• Title/Summary/Keyword: plant-based protein

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Virus-induced Silencing of the WRKY1 Transcription Factor that Interacts with the SL1 Structure of Potato virus X Leads to Higher Viral RNA Accumulation and Severe Necrotic Symptoms

  • Park, Sang-Ho;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.28 no.1
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    • pp.40-48
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    • 2012
  • $Potato$ $virus$ $X$ (PVX) replication is precisely regulated by regulatory viral sequences and by viral and/or host proteins. In a previous study, we identified a 54-kDa cellular tobacco protein that bound to a region within the first 46 nucleotides (nt) of the 5' non-translated region (NTR) of the viral genome. Optimal binding was dependent upon the presence of an ACCA sequence at nt 10-13. To identify host factors that bind to 5' NTR elements including AC-rich sequences as well as stemloop 1 (SL1), we used northwestern blotting and matrixassisted laser desorption/ionization time-of-flight mass spectrometry for peptide mass fingerprinting. We screened several host factors that might affect PVX replication and selected a candidate protein, $Nicotiana$ $tabacum$ WRKY transcription factor 1 (NtWRKY1). We used a $Tobacco$ $rattle$ $virus$ (TRV)-based virus-induced gene silencing (VIGS) system to investigate the role of NtWRKY1 in PVX replication. Silencing of $WRKY1$ in $Nicotiana$ $benthamiana$ caused lethal apical necrosis and allowed an increase in PVX RNA accumulation. This result could reflect the balancing of PVX accumulation in a systemic $N.$ $benthamiana$ host to maintain PVX survival and still produce a suitable appearance of mosaic and mottle symptoms. Our results suggest that PVX may recruit the WRKY transcription factor, which binds to the 5' NTR of viral genomic RNA and acts as a key regulator of viral infection.

Effect of Seeding Date on Forage Yield and Chemical Composition of Echinochloa crusgalli var. Frumentacea(Roxb) Wight in Jeju Region (제주지역에서 파종기에 따른 청예피의 사료수량 및 조성분 변화)

  • 조남기;강영길;송창길;고영순;조영일
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.21 no.4
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    • pp.217-224
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    • 2001
  • In order to identify the growth characterization, the yield and chemical composition of Jeju barnyard grass (Echinochloa crusgalli var. fiurnentacea(Roxb) Wight) based on seeding date in Jeju region, seeding carried out the 10-day intervals from March 27 to September 30 in 2000, respectively. Plant height was 143.2 cm, showing the highest on seeding date, April 6 among that of any other seeding date. On the other hand, those of early and late seeding gradually decrease. Plant height was 119.2 an in May 16 seeding. The results of stem diameter, number of withering leaves, number of leaves and fresh weight per plant were similar to those of the plant heights. The yield of fresh, dry matter forage, crude protein and TDN found the highest on April 6 seeding, 63.5 MT/ha, 13.9 MTha, 1.1 MT/ha, and 7.6 MT/ha, respectively. In early and late seeding, the yield was gradually decreased. In seeding May 16, the yield found .38.2 MTIha in fresh forage, 6.2 MTha in dry matter forage, 0.7 MT/ha in crude protein and 3.7 MTha in TDN, respectively. According to delaying the seeding date, March 27 to May 16, the contents of crude protein (from 7.9 to 10.8%), ether extract (from 4.6 to 6.0%), nitrogen free extract (from 45.1 to 46.5%), and TDN (from 54.2 to 60.8%) were gradually increased, respectively. On the other hand, those of crude fiber (from 28.9 to 25.6%) and crud ash (from 13.5 to 11.2%) were decreased. These results showed that April 6 was the optimum seeding date with the sole object of feed production of Jeju barnyard grass under the environmental condition like as atmospheric phenomena and soil in Jeju region. (Key words : Jeju barnyard grass, Seeding date, Forage yield, Chemical composition)

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A Systematic Study on the Selected Amentiferous Plant Taxa -On the Genus Quercus- (유이화서 식물군의 계통분류학적 연구 -참나무과를 중심으로-)

  • Lee, Yoo Sung
    • Journal of Plant Biology
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    • v.27 no.2
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    • pp.61-72
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    • 1984
  • The concept of natural grouping of plant designated as the "Amentiferous" is no longer given serious credence, and many of the families included in this grouping have been dispersed in diverse order. Because a review of taxonomic treatments of amentiferous taxa reveals diverse classifications, it has become necessary to investigate new characteristics and attempt to determine the significance of these characteristics in terms of amentiferous taxonomy. Protein analyses by isoelectrofocusing(IEF) and rocket immunoelectrophoresis(RIE) have proved to be useful in the delicitation of Quercus taxa using pollen extracts from selected taxa. When Quercus pollen extracts were separated by electrophoresis based on their isoelectric points in a stable pH gradient and substrates for estrase activity were stained, ten bands were revealed between pH 5-14. Within Lepidobalanus grouping, a great diversity in the pollen protein zymograms was observed with some segregation corresponding to the designated taxonomic sections. Two taxa of Cyclobalanopsis produced a zymogram that is somewhat similar to taxa included within the section Prinus of Lepidobalanus, and less similar to taxa within the section Cerris of the same subgenus. Three tested taxa of the Cerris are in the similar zymogram each other, being segregable from the taxa of Prinus. Quantitative and qualitative analyses for serological relationships within and among th Quercus were also employed. To calculate the degree of protein similarity, total rocket heights obtained from RIE provided an index of serological correspondence(SC). It is reconfirmed that the Quercus is distantly separated from the Fagus according to SC. Comparative data from rocket number and SC in the tested taxa of Quercus also indicate that Lepidobalanus is separable from Cyclobalanopsis. Within the Lepidobalanus Q. acutissima and Q. acutissima x variabilis are almost homogeneors and distinguishable from the other tested taxa of same subgenus. Although the number of taxa tested has been limited, the overall serological evidence best reflects the classification proposed by Redher(1940) and Melchior (1964), having the genus Quercus subdivided into three subgunera: Erythrobalanus, Lepidobalanus, and Cyclobalanopsis.alanopsis.

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Genome Mapping Technology And Its Application In Plant Breeding (작물 육종에서 분자유전자 지도의 이용)

  • 은무영
    • Proceedings of the Botanical Society of Korea Conference
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    • 1995.07a
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    • pp.57-86
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    • 1995
  • Molecular mapping of plant genomes has progressed rapidly since Bostein et al.(1980) introduced the idea of constructing linkage maps of human genome based on restriction fragment length polymorphism (RFLP) markers. In recent years, the development of protein and DNA markers has stimulated interest for the new approaches to plant improvement. While classical maps based on morphological mutant markers have provided important insights into the plant genetics and cytology, the molecular maps based on molecular markers have a number of inherent advatages over classical genetic maps for the applications in genetic studies and/or breeding schemes. Isozymes and DNA markers are numerous, discrete, non-deleterious, codominant, and almost entirely free of environmental and epistatic interactions. For these reasons, they are widely used in constructing detailed linkage maps in a number of plant species. Plant breeders improve crops by selecting plants with desirable phenotypes. However a plant's phenotyes is often under genetic control, positioning at different "quantitative trait loci" (QTLs) together with environmental effects. Molecular maps provide a possible way to determine the effect of the individual gene that combines to produce a quantitative trait because the segregation of a large number of markers can be followed in a single genetic cross. Using market-assisted selection, plants that contain several favorable genes for the trait and do not contain unfavourable segments can be obtained during early breeding processes. Providing molecular maps are available, valuable data relevant to the taxonomic relationships and chromosome evolution can be accumulated by comparative mapping and also the structural relationships between linkage map and physical map can be identified by cDNA sequencing. After constructing high density maps, it will be possible to clone genes, whose products are unknown, such as semidwarf and disease resistance genes. However, much attention has to be paid to level-up the basic knowledge of genetics, physiology, biochemistry, plant pathology, entomology, microbiology, and so on. It must also be kept in mind that scientists in various fields will have to make another take off by intensive cooperation together for early integration and utilization of these newly emerging high-techs in practical breeding. breeding.

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Generation of Antibodies Against Rice stripe virus Proteins Based on Recombinant Proteins and Synthetic Polypeptides

  • Lian, Sen;Jonson, Miranda Gilda;Cho, Won-Kyong;Choi, Hong-Soo;Je, Yeon-Ho;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.27 no.1
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    • pp.37-43
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    • 2011
  • Rice stripe virus (RSV) is one of serious epidemic pathogens for rice species grown in many Asian countries. Therefore, it is necessary to produce a diagnostic detection kit applicable in fields for RSV detection. In this study, RSV proteins that were derived from recombinant proteins and synthetic polypeptides as antigens were generated and were raised in rabbits for antiserum production. Among seven proteins in RSV, genes that code for NCP and NS3 proteins were cloned and subcloned into vector carrying His-tag protein and were expressed in E. coli. Of two recombinant proteins, only anti-NCP displayed stable hybridization signals in western blot analysis. Alternately, synthetic RSV polypeptides for CP, NCP, NS3 and NSvc4 we also generated and only antibodies against CP and NCP were very effective to detect RSV in both RSV infected rice and weed plants. However, antibodies against NS3 and NSvc4 showed weak specific bands as well as strong non-specific background due to the difference of viral proteins produced in the infected leaves. In summary, the antibodies generated against RSV proteins produced in this study will be useful for various assays such as for RSV diagnostic detection, immunoprecipitation, protein purification, and western blot analysis.

Studies on Protein Profiles and Isozymes in Germinating Seeds (종자발아에 있어서 Protein Profile과 Isozyme에 관한 연구)

  • 권오용
    • Journal of Plant Biology
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    • v.17 no.4
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    • pp.143-156
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    • 1974
  • The purpose of this experiment was to study one side of germination physiology based on that protein profiles and protease relating to protein metabolism, that peroxidase, catalase, $\alpha$-amylase, $\beta$-amylase, and malate dehydrogenase involved in the carbohydrate metabolism of seed germination. All these experiments were divided into the two groups with and without acetone treatment, and were carried out. The protein bands of each germinating stage between the groups treated with and without acetone showed certain basic pattern in polyacrylamide gel disc electrophoresis. However, there was a little difference in the number of protein band, optical density, and migration velocity between two groups. The isozyme bands of peroxidase, and catalase between two groups in polyacrylamide gel disc electrophoresis did not show the numeral difference, but the optical density of certain germinating stage treated with acetone was higher than the group untreated with it and it showed their enzyme activity. The $\alpha$-amylase and $\beta$-amylase activities which involved in starch metabolism of seed germination were higher in the treated group than the other. On one hand, the protease activity of hydrolase occurred in the seeds for germination was also higher, more or less in the treated group than in the other. The isozyme band pattern of malate dehydrogenase in TCA cycle of energy metabolism pathway was very different between two groups growing for 72 hours with and without acetone treatment in cellulose acetate electrophoresis. It indicated that two isozyme bands of malate dehydrogenase was high. Consequently these experimental results mentioned above indicated that acetone treatment before sowing had an effect on dissolving certain complexed lipid substance involved in the seed coats, the activity of carbohydrate hydrolase increased with water absorption which was most comfortable in its germination, dissolved glycerin and fatty acid became certain energy source, and they stimulated the acceleration of respiration metabolism.

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GENOTYPIC AND PHENOTYPIC CORRELATIONS IN A SOYBEAN CROSS

  • Shin-Han Kwon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.1 no.1
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    • pp.42-45
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    • 1963
  • In a plant breeding program, an efficient selection of desired characters in a population is important. Generally, many agronomic characters in a given population are determined by polygenes and quantitatively inherited. In practice, the genetic relationship between two observed characters which are undoubtedly subjected to the environmental influence is difficult to identify. In recent years, many workers have attempted to understant the genetic relationship between characters in terms of genotypic correlation, and the knowledge thus gained should furnish many important and useful information for the planning of breeding, selection, and interpretation of the result. The genotypic correlation is the result of pleiotropy, linkage of genes(2, 3, 5, 6, 8) and natural or artificial selection(4). The purposes of this study were to estimate genotyric and phenotypic correlations between all possible pairs of nine characters. and to seek certain characters which may be useful as indicators of certain important agronomic characters. Weber and Moorthy(10), Johnson et al. (5) and Sheth(7) found that in general, the genotypic correlations were higher than the phenotypic correlations. Weiss et al. (11) obtained significant positive correlations between maturity and oil content, maturity and low protein content, and high protein content and low oil content. Weber and Moorthy(10) reported the positive genotypic correlations between flowering and maturity, yield and maturity, yield and plant height, yield and seed weight, and negative genotypic correlations between maturity and oil content, and oil content and seed weight. Johnson et al. (5) studied the genotypic and phenotypic correlations among 24 characters and concluded that selection based entirely on a long fruiting period, lateness, heavy seed, low protein, high oil and resistance to lodging would be effective in increasing yield. Sheth(7) found the following positive associations among characters; height and maturity, yield and lodging, low protein content and high oil content, and yield and low protein content. Hanson et al.(1) also reported high negative correlation between seed yield and protein content.

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Gene Identification and Molecular Characterization of Solvent Stable Protease from A Moderately Haloalkaliphilic Bacterium, Geomicrobium sp. EMB2

  • Karan, Ram;Singh, Raj Kumar Mohan;Kapoor, Sanjay;Khare, S.K.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.2
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    • pp.129-135
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    • 2011
  • Cloning and characterization of the gene encoding a solvent-tolerant protease from the haloalkaliphilic bacterium Geomicrobium sp. EMB2 are described. Primers designed based on the N-terminal amino acid sequence of the purified EMB2 protease helped in the amplification of a 1,505-bp open reading frame that had a coding potential of a 42.7-kDa polypeptide. The deduced EMB2 protein contained a 35.4-kDa mature protein of 311 residues, with a high proportion of acidic amino acid residues. Phylogenetic analysis placed the EMB2 gene close to a known serine protease from Bacillus clausii KSM-K16. Primary sequence analysis indicated a hydrophobic inclination of the protein; and the 3D structure modeling elucidated a relatively higher percentage of small (glycine, alanine, and valine) and borderline (serine and threonine) hydrophobic residues on its surface. The structure analysis also highlighted enrichment of acidic residues at the cost of basic residues. The study indicated that solvent and salt stabilities in Geomicrobium sp. protease may be accorded to different structural features; that is, the presence of a number of small hydrophobic amino acid residues on the surface and a higher content of acidic amino acid residues, respectively.

Prediction of subcellular localization of proteins using pairwise sequence alignment and support vector machine

  • Kim, Jong-Kyoung;Raghava, G. P. S.;Kim, Kwang-S.;Bang, Sung-Yang;Choi, Seung-Jin
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2004.11a
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    • pp.158-166
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    • 2004
  • Predicting the destination of a protein in a cell gives valuable information for annotating the function of the protein. Recent technological breakthroughs have led us to develop more accurate methods for predicting the subcellular localization of proteins. The most important factor in determining the accuracy of these methods, is a way of extracting useful features from protein sequences. We propose a new method for extracting appropriate features only from the sequence data by computing pairwise sequence alignment scores. As a classifier, support vector machine (SVM) is used. The overall prediction accuracy evaluated by the jackknife validation technique reach 94.70% for the eukaryotic non-plant data set and 92.10% for the eukaryotic plant data set, which show the highest prediction accuracy among methods reported so far with such data sets. Our numerical experimental results confirm that our feature extraction method based on pairwise sequence alignment, is useful for this classification problem.

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Expression of Lily mottle virus Coat Protein and Preparation of IgY Antibody against the Recombinant Coat Protein

  • Yoo, Ha Na;Jung, Yong-Tae
    • Horticultural Science & Technology
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    • v.32 no.4
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    • pp.544-549
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    • 2014
  • Lily symptomless virus (LSV), Lily mottle virus (LMoV), and Cucumber mosaic virus (CMV) are the most prevalent viruses infecting lilies in Korea. Leaf and bulb samples showing characteristic symptoms of virus infection were collected in 2012, and 80 field samples were analyzed by reverse transcription polymerase chain reaction (RT-PCR). The infection frequencies were 79% for LMoV, 5% for LSV, and 3% for CMV. The LMoV coat protein gene was amplified and cloned into the pET21d(+) expression vector to develop serological diagnostic tools to detect LMoV. The resulting carboxy-terminal His-tagged coat proteins were expressed in Escherichia coli strain BL21 (DE3) by induction with IPTG. The recombinant proteins were purified using Ni-NTA agarose beads and used as an antigen to produce polyclonal antibodies in laying hens. The resulting egg yolk immunoglobulin (IgY) specifically recognized LMoV from infected plant tissues in immunoblotting assays and had comparable sensitivity to that of a mammalian antibody. In addition, method of immunocapture RT-PCR using this IgY was developed for sensitive, efficient, and rapid detection of LMoV. Based on these results, large-scale bulb tests and detection of LMoV in epidemiological studies can be performed routinely using this IgY. This is the first report of production of a polyclonal IgY against a plant virus and its use for diagnosis.