• Title/Summary/Keyword: ITS rDNA

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Report of the Genus Rhodobates (Lepidoptera, Tineidae) New to Korea

  • Sohn, Jae-Cheon
    • Animal Systematics, Evolution and Diversity
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    • v.38 no.3
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    • pp.140-143
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    • 2022
  • A tineid genus, Rhodobates Ragonot, 1895 in Myrmecozelinae is reported for the first time from Korea, with a congener, R. cupulatus Li and Xiao, 2006. The voucher specimens of the species comprise four males and four females collected from Seoul City, Sejong City, Gangwon-do, Chungcheongbuk-do, and Chungcheongnam-do. Among the previously-known species of Tineidae in Korea, Rhodobates cupulatus is similar to Psychoides gosari Kim and Bae in having the uniform coloration over the body and wings, but differs from the latter in the much larger body size. External appearance and genital features of R. cupulatus are redescribed and illustrated. A COI sequence of R. cupulatus is provided for the first time and compared in the world database of DNA barcodes. The Korean records of the species represent the first evidence of its occurrence out of the type locality. Circumstances of its collecting in Korea suggest that it is possibly feeding on dead woods in damp environments.

Preliminary Application of Molecular Monitoring of the Pacific Herring (Clupea pallasii) Based on Real-time PCR Assay Utilization on Environmental Water Samples

  • Kim, Keun-Yong;Heo, Jung Soo;Moon, Seong Yong;Kim, Keun-Sik;Choi, Jung-Hwa;Yoo, Joon-Taek
    • Korean Journal of Ecology and Environment
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    • v.54 no.3
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    • pp.209-220
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    • 2021
  • Pacific herring, Clupea pallasii, a keystone species with significant ecological and commercial importance, is declining globally throughout much of its range. While traditional fishing equipment methods remain limited, new sensitive and rapid detection methods should be developed to monitor fisheries resources. To monitor the presence and quantity of C. pallasii from environmental DNA (eDNA) extracted from seawater samples, a pair of primers and a TaqMan® probe specific to this fish based on mitochondrial cytochrome b (COB) sequences were designed for the real-time PCR (qPCR) assay. The combination of our molecular markers showed high specificity in the qPCR assay, which affirmed the success of presenting a positive signal only in the C. pallasii specimens. The markers also showed a high sensitivity for detecting C. pallasii genomic DNA in the range of 1 pg~100 ng rxn-1 and its DNA plasmid containing COB amplicon in the range of 1~100,000copies rxn-1, which produced linear standard calibration curves (r2=0.99). We performed a qPCR assay for environmental water samples obtained from 29 sampling stations in the southeastern coastal regions of South Korea using molecular markers. The assay successfully detected the C. pallasii eDNA from 14 stations (48.2%), with the highest mean concentration in Jinhae Bay with a value of 76.09±18.39 pg L-1 (246.20±58.58 copies L-1). Our preliminary application of molecular monitoring of C. pallasii will provide essential information for efficient ecological control and management of this valuable fisheries resource.

Isolation, Identification and Characterization of a Antidementia Acetylcholinesterase Inhibitor-Producing $Yarrowia$ $lipolytica$ S-3

  • Kang, Min-Gu;Yoon, Min-Ho;Choi, Young-Jun;Lee, Jong-Soo
    • Mycobiology
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    • v.40 no.1
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    • pp.42-46
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    • 2012
  • This report describes the isolation and identification of a potent acetylcholinesterase (AChE) inhibitor-producing yeasts. Of 731 species of yeast strain, the S-3 strain was selected as a potent producer of AChE inhibitor. The selected S-3 strain was investigated for its microbiological characteristics. The S-3 strain was found to be short-oval yeast that did not form an ascospore. The strain formed a pseudomycelium and grew in yeast malt medium containing 50% glucose and 10% ethanol. Finally, the S-3 strain was identified by its physiological characteristics and 26S ribosomal DNA sequences as $Yarrowia$ $lipolytica$ S-3.

Evaluation of Xenotropic Murine Leukemia Virus and its R426Q Polymorphism in Patients with Prostate Cancer in Kerman, Southeast of Iran

  • Reza, Malekpour Afshar;Fahimeh, Gadari;Reza, Mollaie Hamid
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.3669-3673
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    • 2012
  • A role for the xenotropic murine leukemia virus (XMRV) in prostate cancer development has been postulated. To answer questions regarding the prevalence of XMRV in Iranian patients with prostate cancer and its association with the RNASEL R462Q polymorphism, we here investigated a series of cases in Kerman, in the Southeast of Iran, and sought to verify the association with the R462Q using Real Time PCR Method. Prostate tissue specimens of 200 patients with prostate cancer were genotyped for R462Q by real time polymerase chain reaction allelic discrimination and were screened for XMRV proviral DNA by real time polymerase chain reaction specific for the envelope gene. Of 200 patients in this study 8 (4%) cases were positive for XMRV, the QQ allele being the most frequenct regarding the R426Q polymorphism while in negative patients it was the RQ allele. There was significant correlation between high pathological scores and XMRV positive samples. No significant relationship was found between age groups and XMRV results. XMRV was only found in patients with QQ and RQ alleles, not RR. XMRV is detectable in tumor prostate tissue from some patients with prostate cancer, independent of R462Q.

Molecular cloning and characterization of 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (CaHDR) from Camptotheca acuminata and its functional identification in Escherichia coli

  • Wang, Qian;Pi, Yan;Hou, Rong;Jiang, Keji;Huang, Zhuoshi;Hsieh, Ming-shiun;Sun, Xiaofen;Tang, Kexuan
    • BMB Reports
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    • v.41 no.2
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    • pp.112-118
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    • 2008
  • Camptothecin is an anti-cancer monoterpene indole alkaloid. The gene encoding 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (designated as CaHDR), the last catalytic enzyme of the MEP pathway for terpenoid biosynthesis, was isolated from camptothecin-producing Camptotheca acuminata. The full-length cDNA of CaHDR was 1686 bp encoding 459 amino acids. Comparison of the cDNA and genomic DNA of CaHDR revealed that there was no intron in genomic CaHDR. Southern blot analysis indicated that CaHDR belonged to a low-copy gene family. RT-PCR analysis revealed that CaHDR expressed constitutively in all tested plant organs with the highest expression level in flowers, and the expression of CaHDR could be induced by 100 ${\mu}M$ methyl-jasmonate (MeJA), but not by 100 mg/L salicylic acid (SA) in the callus of C. acuminata. The complementation of CaHDR in Escherichia coli ispH mutant MG1655 demonstrated its function.

Isolation of Fungal Pathogens to an Edible Mushroom, Pleurotus eryngii, and Development of Specific ITS Primers

  • Kim, Sang-Woo;Kim, Sinil;Lee, Hyun-Jun;Park, Ju-Wan;Ro, Hyeon-Su
    • Mycobiology
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    • v.41 no.4
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    • pp.252-255
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    • 2013
  • Fungal pathogens have caused severe damage to the commercial production of Pleurotus eryngii, the king oyster mushroom, by reducing production yield, causing deterioration of commercial value, and shortening shelf-life. Four strains of pathogenic fungi, including Trichoderma koningiopsis DC3, Phomopsis sp. MP4, Mucor circinelloides MP5, and Cladosporium bruhnei MP6, were isolated from the bottle culture of diseased P. eryngii. A species-specific primer set was designed for each fungus from the ITS1-5.8S rDNA-ITS2 sequences. PCR using the ITS primer set yielded a unique DNA band for each fungus without any cross-reaction, proving the validity of our method in detection of mushroom fungal pathogens.

Phylogenetic Study of Ganoderma spp. Based on the DNA Sequences in ITS II Region (ITS II 영역의 DNA 염기서열 분석에 의한 불로초(Ganoderma)속의 계통분류학적 고찰)

  • Park, Dong-Suk;Go, Seung-Joo;Ryu, Jin-Chang;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.27 no.1 s.88
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    • pp.39-43
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    • 1999
  • The internal transcribed spacer II regions (ITS II) of the ribosomal DNA gene repeat from Ganoderma spp. were amplified using polymerase chain reaction (PCR) and sequenced. Sequences from 9 species including Ganoderma lucidum, G. tsugae, G. pfeifferi, G. resinaceum, G. australe-applanatum, G. oregonense, G. neo-japonicum, G. applanatum and Inonotus xeranticus as an out-group were compared. The spacer regions of them were $247{\sim}257$ nucleotides in length and contained partial sequences of 5.8S and 25S gene. The reciprocal homologies of each ITS II sequence of the species were in the range of $70{\sim}100%$ except outgroup species, I. xeranticus. According to the analysis of ITS II sequences, Ganoderma spp. constructed 5 clusters. Ganoderma lucidum isolates were to be divided into two groups. One group was consisted of isolates from South Korea. The other group comprised isolates from UK. G. lucidum isolates belonging to the group I were closely related with G. tsugae. These results suggested that G. lucidum from Korea should be G. tsugae, otherwise G. tsugae was to be synonym of G. lucidum.

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Natural hybridization of Iris species in Mt. Palgong-san, Korea (팔공산 금붓꽃 계열의 자연 잡종 현상)

  • Son, OGyeong;Son, Sung-Won;Suh, Gang-Uk;Park, SeonJoo
    • Korean Journal of Plant Taxonomy
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    • v.45 no.3
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    • pp.243-253
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    • 2015
  • Series Chinensis, Genus Iris, endemic to the far regions of East Asia, consists of four species and related varieties. This series is divided into two major groups (I. rossii and I. minutiaurea complex). In this study, the ITS region and matK gene sequences within nuclear ribosomal DNA and plastid DNA were analyzed in order to investigate the phylogenetic relationships among the I. minutiaurea complex (I. minutiaurea, I. odaesanensis, and I. koreana) and the taxonomic identities of a putative hybrid in Mt. Palgong. In the internal transcribed spacer (ITS1, 5.8S, and ITS2) region, a total of 106 cloned genomic sequences from three taxa were obtained to study the intragenomic polymorphisms of the ITS regions. Three taxa revealed high levels of intragenomic polymorphisms, indicative of incomplete nrDNA concerted evolution. This incomplete ITS concerted evolution in the series Chinensis may be linked to the recent species divergence and frequent interspecies hybridization of the series Chinensis. In the matK gene, three taxa were fairly separated by eleven variable sites. In eight individuals collected on Mt. Palgong, putative hybrids between I. odaesanensis and I. minutiaurea were clustered in the I. minutiaurea clade in the NJ (neighbor-joining) tree based on the matK gene. However, in the ITS tree, some of them were clustered in the I. odaesanensis clade and others were clustered in the I. minutiaurea clade. Therefore, the individuals on Mt. Palgong were formed by the hybridization between two taxa (I. odaesanensis and I. minutiaurea) and not through the lineage of I. koreana.

Optimization of artificial cultivation of Tremella fuciformis in closed culture bottle (흰목이버섯 대량생산을 위한 용기내 재배 최적화 연구)

  • Choi, Sung Woo;Chang, Hyun-You;Yoon, Jeong Weon;Lee, Chan
    • Journal of Mushroom
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    • v.6 no.1
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    • pp.20-26
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    • 2008
  • The stromatal forms of T. fuciformis and the mycelia of Hypoxylon sp. were collected. The DNA sequence in the ITS region of the 5.8S ribosomal genes of isolated strain KG103 was very similar to that of T. fuciformis AF042409 with a homology of over 98% in the EMBL/GenBank database through BLAST searching. A second isolate, No KG201, one of the symbiotic strains for cultivating T. fuciformis also exhibited high homology with Annulohhypoxylon stygium AJ390406. Potato Dextrose Medium exhibited the best mycelial growth of 14 mm/14 days and 85 mm/14 days for T. fuciformis and its symbiotic fungi, respectively. Optimum culture conditions for the micelial growth were pH 5 at $25^{\circ}C$. For the optimization of artificial cultivation of T. fuciformis in bottle with sawdust medium, several conditions such as type of sawdust, supplements, pH, moisture content, and incubation temperature were investigated. T. fuciformis and symbiotic fungi showed fast mycelial growth on corn cob media (77 and 52%) followed by oak tree sawdust and cotton seed meal. The optimal temperature for mycelial growth of T. fuciformis and symbiotic fungi on corn cob media was $25^{\circ}C$ at 55% of moisture content.

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Identification of Phellinus linteus by Comparison of Colony Shapes and Using PCR techniques (목질진흙버섯(Phellinus linteus)의 균총형태 비교 및 PCR 기법을 이용한 동정)

  • Kong, Won-Sik;Kim, Dong-Hyun;You, Chang-Hyun;Kim, Young-Ho;Kim, Kyung-Soo;Kim, Kwang-Ho
    • The Korean Journal of Mycology
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    • v.26 no.4 s.87
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    • pp.466-477
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    • 1998
  • Twenty-two Phellinus strains were characterized using colony morphologies and polymerase chain reaction (PCR) to divide into Phellinus linteus. There were some differences in mycelial growth and colony shapes among the strains when they were grown on various media such as PDA, MCM, MEA and YM. Phellinus linteus was slowly growing, formed golden-yellow colony, and produced blue pigment on PDA media. When the regions of internal transcribed spacer (ITS) were amplified from ribosomal RNA (rRNA) coding genes of P. igniarius and P. linteus strains by means of PCR, two types of band (700 bp and 800 bp) were appeared, respectively. For the amplified intergenic region I (IGRI), P. igniarius strains showed a different band among 500, 600, 700 and 800 bp according to the strains, whereas P. linteus strains did one specific band of 700 bp. By polymorphism analysis after digesting the amplified products with 6 different restriction enzymes, a band specific to P. linteus was generated when the products for ITS region were digested with HaeIII, suggesting that the enzyme digestion could provide effective method to distinguish between P. igniarius and P. linteus. And also, the analysis of genetic relationship showed that the genetic similarities were 89% and 95% in P. igniarius and P. linteus strains, respectively. Random amplification polymorphic DNA (RAPD) analysis using multiple primer sets and arbitrarily primed PCR (AP-PCR) with ITS3 primer could also result in a reproducible way to identify P. linteus strains.

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