• Title/Summary/Keyword: cauliflower

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Mass production and application of activation tagged hairy root lines for functional genomic of secondary metabolism in ginseng

  • Choi, Dong-Woog;Chung, Hwa-Jee;Ko, Suk-Min;In, Dong-Soo;Song, Ji-Sook;Woo, Sung-Sick;Liu, Jang R.
    • Journal of Plant Biotechnology
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    • v.36 no.3
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    • pp.294-300
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    • 2009
  • Activation tagging that uses T-DNA vectors containing multimerized transcriptional enhancers from the cauliflower mosaic virus (CaMV) 35S gene is a powerful tool to determine gene function in plants. This approach has been successfully applied in screening various types of mutations and cloning the corresponding genes. We generated an activation tagged hairy root pool of ginseng (Panax ginseng C.A. Meyer) in an attempt to isolate genes involved in the biosynthetic pathway of ginsenoside (triterpene saponin), which is known as the major active ingredient of the root. Quantitative and qualitative variation of ginsenoside in activation tagged hairy root lines were profiled using LC/MS. Metabolic profiling data enabled selection of a specific hairy root line which accumulated ginsenoside at a higher level than other lines. The relative expression level of several genes of triterpene biosynthetic pathway in the selected hairy root line was determined by real time RT-PCR. Overall results suggest that the activation tagged ginseng hairy root system described in this study would be useful in isolating genes involved in a complex metabolic pathway from genetically intractable plant species by metabolic profiling.

Overexpression of Cuphea viscosissima CvFatB4 enhances 16:0 fatty acid accumulation in Arabidopsis

  • Yeon, Jinouk;Park, Jong-Sug;Lee, Sang Ho;Lee, Kyeong-Ryeol;Yi, Hankuil
    • Journal of Plant Biotechnology
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    • v.46 no.4
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    • pp.282-290
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    • 2019
  • Cuphea viscosissima plants accumulate medium-chain fatty acids (MCFAs), i.e., those containing 8 ~ 14 carbons, in their seeds, in addition to the longer carbon chain fatty acids (≥16 carbons) found in a variety of plant species. Previous studies have reported the existence of three C. viscosissima MCFA-producing acyl-acyl carrier protein (ACP) thioesterases with different substrate specificities. In this study, CvFatB4, a novel cDNA clone encoding an acyl-ACP thioesterase (EC 3.1.2.14), was isolated from developing C. viscosissima seeds. Sequence alignment of the deduced amino acid sequence revealed that four catalytic residues for thioesterase activity are conserved and a putative N-terminal chloroplast transit peptide is present. Overexpression of CvFatB4 cDNA, which was under the control of the cauliflower mosaic virus 35S promoter, in Arabidopsis thaliana led to an increase in 16:0 fatty acid (palmitate) levels in the seed oil at the expense of 18:1 and other non-MCFAs.

Plant Protein Improvement by Synthetic Gene (합성유전자를 이용한 식물단백질의 향상)

  • 김태금;양문식
    • KSBB Journal
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    • v.7 no.3
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    • pp.155-160
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    • 1992
  • To improve the nutritional quality of plant proteins, a synthetic gene, called HEAAE (high essential amino acid encoding)-DNA, was introduced and expressed in tobacco plants. The synthetic gene, which is 292 basepair-long, codes for a protein composed of about 80% essential amino acids. To improve its expression level in plants, Cauliflower Mosaic Virus (CaMV) 355 and CaMV duplicate 35S promoters which are known as strong promoters were used with Nopaline Synthase promoter as a control. Transformed and regenerated tobacco plants were subject to analysis for introduction and expression of this gene. Integration of the gene into the plant genome and its expression into mRNAs and its proteins have been demonstrated using Southern, northern blot analysis and amino acid analysis. The differences of expression levels among CaMV duplicate 35S, CaMV 35S and Nopaline Synthase promoters are significant in term of mRNAs, but not in terms of proteins.

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Increase in Linolenate Contents by Expression of the fad3 Gene in Transgenic Tobacco Plants

  • Kang, Young-Hwi;Min, Bok-Kee;Park, Hee-Sung;Lim, Kyung-Jun;Huh, Tae-Lin;Lee, Se-Yong
    • BMB Reports
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    • v.29 no.4
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    • pp.308-313
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    • 1996
  • An 1.4 kb of the fad3 cDNA encoding microsomal linoleic acid desaturase catalyzing the conversion of linoleic acid (18:2, ${\omega}-6$) to linolenic acid (18:2, ${\omega}-3$) was introduced into tobacco plants by the Agrobacterium-mediated plant transformation, Among the transgenic tobacco plants conferring kanamycin resistance, five transformants showing increment in unsaturated fatty acid contents were selected and further analyzed for the transgenecity, In genomic Southern blot analyses, copy numbers of the integrated fad3 DNA in chromosomal DNA of the five transgenic tobacco plants were varied among the transgenic lines. By Northern blot analyses, the abundancy of the fad3 mRNA transcript directed by Cauliflower Mosaic Virus 35S promoter was consistent with the relative copy number of the fad3 DNA integrated in the chromosome of transgenic tobacco plants. When compared with the wild type, accumulation of linolenic acid in transgenic tobacco roots was elevated 3.7- to 4.7-fold showing a corresponding decrease in the linoleic acid contents; however, slight increments for linolenic acid were noticed in transgenic leaf tissues. These results indicated that the elevated level of fad3 expression is achieved in transgenic tobacco plants.

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Quantitative Analysis of Two Genetically Modified Maize Lines by Real-Time PCR

  • Lee Seong-Hun;Kang Sang-Ho;Park Yong-Hwan;Min Dong-Myung;Kim Young-Mi
    • Journal of Microbiology and Biotechnology
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    • v.16 no.2
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    • pp.205-211
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    • 2006
  • A quantitative analytical method to detect new lines of genetically modified (GM) maize, NK603 and TC1507, has been developed by using a real-time polymerase chain reaction (PCR). To detect these GM lines, two specific primer pairs and probes were designed. A plasmid as a reference molecule was constructed from an endogenous DNA sequence of maize, a universal sequence of a cauliflower mosaic virus (CaMV) 35S promoter used in most GMOs, and each DNA sequence specific to the NK603 and TC1507 lines. For the validation of this method, the test samples of 0, 0.1, 0.5, 1.0, 3.0, 5.0, and 10.0% each of the NK603 and TC1507 GM maize were quantitated. At the 3.0% level, the biases (mean vs. true value) for the NK603 and TC1507 lines were 3.3% and 15.7%, respectively, and their relative standard deviations were 7.2% and 5.5%, respectively. These results indicate that the PCR method developed in this study can be used to quantitatively detect the NK603 and TC1507 lines of GM maize.

Treatment of Various Auricular Diseases Using Magnetic Disks (자석 압박을 이용한 다양한 이개 질환의 치료)

  • Seo, Hyo Seuk;Hong, Yoon Gi;Chang, Choong Hyun
    • Archives of Plastic Surgery
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    • v.34 no.3
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    • pp.371-376
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    • 2007
  • Purpose: The earlobe is one of the areas which are most vulnerable to trauma. Various auricular diseases need compression treatment. We report a new compression method using magnetic disks. Methods: Seventeen patients with auricular diseases were treated from October 2002 to September 2006. The mean age was 29.1 years. The diseases details were osteochondroma in 2 patients; cauliflower's ears in 2 patients; acute otohematoma in 1 patient; and hypertrophic scars in 11 patients. The most common cause of their disease was ear piercing. The mean follow-up period was 8.9 months. All surgical procedures were performed under local anesthesia. To compress immediately, a pair of magnetic disks was applied to the anterior and posterior surface of the earlobe. Results: The results were generally good. Major complications, such as recurrence, necrosis, dehiscence, or infection, did not occur. Conclusion: A pair of magnetic disks are useful compression tool in various auricular diseases.

Incidence, Pathogenicity of Clubroot Fungus(Plasmodiophora brassicae) and Varietal Resistance in Chinese Cabbage (배추 무사마귀병의 발생상황과 병원균(Plasmodiophora brassicae)의 병원성 및 배추품종의 병저항성)

  • 김두욱;오정행
    • Korean Journal Plant Pathology
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    • v.13 no.2
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    • pp.95-99
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    • 1997
  • To obtain a basic information of breeding for resistance to clubroot in Chinese cabbage, disease incidence, pathogenicity, and varietal response to the pathogen were studied. Incidence of clubroot was observed at 3 districts in Gyeonggi-Do, 2 districts in Kangwon-Do, and 1 district each in Gyeongnam, Geongbuk and Jeonbuk, respectively. Disease infection rate and diseased ara were most severe in northern part of Gyeonggi-Do. The isolates of clubroot collected from 8 different districts were not different in their virulence one another in view of their infection rate and disease severity in Chinese cabbage. The clubroot fungus had a wide host range for the cruciferous vegetables. Disease severity was high in rape, turnip and mustard, moderate in Chinese cabbage and broccoli, and low in kale and cauliflower. All of Korean hybrids of Chinese cabbage tested were highly susceptible to clubroot, but Japanese varieties were resistant to the highly pathogenic isolate (EJ-93) which was isolated from the Chinese cabbage in Korea. The hybrid(F1) between clubroot resistant line(930WG) and the susceptible line(332MS) showed completely resistant reaction, which indicated that clubroot resistance was governed by a dominant gene.

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Construction of a Plant Expression Vector for the Coat Protein Gene of Cucumber Mosaic Virus-As Strain for Plant Transformation (오이 모자이크 바이러스 As계통 외피단백질 유전자의 식물체 형질질환을 위한 발현벡타의 구축)

  • 류기현;박원목
    • Korean Journal Plant Pathology
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    • v.11 no.1
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    • pp.66-72
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    • 1995
  • The coat protein (CP) gene of cucumber mosaic virus-As (CMV-As) strain was engineered for expression in the plant by using the cauliflower mosaic virus 35S transcript regulatory sequences. The CP gene was cloned into an Agrobacterium-derived binary vector. A chimeric gene was constructed by the cDNA of CMV-As CP and plant expression vector pBI121. The clone, pCMAS66, was first introduced into the phagemid vector pSPORT1 for situating sense orientation for translation and making restriction sites in order to re-introduce plant expression vector, pHI121. The resulting subclone pCASCP02 and plant expression vector pBI121 were treated with BamHI-SacI for excising the target gene and removing GUS gene, respectively. After Agrobacterium transformation by freeze-thaw technique, the clone, pCMASCP121-123 which contains sense orientation of the target gene, was selected and confirmed by restriction endonuclease analysis. The CMV-As CP gene was introduced into A. tumefaciens. The results on tobacco plant transformation with the vector system revealed that the system could be successfully introduced and showed high frequency of selection to putative transformations.

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Molecular and Biological Characterization of an Isolate of Cucumber mosaic virus from Glycine soja by Generating its Infectious Full-genome cDNA Clones

  • Phan, Mi Sa Vo;Seo, Jang-Kyun;Choi, Hong-Soo;Lee, Su-Heon;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.30 no.2
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    • pp.159-167
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    • 2014
  • Molecular and biological characteristics of an isolate of Cucumber mosaic virus (CMV) from Glycine soja (wild soybean), named as CMV-209, was examined in this study. Comparison of nucleotide sequences and phylogenetic analyses of CMV-209 with the other CMV strains revealed that CMV-209 belonged to CMV subgroup I. However, CMV-209 showed some genetic distance from the CMV strains assigned to subgroup IA or subgroup IB. Infectious full-genome cDNA clones of CMV-209 were generated under the control of the Cauliflower mosaic virus 35S promoter. Infectivity of the CMV-209 clones was evaluated in Nicotiana benthamiana and various legume species. Our assays revealed that CMV-209 could systemically infect Glycine soja (wild soybean) and Pisum sativum (pea) as well as N. benthamiana, but not the other legume species.

Analysis of Flavonoid 3',5'-Hydroxylase Gene in Transgenic Petunia (Petunia hybrida) Plants (형질 전환된 페튜니아 식물체에서의 Flavonoid 3',5' -Hydroxylase 유전자의 분석)

  • 김영희
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.5
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    • pp.323-327
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    • 1998
  • The flavonoid biosynthetic pathway has been studied as a genetic model system, particularly in Petunia hybrida. In order to study the flavonoid biosynthetic pathway, we constructed a fusion gene system between Cauliflower Mosaic Virus (CaMV) 35S promoter and eggplant flavonoid 3', 5'-hydroxylase in pBI 121 plasmid. An optimal condition for plant regeneration was observed when internode explants were cultured on MS medium supplemented with IAA 0.2 mg/L plus BA 3 mg/L. For plant transformation internode explants of Petunia hybrida were precultured on BM medium supplemented with IAA 0.2 mg/L plus BA 3 mg/L. Putative transgenic plants were selected on medium containing kanamycin 50 mg/L plus cefotaxim 300 mg/L. Putative selected transformants were confirmed by amplification of selectable marker gene (nptII) by polymerase chain reaction (PCR) and Southern hybridization of flavonoid 3',5'-hydroxylase gene.

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