• Title/Summary/Keyword: Agrobacterium tumefaciens.

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Isolation of Coat Protein Gene from Cucumber Mosaic Virus and Its Introduction into Tobacco (오이 모자이크 바이러스 외피 단백질 유전자 분리 및 담배로의 형질전환)

  • 손성한;김경환;김영태;박종석;김주곤;이광웅;황영수
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.3
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    • pp.149-155
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    • 1995
  • The coat protein (CP) gene was cloned from RNA genome of the Cucumber Mosaic Virus strain ABI (CMV-ABI) isolated in Korea. The comparisons of the nucleotide sequence of the cloned CP gene and its deduced amino acid sequences with other CP genes revealed that the CMV-ABI belongs to subgroup I (type I), CMV-ABI developed the typical mosaic symptom in infected plants. Tobacco plants (Samsun and NC82) were transformed by leaf-disc transformation via Agrobacterium, temefaciens LB4404 harboring pVCP, witch CMV-ABI CP gene was inserted into the pBI121, and a number of mature transgenic tobacco plants were developed. Southern and PCR analysis of genomic DNA from the transgenic plants showed that the CP gene was integrated into the genomes of the most of the transgenic plant. Result of the segregation patterns of resistance in T1 seedlings of the plants to kanamycin showed that the transgenic plants containing l,2 and 3 copies of CP gene were50%, 39% and 11% of the total transgenic plants, respectively.

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Transgenic Expression of MsHsp23 Confers Enhanced Tolerance to Abiotic Stresses in Tall Fescue

  • Lee, Ki-Won;Choi, Gi-Jun;Kim, Ki-Yong;Ji, Hee-Jung;Park, Hyung-Soo;Kim, Yong-Goo;Lee, Byung-Hyun;Lee, Sang-Hoon
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.6
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    • pp.818-823
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    • 2012
  • Tall fescue (Festuca arundinacea Schreb.) is an important cool season forage plant that is not well suited to extreme heat, salts, or heavy metals. To develop transgenic tall fescue plants with enhanced tolerance to abiotic stress, we introduced an alfalfa Hsp23 gene expression vector construct through Agrobacterium-mediated transformation. Integration and expression of the transgene were confirmed by polymerase chain reaction, northern blot, and western blot analyses. Under normal growth conditions, there was no significant difference in the growth of the transgenic plants and the non-transgenic controls. However, when exposed to various stresses such as salt or arsenic, transgenic plants showed a significantly lower accumulation of hydrogen peroxide and thiobarbituric acid reactive substances than control plants. The reduced accumulation of thiobarbituric acid reactive substances indicates that the transgenic plants possessed a more efficient reactive oxygen species-scavenging system. We speculate that the high levels of MsHsp23 proteins in the transgenic plants protect leaves from oxidative damage through chaperon and antioxidant activities. These results suggest that MsHsp23 confers abiotic stress tolerance in transgenic tall fescue and may be useful in developing stress tolerance in other crops.

Fungal pathogen protection in transgenic lettuce by expression of a apoptosis related Bcl-2 gene (Apoptosis 관련 Bcl-2유전자의 도입을 통한 곰팡이 저항성 형질전환 상추의 육성)

  • Seo, Kyung-Sun;Min, Byung-Whan
    • Journal of Plant Biotechnology
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    • v.38 no.3
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    • pp.209-214
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    • 2011
  • Transgenic lettuce plants were successfully obtained from hypocotyl explants inoculated with Agrobacterium tumefaciens, which harbored a binary vector plasmid with Bcl-2 gene, related to apoptosis. After culture and selection on MS medium a number of kanamycin-resistant plantlets were regenerated. Polymerase chain reaction, Southern blot analysis and Northern blot analysis were used to identify and characterize the transgenic plants with the integrated Bcl-2 gene. Over 100 transgenic plants have been established in soil and flowered in the greenhouse. T1 progeny of 100 transgenic lettuce inbred lines were inoculated with Sclerotinia sclerotiorum. Expression of the Bcl-2 peptide in transgenic lettuce plants provides high levels of field resistance against Sclerotinia sclerotiorum, causal agent of the agronomically important fungal disease of lettuce.

Factors and Cause of Rusty-Ginseng Occurrence (인삼 적변의 발생 요인과 원인)

  • Park Hong-Woo;Lim Tai-Kyo;Choi Chun-Hwan;Choi Jae-Eul
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.5
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    • pp.396-400
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    • 2006
  • In the preliminary experiment, the factors(soil moisture content, type of organic fertilizers, wound of ginseng, and concentration of rusty-root causing bacteria) inducing the rusty-root of ginseng were investigated. The Hue values were measured for the degree of rustiness affected by the factors. The rustiness was severe on the wounded-ginseng and on the ginseng inoculated with Agrobacterium tumefaciens (CG20126). The Hue values of the control, the non-wounded ginseng inoculated with CG20126, the wounded-ginseng, and the wounded-ginseng inoculated with CG20126 were 113.3, 108.1, 85.8, and 57.5, respectively. The Hue value of the horticulture bed-soil was 56.8 whereas the value of the paddy bed-soil counterpart was 64.7. The Hue values on the ginseng roots grown in the soil containing 10%, 20%, 50%, and 70% of moisture, were 96.2, 85.9, 78.0, and 75.7, respectively. The organic fertilizer increased the rustiness of ginseng and the range of Hue values was 35.2-27.8. The increased concentrations of A. tumefaciens CG20126 increased the rustiness of ginseng. The concentration of A. tumefaciens CG20126 at $10^{2},\;10^{4},\;10^{6},\;and\;10^{8}cells/m{\ell}$ resulted in the Hue values of 62.8, 63.3, 55.6, and 48.8, respectively.

Salt Tolerance in Transgenic Pea (Pisum sativum L.) Plants by P5CS Gene Transfer

  • Najafi F.;Rastgar-jazii F.;Khavari-Nejad R. A.;Sticklen M.
    • Journal of Plant Biotechnology
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    • v.7 no.4
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    • pp.233-240
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    • 2005
  • Slices of embryonic axis of mature pea (Pisum sativum L. cv. Green Arrow) seeds were used as explant. Transformation of explants was done via Agrobacterium tumefaciens bearing vector pBI-P5CS construct. The best results for inoculation of explants were obtained when they were immersed for 90 s at a concentration of $6{\times}10^8$ cell $ml^(-1)$ of bacterial suspension. Transformed pea plants were selected on $50\;mg\;l^(-1)$ kanamycin and successful transformants were confirmed by PCR and blotting. Transgenic plants were further analyzed with RT-PCR to confirm the expression of P5CS. Transgenic plants and non-transgenic plants were treated with different concentrations of NaCl 0 (control), 100, 150 and 200 mM in culture medium. Measurement of proline content indicated that transgenic plants produced more amino acid proline in response to salt in comparison with non-transgenic plants. Photosynthetic efficiency in transgenic plants under salt-stress was more than that of non-transgenic plants.

Production of Bacterial Quorum Sensing Antagonists, Caffeoyl- and Feruloyl-HSL, by an Artificial Biosynthetic Pathway

  • Kang, Sun-Young;Kim, Bo-Min;Heo, Kyung Taek;Jang, Jae-Hyuk;Kim, Won-Gon;Hong, Young-Soo
    • Journal of Microbiology and Biotechnology
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    • v.27 no.12
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    • pp.2104-2111
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    • 2017
  • A new series comprising phenylacetyl-homoserine lactones (HSLs), caffeoyl-HSL and feruloyl-HSL, was biologically synthesized using an artificial de novo biosynthetic pathway. We developed an Escherichia coli system containing artificial biosynthetic pathways that yield phenylacetyl-HSLs from simple carbon sources. These artificial biosynthetic pathways contained the LuxI-type synthase gene (rpaI) in addition to caffeoyl-CoA and feruloyl-CoA biosynthetic genes, respectively. Finally, the yields for caffeoyl-HSL and feruloyl-HSL were $97.1{\pm}10.3$ and $65.2{\pm}5.7mg/l$, respectively, by tyrosine-overproducing E. coli with a $\text\tiny{L}$-methionine feeding strategy. In a quorum sensing (QS) competition assay, feruloyl-HSL and p-coumaroyl-HSL antagonized the QS receptor TraR in Agrobacterium tumefaciens NT1, whereas caffeoyl-HSL did not.

Newly Recorded Problematic Plant Diseases in Korea and Their Causal Pathogens

  • Kwon, Jin-Hyeuk
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.25-27
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    • 2003
  • Since 1993, a total of 50 problematic plant diseases unrecorded in Korea were surveyed in Gyeongnam province. Totally 34 new host plants to corresponding pathogens investigated in this study were 5 fruit trees, 9 vegetables, 12 ornamental plants, 3 industrial crops, and 5 medicinal plants. Among the newly recorded fruit tree diseases, fruit rot of pomegranate caused by Coniella granati and Rhizopus soft rot of peach caused by Rhizopus nigricans damaged severely showing 65.5% and 82.4% infection rate. Among the vegetable diseases, corynespora leaf spot of pepper caused by Corynespora cassiicola and the crown gall of pepper caused by Agrobacterium tumefaciens, powdery mildew of tomato caused by Oidiopsis taurica were the most severe revealing 47.6%, 84.7%, and 54.5% infection rate in heavily infected fields, respectively. In ornamental plants, collar rot of lily caused by Sclerotium rolfsii, gray mold of primula caused by Botrytis cinerea, soot leaf blight of dendrobium caused by Pseudocercospora dendrobium, sclerotinia rot of obedient plant caused by Sclerotinia sclerotiorum showed 32.7 to 64.8% disease incidence. On three industrial plants such as sword bean, broad bean, and cowpea, eight diseases were firstly found in this study. Among the diseases occurring on broad bean, rust caused by Uromyces viciae-fabae and red spot caused by Botrytis fabae were the major limiting factor for the cultivation of the plant showing over 64% infection rate in fields. In medicinal plants, anthracnose of safflower caused by Collectotrichum acutatum was considered the most severe disease on the plant and followed by collar rot caused by Sclerotium rolfsii.(중략)

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Transgenic Lettuce Expressing Chalcone Isomerase Gene of Chinese Cabbage Increased Levels of Flavonoids and Polyphenols

  • Han, Eun-Hyang;Lee, Ji-Sun;Lee, Jae-Woong;Chung, In-Sik;Lee, Youn-Hyung
    • Horticultural Science & Technology
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    • v.29 no.5
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    • pp.467-473
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    • 2011
  • Flavonoid are large group of the polyphenolic compounds which are distinguished by an aromatic or phenolic ring structure and the phenolic compounds are induced by microbial infection, ultraviolet radiation, temperature and chemical stress. They are known for their antioxidant activity, anti-allergic, anti-inflammatory, anti-microbial and anti-cancer activities. In this study, changes in flavonoid content were investigated using heterologous chalcone isomerase (CHI) expression system. Also, phenolic compounds level was measured to examine the relation between flavonoids and phenols contents. Explants of lettuce (Lactuca sativa L.) were transformed with Agrobacterium tumefaciens LBA 4404 strain containing pFLH-CHI (derived from pPZP2Ha3) vector constructed with CHI gene from Brassica rapa. The putative transgenic plants were confirmed by genomic DNA PCR analysis. Also the transcription levels of the gene were analyzed by semi-quantitative RT-PCR with gene specific primers. The total flavonoid contents were increased at $T_0$ and $T_1$ generations over 1.4 and 4.0 fold, respectively. Total phenol contents also increased at $T_1$ generation. These results indicate that CHI gene plays an important role to regulate the accumulation of flavonoids and its component changes.

Agroinfiltration-based Potato Virus X Replicons to Dissect the Requirements of Viral Infection

  • Park, Sang-Ho;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.22 no.4
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    • pp.386-390
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    • 2006
  • Extensive research of the Potato virus X(PVX) has been performed in in vitro transcription system using the bacteriophage T7 promoter. We constructed an efficient T-DNA based binary vector, pSNU1, and modified vectors carrying PVX replicons. The suitability of the construct to transiently express PVX RNA using Agrobacterium tumefaciens was tested by analysis of infectivity in plants. The expressed PVX RNA was infectous and systemically spread in three plant species including Nicotiana benthamiana, N. tabacum cv. Xanthi-nc, and Capsicum annuum cv. Chilsungcho. The PVX full length construct, pSPVXp31, was caused severe mosaic symptoms on N. benthamiana, severe necrotic lesions on C. annuum while milder symptoms and delayed mosaic symptoms were appeared on the systemic leaves on N. tabaccum. RT-PCR analysis confirmed the presence of PVX RNAs on both inoculated and systemic leaves in all three plant species tested. Our results indicated that PVX replicons were efficiently expressed PVX RNA in at least three tested species. Further investigation win be needed to elucidate the mechanism of PVX replication, translation, movement and assembly/disassembly processes.

lacZ- and aph-Based Reporter Vectors for In Vivo Expression Technology

  • Baek, Chang-Ho;Kim, Kun-Soo
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.872-880
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    • 2003
  • Three vectors, pSG1, 2, and 3, which facilitate in vivo expression technology (IVET) in Gram-negative bacteria, were developed. Vectors pSG1and 2 are derivatives of ColE1, and pSG3 is a derivative of an R6K replicon. These vectors contain oriT sites that allow mobilization when the RK2 Tra functions are provided in trans. These vectors contain promoterless lacZ (pl-lacZ) and promoterless aph (pl-aph) transcriptionally fused together, which allow qualitative and quantitative measurements of the expression of genes in the genome of bacterial cells. pSG1 and 3 contain gentamicin-resistance genes, and pSG2 carries a streptomycin-/spectinomycin-resistance gene, allowing for selection of recombinants generated by a single crossover between a library fragment cloned into a pSG vector and the identical region in the genome of a bacterial species from which the library fragment originated. These vectors were successfully applied to the generation of random fusions at high rates in the genomes of four representative Gram-negative bacteria. In addition, the expression level of ${\beta}-galactosidase$ and the degree of resistance to kanamycin in cells with fusions generated by these vectors were found to be linearly correlated, proving that these vectors can be used for IVET.