• Title/Summary/Keyword: environmental DNA

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A kinetic study of 4-chlorophenol biodegradation by the novel isolated Bacillus subtilis in batch shake flask

  • Sandhibigraha, Sudhansu;Chakraborty, Sagnik;Bandyopadhyay, Tarunkanti;Bhunia, Biswanath
    • Environmental Engineering Research
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    • v.25 no.1
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    • pp.62-70
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    • 2020
  • Here in this work, a 4-chlorophenol (4-CP)-degrading bacterial strain Bacillus subtilis (B. subtilis) MF447840.1 was isolated from the drain outside the Hyundai car service center, Agartala, Tripura, India. 16S rDNA technique used carried out for genomic recognition of the bacterial species. Isolated bacterial strain was phylogenetically related with B. subtilis. This strain was capable of breaking down both phenol and 4-CP at the concentration of 1,000 mg/L. Also, the isolated strain can able to metabolize five diverse aromatic molecules such as 2-chlorophenol, 2,4-dichlorophenol, 2,4,6-trichlorophenol, 4-nitrophenol, and pentachlorophenol for their growth. An extensive investigation was performed to portray the kinetics of cell growth along with 4-CP degradation in the batch study utilizing 4-CP as substrate. Various unstructured models were applied to evaluate the intrinsic kinetic factors. Levenspiel's model demonstrates a comparatively enhanced R2 value (0.997) amongst every analyzed model. The data of specific growth rate (μ), saturation constant (KS), and YX/S were 0.11 h-1, 39.88 mg/L, along with 0.53 g/g, correspondingly. The isolated strain degrades 1,000 mg/L of 4-CP within 40 h. Therefore, B. subtilis MF447840.1 was considered a potential candidate for 4-CP degradation.

A Novel MAP Kinase Gene in Cotton (Gossypium hirsutum L.), GhMAPK, is Involved in Response to Diverse Environmental Stresses

  • Wang, Meimei;Zhang, Ying;Wang, Jian;Wu, Xiaoliang;Guo, Xingqi
    • BMB Reports
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    • v.40 no.3
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    • pp.325-332
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    • 2007
  • The mitogen-activated protein kinase (MAPK) cascade is one of the major and evolutionally conserved signaling pathways and plays pivotal role in the regulation of stress and developmental signals in plants. Here, a novel gene, termed Gossypium hirsutum MAPK (GhMAPK), was isolated from cotton. The full-length cDNA of GhMAPK encodes for a 372 amino acid protein that contains all 11 of the MAPK conserved subdomains and the phosphorylationactivation motif, TEY. Amino acid sequence alignment revealed that GhMAPK shared high identity with group-C MAPK in plants and showed 83~89% similarities with MAPKs from Arabidopsis, apricot, pea, petunia, and tobacco. Southern blot analysis indicated that the GhMAPK belonged to a multygene family in cotton. Two introns were found within the region of genomic sequence. Northern blot analysis revealed that the transcripts of GhMAPK accumulated markedly when the cotton seedlings were subjected to various abiotic stimuli such as wounding, cold (4$^{\circ}C$), or salinity stress; Furthermore, GhMAPK was upregulated by the exogenous signaling molecules, such as salicylic acid (SA) and hydrogen peroxide ($H_2O_2C$), as well as pathogen attacks. These results indicate that the GhMAPK, which has a high degree of identity with group-C plant MAPKs, may also play an important role in response to environmental stresses.

Binding Pattern Elucidation of NNK and NNAL Cigarette Smoke Carcinogens with NER Pathway Enzymes: an Onco-Informatics Study

  • Jamal, Qazi Mohammad Sajid;Dhasmana, Anupam;Lohani, Mohtashim;Firdaus, Sumbul;Ansari, Md Yousuf;Sahoo, Ganesh Chandra;Haque, Shafiul
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.13
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    • pp.5311-5317
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    • 2015
  • Cigarette smoke derivatives like NNK (4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone) and NNAL (4-(methylnitrosamino)-1-(3-pyridyl)-1-butan-1-ol) are well-known carcinogens. We analyzed the interaction of enzymes involved in the NER (nucleotide excision repair) pathway with ligands (NNK and NNAL). Binding was characterized for the enzymes sharing equivalent or better interaction as compared to +Ve control. The highest obtained docking energy between NNK and enzymes RAD23A, CCNH, CDK7, and CETN2 were -7.13 kcal/mol, -7.27 kcal/mol, -8.05 kcal/mol and -7.58 kcal/mol respectively. Similarly the highest obtained docking energy between NNAL and enzymes RAD23A, CCNH, CDK7, and CETN2 were -7.46 kcal/mol, -7.94 kcal/mol, -7.83 kcal/mol and -7.67 kcal/mol respectively. In order to find out the effect of NNK and NNAL on enzymes involved in the NER pathway applying protein-protein interaction and protein-complex (i.e. enzymes docked with NNK/NNAL) interaction analysis. It was found that carcinogens are well capable to reduce the normal functioning of genes like RAD23A (HR23A), CCNH, CDK7 and CETN2. In silico analysis indicated loss of functions of these genes and their corresponding enzymes, which possibly might be a cause for alteration of DNA repair pathways leading to damage buildup and finally contributing to cancer formation.

Application of Multiparametric Flow Cytometry (FCM) to Enumerate the Diagnosis of Pseudomonas aeruginosa and Escherichia coli

  • Hwang, Myoung-Goo;Oh, Jung-Woo;Katayama, Hiroyuki;Ohgaki, Shinichiro;Cho, Jin-Kyu
    • Environmental Engineering Research
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    • v.17 no.1
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    • pp.35-39
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    • 2012
  • In this study, multiparametric flow cytometry (FCM) was installed to enumerate the diagnosis of Pseudomonas aeruginosa ATCC 10145 and Escherichia coli K12 (IFO 3301). The nucleic acids (DNA/RNA) were double stained by a LIVE/DEAD bacLight viability kit, involving green SYTO 9 and red propidium iodide (PI), based on the permeability of two chemicals according to the integrity of plasma membrane. As the results showed, the gate for dead bacteria was defined as the range of $0.2{\times}10^0$ to $6.0{\times}10^1$ photo multiplier tube (PMT) 2 fluorescence (X-axis) and $2.0{\times}10^0$ to $2.0{\times}10^2$ PMT 4 fluorescence (Y-axis), and the gate for live bacteria was defined as the range of $6.0{\times}10^0$ to $6.0{\times}10^2$ PMT 2 fluorescence (X-axis) and $2.0{\times}10^0$ to $4.0{\times}10^2$ PMT 4 fluorescence (Y-axis). In the comparison of the number of the tested bacteria detected by FCM (viability assessment) and plate culture (cultivability assessment), the number of bacteria detected by FCM well represented the number of bacteria that was detected by the colony forming unit (CFU) counting method when bacteria were exposed to isopropyl alcohol and silver/copper cations. Consequently, it is concluded that the application of FCM to monitor the functional effect of disinfectants on the physiological status of target bacteria can offer more rapid and reliable data than the plate culture colony counting method.

Occurrence of Pythium Blight Caused by Pythium aphanidermatum on Chewing Fescue (Pythium aphanidermatum에 의한 Chewing Fescue에 잎마름병 발생)

  • Chang, Taehyun;Lee, Yong Se
    • Weed & Turfgrass Science
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    • v.2 no.3
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    • pp.306-311
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    • 2013
  • Pythium blight occurred by Pythium aphanidermatum on chewing fescue cv. "Jamestowm II" from early June, 2010 and 2011 at the test field in Daegu University in Gyeongbuk Province, Korea. Disease symptoms on the turfgrass were leaf blights dying from the leaf tip and root rot, which appeared patches of brown to dark brown color or gray brown color in the field. The pathogens (40-1 isolate) of Pythium blight was isolated from the diseased leaf and crown tissue and cultured on potato-dextrose agar (PDA) for identification. Lobulate sporangia were inflated, complex structures, and filamentous sporangia were usually indistinguishable from vegetative hyphae. Sequences of ribosomal RNA gene of the fungus were homologous with similarity of 100% to those of P. aphanidermatum isolates in GenBank database. Pathogenicity was also confirmed on the chewing fescue, creeping betgrass and Kentucky bluegrass by Koch's postulates. This is the first report of Pythium blight on chewing fescue caused by P. aphanidermatum in Korea.

Efficiency to Discovery Transgenic Loci in GM Rice Using Next Generation Sequencing Whole Genome Re-sequencing

  • Park, Doori;Kim, Dongin;Jang, Green;Lim, Jongsung;Shin, Yun-Ji;Kim, Jina;Seo, Mi-Seong;Park, Su-Hyun;Kim, Ju-Kon;Kwon, Tae-Ho;Choi, Ik-Young
    • Genomics & Informatics
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    • v.13 no.3
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    • pp.81-85
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    • 2015
  • Molecular characterization technology in genetically modified organisms, in addition to how transgenic biotechnologies are developed now require full transparency to assess the risk to living modified and non-modified organisms. Next generation sequencing (NGS) methodology is suggested as an effective means in genome characterization and detection of transgenic insertion locations. In the present study, we applied NGS to insert transgenic loci, specifically the epidermal growth factor (EGF) in genetically modified rice cells. A total of 29.3 Gb (${\sim}72{\times}coverage$) was sequenced with a $2{\times}150bp$ paired end method by Illumina HiSeq2500, which was consecutively mapped to the rice genome and T-vector sequence. The compatible pairs of reads were successfully mapped to 10 loci on the rice chromosome and vector sequences were validated to the insertion location by polymerase chain reaction (PCR) amplification. The EGF transgenic site was confirmed only on chromosome 4 by PCR. Results of this study demonstrated the success of NGS data to characterize the rice genome. Bioinformatics analyses must be developed in association with NGS data to identify highly accurate transgenic sites.

Development of succinate producing Cellulomonas flavigena mutants with deleted succinate dehydrogenase gene

  • Lee, Heon-Hak;Jeon, Min-Ki;Yoon, Min-Ho
    • Korean Journal of Agricultural Science
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    • v.44 no.1
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    • pp.30-39
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    • 2017
  • This study was performed to produce succinic acid from biomass by developing mutants of Cellulomonas flavigena in which the succinate dehydrogenase gene (sdh) is deleted. For development of succinate producing mutants, the upstream and downstream regions of sdh gene from C. flavigena and antibiotic resistance gene (neo, bla) were inserted into pKC1139, and the recombinant plasmids were transformed into Escherichia coli ET12567/pUZ8002 which is a donor strain for conjugation. C. flavigena was conjugated with the transformed E. coli ET12567/pUZ8002 to induce the deletion of sdh in chromosome of this bacteria by double-crossover recombination. Two mutants (C. flavigena H-1 and H-2), in which sdh gene was deleted in the chromosome, were constructed and confirmed by PCR. To estimate the production of succinic acid by the two mutants when the culture broth was fermented with biomass such as CMC, xylan, locust gum, and rapeseed straw; the culture broth was analyzed by HPLC analysis. The succinic acid in the culture broth was not detected as a fermentation products of all biomass. One of the reasons for this may be the conversion of succinic acid to fumaric acid by sdh genes (Cfla_1014 - Cfla_1017 or Cfla_1916 - Cfla_1918) which remained in the chromosomal DNA of C. flavigena H-1 and H-2. The other reason could be the conversion of succinyl-CoA to other metabolites by enzymes related to the bypass pathway of TCA cycle.

Occurrence of Sweet-potato Whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae) and Its Response to Insecticide in Gyeonggi Area (경기지역에서 담배가루이의 발생 및 약제반응)

  • Lee, Young-Su;Kim, Jin-Young;Hong, Soon-Sung;Park, Jungan;Park, Hong-Hyun
    • Korean journal of applied entomology
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    • v.51 no.4
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    • pp.377-382
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    • 2012
  • This study investigated the occurrence of sweet potato whitefly, Bemisia tabaci affecting cucumber, eggplant and red pepper, as well as sweet potato species, and its response to insecticides in Gyeonggi province from 2010 to 2011. Sweet potato whitefly is widespread throughout the southern part of Gyeonggi province. Most regional populations of B. tabaci belong to biotype Q having been reported in the south Korea since 2005, but in Goyang mixed populations of two biotypes (B and Q) were found. Survey results of Tomato Yellow Leaf Curl Virus (TYLCV) disease that was vectored by B. tabaci indicated that this virus disease was not spread throughout the Gyeonggi province. Biotype Q of B. tabaci was found to be resistant to neonicotinoid insecticides, whereas biotype B was highly susceptible to them.

New ecological health assessment approaches of an urban stream using molecular and physiological level biomarkers and bioindicators

  • Kim, Ja-Hyun;Yeom, Dong-Hyuk;Kim, Joon-Ha;An, Kwang-Guk
    • Animal cells and systems
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    • v.16 no.4
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    • pp.329-336
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    • 2012
  • This study evaluated ecological health, using various biomarkers and bioindicators, of pale chub (Zacco platypus) as a sentinel species, in Daejeon Stream, South Korea, during AprilMay 2011. The biomarkers and bioindicators were compared among three sites of control: Reference ($C_z$), transition ($T_z$), and the urban zones ($U_z$); and the 7-Ethoxyresorufin-O-deethylase (EROD) activity, DNA damage, acetylcholinesterase (AChE) activity, and vitellogenin (VTG) concentrations were more significantly increased in the $U_z$ than in the $C_z$. Also, physiological markers such as condition factor, liver somatic index, visceral somatic index, and gonad somatic index were significantly increased in the $U_z$ than in the $C_z$. For the health assessments, three categorized parameters of blood chemistry, molecular biomarkers, and physiological bioindicators were standardized and calculated as a star-plot, representing values of Integrated Health Response (IHR). Values of IHR had more significant (P<0.05) increases in the $U_z$ than any other zones, indicating an impairment of ecological health by organic matter, nutrients (N, P), and toxic chemicals. This study is based on low levels of biological organization approach of molecular and physiological biomarkers and bioindicators, so further study of high-levels of biological organization approach such as community and population is required for overall range of health assessments. The approach of IHR values, however, may be useful in providing early warning of future impacts on ecological health.

Zonate Leaf Spot of Acer negundo Caused by Cristulariella moricola in Korea (Cristulariella moricola에 의한 네군도단풍 둥근무늬낙엽병)

  • Cho, Sung-Eun;Park, Ji-Hyun;Lee, Sang-Hyun;Lee, Hyang-Burm;Shin, Hyeon-Dong
    • Research in Plant Disease
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    • v.18 no.1
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    • pp.45-48
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    • 2012
  • In early August of 2011, following a prolonged period of cool and moist weather, several trees of boxelder maple (Acer negundo) planted in Mt. Soyo located in Dongducheon, Korea, were found to be heavily damaged by premature defoliation with zonate leaf spot symptoms. Numerous number of cone-shaped, white sporophores (conidia) were observed on lesions of the abaxial leaf surface. The morphological characteristics of conidia are of typical Cristulariella moricola, which was supported by analyzing sequences of internal transcribed spacer region of ribosomal DNA. Pathogenicity of the fungus was proved by artificial inoculation in the condition of relative humidity 100% and $18{\pm}2^{\circ}C$. This is the first report of the occurrence of zonate leaf spot caused by infection of C. moricola on A. negundo in Korea.