• Title/Summary/Keyword: plant-based protein

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Proteome Profiling Unfurl Differential Expressed Proteins from Various Explants in Platycodon Grandiflorum

  • Kim, Hye-Rim;Kwon, Soo-Jeong;Roy, Swapan Kumar;Cho, Seong-Woo;Kim, Hag-Hyun;Cho, Kab-Yeon;Boo, Hee-Ock;Woo, Sun-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.1
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    • pp.97-106
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    • 2015
  • Platycodon grandiflorum, commonly known as Doraji in Korea, has a wide range of pharmacologic properties, such as reducing adiposity and hyperlipidemia, and antiatherosclerotic effects. However, the mechanisms underlying these effects remain unclear. In order to profile proteins from the nodal segment, callus, root and shoot, high throughput proteome approach was executed in the present study. Two dimensional gels stained with CBB, a total of 84 differential expressed proteins were confirmed out of 839 protein spots using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 58 differential expressed protein spots (${\geq}$ 2-fold) were analyzed using MASCOT search engine according to the similarity of sequences with previously characterized proteins along with the UniProt database. Out of 58 differential expressed protein, 32 protein spots were up-regulated such as ribulose-1,5-bisphosphate carboxylase, endoplasmic oxidoreductin-1, heat stress transcription factor A3, RNA pseudourine synthase 4, cysteine proteinase, GntR family transcriptional regulator, E3 xyloglucan 6-xylosyltransferase, while 26 differential protein spots were down-regulated such as L-ascorbate oxidase precursor, late embryogenesis abundant protein D-34, putative SCO1 protein, oxygen-evolving enhancer protein 3. However, frequency distribution of identified proteins using iProClass databases, and assignment by function based on gene ontology revealed that the identified proteins from the explants were mainly associated with the nucleic acid binding (17%), transferase activity (14%) and ion binding (12%). In that way, the exclusive protein profile may provide insight clues for better understanding the characteristics of proteins and metabolic activity in various explants of the economically important medicinal plant Platycodon grandiflorum.

Isolation of an Rx homolog from C. annuum and the evolution of Rx genes in the Solanaceae family

  • Shi, Jinxia;Yeom, Seon-In;Kang, Won-Hee;Park, Min-Kyu;Choi, Do-Il;Kwon, Jin-Kyung;Han, Jung-Heon;Lee, Heung-Ryul;Kim, Byung-Dong;Kang, Byoung-Cheorl
    • Plant Biotechnology Reports
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    • v.5 no.4
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    • pp.331-344
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    • 2011
  • The well-conserved NBS domain of resistance (R) genes cloned from many plants allows the use of a PCR-based approach to isolate resistance gene analogs (RGAs). In this study, we isolated an RGA (CapRGC) from Capsicum annuum "CM334" using a PCR-based approach. This sequence encodes a protein with very high similarity to Rx genes, the Potato Virus X (PVX) R genes from potato. An evolutionary analysis of the CapRGC gene and its homologs retrieved by an extensive search of a Solanaceae database provided evidence that Rx-like genes (eight ESTs or genes that show very high similarity to Rx) appear to have diverged from R1 [an NBS-LRR R gene against late blight (Phytophthora infestans) from potato]-like genes. Structural comparison of the NBS domains of all the homologs in Solanaceae revealed that one novel motif, 14, is specific to the Rx-like genes, and also indicated that several other novel motifs are characteristic of the R1-like genes. Our results suggest that Rx-like genes are ancient but conserved. Furthermore, the novel conserved motifs can provide a basis for biochemical structural. function analysis and be used for degenerate primer design for the isolation of Rx-like sequences in other plant species. Comparative mapping study revealed that the position of CapRGC is syntenic to the locations of Rx and its homolog genes in the potato and tomato, but cosegregation analysis showed that CapRGC may not be the R gene against PVX in pepper. Our results confirm previous observations that the specificity of R genes is not conserved, while the structure and function of R genes are conserved. It appears that CapRGC may function as a resistance gene to another pathogen, such as the nematode to which the structure of CapRGC is most similar.

Development of a Species-specific PCR Assay for Three Xanthomonas Species, Causing Bulb and Flower Diseases, Based on Their Genome Sequences

  • Back, Chang-Gi;Lee, Seung-Yeol;Lee, Boo-Ja;Yea, Mi-Chi;Kim, Sang-Mok;Kang, In-Kyu;Cha, Jae-Soon;Jung, Hee-Young
    • The Plant Pathology Journal
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    • v.31 no.3
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    • pp.212-218
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    • 2015
  • In this study, we developed a species-specific PCR assay for rapid and accurate detection of three Xanthomonas species, X. axonopodis pv. poinsettiicola (XAP), X. hyacinthi (XH) and X. campestris pv. zantedeschiae (XCZ), based on their draft genome sequences. XAP, XH and XCZ genomes consist of single chromosomes that contain 5,221, 4,395 and 7,986 protein coding genes, respectively. Species-specific primers were designed from variable regions of the draft genome sequence data and assessed by a PCR-based detection method. These primers were also tested for specificity against 17 allied Xanthomonas species as well as against the host DNA and the microbial community of the host surface. Three primer sets were found to be very specific and no amplification product was obtained with the host DNA and the microbial community of the host surface. In addition, a detection limit of $1pg/{\mu}l$ per PCR reaction was detected when these primer sets were used to amplify corresponding bacterial DNAs. Therefore, these primer sets and the developed species-specific PCR assay represent a valuable, sensitive, and rapid diagnostic tool that can be used to detect three specific pathogens at early stages of infection and may help control diseases.

Two- Dimensional Electrophoresis Analysis of Proteins; Bacillus subtilis LTD and Its Antifungal Activity Deficient Mutant

  • Lee, Young-Keun;Dinh, Le Thi;Jang, Yu-Sin;Chung, Hye-Young;Chang, Hwa-Hyoung
    • Korean Journal of Environmental Biology
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    • v.22 no.4
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    • pp.487-493
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    • 2004
  • To investigate the antifungal activity related protein in pesticidal bacteria, a bacterial strain LTD was isolated from soil collected at Gimje in Jeonbuk province, Korea, and identified as Bacillus subtilis LTD based on a API50 CHB kit and 168 rDNA sequencing. It has an antifungal activity against 9 plant pathogenic fungi in a paper disc assay. The antifungal activity- deficient mutant, B. subtilis mLTD was induced at a 5 kGy dose of $^{60}Co$ gamma radiation. Using the two-dimensional electrophoresis and the matrix assisted laser desorption ionization time-of-flight mass spectrometry, the comparison analysis of proteins between the wild and mutant were performed. A major intracellular serine proteinase IspA (MW: 32.5 kDa), a NAD (P) H dehydrogenase (MW: 20.0 kDa), and a stage II sporulation protein AA, SpoIIAA (MW: 14.3kDa) were detected only in the B. subtilis LTD. These results suggested that the functions of these proteins found only in the B. subtilis LTD could. be closely related to the antifungal activity against plant pathogenic fungi.

Identification of the Arabidopsis thaliana cell growth defect factor suppressing yeast cell proliferation

  • Kim, Kyung-Min;Uchimiya, Hirofumi;Sohn, Jae-Keun
    • Current Research on Agriculture and Life Sciences
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    • v.30 no.1
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    • pp.1-11
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    • 2012
  • We identified cdf based on screening of the Arabidopsis cDNA library for functional suppressors of the AtBI-1 (a gene described to suppress the cell death induced by Bax gene expression in yeast). The cdf was located on Chr. V and was composed of 5 exons and 4 introns. It encodes a protein of 258 amino acid residues with a molecular weight of 28.8 kDa. The protein has 3 transmembrane domains in the C-terminal region. The cdf has one homologue, named cdf2, which was found in Arabidopsis. Like cdf, cdf2 also induced growth defect in yeast. The effect of the cell growth defect factor was somewhat lower than Bax. cdf could arrest the growth of yeast. Its localization to the nucleus was essential for the suppression of yeast cell proliferation. Morphological abnormality of intracellular network, which is a hallmark of AtBI-1, was attenuated by expression of cdf.

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Outbreak of Cucumber mosaic virus and Tomato spotted wilt virus on Bell Pepper Grown in Jeonnam Province in Korea

  • Mun, Hye-Yeon;Park, Mi-Ri;Lee, Hyang-Burm;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.24 no.1
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    • pp.93-96
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    • 2008
  • In August 2006, a severe disease incidence showing mosaic and/or necrotic symptoms on two bell pepper varieties including red-colored 'Special' and yellow-colored 'Fiesta' was observed in a greenhouse located in Gwangyang, Jeonnam province, Korea. To identify causal viruses, total RNAs were extracted from 11 fruit samples with and without symptoms. Specific oligonucleotide primers for Cucumber mosaic virus (CMV), Pepper mottle virus (PepMoV), Tomato spotted wilt virus (TSWV) and Pepper mild mottle virus (PMMoV) were designed based on the sequences available on GenBank. Database comparisons of the deduced amino acid sequences of each sequence produced 100% and 98% matches with nucleocapsid protein gene of TSWV (Acc. No. ABE11605) and coat protein gene of CMV (Acc. No. DQ018289), respectively, suggesting that the symptoms on bell pepper fruits might be caused by the infection of CMV and TSWV. To our knowledge this is the first report of necrotic as well as mosaic virus disease on bell pepper fruits by the infection of CMV and TSWV in Jeonnam province, Korea.

Analysis of Phylogenetic Relationships among Medicago Species by Proteins Banding Patterns and RFLP Markers

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    • Korean Journal of Plant Resources
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    • v.10 no.3
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    • pp.250-257
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    • 1997
  • The relationship of nine Medicago species belonging to four subgenera were analyzed by using SDS-PAGE and restriction fragment length polymorphism (RELP) methodologies. Sixty-eight bands of alcohol and salt soluble proteins and 85-133 RFLP markers were used to estimate the genetic distance among the species. These species were clustered together at around 0.1 to 0.4 level of distance for both kind of markers, indicating that Medicago species have a large genetic similarity. A combined cluster diagram, at a dissimilarity level of 0.3, differentiated nine species in four groups: group 1, M. littoralis , M. truncatulam, M.scutellata and M. rigidula; group 2, M. sativa ; group 3, M. lupulina ; group 4, M. orbicularis, M. radiata and M. minima. All of them, but except for M. minima. corrensponded to the existing four subgenera of the genus Medicago classified by Lesins and Lesins(1979).The most similar species were M. littoralis and M. trucatula and the most dissimilar one was M. lupulina. In separate cluster diagrams based on RFLP and protein markers, some differences were observed. In the case of RFLP or DNA markers, M. sativa (alfalfa) was distantly clustered with other Medicago species. But in the case of protein markers, M. sativa was closely clustered with M. scutellata, M. littorulis and M. truncatula.

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Production and Secretion of Human Interleukin-18 in Transgenic Tobacco Cell Suspension Culture

  • Sharma, Niti;Kim, Tae-Geum;Yang, Moon-Sik
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.2
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    • pp.154-159
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    • 2006
  • Interleukin-18 (IL-18), otherwise known as interferon-gamma-inducing factor (IGIF), is one of several well characterized and important cytokines that contribute to host defenses. The complementary DNA (cDNA) of mature human interleukin-18 gene (hIL-18) was fused with the signal peptide of the rice amylase 1A gene (Ramy1A) and introduced into the plant expression vector under the control of a duplicated CaMV 35S promoter. The recombinant plasmid was transformed into tobacco (Nicotiana tabacum L. cv Havana) using the Agrobacterium-mediated transformation method. The integration of the hlL-18 gene into the genome of transgenic tobacco plants was confirmed by polymerase chain reaction (PCR) amplification and its expression was observed in the suspension cells that were derived from the transgenic plant callus by using Northern blot analysis. The hlL-18 protein was detected in the extracts of the transgenic callus and in the medium of the transgenic tobacco suspension culture by using immunoblot analysis. Based upon enzyme-linked immunosorbant assay (ELISA) results, the expression level of the hlL-18 protein approximated $166{\mu}g/L$ in the suspension culture medium. Bioassay results from the induction of $interferon-{\gamma}$ from a KG-1 cell line indicated that the hlL-18 secreted into the suspension culture medium was bioactive.

Diversity Evaluation of Xylella fastidiosa from Infected Olive Trees in Apulia (Southern Italy)

  • Mang, Stefania M.;Frisullo, Salvatore;Elshafie, Hazem S.;Camele, Ippolito
    • The Plant Pathology Journal
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    • v.32 no.2
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    • pp.102-111
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    • 2016
  • Olive culture is very important in the Mediterranean Basin. A severe outbreak of Olive Quick Decline Syndrome (OQDS) caused by Xylella fastidiosa infection was first noticed in 2013 on olive trees in the southern part of Apulia region (Lecce province, southern Italy). Studies were carried out for detection and diversity evaluation of the Apulian strain of Xylella fastidiosa. The presence of the pathogen in olive samples was detected by PCR amplifying the 16S rDNA, gyrase B subunit (gyrB) and HL hypothetical protein genes and single nucleotide polymorphisms (SNPs) assessment was performed to genotype X. fastidiosa. Twelve SNPs were recorded over gyrB and six SNPs were found for HL gene. Less variations were detected on 16S rDNA gene. Only gyrB and HL provided sufficient information for dividing the Apulian X. fastidiosa olive strains into subspecies. Using HL nucleotide sequences was possible to separate X. fastidiosa into subspecies pauca and fastidiosa. Whereas, nucleotide variation present on gyrB gene allowed separation of X. fastidiosa subsp. pauca from the other subspecies multiplex and fastidiosa. The X. fastidiosa strain from Apulia region was included into the subspecies pauca based on three genes phylogenetic analyses.

Interactions Between Genetic Risk Score and Healthy Plant Diet Index on Cardiometabolic Risk Factors Among Obese and Overweight Women

  • Fatemeh Gholami;Mahsa Samadi;Niloufar Rasaei;Mir Saeid Yekaninejad;Seyed Ali Keshavarz;Gholamali Javdan;Farideh Shiraseb;Niki Bahrampour;Khadijeh Mirzaei
    • Clinical Nutrition Research
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    • v.12 no.3
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    • pp.199-217
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    • 2023
  • People with higher genetic predisposition to obesity are more susceptible to cardiovascular diseases (CVDs) and healthy plant-based foods may be associated with reduced risks of obesity and other metabolic markers. We investigated whether healthy plant-foods-rich dietary patterns might have inverse associations with cardiometabolic risk factors in participants at genetically elevated risk of obesity. For this cross-sectional study, 377 obese and overweight women were chosen from health centers in Tehran, Iran. We calculated a healthy plant-based diet index (h-PDI) in which healthy plant foods received positive scores, and unhealthy plant and animal foods received reversed scores. A genetic risk score (GRS) was developed based on 3 polymorphisms. The interaction between GRS and h-PDI on cardiometabolic traits was analyzed using a generalized linear model (GLM). We found significant interactions between GRS and h-PDI on body mass index (BMI) (p = 0.02), body fat mass (p = 0.04), and waist circumference (p = 0.056). There were significant gene-diet interactions for healthful plant-derived diets and BMI-GRS on high-sensitivity C-reactive protein (p = 0.03), aspartate aminotransferase (p = 0.04), alanine transaminase (p = 0.05), insulin (p = 0.04), and plasminogen activator inhibitor 1 (p = 0.002). Adherence to h-PDI was more strongly related to decreased levels of the aforementioned markers among participants in the second or top tertile of GRS than those with low GRS. These results highlight that following a plant-based dietary pattern considering genetics appears to be a protective factor against the risks of cardiometabolic abnormalities.