 서로 다른것으로 나타났다. DNA변이(變異)는 같은 형태내(形態內)의 개체간(個體間)에도 나타나나 각 형태별(形態別)로 뚜렷하게 구분(區分)되어 형태별(形態別)로 특징적(特徵的)인 band들이 관찰(觀察)되었다. 이러한 특징적(特徵的)인 band들로 두 형태(形態)를 구분(區分)할 수 있었다.

• 한국균학회지
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• 제33권1호
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• pp.1-10
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• 2005

본 연구는 강원도 지역에서 채집한, 구름버섯균으로 예상되는 19종의 실험균주의 외부형태와 내부구조의 특징을 파악하여 동정하고, 구름버섯균(Trametes versicolor)의 형태적 특징과 rDNA상의 ITS염기서열간의 상관관계 유무를 확인하기 위하여 Trametes versicolor와 동일 속(Genus)에 속하며 계통학적 유연관계가 가까운 Trametes villosa KCTC06866, Trametes suaveolens KCTC 26205, Trametes hirusta KCTC26200, Trametes versicolor KCTC16781 및 KCTC26203의 4종-5균주와 함께 rDNA의 Internal Transcribed Spacers(ITS1 과 ITS2)영역의 염기서열을 비교 분석하였다. 분석결과, 채집된 실험균주는 Trametes versicolor KCTC16781 및 KCTC26203과의 염기차이가 $0{\sim}6$개로 매우 유사한 근연한 양상을 나타내었고 계통도를 분석한 결과, 구름버섯으로 예상된 실험균주들과 Trametes versicolor 종들이 단계통군을 이루어 실험균주는 모두 동일종으로 확인되었으며, Trametes hirusta KCTC26200, Trametes suaveolens KCTC26205 그리고 Trametes villosa KCTC06866는 측계통을 이루었다. 채집된 19종의 균주는 외형상 다양한 채색과 무늬를 갖고 있으나 ITS1과 ITS2 영역의 염기서열을 분석한 계통수에서는 Trametes versicolor간의 색깔이나 무늬는 ITS1 및 ITS2의 염기서열과는 직접적인 유연관계가 없었다.

• 식물병연구
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• 제29권2호
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• pp.130-136
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• 2023

Citrus melanoses caused by Diaporthe citri, has been one of the serious diseases in many citrus orchards of Jeju Island. To protect melanose in citrus farms, a fast and exact diagnosis method is necessary. In this study, diseased leaves and dieback twigs were collected from a total of 49 farms within March to April in 2022. A total of 465 fungal isolates were obtained from a total of 358 isolated plant samples. Among these fungal isolates, 40 representatives of D. citri isolates which were isolated from 22 twigs and 18 leaves on 23 farms were found based on cultural characteristics on potato dextrose agar and conidial morphology. Additionally, the molecular assay was carried out and compared with those by morphological diagnosis. All isolates were identified as D. citri by analyzing the sequences at the internal transcribed spacer (ITS) rDNA region using primers of ITS1/ITS4 or at β-tubulin using primer Btdcitri-F/R. Therefore, based on the present study, where the results of morphological identification of conidial type were consistent with DNA sequence analysis of certain gene, choosing a suitable method for a fast diagnosis of citrus melanose was suggested.

• Journal of Microbiology and Biotechnology
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• 제8권6호
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• pp.650-655
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• 1998

The chromosomal DNA fragment from Phaffia rhodozyma CBS 6938 which is able to autonomously replicate in the yeast Saccharomyces cerevisiae was cloned on an integrative URA3 plasmid. Its minimal fragment exhibiting autonomously replicating activiy in the S. cerevisiae gave a higher frequency transformation efficiency than that found for centromere-based plasmid, and enabled extrachromosoma1ly stable transmission of the plasmids in one copy per yeast cell under non-selective culture condition. The 836-bp DNA element lacked an ORF and did not contain any acceptable match to an ARS core consensus. Sequence analysis, however, displayed a cluster of three hairpin-Ioop-sequences with individual $\triangle {G_{25}}^{\circ}C$ free energy value of -10.0, -17.5, and -17.0 kcal. $mor^{-l}$as well as a 9-bp sequence with two base pair mismatches to the S. cerevisiae/E. coli gyrase-binding site. This 836-bp sequence also included one 7-bp sequence analogous to the core consensus of centromeric DNA element III (CDEIII) of S. cerevisiae, but CDEIII-like 7 bp sequence alone did not give a replicative function in this yeast.

• Fisheries and Aquatic Sciences
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• 제2권1호
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• pp.66-77
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• 1999

The sequences coding for the 5.8S rDNA and the internal transcribed spacers (ITS1 and ITS 2) from the isolates of nine isolates of Gyrodinium impudicum and two isolates of Gymnodinium sanguineum species were amplified, sequenced and compared with the previously known Alexandrium species and Gymnodinium catenatum. The genetic distance analyses based on the sequence alignment indicated that Gymnodinium catenatum and Gyrodinium impudicum species were some related, Alexandrium species was distant. G. catenatum and G. sanguineum were quite separate, but these two species belonged to the same genus. G. impudicum and G. catenatum forming the closet cluster showed some variation in the alignment of ITS regions. The length of ITS1 varied more than that of ITS2 and the length of ITS1 and ITS2 was different for each G. impudicum, Gymnodinium and Alexandrium species. Also, the length of ITS1 was shorter than that of ITS2. However, on the sequences of G. sanguineum, the length of ITS1 was longer about 23 nucleotides than that of ITS2. The phylogenetic analysis and rDNA similarity of G. impudicum and G. catenatum $(59\%)$ is higher than the that of G. catenatum and G. sanguineum $(55\%)$. It was thought that the phylogenetic analysis and the genetic distance revealed that G. impudicum and G. catenatum were clearly different species and G. impudicum may belong to the genus of Gymnodinium.

• Journal of Microbiology
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• 제38권2호
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• pp.66-73
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• 2000

To investigate the genetic relationship among 12 species belonging to the Fusarium section Martiella, Dlaminia, Gibbosum, Arthrosporiella, Liseola and Elegans, the internal transcribed spacer(ITS) regions of ribosomal DNA (rDNA) were amplified with primer pITS1 and pITS4 using the polymerase chain reaction(PCR). After the amplified products were digested with 7 restriction enzymes, restriction fragment length polymorphism (RFLP) patterns were analyzed. The partial nucleotide sequences of the ITS region were determined and compared. Little variation was observed in the size of the amplified product having sizes of 550bp or 570bp. Based on the RFLP analysis, the 12 species studied were divided into 5 RFLP types. In particular, strains belonging to the section Martiella were separated into three RFLP types. Interestingly, the RFLP type of F. solani f. sp. piperis was identical with that of isolates belonging to the section Elegans. In the dendrogram derived from RFLP analysis of the ITS region, the Fusarium spp. examined were divided into two major groups. In general, section Martiella excluding F. solani f. sp. piperis showed relatively low similarity with the other section. The dendrogram based on the sequencing analysis of the ITS2 region also gave the same results as that of the RFLP analysis. As expected, 5.8S, a coding region, was highly conserved, whereas the ITS2 region was more variable and informative. The difference in the ITS2 region between the length of F. solani and its formae speciales excluding F. solani f. sp. piperis and that of other species was caused by the insertion/deletion of nucleotides in positions 143-148 and 179-192.

• 원예과학기술지
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• 제35권3호
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• pp.344-360
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• 2017

Zoysia 속 잔디는 학교운동장 및 공원, 골프장, 스포츠경기장과 같이 다양한 장소에 식재되고 있는 중요한 잔디이다. 해안가에서 자생하는 Zoysia 속 들잔디와 갯잔디는 외부 형태적 특성이 유사하여 외부 형태적 분류 뿐 만 아니라 분자생물학적 분류도 필요하다. 본 연구에서는 nrDNA-ITS(Internal Transcribed Spacer)의 DNA 바코드 분석을 통해서 자생하는 들잔디와 갯잔디의 분자생물학적 신속한 분류체계를 확립하고자 하였다. 이를 위해 난지형 잔디인 Zoysia 속 들잔디(Z. japonica) 및 갯잔디(Z. sinica)와 한지형 대표 잔디인 크리핑 벤트그라스(A. stolonifera) 및 켄터키 블루그라스(P. pratensis)의 nrDNA-ITS 염기서열을 확보하였다. 확보된 들잔디및 갯잔디, 크리핑 벤트그라스, 켄터키 블루그라스의 ITS 염기서열 전체 구간은 각 686bp와 687bp, 683bp, 681bp으로 확인되었으며, nrDNA-ITS 내부 염기서열구간 분석 결과, ITS1의 크기는 248-249bp, ITS2는 270̵-274bp, 5.8S rDNA는 163-164bp의 차이로, 각 4종의 잔디가 ITS 염기서열을 이용하여 식별되었다. 특히, 들잔디와 갯잔디 nrDNA-ITS 염기서열은 19 염기(2.8%) 차이를 나타냈으며, ITS1과 ITS2의 G + C 함량은 55.4-63.3% 임을 확인하였다. 이러한 들잔디와 갯잔디의 ITS 염기서열 차이를 바탕으로 CAPS 마커로 전환하여 대조구 및 수집된 자생 Zoysia 속 잔디 영양체 62개체를 분석한 결과, 외부형태학적 분류법으로 들잔디 개체, 갯잔디 개체로 동정되었지만, ITSCAPS 마커를 이용한 분자생물학적 분류법으로 들잔디 36개체와 갯잔디 22개체 뿐만 아니라 들잔디와 갯잔디간의 자연교배종 4개체도 식별하였다. 이상의 결과에서 들잔디와 갯잔디는 ITS 염기서열 및 ITS 기반 CAPS를 통하여 식별할 수 있을 것으로 판단된다.