• Title/Summary/Keyword: tubulin

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Diaporthe phaseolorum var. caulivora, a Causal Agent for Both Stem Canker and Seed Decay on Soybean

  • Sun, Su-li;Van, Kyu-Jung;Kim, Moon-Young;Min, Kyung-Hun;Lee, Yin-Won;Lee, Suk-Ha
    • The Plant Pathology Journal
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    • v.28 no.1
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    • pp.55-59
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    • 2012
  • Northern stem canker caused by $Diaporthe$ $phaseolorum$ var. $caulivora$ ($Dpc$) has become a serious disease in soybean. The objectives of this study were to survey the existence of $Dpc$ on soybean in Korea, and to examine the potential pathogenicity of $Dpc$ in seed decay. One such isolate, SSLP-4, isolated from a field-grown plant of the Korean soybean cultivar Danbaekkong, was identified as $Dpc$, based on its morphological and molecular characteristics by sequences of internal transcribed spacer (ITS), translation elongation factor (TEF) 1-${\alpha}$ and ${\beta}$-tubulin regions, as well as pathogenic analyses. Moreover, morphological and molecular analyses revealed that isolate SSLP-4 was nearly identical to $Dpc$ strains from the United States. Pathogenicity tests on hypocotyls of soybean seedlings and detached leaves resulted in typical symptoms of soybean northern stem canker and inoculation on plants at R5-R7 stage caused seed decay. All results suggest that the $Dpc$ strain SSLP-4 can cause both stem canker and seed decay on soybean. Thus, the SSLP-4 isolate has the potential to contribute greatly to understanding of host plant resistance mechanisms, both at vegetative and reproductive growth stages in soybean.

Selection of Reliable Reference Genes for Real-time qRT-PCR Analysis of Zi Geese (Anser anser domestica) Gene Expression

  • Ji, Hong;Wang, Jianfa;Liu, Juxiong;Guo, Jingru;Wang, Zhongwei;Zhang, Xu;Guo, Li;Yang, Huanmin
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.3
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    • pp.423-432
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    • 2013
  • Zi geese (Anser anser domestica) belong to the white geese and are excellent layers with a superior feed-to-egg conversion ratio. Quantitative gene expression analysis, such as Real-time qRT-PCR, will provide a good understanding of ovarian function during egg-laying and consequently improve egg production. However, we still don't know what reference genes in geese, which show stable expression, should be used for such quantitative analysis. In order to reveal such reference genes, the stability of seven genes were tested in five tissues of Zi geese. Methodology/Principal Findings: The relative transcription levels of genes encoding hypoxanthine guanine phosphoribosyl transferase 1 (HPRT1), ${\beta}$-actin (ACTB), ${\beta}$-tubulin (TUB), glyceraldehyde-3-phosphate-dehydrogenase (GADPH), succinate dehydrogenase flavoprotein (SDH), 28S rRNA (28S) and 18S rRNA (18S) have been quantified in heart, liver, kidney, muscle and ovary in Zi geese respectively at different developmental stages (1 d, 2, 4, 6 and 8 months). The expression stability of these genes was analyzed using geNorm, NormFinder and BestKeeper software. Conclusions: The expression of 28S in heart, GAPDH in liver and ovary, ACTB in kidney and HPRT1 in muscle are the most stable genes as identified by the three different analysis methods. Thus, these genes are recommended for use as candidate reference genes to compare mRNA transcription in various developmental stages of geese.

Plasma Phosphoproteome and Differential Plasma Phosphoproteins with Opisthorchis Viverrini-Related Cholangiocarcinoma

  • Kotawong, Kanawut;Thitapakorn, Veerachai;Roytrakul, Sittiruk;Phaonakrop, Narumon;Viyanant, Vithoon;Na-Bangchang, Kesara
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.3
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    • pp.1011-1018
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    • 2015
  • This study was conducted to investigate the plasma phosphoproteome and differential plasma phosphoproteins in cases of of Opisthorchis viverrini (OV)-related cholangiocarcinoma (CCA). Plasma phosphoproteomes from CCA patients (10) and non-CCA subjects (5 each for healthy subjects and OV infection) were investigated using gel-based and solution-based LC-MS/MS. Phosphoproteins in plasma samples were enriched and analyzed by LC-MS/MS. STRAP, PANTHER, iPath, and MeV programs were applied for the identification of their functions, signaling and metabolic pathways; and for the discrimination of potential biomarkers in CCA patients and non-CCA subjects, respectively. A total of 90 and 60 plasma phosphoproteins were identified by gel-based and solution-based LC-MS/MS, respectively. Most of the phosphoproteins were cytosol proteins which play roles in several cellular processes, signaling pathways, and metabolic pathways (STRAP, PANTHER, and iPath analysis). The absence of serine/arginine repetitive matrix protein 3 (A6NNA2), tubulin tyrosine ligase-like family, member 6, and biorientation of chromosomes in cell division protein 1-like (Q8NFC6) in plasma phosphoprotein were identified as potential biomarkers for the differentiation of healthy subjects from patients with CCA and OV infection. To differentiate CCA from OV infection, the absence of both serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit beta isoform and coiled-coil domain-containing protein 126 precursor (Q96EE4) were then applied. A combination of 5 phosphoproteins may new alternative choices for CCA diagnosis.

A New Record of Epicoccum draconis Isolated from the Soil in Korea

  • Ayim, Benjamin Yaw;Das, Kallol;Cho, Young-Je;Lee, Seung-Yeol;Jung, Hee-Young
    • The Korean Journal of Mycology
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    • v.48 no.1
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    • pp.39-45
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    • 2020
  • A fungal isolate US-18-11 was isolated from the soil in Uiseong, Korea. The mycelium growth measured after 7 days of incubation at 22℃ on malt extract agar (MEA) and oatmeal agar (OA) media was 42-43 mm and 41-44 mm in diameter, respectively. The fungal colony formed white to dull green aerial mycelia that were floccose with regular margins and olivaceous black with leaden gray patches on the reverse side. The conidia were hyaline to brown in color, ellipsoidal to ovoid, guttulate, abundant, globose, solitary, or confluent measuring 3.2-7.2×1.1-2.3 ㎛. A BLAST search of the large subunit (LSU), internal transcribed spacer (ITS) region, second largest subunit of DNA-directed RNA polymerase II (RPB2) and β-tubulin (TUB2) gene sequences revealed that the isolate US-18-11 has similarities of 99, 100, 97, and 99% with those of Epicoccum draconis CBS 186.83, respectively. A neighbor-joining phylogenetic tree constructed based on the concatenated dataset of above-mentioned sequences showed that isolate US-18-11 clustered with Epicoccum draconis CBS 186.83 in the same clade. Based on the results of morphological, cultural, and phylogenetic analysis, the isolate US-18-11 was identical to the previously described E. draconis CBS 186.83. To our knowledge, this is the first report of E. draconis in Korea.

Differential Gene Expression of Soybean[Glycine max(L.) Merr.] in Response to Xanthomonas axonopodis pv. glycines by Using Oligonulceotide Macroarray

  • Van, Kyujung;Lestari, Puji;Park, Yong-Jin;Gwag, Jae-Gyun;Kim, Moon-Young;Kim, Dong-Hyun;Heu, Sung-Gi;Lee, Suk-Ha
    • Journal of Crop Science and Biotechnology
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    • v.10 no.3
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    • pp.147-158
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    • 2007
  • Xanthomonas axonopodis pv. glycines(Xag) is a pathogen that causes bacterial leaf pustule(BLP) disease in soybeans grown in Korea and the southern United States. Typical and early symptoms of the disease are small, yellow to brown lesions with raised pustules that develop into large necrotic lesions leading to a substantial loss in yield due to premature defoliation. After Xag infects PI 96188, only pustules without chlorotic haloes were observed, indicating the different response to Xag. To identify differentially expressed genes prior to and 24 hr after Xag inoculation to PI 96188 and BLP-resistant SS2-2, an oligonucleotide macroarray was constructed with 100 genes related to disease resistance and metabolism from soybean and Arabidopsis. After cDNAs from each genotype were applied on the oligonucleotide macroarrays with three replicates and dye swapping, 36 and 81 genes were expressed as significantly different between 0 hr and 24 hr in PI 96188 and SS2-2, respectively. Six UniGenes, such as the leucine-rich repeat protein precursor or 14-3-3-like protein, were selected because they down-regulated in PI 96188 and up-regulated in SS2-2 after Xag infection, simultaneously. Using tubulin and cDNA of Jangyeobkong(BLP-susceptible) as controls, the oligonucleotide macroarray data concurred with quantitative real-time RT-PCR(QRT RT-PCR) results in most cases, supporting the accuracy of the oligonucleotide macroarray experiments. Also, QRT RT-PCR data suggested six candidate genes that might be involved in a necrotic response to Xag in PI 96188.

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Chromosome and Spindle Configuration of Mouse Oocytes after Vitrification at the Mature Stage (마우스 성숙난자의 유리화 동결법에 따른 동결 융해 후의 염색체와 방추사의 분석)

  • ;;;;Gary B. Anderson
    • Korean Journal of Animal Reproduction
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    • v.25 no.3
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    • pp.287-292
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    • 2001
  • Selection of oocyte cryopreseivation method is a prerequisite factor for developing an effective bank system. To develop an effective vitrification method, we examined whether damages in spindle and chromosome morphology induced by vitrification. Intact cumulus-enclosed oocytes were vitrified with DPBS with 5.5 M ethylene glycol and 1.0 M sucrose, and loaded onto eletron microscopic copper grid for storing in liquid nitrogen. Intact vitrified and thawed oocytes were immunostaining for tubulin and karyotying for chromosome. Vitrfied and thawed oocytes had a higher rate of chromosome (32.8% vs. 19.6%) and spindle (32.3% vs. 20.2%) abnormalities compared with fresh oocytes. Mouse oocytes after vitrification at the mature stage showed increased incidence of chromosomal and spindle abnormalities.

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Unrecorded fungi isolated from Lentinula edodes cultivation houses in Korea (표고 재배사에서 분리한 국내 미기록 진균 보고)

  • Ahn, Geum Ran;Noh, Hyeungjin;Kim, Jun Young;Ko, Han Gyu;Kim, Seong Hwan
    • Journal of Mushroom
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    • v.18 no.1
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    • pp.72-78
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    • 2020
  • During the monitoring of fungal pests in 2016 and 2017, Acrodontium crateriforme, Naganishia friedmannii, Pestalotiopsis trachicarpicola, Penicillium wollemiicola, and Trichoderma thailandicum were isolated from indoor air, mushroom flies (Phytosciara flavipes), and media materials in the cultivation houses of oak wood mushroom (Lentinula edodes) located in Seocheon, Jangheung, Buyeo, and Yeoju, Korea. These fungal species were identified based on their morphological characteristics after their growth on PDA and subsequent molecular analyses of the 26S rDNA, 28S rDNA, β-tubulin gene, and translation elongation factor 1-α gene using PCR amplification and nucleotide sequencing were performed. The results showed that these fungi were previously undocumented in Korea. This study reports descriptions of their taxonomical and known properties.

The Effect of Cytochalasin B on Cytoskeletal Stability of Mouse Oocyte Frozen by Vitrification (생쥐 난자의 초자화동결 시 Cytochalasin B가 Cytoskeleton에 미치는 영향)

  • Paik, Wong-Young;Choi, Won-Jun;Kim, Se-Na;Lee, Jong-Hak
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.4
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    • pp.229-236
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    • 2002
  • Objective : The purpose of this study was to evaluate the effect of Cytochalasin B (CCB) on the cytoskeletal stability of mouse oocyte frozen by vitrification. Methods : Mouse oocytes retrieved from cycle stimulated by PMSG and hCG were treated by CCB and then vitrified in EFS-30. These oocytes were placed onto an EM grid and submerged immediately in liquid nitrogen. Thawing of the oocytes was carried out at room temperature for 5 seconds, then the EM grid was placed into 0.75 M, 0.5 M and 0.25 M sucrose at $37^{circ}C$ for 3 minutes, each. These oocytes were fixed in 4% formaldehyde for an hour and then washed in PPB for 15 minutes 3 times, then incubated in PPB containing anti-tubulin monoclonal antibody at $4^{circ}C$ overnight. And then, the oocytes were incubated with FITC-conjugated anti-mouse IgG and propidium iodide (PI) for 45 minutes. Pattern of microtubules and microfilaments of oocytes were evaluated with a confocal microscope. Results: The rate of oocytes containing normal microtubules and microfilaments was significantly decreased after vitrification. The rate of oocyte containing normal microtubules in CCB treated group was higher than those in non-treated group (53.7% vs. 48.9%), but the difference was not significant. The rate of oocyte containing normal microfilaments in CCB treated group was significantly higher than those in non-treated group (64.5% vs. 38.3%, p<0.05). Conclusion: Microfilaments stability could be improved by CCB treatment prior to vitrification. It is suggested that CCB treatment prior to vitrification improve stability of cytoskeleton and then increase success rate in IVF-ET program using vitrification and thawing oocyte.

Kretzschmaria quercicola sp. nov., an Undescribed Fungus from Living Oak in Mt. Daeryong, Korea

  • Yun, Ji Ho;Jo, Jong Won;Lee, Jin Heung;Han, Sang Kuk;Kim, Dae Ho;Lee, Jong Kyu
    • Mycobiology
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    • v.44 no.2
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    • pp.112-116
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    • 2016
  • We encountered an unfamiliar ascomycete fruiting body, fitting characteristics of the genus Kretzschmaria, which features in a stipitate ascigerous stroma with carbonaceous interior and disintegrating perithecia. In this study, we report and characterize a new species of the decaying fungus. Compared to other species, one of the notable features of this specimen (TPML150908-046) is its stromatal size (up to 15 cm). Although TPML150908-046 is morphologically similar to K. milleri and K. sandvicensis, it differs sharply from both species in apical ring size (TPML150908-046, $6.5{\sim}10.5{\mu}m$; K. milleri, $11{\sim}16{\mu}m$) and ascospore width (TPML150908-046, $10.5{\sim}17{\mu}m$; K. sandvicensis, $8.5~11.5{\mu}m$). Phylogenetic trees based on ${\beta}$-tubulin, ITS, and RPB2 sequences showed that our collection clustered with K. sandvicensis, with the respective similarities for these sequences being 95.6%, 91.3%, and 97.7%, signifying it as another species. With these results, we report it as a new species, which we call Kretzschmaria quercicola sp. nov.

Incidence, Molecular Characteristics and Pathogenicity of Gibberella fujikuroi Species Complex Associated with Rice Seeds from Asian Countries

  • Jeon, Young-Ah;Yu, Seung-Hun;Lee, Young Yi;Park, Hong-Jae;Lee, Sokyoung;Sung, Jung Sook;Kim, Yeon-Gyu;Lee, Ho-Sun
    • Mycobiology
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    • v.41 no.4
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    • pp.225-233
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    • 2013
  • Gibberella fujikuroi species complex (GFSC) was isolated from rice (Oryza sativa L.) seed samples from ten Asian countries and investigated for incidence of GFSC, molecular characteristics, and pathogenicity. Regardless of geographic origin, GFSC was detected with incidences ranging from 3% to 80%. Four species, Fusarium fujikuroi, F. concentricum, F. proliferatum, and F. verticillioides, were found to show an association with rice seeds, with F. fujikuroi being the predominant species. In phylogenetic analyses of DNA sequences, no relationship was found between species, isolates, and geographic sources of samples. Unidentified fragments of the ${\beta}$-tubulin gene were observed in ten isolates of F. fujikuroi and F. verticillioides. With the exception of three isolates of F. fujikuroi, F. fujikuroi, F. proliferatum, and F. verticillioides were found to have FUM1 (the fumonisin biosynthetic gene); however, FUM1 was not found in isolates of F. concentricum. Results of pathogenicity testing showed that all isolates caused reduced germination of rice seed. In addition, F. fujikuroi and F. concentricum caused typical symptoms of bakanae, leaf elongation and chlorosis, whereas F. proliferatum and F. verticillioides only caused stunting of seedlings. These findings provide insight into the characteristics of GFSC associated with rice seeds and might be helpful in development of strategies for management of bakanae.