• Korean Journal Plant Pathology
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• v.12 no.1
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• pp.91-98
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• 1996

벼도열병균 14개 균주와 벼 이외 화본과 식물 도열병균 12개 균주를 대상으로 rDNA의 ITS II 부위를 증폭하여 그들의 핵산 구조 차이를 분석함으로 도열병균 균주간 분류를 시도하였다. 5.8S rDNA의 3`-말단 부위와 28S rDNA의 5`-말단 부위의 sequence 중 5`-CCCGGGAATTCGCATCGATCGATCGAATGAAGA-ACGCAGC-3`와 5`-CCCGGGATCCTCCGCTTATT-GATATGC-3`를 이용하여 PCR 증폭을 하였을 때 벼도열병균 14개 균주는 동일한 길이의 단일 밴드를 형성하였으며 벼 이외 화본과 식물 도열병균에서는 레드톱 식물로부터 분리한 도열병균만이 나머지 균주보다 38bp가 더 큰 길이를 가진 밴드를 형성하였다. PCR로 증폭된 DNA를 HaeIII와 MspI 제한효소로 절단하였을 때 벼도열병균 레이스간에는 제한효소 절단에 의한 전기영동 밴드 형태 차이를 관찰할 수 없었으나, 벼 이외 화본과 식물 도열병균 12개 균주는 3군으로 구분할 수 있었다. 벼도열병균 90=054와 강아지풀에서 분리한 도열병균 G90-5, 기장에서 분리한 G88-4, 바랭이에서 분리한 G88-5 그리고 레드톱에서 분리한 RT 균주의 ITS II 부위의 DNA 염기서열 비교 분석에 의하면 G88-4와는 다른 HaeIII와 MspI 제한효소 위치를 가지고 있었기에 제한효소 절단에 의한 전기영동 형태가 상이하였다. 또한 RT균주는 HaeIII와 MspI위치가 존재하지 않았다.

• Korean Journal of Medicinal Crop Science
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• v.18 no.6
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• pp.379-388
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• 2010

Adenophora racemosa is recently reported as a new Korean endemic plant species. However, the phylogenetic relationship of this genus has been controversial due to the morphological similarity and frequent morphological change of aerial parts. To verify the phylogenetic position of Adenophora racemosa and phylogenetic relationship of genus Adenophora, we analyzed the internal transcribed spacer (ITS) sequence of nuclear ribosomal DNA (nrDNA) and random amplified polymorphic DNA (RAPD) using 21 individual of 6 Adenophora species, A. verticillata, A. divaricata, A. racemosa, A. remotiflora, A. stricata and A. tetraphylla. In comparative analysis of the nrDNA-ITS sequences, we could not found not only any species specific nucleotide sequence but also could not estimated their inter or intra species. In the phylogenic analysis based on the RAPD derived DNA polymorphism, Adenophora species were classified into four groups by clustering analysis of the UPGMA. These results suggest that the DNA fingerprinting based on RAPD is more suitable than nrDNA-ITS sequence for the phylogenetic analysis of Adenophora species.

• ALGAE
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• v.37 no.3
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• pp.185-204
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• 2022

The genus Coolia A. Meunier 1919 has a global distribution and is a common member of epiphytic dinoflagellate assemblages in neritic ecosystems. Coolia monotis is the type species of the genus and was the only known species for 76 years. Over the past few decades, molecular characterization has unveiled two species complexes that group morphologically very similar species, so their limits are often unclear. To provide new knowledge on the biogeography and species composition of the genus Coolia, 16 strains were isolated from Bahía de La Paz, Gulf of California. The species were identified by applying morphological and molecular approaches. The morphometric characteristics of all isolated Coolia species were consistent with the original taxa descriptions. Phylogenetic analyses (large subunit [LSU] rDNA D1 / D2 and internal transcribed spacer [ITS] 1 / 5.8S / ITS2) revealed a species assemblage comprising Coolia malayensis, C. palmyrensis, C. tropicalis, and the C. cf. canariensis lineage. This is the first report of Coolia palmyrensis and C. cf. canariensis in Mexico and C. tropicalis in the Gulf of California. Our results strengthen the biogeographical understanding of these potentially harmful epiphytic dinoflagellate species.

• Mycobiology
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• v.51 no.2
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• pp.109-113
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• 2023

Auricularia is one of the broadly cultivated edible mushrooms in Korea. Most of the Korean Auricularia strains used for cultivation and breeding are known as A. auricula-judae. Recently, this species has been reported to belong to a species complex. Therefore, this study was carried out to genetically clarify the bred and cultivated Korean A. auricula-judae strains. The internal transcribed spacer (ITS) and IGS1 rDNA region sequences were determined from 10 A. auricula-judae strains by PCR and sequencing. Variation in the nucleotide sequence and sequence length of the two rDNA regions were found among the seven A. auricula-judae strains. A maximum-likelihood (ML) phylogenetic tree based on the ITS sequences clearly placed all the 10 Korean A. auricula-judae strains in the A. heimuer clade of the A. auriculajudae complex. A. heimuer is diverged from A. auricula-judae. An ML phylogenetic tree based on the IGS1 sequences revealed the close relationship between Korean A. heimuer strains to Chinese A. heimuer strains. But each strain could be distinguishable by the IGS1 sequence. Furthermore, progeny strains in the seven Korean strains could be differentiated from their parental strains by the IGS1 sequence based phylogenetic tree. Our results are expected to be used to complement the distinction of domestic Auricularia cultivars.

• ALGAE
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• v.37 no.4
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• pp.281-291
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• 2022

Quantifying the abundance of Chattonella species is necessary to effectively manage the threats from ichthyotoxic raphidophytes, which can cause large-scale mortality of aquacultured fish in temperate waters. The identification and cell counting of Chattonella species have been conducted primarily on living cells without fixation by light microscopy because routine fixatives do not retain their morphological features. Species belonging to the Chattonella marina complex, including C. marina and C. marina var. ovata, had high genetic similarities and the lack of clear morphological delimitations between the species. To estimate the abundance of C. marina complex in marine plankton samples, we developed a protocol based on the droplet digital polymerase chain reaction (ddPCR) assay, with C. marina complex-specific primers targeting the internal transcribed spacer (ITS) region of the rDNA. Cell abundance of the C. marina complex can be determined using the ITS copy number per cell, ranging from 25 ± 1 for C. marina to 112 ± 7 for C. marina var. ovata. There were no significant differences in ITS copies estimated by the ddPCR assay between environmental DNA samples from various localities spiked with the same number of cells of culture strains. This approach can be employed to improve the monitoring efficiency of various marine protists and to support the implementation of management for harmful algal blooms, which are difficult to analyze using microscopy alone.

• The Korean Journal of Mycology
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• v.27 no.5 s.92
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• pp.337-340
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• 1999

The context and upper surface of Phellinus basidiocarp become blackened, rimose and woody. The basidiocarp is sessile, dimidiate and elongate. The basidiospores are pigmented and ovoid to globose. Hymenial setae are $17{\sim}35{\times}6{\sim}8{\mu}m$. Nineteen isolates of Phellinus species, including Phellinus linteus, were used for sequencing of the internal transcribed spacer (ITS) region of the nuclear rDNA. Based on these sequence data, specific primers were designed for identification of Phellinus linteus isolates in Korea. The specific primers were within the ITS1 and ITS2 regions and were nested within the universal primers flanking the spacer regions. A total of four primers (the universal primers ITS-1F and ITS-4, and the specific primers PL-F and PL-R) were used for detection of Phellinus linteus collected in Korea. The length of the four amplification products of Phellinus linteus DNA were 800 bp (ITS-1F/ITS-4), two bands of about 720 bp (ITS-1F/PL-R and PL-F/ITS-4), and 610 bp (PL-F/PL-R). Among 23 isolates of Phellinus species collected in Korea, Thirteen isolates were identified as Phellinus linteus based on the presence of the four bands. The other species produced only the single ITS-1F/ITS-4 product.

• The Korean Journal of Mycology
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• v.45 no.3
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• pp.246-250
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• 2017

Sclerotium rot was observed on mungbean (Phaseolus radiatus L.) plants cultivated in the exhibition field of Gyeongsangnam-do Agricultural Research and Extension Services in September 2015. The progression of rot was initially observed as water-soaked lesions on several parts of the affected plant. Severely infected plants were blighted and eventually died. White mycelial mats spread over the lesions and numerous sclerotia formed on stems near the soil line. The sclerotia were globoid in shape, 1~3 mm in size, and white to brown in color. The optimum temperature for mycelial growth and sclerotia formation on potato dextrose agar (PDA) was $30^{\circ}C$ and the hyphal width was $4{\sim}8{\mu}m$. Typical clamp connections were observed on the hyphae of fungus grown on PDA. For molecular identification, the complete internal transcribed spacer (ITS) ribosomal DNA (rDNA) of the causal fungus was sequenced and analyzed. Based on the mycological characteristics, ITS rDNA sequence analysis, and pathogenicity to host plants, the fungus was identified as Sclerotium rolfsii. This is the first report of Sclerotium rot on mungbean caused by S. rolfsii in Korea.

• The Korean Journal of Mycology
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• v.43 no.4
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• pp.281-285
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• 2015

This study was conducted to examine changes in the fungal community on fallen leaves of pear by treatment with lime sulfur. Although the lime sulfur could reduce the primary inoculum of several pathogens on spring season, the effect of lime sulfur has not been well determined scientifically. Fallen leaves infected by pear diseases in pear orchards in Naju were collected and treated with lime sulfur or water as a control. To determine the fungal diversity from each treatment, rDNA internal transcribed spacer (ITS) regions were analyzed after extraction of fungal genomic DNA from lime sulfur-treated or water-treated fallen leaves, respectively. The most common fungal species were Ascomycota and Basidiomycota in both treated leaves. However, the population dynamics of several fungal species including Alternari sp., Cladosporium sp., and Phomopsis sp., which are known as pear pathogens for skin sooty dapple disease, were quite different from each treated leaves. These results indicated that lime sulfur treatment led to changes of fungal communities on pear fallen leaves and could be applicable as a dormant spray.

• The Korean Journal of Mycology
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• v.44 no.3
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• pp.201-205
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• 2016

In August 2015, leaf spot symptoms were observed on Korean gondre thistle (Cirsium setidens) in Youngwol, Korea. During the early stage, the symptoms appeared as one or more small gray-brown to brown spots on plant leaves. The spots showed extensive enlargement over time and eventually became large dark brown to black lesions on the whole leaf. Stemphylium species were consistently isolated from affected leaves. All isolates were identified as S. lycopersici, S. solani, or S. xanthosomatis based on morphological and cultural characteristics. The isolates were confirmed as S. lycopersici based on a multilocus sequence analysis using the ribosomal internal transcribed spacer (ITS) region, elongation factor 1, GAPDH (glyceraldehyde-3-phosphate dehydrogenase), and the noncoding region between the vacuolar membrane ATPase catalytic subunit A gene and a gene involved in vacuolar biogenesis. Pathogenicity was tested by spore suspension inoculation on wounded or unwounded gondre leaves. The lesions were observed on inoculated leaves within 3 days after inoculation, regardless of wound. To our knowledge, this is the first report of the leaf spot on gondre thistle caused by S. lycopersici in Korea or elsewhere.

• Research in Plant Disease
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• v.18 no.2
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• pp.73-79
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• 2012

Phytophthora katsurae is a fungal pathogen responsible for chestnut ink disease. We designed two duplex primer sets (SOPC 1F/1R+KatI 3F/5R, SOPC 1-1F/1-1R+KatI 3F/5R) to detect P. katsurae. SOPC 1F/1R and SOPC 1-1F/1-1R primer pairs were designed for sequence characteristic amplification regions (SCAR) marker, and KatI 3F/5R primer pair was used for P. katsurae-specific primer designed from internal transcribed spacer (ITS) region. To assess the sensitivity of duplex PCR, genomic DNA was serially diluted 10-fold to make the final concentrations from 1 mg/ml to 1 ng/ml. The sensitivity for two primer sets were 1 ${\mu}g/ml$ and 100 ng/ml, respectively. To find detection limits for zoospores of P. katsurae, each zoospore suspension was serially diluted 10-fold to make the final concentrations from $1{\times}10^6$ to $1{\times}10^2$ cells/ml, and then DNA was extracted. The limits of detection for all of two primer sets were $1{\times}10^5$ cells/ml. All of two primer sets were specific to P. katsurae in PCR detection and did not produce any P. katsurae-specific PCR amplicons from other 16 Phytophthora species used as the control. This study shows that duplex PCR using two primer sets might be a useful tool for rapid and efficient detection of P. katsurae.