• Title/Summary/Keyword: Molecular techniques

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Rhizospheric fungi of Panax notoginseng: diversity and antagonism to host phytopathogens

  • Miao, Cui-Ping;Mi, Qi-Li;Qiao, Xin-Guo;Zheng, You-Kun;Chen, You-Wei;Xu, Li-Hua;Guan, Hui-Lin;Zhao, Li-Xing
    • Journal of Ginseng Research
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    • v.40 no.2
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    • pp.127-134
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    • 2016
  • Background: Rhizospheric fungi play an essential role in the plantesoil ecosystem, affecting plant growth and health. In this study, we evaluated the fungal diversity in the rhizosphere soil of 2-yr-old healthy Panax notoginseng cultivated in Wenshan, China. Methods: Culture-independent Illumina MiSeq and culture-dependent techniques, combining molecular and morphological characteristics, were used to analyze the rhizospheric fungal diversity. A diffusion test was used to challenge the phytopathogens of P. notoginseng. Results: A total of 16,130 paired-end reads of the nuclear ribosomal internal transcribed spacer 2 were generated and clustered into 860 operational taxonomic units at 97% sequence similarity. All the operational taxonomic units were assigned to five phyla and 79 genera. Zygomycota (46.2%) and Ascomycota (37.8%) were the dominant taxa; Mortierella and unclassified Mortierellales accounted for a large proportion (44.9%) at genus level. The relative abundance of Fusarium and Phoma sequenceswas high, accounting for 12.9% and 5.5%, respectively. In total,113 fungal isolates were isolated from rhizosphere soil. They were assigned to five classes, eight orders (except for an Incertae sedis), 26 genera, and 43 species based on morphological characteristics and phylogenetic analysis of the internal transcribed spacer. Fusarium was the most isolated genus with six species (24 isolates, 21.2%). The abundance of Phoma was also relatively high (8.0%). Thirteen isolates displayed antimicrobial activity against at least one test fungus. Conclusion: Our results suggest that diverse fungi including potential pathogenic ones exist in the rhizosphere soil of 2-yr-old P. notoginseng and that antagonistic isolates may be useful for biological control of pathogens.

Impact of phosphorus application on the indigenous arbuscular mycorrhizal fungi, soybean growth and yield in a 5-year phosphorus-unfertilized crop rotation

  • Higo, Masao;Sato, Ryohei;Serizawa, Ayu;Gunji, Kento;Suzuki, Daisuke;Isobe, Katsunori
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.351-351
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    • 2017
  • Arbuscular mycorrhizal fungi (AMF) are particular soil fungi that benefit many crops and require a symbiosis with plant roots to survive. In our previous study, there was a positive correlation between AMF root colonization and soybean grain yield in a four-year consecutive winter cover crop-soybean rotational system without phosphorus fertilizer. It is suggested that higher AMF root colonization can be a better solution for improving soybean growth and grain yield in P-limited soil. Our purpose in this study was to test the hypothesis that a P application is the main factor improving soybean growth, P nutrition and grain yield, and the benefit from AMF to soybean P uptake and growth in a P-limited soil. Impact of a P application on AMF root colonization and communities in soybean roots and their potential contribution to soybean growth and P nutrition under a five-year P-unfertilized crop rotational system were investigated over two-years. In this study, four cover crop treatments included 1) wheat (Triticum aestivum); 2) red clover (Trifolium pratense); 3) rapeseed (Brassica napus); and 4) fallow in the crop rotation. The amount of triple superphosphate as a P fertilizer applied rate after cultivation of cover crops was 120 and $360k\;ha^{-1}$ in 2014 and 2015, respectively. Soybean roots were sampled at full-flowering and analyzed for AMF communities using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and quantitative real-time PCR (qPCR) techniques. The AMF root colonization in the soybean roots at full bloom stage was significantly influenced by cover crop and P application throughout the two-year rotation. The two-year rotation of different cover crops or fallow impacted the molecular diversity of AMF communities colonizing roots of soybean. Redundancy analysis (RDA) indicated that AMF communities colonizing roots of soybean were significantly different among cover crop rotations. The AMF communities colonizing roots of soybean were clearly influenced by a P application in the two-year trial. Moreover, a P application may have positively impacts on the AMF communities under P-deficit soil due to the continuous cover crop-soybean rotational system without a P fertilizer.

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Invasion of Korean Pine Seedlings Originated from Neighbour Plantations into the Natural Mature Deciduous Broad-leaved Forest in Gwangneung, Korea (광릉 천연활엽수 성숙림에서 주변 인공림으로부터 잣나무 치수의 침입 정착)

  • Kang, Ho Sang;Lim, Jong-Hwan;Chun, Jung Hwa;Lee, Im Kyun;Kim, Young Kul;Lee, Jae Ho
    • Journal of Korean Society of Forest Science
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    • v.96 no.1
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    • pp.107-114
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    • 2007
  • Establishments of the seedlings inside the natural forest from adjacent artificial forests would be an important factor in forest stand dynamics. This study was conducted to see the invasion of Korean pine (Pinus koraiensis) seedlings which is not native in this region, into the natural deciduous broad-leaved forest in Gwangneung, Korea. There is no mother tree at the I ha study site while the number of naturally regenerated P. koraiensis seedlings was 345 trees and 56% of them were clumped with more than two seedlings at each point. Applying the image segmentation method to IKONOS satellite image of January, 2003, the distance from the center of 1 ha study site to the nearest mother tree and plantation of Korean pine were 200 m and 270 m, respectively. The average height and root-collar diameter of the seedlings were 34 em and 7 mm, respectively and the age of 207 seedlings (60%) were below 5 years old. Most abundant range of soil moisture gradient and LAl (leaf area index) were from 16 to 20% and those of LAI were from 3.1 to 3.5. To understand the dynamics and seed dispersal pattern of Korean pine in the Gwangneung natural deciduous broad-leaved forests, additional studies not only long-term monitoring of growth and mortality of naturally regenerated Korean pine seedlings but also application of stable isotope analysis and molecular genetic techniques was recommended.

High Frequency of TP53 but not K-ras Gene Mutations in Bolivian Patients with Gallbladder Cancer

  • Asai, Takao;Loza, Ernesto;Roig, Guido Villa-Gomez;Ajioka, Yoichi;Tsuchiya, Yasuo;Yamamoto, Masaharu;Nakamura, Kazutoshi
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.13
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    • pp.5449-5454
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    • 2014
  • Although genetic characteristics are considered to be a factor influencing the geographic variation in the prevalence of gallbladder cancer (GBC), they have not been well studied in Bolivia, which has a high prevalence rate of GBC. The purpose of this study was to examine the frequency of TP53 and K-ras mutations in Bolivian patients with GBC and to compare them with our previous data obtained in other high-GBC-prevalence countries, namely Japan, Chile, and Hungary. DNA was extracted from cancer sites in paraffin-embedded tissue from 36 patients using a microdissection technique. TP53 mutations at exons 5 to 8 and K-ras mutations at codons 12, 13 and 61 were examined using direct sequencing techniques. The data obtained were compared with those in the other high-GBC-prevalence countries. Of the 36 patients, 18 (50.0%) had a TP53 mutation (one mutation in each of 17 patients and three mutations in one patient), and only one (2.8%) had a K-ras mutation. Of the 20 TP53 mutations, 12 were of the transition type (60.0%). This rate was significantly lower than that in Chile (12/12, P<0.05). In addition, three mutations were of the CpG transition type (15.0%), which is a feature of endogenous mutation. All three were found in the hot spot region of the TP53 gene. In contrast, G:C to T:A transversion was found in Bolivia, suggesting the presence of exogenous carcinogens. Our findings suggest that the development of GBC in Bolivia is associated with both exogenous carcinogens and endogenous mechanisms. The identification of an environmental risk factor for GBC is needed to confirm these findings.

The Study on Phase Separation Development by Curing Reaction Rate for Unsaturated Polyester/Polyvinylacetate Semi-IPN (Unsaturated Polyester/Polyvinylacetate Semi-IPN의 경화반응속도에 따른 상분리현상 연구)

  • Chang, Won-Young;Kim, Moo-Sool;Kim, Jin-Hwan;Nam, Jae-Do
    • Polymer(Korea)
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    • v.25 no.1
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    • pp.78-89
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    • 2001
  • Morphological changes of unsaturated polyester/polyvinylacetate semi-IPN were studied while the phase separation and the cure reaction occurred in a competing fashion. The light scattering and thermal analysis techniques were used to investigate the phase separation rates and mechanical properties resultantly induced by molecular diffusion of thermoplastic polymer during the curing process of thermosetting polymer. The reaction activation energy was calculated by using Flynn-Wall method and the semi-IPN structure exhibited various phase-separation morphological characteristics. When PVAc composition was 10 wt%, the phase separation was not observed during the curing reaction, but the phase separation occurred in a similar fashion to nucleation and growth(NG) mechanism at room temperature. On the other hand, when PVAc composition was over 11.65 wt%, the phase separation was generated in the middle of the curing process. Consequently, the phase separation seemed to influence the curing reaction rate, which was also supported by the changing activation energy with conversion and PVAc composition. Finally, the total scattered intensity was measured at various temperature, and subsequently the diffusion rates of phase separation R(${\beta}m$) were evaluated.

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Identification of Petroselinic Acid (Cis-6-octadecenoic Acid) in the Seed Oils of Some of the Family Umbelliferae (Panax schinseng, Aralia continentalis and Acanthopanax sessiliflorus) by GC-MS, IR, $^1H-and$ $^13C-NMR$ Spectroscopic Techniques

  • Kim, Seong-Jin
    • Journal of the Korean Applied Science and Technology
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    • v.22 no.4
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    • pp.323-331
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    • 2005
  • Fatty acid compositions of the seed oils of P. schinseng, A. continentalis and A. sessiliflorus, were analyzed by gas chromatography (GC) equipped with a capillary column. A large unusual peak was observed just before the peak corresponding to oleic acid $(cis-9-C_{18:1})$. This unknown fatty acid was isolated by silver ion chromatography and then derivatized into the picolinyl ester. The mass spectrum of the picolinyl ester showed molecular ion at m/z=373 with other diagnostic ions such as m/z=178, 218, 232, 246, 274, 288, 302 and 344. Characteristic absorption peaks at $720\;cm^{-1}$, $1640\;cm^{-1}$ and $3010\;cm^{-1}$ in IR spectrum indicated the presence of cis-configurational double bond in the molecule. The $^1H-NMR$ spectrum of this acid gave two quintets centered at ${\delta}1.638$ (2H, C-3) and ${\delta}1.377$ (2H, C-4), and two multiplets centered at ${\delta}2.022{\sim}2.047$ (2H, C-5) and ${\delta}2.000{\sim}2.022$ (2H, C-8), and multiplet signals of olefinic protons centered at ${\delta}5.3015{\sim}5.3426$ (C-6, J=9.5 Hz) and ${\delta}\;5.3465{\sim}5.3877$ (C-7, J=9.5 Hz). The $^13C-NMR$ spectrum showed 18 carbon resonance signals including an overlapped signal at ${\delta}29.7002$ for C-12 and ${\delta}29.6520$ for C-13 (or they can be reversed), and other highly resolved signals at ${\delta}33.950$, ${\delta}24.558$, ${\delta}26.773$ and ${\delta}27.205$ due to C-2, C-3, C-5 and C-8 of a ${\Delta}^6-octadecenoic$ acid, respectively. From analysis results this unknown fatty acid could be identified as cis-6-octadecenoic acid. The seed oils of P. schinseng and A. sessiliflorus contained petroselinic acid (59.7%, 56.0%), oleic acid (18.3%, 6.1%) and linoleic acid (16.2%, 30.4%) with small amount of palmitic acid (3.0%, 3.1%) while the seed oil of A. continentalis comprised mainly oleic acid (30.2%), petroselinic acid (29.0%), linoleic acid (24.1%) and palmitic acid (13.1%).

A Nationwide Survey on the Prevalence of Intestinal Parasitic Infections in the Republic of Korea, 2004

  • Kim, Tong-Soo;Cho, Shin-Hyeong;Huh, Sun;Kong, Yoon;Sohn, Woon-Mok;Hwang, Seung-Sik;Chai, Jong-Yil;Lee, Soon-Hyung;Park, Yun-Kyu;Oh, Dae-Kyu;Lee, Jong-Koo;Working Groups in National Institute of Health, Working Groups in National Institute of Health;Korea Association of Health Promotion, Korea Association of Health Promotion
    • Parasites, Hosts and Diseases
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    • v.47 no.1
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    • pp.37-47
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    • 2009
  • National surveys on the prevalence of intestinal parasitic infections have been carried out every 5-7 years since 1971 in the Republic of Korea in order to establish control measures. The present nationwide survey was conducted from June to December 2004. The 10% population sampling data of Population and Housing Census by the Korean government in 2000 was used as the survey population. One sample was selected randomly from each of the 22,858 registered subjects, and a total of 20,541 people were ultimately included in this survey. Fecal examinations were performed by the cellophane thick smear and saturated brine flotation techniques. Pinworm infection was examined by cello-tape anal swab method. This survey also included a questionnaire study for a socioeconomic analysis. The total helminth egg positive rate was 3.7%, and the estimated total positive number among nationwide people was 1,780,000. The rates in urban and rural areas were 3.1% and 6.8%, respectively. As the total egg positive rate in the 6th survey in 1997 was 2.4%, the present survey showed that there was a considerable degree of increase in the prevalence rate of intestinal parasitic infections over the 7-year period following the 6th survey. The largest increases occurred in the egg positive rates of Clonorchis sinensis and heterophyids including Metagonimus yokogawai.

Measurement of Shunt Amount Using Radionuclide Angiocardiography: Accuracy According to Level of Shunt and Associated Lesion (방사성 동위원소 심혈관 조영술을 이용한 단락량 측정법: 정확성에 영향을 미치는 인자)

  • Kim, Yang-Min
    • Nuclear Medicine and Molecular Imaging
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    • v.40 no.4
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    • pp.200-204
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    • 2006
  • Purpose: Determination of pulmonary to systemic blood flow ratio (QP/QS) is important for the management of patients with left-to-right shunt. This study was performed to assess the agreement of Qp/Qs ratio using the radionuclide method and oxymetry, to investigate the factors influencing the agreement, and to know how interchangeable the results of each technique. Materials and Methods: We compared the Qp/Qs measured by single-pass radionuclide angiocardiography and oxymetry during catheterization in 207 patients who underwent both studies. In radionuclide method, Qp/Qs was calculated from the pulmonary time-activity curves using a gamma variate fit. The correlation and Bland-Altman analysis were performed according to the levels of shunt and associated lesions. Results: The mean Qp/Qs was $1.83{\pm}0.50$ by radionuclide, and $1.74{\pm}0.51$ by oxymetry. The overall correlation coefficient was 0.86(p<0.001), and Bland-Altman range of agreement encompassing 4SD was 1.05. For atrial septal defect, ventricular septal defect, patent ductus arteriosus, tricuspid and mitral insufficiency, the correlation coefficient was 0.78, 0.90, 0.84, 0.63 and 0.44, and Bland-Altman range was 1.51, 0.74, 0.96, 1.57, and 1.50, respectively. Conclusion: There is good agreement but wide variance between the Qp/Qs ratios by radionuclide method and oxymetry. Associated atrioventricular valvar insufficiency decreases the correlation coefficient and widens the variance. Wide overall variance suggests that Qp/Qs measurements by two techniques should not be used interchangeably.

Factors Influencing Efficient Agrobacterium-mediated Transformation of Panicum spp. (Agrobacterium법에 의한 Panicum속 식물들의 효과적인 형질전환에 영향을 미치는 요인)

  • Seo, Mi-Suk;Takahara, Manabu;Takamizo, Tadashi
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.31 no.1
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    • pp.1-8
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    • 2011
  • Molecular techniques such as genetic transformation are powerful tools that can be used for the genetic modification of warm-season grasses. The P. meyerianum with high regeneration ability was used for establishing an Agrobacterium-mediated transformation system. We investigated various factors affecting Agrobacterium infection by examining GUS gene expression of pCAMBIA1304 vector. Among various concentration of acetosyringone and betaine tested for inoculation and co-cultivation, 10 mg/L acetosyringone and 60 mg/L betaine resulted in the highest transformation frequency in terms of GUS expression. The calli of 4 species of Panicum spp. with excellent tissue culture response were inoculated with Agrobacterium under the optimal infection conditions. The high activity of GUS gene was observed in all species and hygromycin-resistant calli expressing GFP were obtained in P. meyerianum, P. longijubatum, P. stapfianum and guineagrass Noh-PL1. Co-cultivated calli were transferred onto the selection medium containing hygromycin, and the hygromycin resistant calli were selected after 3 months. Hygromycin-resistant plantlets were then successfully regenerated from the calli and grown in a greenhouse. We confirmed stable insertion of hpt gene among the hygromycin-resistant plantlets of P. meyerianum by PCR analysis.

브라디키닌 수용체 발현 및 길항제개발에 관한 연구

  • 정성현;이은순
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.146-146
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    • 1993
  • 신장에서 브라디키닌(Bradikinin, BK)의 생리적 역할을 규명하기 위한 첫 단계로 토끼신장 근위세뇨관의 일차배양세포를 이용, BK 수용체를 ($^3$H) BK를 이용하여 수용체결합실험을 함과 아울러 세포배양과정에서 여러 성장인자들의 BK 수용체발현에 미치는 영향을 관찰하였다. 첫째, 토끼 신장의 피질, 수질 및 근 위세뇨관 둥 각 부위에 대한 BK 수용체결합 실험결과 신수질 부분에 가장 많은 BK 수용체가 발견되었으며 이때 해리항수는 0.52 nM, 그리고 최대결합부위는 mg 단배질당 112.5 fmol이었다. 둘째, serum free 배지에서 insulin, transferrin 그리고 hydrocortisone이 성장인자로 사용될 때 BK 수용체발현이 가장 높았으며 이 중 인슐린에 가장 큰 영향을 받았다. 한편 인슐린의 최적농도를 결정하는 실험의 결과 5 $\mu\textrm{g}$/ml매서 가장 높은 수용체발현을 나타내었으며 그 이상에서는 별다른 차이를 보이지 못했다. 세째, 위의 세 성장인자에 prostaglandin E$_1$이나 triiodothyronine을 첨가시 BK 수용체발현은 오히려 저하되었다. 네째, fetal bovine serum (FBS)과 위의 세 성장인자간의 수용체발현능을 비교한 실험에서 세포배양 후 첫 일주일에는 FBS가 세 성장인자보다 약간 나은 수용체발현능을 나타내었으나 그후 이주째에는 세 성장인자가 BK 수용체발현에 더 적절한 요소임을 보여주었다. 이상의 결과로 보아 토끼 근위세뇨관 상피세포에서의 BK 수용체발현은 세포성장인자로 insulin, transferrin, hydrocortisone 중 insulin에 가장 큰 영향을 받는것으로 보이며, 이들 세가지 성장인자는 serum free 배지에서 세포성장 및 기능에 많은 영향을 주는 것으로 생각된다.prolidine이 $K_{M}$ /K$_{H}$ 비가 가장 높았고 diphenidol이 가장 낮았다. 이상의 결과로 보아 항 histamine제의 muscarinic receptor 차단작용은 이들 약물의 항 alleragy 효과에 필요한 작용이 아니며 본 실험에서 추정된 항 histamine제의 H$_1$-receptor와 muscarinic receptor에 대한 상대적 역가는 이들 약물의 선택과 평가에 중요한 지표가 될수 있을 것으로 생각된다.ing ischemic insults. The nature of the receptor is being explored by molecular genetic techniques, and we have recently cloned two of the major subunits; some of the data will be presented.LIFO, 우선 순위 방식등을 선택할 수 있도록 확장하였다. SIMPLE는 자료구조 및 프로그램이 공개되어 있으므로 프로그래머가 원하는 기능을 쉽게 추가할 수 있는 장점도 있다. 아울러 SMPLE에서 새로이 추가된 자료구조와 함수 및 설비제어 방식등을 활용하여 실제 중형급 시스템에 대한 시뮬레이션 구현과 시스템 분석의 예를 보인다._3$", chain segment, with the activation energy of carriers from the shallow trap with 0.4[eV], in he amorphous regions.의 증발산율은 우기의 기상자료를 이용하여 구한 결과 0.05 - 0.10 mm/hr 의 범위로서 이로 인한 강우손실량은 큰 의미가 없음을 알았다.재발이 나타난 3례의 환자를 제외한 9례 (75%)에서는 현재까지 재발소견을 보이지 않고 있다. 이러한 결과는 다른 보고자들과 유사한 결과를

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