• Title/Summary/Keyword: 18S-rDNA sequencing

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Morphological Characteristics and Molecular Identification of Calonectria ilicicola Causing Soybean Red Crown Rot (콩 검은뿌리썩음병균(Calonectria ilicicola)의 형태적 특징과 유전자 수준에서의 동정)

  • Park, Seong-Woo;Kim, Hong-Sik;Woo, Sun-Hee;Shim, Heong-Kwon;Kim, Heung-Tae
    • Research in Plant Disease
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    • v.12 no.3
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    • pp.178-184
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    • 2006
  • The 18 isolates of soybean red crown rot pathogen(Calonectria ilicicola) were isolated from infected soybean plants showing chlorosis symptom and red perithecia sign in 2002 and 2003 season in Korea. The morphological characteristics of the typical isolate SC03-15, such as macroconidia and macrovesicles in anamorph as well as perithecia, asci and ascospores in teleomorph, were observed from red crown rot pathogens. The perithecia were red-brown globose to ovoid, and $357.9(312.5{\sim}391.5.7){\mu}m$ high and $474.3(419{\sim}535){\mu}m$ diameter. Asci were hyaline and clavate and included 8 ascospores, which were straight to slightly curved, 1(-3)-septate, and $36.9(31.1{\sim}49.4){\times}5.2(4.5{\sim}6.1){\mu}m$. Macroconidia in anamorph were cylindrical, hyaline and in size $73.2(68.1{\sim}77.6){\times}5.8(4.9{\sim}6.8){\mu}m$. Further identification process was performed through molecular method, such as a phylogenetic analysis by rDNA sequencing of ITS and $\beta$-tubuline genes of all isolates tested in this study. All isolates were identified as Calonectria ilicicola by DNA sequencing of ITS and $\beta$-tubuline genes, and morphological characteristics.

Growth hormone and receptor gene mutations in Chinese Banna miniature pig

  • Deng, J.Z.;Hao, L.L.;Li, M.T.;Lang, S.;Zeng, Y.Z.;Liu, S.C.;Zhang, Y.L.
    • Animal cells and systems
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    • v.15 no.4
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    • pp.310-314
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    • 2011
  • The Banna miniature pig (BNMP) is a representative miniature pig breed in China. Even though BNMP dwarfism is obvious, its underlying causative mutations remain unknown. In this study, the BNMP and Large White pig (LWP) serum growth hormone (GH) and insulin-like growth factor (IGF-1) levels were detected by ELISA and compared. BNMP serum IGF-1 levels were significantly lower than LWP levels (P<0.05). The miniature condition may arise from mutations in the GH and GH receptor (GHR) genes. Therefore, GH and GHR cDNA from the BNMP were cloned into a pMD18-T vector by RT-PCR using the total RNA obtained from the BNMP's pituitary and liver tissues. Sequencing results indicated that the open reading frame of the BNMP GH gene is composed of a 26-residue signal peptide and a 191-residue mature peptide. The coding sequence of the BNMP GHR gene contained 639 amino acids, including a signal peptide that is 18 amino acids long. Two amino acid substitutions, A09V and R22Q, were found in the signal peptide of the GH gene. Additionally, the S104P mutation was found in the BNMP's mature GH protein. Four mutations in the cytoplasmic domain of GHR may influence the downstream signal transduction of GHR, which needs further experimental evidence.

Flora of drift plastics: a new red algal genus, Tsunamia transpacifica(Stylonematophyceae) from Japanese tsunami debris in the northeast Pacific Ocean

  • West, John A.;Hansen, Gayle I.;Hanyuda, Takeaki;Zuccarello, Giuseppe C.
    • ALGAE
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    • v.31 no.4
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    • pp.289-301
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    • 2016
  • Floating debris provides substrates for dispersal of organisms by ocean currents, including algae that thrive on plastics. The 2011 earthquake and tsunami in Tohuku, Japan resulted in large amounts of debris carried by the North Pacific Current to North America from 2012 to 2016. In 2015-2016, the plastics in the debris bore a complex biota including pink algal crusts. One sample (JAW4874) was isolated into culture and a three-gene phylogeny (psbA, rbcL, and SSU) indicated it was an unknown member of the red algal class Stylonematophyceae. It is a small pulvinate crust of radiating, branched, uniseriate filaments with cells containing a single centrally suspended nucleus and a single purple to pink, multi-lobed, parietal plastid lacking a pyrenoid. Cells can be released as spores that attach and germinate to form straight filaments by transverse apical cell divisions, and subsequent longitudinal and oblique intercalary divisions produce masses of lateral branches. This alga is named Tsunamia transpacifica gen. nov. et sp. nov. Sequencing of additional samples of red algal crusts on plastics revealed another undescribed Stylonematophycean species, suggesting that these algae may be frequent on drift oceanic plastics.

An Outbreak of Gray Mold Caused by Botrytis cinerea on Kenaf(Hibiscus cannabinus L.)

  • Kwon, Jin-Hyeuk;Kang, Dong-Wan;Lee, Seong-Tae;Shim, Chang-Ki;Kim, Min-Jeong;Kim, Jinwoo
    • Journal of agriculture & life science
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    • v.50 no.2
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    • pp.25-32
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    • 2016
  • A severe outbreak of gray mold on kenaf (Hibiscus cannabinus L.) was observed on kenaf grown in the research field of Gyeongsangnam-do Agricultural Research and Extension Services, Jinju, Korea in 2014. Gray mold appeared on young plants as gray-brown velvety mold covering stems and leaves. Infections that girdled the stem caused wilting above the infected area and developed a canker. The casual fungus formed grayish brown colonies on potato dextrose agar. The conidia were one celled, mostly ellipsoid or ovoid in shape, colorless or pale brown in color, and 6-18 × 4-10 ㎛ in size. The conidiophores were 15-32 ㎛ in length. These measurements and taxonomic characteristics were most similar to those of Botrytis. DNA sequencing and phylogenetic analysis of the complete internal transcribed spacer rRNA gene region confirmed that the fungal isolates were indeed Borytis cinerea. Koch's postulates were supported by pathogenicity tests conducted on healthy plants. On the basis of mycological characteristics and pathogenicity test on host plants, the fungus was identified as Botrytis cinerea. To the best of our knowledge, this is the first report of a gray mold caused by B. cinerea on kenaf in Korea.

Helicobacter pylori vacA Mosaicism and New Primers for vacA Signal Sequence Indigenous to Korea (Helicobacter pylori vacA 대립유전자의 Mosaicism과 Signal Sequence의 한국고유 시발체)

  • Ahn, Yeon-Hwa;Kim, Heung-Ryel;Lee, Ji-Eun;Hwang, Tae-Sook;Choe, Yon-Ho
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.4 no.2
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    • pp.155-160
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    • 2001
  • Purpose: Helicobacter pylori has been known to have diverse vacA allelic types. The purpose of the study was to identify vacA diversity in Korea and design new primers for signal sequence alleles indigenous to Korea. Methods: Fifty antral biopsy specimens, which had been proven to be H. pylori-positive, were examined for vacA status; signal sequence and mid-region. After PCR amplification and DNA sequencing, vacA alleles of Korean H. pylori strains were compared with those from other countries. Results: Among Korean H. pylori strains vacA alleles with all combinations of signal sequence and mid-region were found, with the exception of s1b or s2. vacA genotype s1c/m1 was predominant in Korea. We found that GGGAGCGTTR in s1a and GGGGYTATTG in s1c were the indigenous sequences to Korea and constructed the new Korean specific primers for the vacA signal sequence; VASK-F, VASK-R, S1AK-F, and S1CK-F. Conclusion: This study showed that s1c/m1 is the predominant type of vacA allele in Korea. We designed new primers for the vacA signal sequence.

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Correlation between EGFR Gene Mutations and Lung Cancer: a Hospital-Based Study

  • Kavitha, Matam;Iravathy, Goud;Adi Maha, Lakshmi M;Ravi, V;Sridhar, K;Vijayanand, Reddy P;Chakravarthy, Srinivas;Prasad, SVSS;Tabassum, Shaik Nazia;Shaik, Noor Ahmad;Syed, Rabbani;Alharbi, Khalid Khalaf;Khan, Imran Ali
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7071-7076
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    • 2015
  • Epidermal growth factor receptor (EGFR) is one of the targeted molecular markers in many cancers including lung malignancies. Gefitinib and erlotinib are two available therapeutics that act as specific inhibitors of tyrosine kinase (TK) domains. We performed a case-control study with formalin-fixed paraffin-embedded tissue blocks (FFPE) from tissue biopsies of 167 non-small cell lung carcinoma (NSCLC) patients and 167 healthy controls. The tissue biopsies were studied for mutations in exons 18-21 of the EGFR gene. This study was performed using PCR followed by DNA sequencing. We identified 63 mutations in 33 men and 30 women. Mutations were detected in exon 19 (delE746-A750, delE746-T751, delL747-E749, delL747-P753, delL747-T751) in 32 patients, exon 20 (S786I, T790M) in 16, and exon 21 (L858R) in 15. No mutations were observed in exon 18. The 63 patients with EFGR mutations were considered for upfront therapy with oral tyrosine kinase inhibitor (TKI) drugs and have responded well to therapy over the last 15 months. The control patients had no mutations in any of the exons studied. The advent of EGFR TKI therapy has provided a powerful new treatment modality for patients diagnosed with NSCLC. The study emphasizes the frequency of EGFR mutations in NSCLC patients and its role as an important predictive marker for response to oral TKI in the south Indian population.

Characterization of the Acetolactate synthase (ALS) gene and Molecular Assay of Mutations Associated with Sulfonylurea Herbicide Resistance of Monochoria vaginalis (물달개비의 Acetolactate synthase (ALS) 유전자의 특성과 Sulfonylurea 제초제 저항성과 관련 돌연변의 분자생물학적 접근)

  • Park, Tae-Seon;Park, Hong-Kyu;Ku, Bon-Il;Kim, Young-Doo;Ko, Jae-Kwon;Lee, In-Yong;Park, Jae-Eup
    • The Korean Journal of Pesticide Science
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    • v.13 no.4
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    • pp.290-297
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    • 2009
  • This research aims to contribute the characterization of acetolactate synthase (Ec 4.1.3.18; ALS) and the resistance mechanism by sequence analysis of ALS gene of the sulfonylurea-resistant and -susceptible Monochoria vaginalis. The ALS gene was obtained from susceptible (S) and resistant (R) M. vaginalis to sulfonylurea herbicides (SUs). The 815 bp the fragment and the genomic DNA sequence coding for acetolactate synthase (ALS) of S and R biotypes of M. vaginalis were cloned and sequenced. Nineteen clones were divided greatly into 4 groups as result of sequencing. The first group was not difference to S type, the second group was amino acid of P197S which found point mutations causing substitution of serine for proline at amino acid 197, the third group was observed greatly other part of 6 places than group 1, and the fourth group appeared the intergrade of group 1 and 3. Therefore, it could be assumed what ALS gene of various types can be one plant. The peptide of the 13 amino acid Domain A region for ALS genes from R biotype of M. vaginalis differed from that of the S biotype by one base substitution at proline codon of Domain A. It could also be confirmed that point mutation of serine for proline at amino acid 197.

Characteristics and Partial Purification of a Bacteriocin Produced by Pediococcus damnosus JNU 534 (Pediococcus damnosus JNU 534가 생산하는 박테리오신의 특성 및 정제)

  • Lee, Jae-Won;Han, Su-Min;Yun, Bo-Hyun;Oh, Se-Jong
    • Food Science of Animal Resources
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    • v.31 no.6
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    • pp.952-959
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    • 2011
  • A new bacteriocin-producing lactic acid bacteria (LAB) which has been isolated from kimchi was identified as Pediococcus damnosus by use of API kit and 16S rDNA sequencing, and designated as P. damnosus JNU 534. The bacteriocin produced by P. damnosus JNU 534 markedly inhibited the growth of some of LAB and Listeria monocytogenes, whereas other pathogens including Gram negative bacteria were not susceptible. The production of bacteriocin started at the beginning of exponential phase and reached maximum activity at the early stationary phase. The bacteriocin was stable on the wide pH range of 2-9 and heat treatment up to $100^{\circ}C$ for 15 min. The antimicrobial compound was inactivated by treatments of proteolytic enzymes indicating its proteinaceous in nature. The bacteriocin was purified by 30% ammonium sulfate precipitation followed by hydrophobic interaction column and $C_{18}$ column chromatography. The estimated molecular weight of the bacteriocin using tricine SDS-PAGE was approximately 3.4 kDa and the identified N-terminal amino acid sequence was $NH_2$-ILLEELNV.

Isolation and Characterization of Phosphate Solubilizing Bacteria Pantoea Species as a Plant Growth Promoting Rhizobacteria (식물 생장 촉진 활성을 가진 인산분해 미생물 Pantoea 종의 분리 및 특성 규명)

  • Yun, Chang Yeon;Cheong, Yong Hwa
    • Journal of Life Science
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    • v.26 no.10
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    • pp.1163-1168
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    • 2016
  • Plant growth-promoting rhizobacteria (PGPR) have gained worldwide importance and acceptance due to their agricultural benefits. These microorganisms are potential tools for sustainable agriculture, with effects on plant growth, biofertilization, induced systemic resistance, and biocontrol of plant pathogens. In this study, four different Pantoea species were isolated from field soil, and their plant growth-promoting characteristics were studied. Based on 16S rDNA gene sequencing analyses, the se were grouped into Pantoea ananatis, Pantoea citrea, Pantoea dispersa, Pantoea vagans and named as Pa1, Pc1, Pd1, Pv1, respectively. All of these strains have their ability for solubilization of insoluble phosphate depending on pH decrease at the range around pH 5 at 1days after inoculation and production of plant hormone indole acetic acid (IAA) with 85.3±16.3 μg/ml of Pa1, 183.9±16.8 μg/ml of Pc1, 28.8±17.3 μg/ml of Pd1 and 114.1±16.5 μg/ml of Pv1, respectively. Pa1, Pc1 and Pd1 also have high activity for production of gibberellin (GA3) hormone with 331.1±19.2 μg/ml of Pa1, 288.5±16.8 μg/ml of Pc1, 309.2±18.2 μg/ml of Pd1, but Pv1 does not. Furthermore, all these species have significantly promoted the growth of the lettuce seedling plants at the range around 32~37% for fresh weight and 10~15% for shoot length enhancement, so that these microbe could be used as a potential bio-fertilizer agents.

Development of an Enrichment Culture Growing at Low Temperature used for Ensiling Rice Straw

  • Yang, Hong Yan;Wang, Xiao Fen;Gao, Li Juan;Haruta, Shin;Ishii, Masaharu;Igarashi, Yasuo;Cui, Zong Jun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.711-717
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    • 2008
  • To speed up the conversion of rice straw into feeds in a low-temperature region, a start culture used for ensiling rice straw at low temperature was selected by continuous enrichment cultivation. During the selection, the microbial source for enrichment was rice straw and soil from two places in Northeast China. Lab-scale rice straw fermentation at $10^{\circ}C$ verified, compared with the commercial inoculant, that the selected start culture lowered the pH of the fermented rice straw more rapidly and produced more lactic acid. The results from denatured gradient gel eletrophoresis showed that the selected start culture could colonize into the rice straw fermentation system. To analyze the composition of the culture, a 16S rRNA gene clone library was constructed. Sequencing results showed that the culture mainly consisted of two bacterial species. One (A) belonged to Lactobacillus and another (B) belonged to Leuconostoc. To make clear the roles of composition microbes in the fermented system, quantitative PCR was used. For species A, the DNA mass increased continuously until sixteen days of the fermentation, which occupied 65%. For species B, the DNA mass amounted to 5.5% at six days of the fermentation, which was the maximum relative value during the fermentation. To the authors' best knowledge, this is the first report on ensiling rice straw with a selected starter at low temperature and investigation of the fermented characteristics.