• Title/Summary/Keyword: environmental DNA

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A Study of Potential of Diet Analysis in the Korean Water Deer(Hydropotes inermis argyropus) using Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis(PCR-DGGE) (고라니의 식이물 분석에 있어 Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis(PCR-DGGE)의 이용 가능성 연구)

  • Park, Ji-Eun;Kim, Baek-Jun;Lee, Sang-Don
    • Korean Journal of Environment and Ecology
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    • v.24 no.3
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    • pp.318-324
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    • 2010
  • The aim of this study is to examine feeding habits of the Korean water deer(Hydropotes inermis argyropus) from its rumen contents using a PCR-DGGE method. For this study, rumen contents were collected from water deer causalities by natural death or road-kill in two different sites(Cheorwon, Gangwon province and the Eastern part of Jeonnam province). DNA was extracted from rumen contents of a total of 44 individuals. Two primers, rbcLZ1aF(GC) and rbcL19bR, were used for PCR amplifications of ribulose-1,5-bisphosphate carboxylase large subunit (rbcL) gene. Among 44 samples, twenty-nine samples were successfully amplified by PCRs. The 29 PCR products of partial rbcL gene were applied for PCR-DGGE. Totally, six families of plants were detected from the diet analyses. Five families of plants were found in Cheorwon, Gangwon province, but only three families of plants were found in the Eastern part of Jeonnam province. The PCR-DGGE method will provide us with a potential tool to study feeding habits of ungulates including water deer, even though our results failed to identify the prey plants at the level of species.

Isolation and Identification of Antagonistic Bacteria for Biological Control of Large Patch Disease of Zoysiagrass Caused by Rhizoctonia solani AG2-2 (IV) (들잔디 갈색퍼짐병의 생물학적 방제를 위한 길항 세균의 분리와 동정)

  • Song, Chi-Hyun;Islam, Md. Rezuanul;Chang, Tae-Hyun;Lee, Yong-Se
    • Asian Journal of Turfgrass Science
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    • v.26 no.1
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    • pp.8-16
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    • 2012
  • The objective of this study was to identify bacterial antagonists of R. solani AG2-2 (IV) on zoysiagrass and to evaluate their antifungal activity in vitro and in vivo to select an antagonistic isolate. Antagonistic isolates that inhibit large patch disease caused by R. solani AG2-2 (IV) in zoysiagrass were selected from several soils, and their antagonistic activities were investigated in vitro and in vivo. Of 216 bacterial isolates, 67 inhibited several plant pathogenic fungi. The isolates that inhibited stem-segment colonization by R. solani AG2-2 (IV) in zoysiagrass were tested in a growth chamber. Eleven isolates were active as plant growth promoting isolates. Among them, five plant growth promoting isolates and their concentration dependent efficiency on zoysiagrass following inoculation with R. solani AG2-2 (IV) was evaluated. Isolate H33 was one of the potential antagonistic isolates, and it was further tested against various plant pathogens. H33 not only suppressed the disease caused by R. solani AG2-2 (IV) on zoysiagrass but also promoted leaf weight and leaf height of zoysiagrass under growth chamber and greenhouse conditions. The H33 isolate, which belongs to Streptomyces arenae, was identified through physiological, biochemical, and 16S rDNA studies. Further studies will investigate the cultural characterization of S. arenae H33 and isolation and identification of antifungal substance produced by S. arenae H33.

DNA Adduct Formation and Expression of Ras Gene in the Liver of Rats Treated with Aflatoxins at Various Levels (랫드의 간에서 다양한 농도의 아플라톡신 투여에 의한 DNA Adduct의 형성과 Ras의 발현양상)

  • Kim Tae Myoung;Hue Jin Joo;Li Lan;Kim Dae Joong;Nam Sang Yoon;Yun Young Won;Lee Beom Jun
    • Toxicological Research
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    • v.21 no.4
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    • pp.339-345
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    • 2005
  • Aflatoxins are produced by Aspergillus flavus, parasiticus that grows in improperly stored cereals. Aflatoxin $B_1\;(AFB_1)$ is a potent hepatocarcinogen in a variety of experimental animals including human beings. In spite of a high attention to the hepatocarcinogenecity of aflatoxins, the relative toxicity of other types $(AFB_2,\;AFG_1\;and\;AFG_2)$ of the toxins is not fully clarified. Sprague-Dawley male rats were orally administered with $AFB_1,\;AFB_2,\;AFG_1\;and\;AFG_2$ at the dose of 250, 1250, and $2500\;{\mu}g/kg$ body weight. Animals were then killed at 12, 24 or 48 hrs following aflatoxin adminstration. Subsequently the relative weight of liver was measured and histopathological examination on the liver was performed. Level of 8-OxodG and expression of ras gene in the liver was determined. The relative liver weights at high doses of $AFB_1\;and\;AFG_1$ was significantly low. The treatment of $AFB_1$ at the high dose of $2500\;{\mu}g/kg$ showed vacuolar degeneration and centrilobular hepatic necrosis with inflammatory cells. The pathological changes by $AFB_2\;AFG_1,\;and\;AFG_2$ were not clearly found. The formation of 8-OxodG by $AFB_1$ increased in a dose-dependent manner up to 24 hrs after a single treatment of $AFB_1$ thereafter decreased to the level of the control. The treatments of $AFB_2\;AFG_1,\;and\;AFG_2$ showed an inconsistent pattern in the formation of 8-OxodG in the liver of rats with increasing time. The expression of ras oncogene in the liver by $AFB_1$ at the dose of $1250\;{\mu}g/kg$ was increased twice compared to the control. The treatments of $AFB_2\;AFG_1,\;and\;AFG_2$ at all doses decreased the expression of ras in the liver. These results in the present study indicate that $AFB_1$ among aflatoxins with low comparable levels is the most toxic as determined by early biomarkers such as 8-OxodG formation and ras expression. However, the levels of 8-OxodG and ras as biomarkers were not useful to predict the relative hepatocarcinogenicity of aflatoxins to $AFB_1$ in the present model. Further studies are required to look for other biomarkers to predict carcinogenic potency of aflatoxins.

Evaluation of horizontal gene transfer from genetically modified zoysiagrass to the indigenous microorganisms in isolated GMO field (GMO 격리포장에서의 유전자변형 들잔디로부터 토착미생물로의 수평유전자전달 평가)

  • Bae, Tae-Wung;Lee, Hyo-Yeon;Ryu, Ki-Hyun;Lee, Tae-Hyeong;Lim, Pyung-Ok;Yoon, Pill-Yong;Park, Sin-Young;Riu, Key-Zung;Song, Pill-Soon;Lee, Yong-Eok
    • Journal of Plant Biotechnology
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    • v.34 no.1
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    • pp.75-80
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    • 2007
  • The release of genetically modified organisms ($GMO_{s}$) into the environment has the potential risks regarding the possibility of gene transfer from $GMO_{s}$ to natural organisms and this needs to be evaluated. This study was conducted to monitor the possible horizontal gene transfer from herbicide-resistant zoysiagrass (Zoysia japonica Steud.) to indigenous microorganisms. We have first examined the effect of field-released GM zoysiagrass on the microbial flora in the gut of locust (Locusts mlgratoria). The microbial flora was analyzed through determining the 165 rDHA sequences of microorganisms. The comparison of the microbial flora in the gut of locusts that were captured at the field of GM zoysiagrass and of wild-type revealed that there is no noticeable difference between these two groups. This result indicates that the GM zoysiagrass does not have negative impact on microbial flora in the gut of locust. We then investigated whether the horizontal gene transfer occurred from GM zoysiagrass to microbes in soil, rhizosphere and faecal pellets from locusts by utilizing molecular tools such as Southern hybridization and polymerase chain reaction (PCR). When the total DNAs isolated from microbes in GM zoysiagrass and in wild-type zoysiagrass fields were hybridized with probes for bar or hpt gene, no hybridization signal was detected from both field isolates, while the probes were hybridized with DNA from the positive control. Absence of these genes in the FNAs of soil microorganisms as well as microbes in the gut of locust was further confirmed by PCR. Taken together, our data showed that horizontal gene transfer did not occur in this system. These results further indicate that frequencies of transfer of engineered plant DNA to bacteria are likely to be negligible.

CGMMV Tolerance Test of CGMMV-CP Trangenic Watermelon Rootstock and Establishment of Transgenic Line (CGMMV-CP 형질전환 수박대목의 CGMMV 내성시험 및 계통확보)

  • Park, Sang-Mi;Kwon, Jung-Hee;Lim, Mi-Young;Shin, Yoon-Sup;Her, Nam-Han;Lee, Jang-Ha;Ryu, Ki-Hyun;Harn, Chee-Hark
    • Journal of Plant Biotechnology
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    • v.34 no.1
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    • pp.11-17
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    • 2007
  • Previously developed transgenic watermelon rootstocks (gongdae) inserted by CGMMV-CP were examined to test the virus tolerance levels. In the restricted plastic house, the $T_{3}$ watermelon rootstock showed tolerance to CGMMV until 70 days after inoculation on the leaves while the non-transformed watermelon rootstock became susceptible at 20 days after inoculation. In the field, tolerance efficiency of transgenic rootstocks maintained up to 40% at 71 days after contamination with CGMMV in the soil while all of the non-transformed rootstocks became susceptible at 37 days with the same condition. In the same field, transgenic rootstocks showed more tolerance to CGMMV than the non-transformed rootstocks as those were inoculated on the leaves, but it showed only 10 days delay before being susceptible. Therefore, transgenic rootstocks have a characteristic of delay effect against CGMMV susceptibility, rather than resistance character. From $T_{3}$ rootstocks homozygous for the CGMMV-CP horticulturally favorable individuals were selected for further breeding and a transgenic line was finally obtained at the $BC_{1}T_{5}$. A material transfer experiment was conducted to find out if the DNA, RNA or expressed protein in the transgenic rootstocks could move to the grafted scion (non-transformed watermelon, Super-Kumcheon). PCR, northern, and western blot analysis were performed and no evidence of transferring of those materials from rootstock to scion was ever found.

Establishment of detection methods for approved LMO in Korea (국내 승인 유전자변형 작물의 검출 기법 확립)

  • Seol, Min-A;Lee, Jung Ro;Choi, Wonkyun;Jo, Beom-Ho;Moon, Jeong Chan;Shin, Su Young;Eum, Soon-Jae;Kim, Il Ryong;Song, Hae-Ryong
    • Journal of Plant Biotechnology
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    • v.42 no.3
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    • pp.196-203
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    • 2015
  • AbstractLiving modified organisms (LMO) are one of the most widespread products of modern biotechnology after DNA discovery. Due to the decline of grain self-sufficiency rate and the increase of reliance on LMO imports in Korea, a series of concerns with regard to safety of living modified(LM) crops has been raised. The aim of this study is to establish the detection methods for unintentional release or growing of LMO plants in environmental conditions. To detect LM crop events, general concepts of specific primer design and PCR conditions were provided by the Joint Research Centre (JRC). The certified reference materials of seven LM events (4 soybean, 2 cotton and 1 corn) were obtained from the Institute for Reference Materials and Measurements (IRMM) and the American Oil Chemists' Society (AOCS). Genomic DNA from seven LM events were purified and PCR amplifications were carried out by using individual event-specific primer sets. LM-specific PCR products of all seven events were efficiently amplified by our methods. The results indicate that the established detection method for LMOs is suitable as a scientific tool to monitor whether the crops found in natural environments are LMOs.

Development of herbicide tolerant soybean using Agrobacterium tumefaciens (아그로박테리움을 이용만 제초제 저항성 콩 개발)

  • Lee, Ki-Jong;Park, Hong-Jae;Yi, Bu-Young;Lee, Kyeong-Ryeol;Kim, Myung-Sik;Woo, Hee-Jong;Jin, Yong-Moon;Kweon, Soon-Jong
    • Journal of Plant Biotechnology
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    • v.35 no.1
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    • pp.69-74
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    • 2008
  • This study aims to establish the efficient soybean transformation system and develop soybean [Glycine max (L.) Merill] transformants using cotyledonary node explants. The cotyledonary node of soybean were co-cultivated with Agrobacterium tumefaciens strains (KYRT1, EHA105). These strains contain the binary vector pCAMBIA3301 which carries a herbicide-resistant far gene. Korean cultivars (Danbaekkong, Eunhakong) and foreign cultivars (Jack, Peking) were the most efficient in regenerating cotyledonary node. Therefore, they were chosen for the transformation. Results showed that the T-DNA transfer reached up to 60% and transformation efficiency reached up to 3% in the cotyledonary node explants from Jack cultivar, co-cultivated with EHA105 strain. Histochemical GUS evaluation showed that 12 individual lines, transformed with the 현 gene, have positive response. The transformed soybeans have been confirmed in the $T_0$ generation through phenotypic assay using herbicide $Basta^{(R)}$ and Southern blot analysis.

Isolation of Cysteine Proteinase Gene (PgCysP1) from Panax ginseng and Response of This Gene to Abiotic Stresses (인삼으로부터 Cysteine Proteinase 유전자의 분리 및 환경 스트레스에 대한 반응)

  • Jeong, Dae-Young;Kim, Yu-Jin;Shim, Ju-Sun;Lee, Jung-Hye;In, Jun-Gyo;Lee, Bum-Soo;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.32 no.4
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    • pp.300-304
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    • 2008
  • Cysteine proteinases play an essential role in plant growth and development but also in senescence and programmed cell death. They participate in both anabolic and catabolic processes. In addition, they are involved in signalling pathways and in the response to biotic and abiotic stresses. A cDNA clone encoding cysteine proteinase (CP) gene, designated PgCysP1, was isolated from Panax ginseng C. A. Meyer. Reverse transcriptase (RT)-PCR results showed that PgCysP1 expressed at different level in P. ginseng hairy root. Different stresses such as biotic as well as abiotic stresses triggered a significant induction of PgCysP1. The positive responses of PgCysP1 to the various stimuli suggested that PgCysP1 may help to protect the plant against reactive environmental stresses.

Fusarium Wilt Caused by Fusarium oxysporum on Passionfruit in Korea (Fusarium oxysporum에 의한 패션프루트 시들음병)

  • Joa, Jae-Ho;Choi, In-Young;Choi, Min-Kyung;Heo, Byong-Soo;Jang, Jong-Ok;Shin, Hyeon-Dong
    • Research in Plant Disease
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    • v.24 no.1
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    • pp.75-80
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    • 2018
  • From 2014 to 2016, Fusarium wilt disease was found on fassionfruit in Iksan and Jeju, Korea. Symptoms included wilting of foliage, drying and withering of leaves, and stunting of the plants. The infected plants eventually died during growth. Colonies on potato dextrose agar were pinkish white, and felted with cottony and aerial mycelia with 35 mm after one week. Macroconidia were falcate to almost straight, thin-walled and usually 2-3 septate. Microconidia were usually formed on monophialides of the hyphae and were hyaline, smooth, oval to ellipsoidal, aseptate or medianly 1-septate, very occasionally 2-septate, slightly constricted at the septa, $3-12{\times}2.5-6{\mu}m$. On the basis of the morphological characteristics and phylogenetic analyses of two molecular markers, internal transcribed spacer rDNA and translation elongation factor $1{\alpha}$, the fungus was identified as Fusarium oxysporum. Pathogenicity of a representative isolate was proved by artificial inoculation, fulfilling Koch's postulates. To our knowledge, this is the first report on the occurrence of F. oxysporum on fassionfruit in Korea.

Miscanthus EST-originated Transcription Factor WRKY Expression in Response to Low Temperature in Warm-season Turfgrasses (억새 EST 정보 유래 전사요소 WRKY의 난지형 잔디의 저온 발현 반응성)

  • Chung, Sung Jin;Choi, Young In;Lee, Geung-Joo
    • Weed & Turfgrass Science
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    • v.2 no.4
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    • pp.368-375
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    • 2013
  • Whole genome transcriptomes from Miscanthus species were sequenced and analyzed, which provided 50 different types of transcription factor (TF) involving various developmental processes or environmental stresses. Among the explored TF, WRKY gene family was the major type and one of the WRKY genes, MSIR7180_WRKY4, induced under low temperature environment was selected to investigate how the Miscanthus-originated MSIR7180_WRKY4 TF responds when exposed to low temperature in four warm-season turfgrasses (Z. matrella 'Semil', bermudagrass, St. Augustinegrass, and seashore paspalum). The MSIR7180_WRKY4 was expressed higher during low temperature period in Bermudagrass, but the expression was enhanced in St. Augustinegrass. In contrast, the gene in 'Semil' cultivar was barely expressed and relatively less expressed, but repressed gradually in seashore paspalum, which seems to allow two turfgrasses stay-green longer in the fall season. The results indicate that bermudagrass and St. Augustinegrass adapt to low temperature quickly, but relative tolerance to low or cold temperature at the molecular level needs to be further investigated at different physiological stages and the corresponding genes systematically.