Cultivated hot pepper crops showing severe mosaic symptom were found in Korea in 2004 and their causal agent was identified as Cucumber mosaic virus (CMV). These pepper crops was resistant to the virus in the filled, and they belonged to pathotype 0 (P0) resistant pepper. Resistance screening of selected pepper plants showed that a pepper isolate of CMV was the P0 resistance-breaking virus. This P0 resistance-breaking isolate of CMV, named as Ca-P1, was isolated from leaves of the virus-infected Capsicum annuum cv. Manidda that showed systemic severe mosaic symptom. Ca-P1-CMV could induce systemic mosaic symptoms on P0-susceptible (P0-S) and P0-resistant (P0-R) cultivars whereas an ordinary strain (Fny-CMV) could not infect P0-R. This result suggests that Ca-P1-CMV can overcome P0 resistant pepper cultivars. To analyze its genome sequence, the complete nucleotide sequence of RNA3 of Ca-P1-CMV was determined from the infectious full-length cDNA clone of the virus. RNA3 of Ca-P1-CMV consisted of 2,219 nucleotides. Overall sequence homology of RNA3-encoded two viral proteins (movement protein and coat protein) revealed high similarity (75.2-97.2%) with the known CMV strains. By sequence analysis with known representative strains of CMV, Ca-P1-CMV belongs to a typical member of CMV subgroup IB. The resistance and resistance-breaking mechanisms of pepper and counterpart CMV, respectively, remain to be investigated, which will enrich the genetic resources and accelerate CMV-resistant pepper breeding programs.
Brown patch caused by Rhizoctonia solani AG-1 IB occurred on Kentucky bluegrass during late May through early October 2010 at golf course in Gyeongbuk Province, Korea. Disease symptoms on the turfgrass for spring season were leaf blights dying from the leaf tip, which appeared patches of brown color in the field. However, it appeared patches of dark brown color or gray brown color in fall. The fungus (B-7 isolate) of brown patch was isolated from the diseased leaf tissue and cultured on potato-dextrose agar (PDA) for identification. The young hyphae had acute angular branching and few septa and mature hyphal branches showed about 90-degree angles and development of monilioid cells, which were morphologically identical to Rhizoctonia solani AG-1 IB reported previously. DNA sequences of ribosomal RNA gene (internal transcribed spacer) of the fungus were homologous with similarity of 99% to those of Rhizoctonia solani AG-1 IB isolates in GenBank database, confirming the identity of the causal agent of the disease. Pathogenicity of the fungus was also confirmed on the creeping bentgrass and Kentucky bluegrass by Koch's postulates. This is the first report of brown patch on Kentucky bluegrass caused by Rhizoctonia solani AG-1 IB in Korea.
Park, Mi-Jeong;Park, Jong-Han;Hong, Seung-Beom;Shin, Hyeon-Dong
한국균학회소식:학술대회논문집
/
2015.05a
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pp.55-55
/
2015
Rust is one of the most destructive diseases on economically important plants such as agricultural and horticultural crops, as well as forest trees [1]. Chemical treatment is the most effective means to control rust, but use of the chemical fungicides involves inevitable risks to human health and environment [2]. Unfortunately, biocontrol is currently impracticable for rust disease management [3]. It is necessary to exploit biocontrol agents to help prevent rust diseases. As a fundamental research for future development of biocontrol agents for rusts, biodiversity of hyperparasites occurring on rust fungi was investigated. During 2006-2010, 197 fungal isolates of the rust hyperparasites were collected and isolated from various combinations of mycohosts and plant hosts in many regions of Korea. Based on morphological and molecular data, they were identified as 8 genera and 12 species. Besides, phylogenetic relationships between the hyperparasites and related taxa were inferred. A total of 114 isolates of Pseudovirgaria were obtained from rust pustules of Phragmidium spp. and Pucciniastrum agrimoniae infecting rosaceous plants. Phylogenetic analysis using multigene sequences revealed a high level of genetic variability among many isolates of Pseudovirgaria and close correlation between the isolates and mycohosts. Only two species of Pseudovirgaria, P. hyperparasitica and P. grisea are often difficult to distinguish by their morphological similarity, but on the molecular basis they were clearly differentiated from each other. There had been no previous record of P. grisea outside Europe, but the present study has proved its presence in Korea. Among six distinct groups (five of P. hyperparasitica and one of P. grisea) within the Pseudovirgaria isolates, each lineage of P. hyperparasitica was closely associated with specific mycohosts and thus might have cospeciated with their mycohosts, which probably led to coevolution. Although P. grisea possesses a host preference for Phragmidium species occurring on Rubus, it was not specific for a mycohost. P. grisea seems to evolve in the direction of having a broad mycohost range. Seventeen isolates of Verticillium-like fungi were isolated from rust sori. Based on morphological data and DNA sequence analysis, the isolates were identified as three Lecanicillium species, viz. L. attenuatum, Lecanicillium sp. 1, Lecanicillium sp. 2, and V. epiphytum. The unidenified two species of Lecanicillium appear to be previously unknown taxa. Sixty-six isolates of miscellaneous hyphomycetes belonging to 6 species of 5 genera were obtained from pustules of rust fungi. On the basis of morphological and molecular analyses, the miscellaneous hyphomycetes growing on rusts were identified as Acrodontium crateriforme, Cladophialophora pucciniophila, Cladosporium cladosporioides, Phacellium vossianum, Ramularia coleosporii, and R. uredinicola.
Kim, Nam-Hoon;Choi, Hong-Il;Kim, Kyung Hee;Jang, Woojong;Yang, Tae-Jin
Journal of Ginseng Research
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v.38
no.2
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pp.130-135
/
2014
Background: Panax ginseng, the most famous medicinal herb, has a highly duplicated genome structure. However, the genome duplication of P. ginseng has not been characterized at the sequence level. Multiple band patterns have been consistently observed during the development of DNA markers using unique sequences in P. ginseng. Methods: We compared the sequences of multiple bands derived from unique expressed sequence tagsimple sequence repeat (EST-SSR) markers to investigate the sequence level genome duplication. Results: Reamplification and sequencing of the individual bands revealed that, for each marker, two bands around the expected size were genuine amplicons derived from two paralogous loci. In each case, one of the two bands was polymorphic, showing different allelic forms among nine ginseng cultivars, whereas the other band was usually monomorphic. Sequences derived from the two loci showed a high similarity, including the same primer-binding site, but each locus could be distinguished based on SSR number variations and additional single nucleotide polymorphisms (SNPs) or InDels. A locus-specific marker designed from the SNP site between the paralogous loci produced a single band that also showed clear polymorphism among ginseng cultivars. Conclusion: Our data imply that the recent genome duplication has resulted in two highly similar paralogous regions in the ginseng genome. The two paralogous sequences could be differentiated by large SSR number variations and one or two additional SNPs or InDels in every 100 bp of genic region, which can serve as a reliable identifier for each locus.
Over the past 20 years, global warming has transformed the marine ecosystem of the Jeju Island into a subtropical zone making it conducive to the production of tropical fishes. Recently, the balloon fish (Diodon holoanthus) has been found off the coast of the Jeju Island. In this study, we analyzed the diversity of its intestinal microorganisms as a representative for the surrounding environment. In addition, the isolates were evaluated for their antibacterial activity. A total of 161 strains of various species were identified and isolated using 16S ribosomal RNA gene sequence analysis. They were separated into three groups, of which Phylum Proteobacteria was found to be the most dominant with 91% sequence similarity. This includes the class γ-proteobacteria that is made up of twelve genera and twenty-four hundred species. The second group comprised strains of the genus Vibrio, made up of 35% Photobacteria, 32% Shewanella, and 6% Psychrobacter. It was also determined that 4% of the isolates were Acinetobacter, 3% were Enterovibrio, while Moraxella_g2 accounted for 1% of the total isolates. Class α-proteobactera includes five genera and five species; Brevundimonas, Allorhizobium, Pseudoceanicola and Erythrobcter, each accounting for 1% of the total isolates. The Firmicute strains belonged to six genera and ten species. 5% of the strains were Terribacillus, while Paenibacillus, Salinicoccus, Staphylococcus and Streptococcus accounted for 1% each of the total isolates. Actinobacteria accounted for the final phylum with strains belonging to three genera and ten species with Janibacter, Micrococcus and Isoptericola each accounting for 1% of the total isolates.
Orf virus (ORFV), a member of genus Parapoxvirus (family-Poxviridae), a causative agent of contagious ecthyma in sheep and goat leading to a condition commonly known as vesicular dermatitis. Recently, twelve goats from Iksan in Jeonbuk province were observed with clinical signs like necrotic vesicular lesions around the mucosa of mouth, nasal cavity, eye, ear, teats, abdomen and groin. Based on these clinical symptoms, contagious ecthyma infection was suspected. The skin scrapping was collected from lesions for isolation of DNA and subsequent PCR amplification of ORFV specific 235 bp region of B2L gene. All of the samples were found positive by PCR analysis. Sequencing and further phylogenetic analysis of the PCR product revealed 100% identity to Japan isolate of ORFV (Okinawa, GenBank accession number AB080769), and showed 99.6% of similarity to New Zealand strain (NZ-2, GenBank accession number U06671). It was concluded that ORFV strain detected in the present study is homologous to Japan isolate and New Zealand strain. The PCR test based on amplification of B2L gene is a highly useful tools for rapid and specific diagnosis of contagious ecthyma.
Seo, Kyoung-In;Yoo, Young-Bok;Jang, Kab-Yeul;Shin, Pyung-Gyun;Oh, YounLee;Kim, Kwang-Ho;Kong, Won-Sik
The Korean Journal of Mycology
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v.41
no.1
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pp.14-20
/
2013
We already reported four groups which contains some similar strains based on URP-PCR in the previous paper. The objective of this study was to confirm those strains by the amplified fragment length polymorphism (AFLP) and vegetative compatibility group (VCG). AFLP analysis showed no difference among these strains except ASI 2595 and 2183 in Weonhyeong group and ASI 2829 in Suhan group. They showed specific DNA bands only in the result of P + AG/M + AAG and P + GT/M + ATG primer combinations out of eight different combinations. The AFLP primers produced a total of 330 fragments between 80 and 1000 bp in length for 31 Pleurotus ostreatus strains. At a genetic similarity of 0.96, the UPGMA analysis separated the isolates into four distinct clusters. Each group was classified by similar strains. Confrontation test by vegetative compatibility groups (VCGs) also showed distinct line between strains from different groups, but no line between similar strains within the cluster. Our results indicate that most of similar strains was not distinctness. Thus, similar strains are considered to be very close on the genealogy of their parent or same strain with different name.
To determine evolution and genotype of new chromosomal AmpC $\beta$-lactamases among clinical isolates of Enterobacter species, we performed antibiotic susceptibility testing, pI determination, sequencing, and phy-logenetic analysis using developed expression/secretion vector. Six isolates have shown to produce AmpC $\beta$-lactamases. Six genes of AmpC $\beta$-lactamases that are responsible for the resistance to cephamycins (cefoxitin and cefotetan), amoxicillin, cephalothin, and amoxicillin-clavulanic acid were cloned and characterized in pMSG12119. Insert fragment containing the ampC genes was sequenced and found to have an open reading frame coding for 381-amino-acid $\beta$-lactamase. The nucleotide sequence of four ampC genes ($bla_EcloK992004.l$, $bla_EcloK995120.1$, $bla_EcloK99230$, and $bla_EareK9911729$) shared considerable homology with that of chromosomal ampC gene ($bla_EcloMHN1$) of E. cloacae MHN1 (more than 99.6% identity). The sequences of two ampC genes ($bla_EcloK9973$ and $bla_EcloK9914325$) showed close similarity to the chromosomal ampC gene ($bla_EcloQ908R$) of E. clo-acae 908R (99.7% identity). The results from phylogenetic analysis suggested that six ampC genes could be originated from $bla_EcloMHN1$ / or $bla_EcloQ908R$ / MIC patterns and exact pI values of six transformants indicated that the developed expression/secretion vector (pMSG1219) was suitable for the characterization of foreign genes in E. coli strain.
Kim, Byung-Yong;Ahn, Jae-Hyung;Song, Jaekyeong;Kim, Myung-Sook;Weon, Hang-Yeon
Korean Journal of Soil Science and Fertilizer
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v.45
no.6
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pp.1100-1107
/
2012
Bacterial endophytes may be important factors in plant growth and ecologically relevant functions in rice. Using pyrosequencing technology, we analyzed the composition of endophytic bacterial communities that colonized the roots of rice cultivated in long-term fertilized (APK) and non-fertilized (NF) paddy soils. A total of 1,900 reads were obtained from 2 samples. All sequences were classified into 177 OTUs (APK sample) or 72 OTUs (NF sample) at a 97% similarity cut-off. Twenty-two OTUs were shared between the 2 samples, and these were also the most dominant OTUs in both samples. Proteobacteria was the most dominant phylum with 90.2%, followed by Actinobacteria (7.1%) and Bacteroidetes (1.1%). Furthermore, Pseudomonas was the most abundant genus in both samples. We observed clear differences in the structure of the endophytic bacterial community structure between the 2 samples. Notably, the distributions of Alphaproteobacteria and Gammaproteobacteria were markedly different. The diversity index of the APK sample was higher than that of the NF sample. These findings showed that the endophytic bacterial community of rice roots was affected by the presence of fertilizers in the rice field soil.
Park, Kyoung-Joo;Cho, Kyoung-Sook;Kim, Jeong-Bo;Lee, Min-Gyu;Lee, Byung-Hun;Hong, Young-Ki;Kim, Joong-Kyun
Environmental Engineering Research
/
v.10
no.5
/
pp.213-226
/
2005
To treat wastewater efficiently by a one-step process of nitrogen removal, a new bacterial strain producing $N_2$ gas from ${NH_4}^+$ under an aerobic condition was isolated and identified. The cell was motile and a Gram-negative rod, and usually occurred in pairs. By 16S-rDNA analysis, the isolated strain was identified as Enterobacter asburiae with 96% similarity. The isolate showed that the capacity of $N_2$ production under an oxic condition was approximately three times higher than that under an anoxic condition. Thus, the consumption of ${NH_4}^+$ by the isolate was significantly different in the metabolism of $N_2$ production under the two different environmental conditions. The optimal conditions of the immobilized isolate for $N_2$ production were found to be pH 7.0, $30^{\circ}C$ and C/N ratio 5, respectively. Under all the optimum reaction conditions, $N_2$ production by the immobilized isolate resulted in reduction of ORP with both the consumption of DO and the drop of pH. The removal efficiencies of $COD_{Cr}$, and TN were 56.1 and 60.9%, respectively. The removal rates of $COD_{Cr}$, and TN were the highest for the first 2.5 hrs with the removal $COD_{Cr}/TN$ ratios of 32.1, and afterwards the rates decreased as reaction proceeded. For application of the immobilized isolate to a practical process of ammonium removal, a continuous operation was executed with a synthetic medium of a low C/N ratio. The continuous bioreactor system exhibited a satisfactory performance at 12.1 hrs of HRT, in which the effluent concentrations of ${NH_4}^+$-N was measured to be 15.4 mg/L with its removal efficiency of 56.0%. The maximum removal rate of ${NH_4}^+$-N reached 1.6 mg ${NH_4}^+$-N/L/hr at 12.1 hrs of HRT(with N loading rate of $0.08\;Kg-N/m^3$-carrier/d). As a result, the application of the immobilized isolate appears a viable alternative to the nitrification-denitrification processes.
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