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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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Journal of Plant Biotechnology
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Journal DOI :
The Korean Society of Plant Biotechnology
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Volume & Issues
Volume 34, Issue 4 - Dec 2007
Volume 34, Issue 3 - Sep 2007
Volume 34, Issue 2 - Jun 2007
Volume 34, Issue 1 - Mar 2007
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Inhibition of Starch Biosynthesis by Antisense Expression of cDNAs Encoding ADP-Glucose Pyrophosphorylase Small Subunit in Sweetpotato
Min, Sung-Ran ; Bae, Jung-Myung ; Harn, Chee-Hark ; Jeong, Won-Joong ; Lee, Young-Bok ; Liu, Jang-Ryol ;
Journal of Plant Biotechnology, volume 34, issue 4, 2007, Pages 277~283
DOI : 10.5010/JPB.2007.34.4.277
Embryogenic calluses derived from shoot apical meristem explants of sweetpotato were subjected to particle bombardment to generate transgenic plants for antisense expression of cDNAs encoding two different AGPase small subunit (ibAGP1 and ibAGP2). Plants were generated via somatic embryogenesis. PCR and Southern analysis demonstrated that the incorporation of ibAGP1 and ibAGP2 into the genome in an antisense orientation. Immunoblot analysis confirmed reduced levels of AGPase small subunit in transgenic plant leaves. Plants with both ibAGP1 and ibAGP2 produced a lower level of the protein than plants with ibAGP1 alone. iodine test demonstrated that transgenic plant leaves and storage root accumulated reduced amounts of starch. Iodine staining of leaf tissues indicated that transgenic plants accumulated less amount of starch than control. In accordance with western blot analysis, plants with both ibAGP1 and ibAGP2 accumulated a lower amount of starch than plants with ibAGP1 alone. Both transgenic plants exhibited a severely retarded growth, resulting in bare survival. It is suggested that disrupted expression of the gene encoding AGPase small subunit is lethal to the growth of sweetpotato contrast to other species including potato.
Mass Production of Gain-of-Function Mutants of Hair Roots in Ginseng
Ko, Suk-Min ; In, Dong-Soo ; Chung, Hwa-Jee ; Choi, Dong-Woog ; Liu, Jang-Ryol ;
Journal of Plant Biotechnology, volume 34, issue 4, 2007, Pages 285~291
DOI : 10.5010/JPB.2007.34.4.285
This study describes conditions for the mass production of activation-tagged mutant hairy root lines of ginseng by cocultivation with Agrobacterium rhizogenes. Because it is not currently possible to produce progeny from transgenic ginseng, a loss-of-function approach for functional genomics cannot be appliable to this species. A gain-of-function approach is alternatively the choice and hairy root production by cocultivation of A. rhizogenes would be most practical to obtain a large number of mutants. Various sources of explants were subjected to genetic transformation with various strains of A. rhizogenes harboring the activation-tagging vector pKH01 to determine optimum conditions for the highest frequency of hairy root formation on explants. Petiole explants cocultivated with A. rhizogenes R1000 produced hairy roots at a frequency of 85.9% after 4 weeks of culture. Conditions for maximum growth or branching rate of hairy roots were also investigated by using various culture media. Petiole explants cultured on half strength Schenk and Hildebrandt medium produced vigorously growing branched roots at a rate of 2.6 after 4 weeks of culture. A total of 1,989 lines of hairy root mutants were established in this study. These hairy root lines will be useful to determine functions of genes for biosynthesis of ginsenosides.
An Efficient Plant Regeneration and Transformation System of Robinia pseudoacacia var. umbraculifera for Phytoremediation
Kwon, Hye-Jin ; Woo, Seong-Min ; Seul, Eun-Jun ; Kim, Teh-Ryung ; Shin, Dong-Un ; Kim, Hag-Hyun ;
Journal of Plant Biotechnology, volume 34, issue 4, 2007, Pages 293~298
DOI : 10.5010/JPB.2007.34.4.293
Robinia pseudoacacia var. umbraculifera, commonly called umbrella black locust were regenerated after co-cultivation of internode segments with Agrobacterium tumefaciens which included yeast cadmium factor 1 (YCF 1) gene. The tolerance to cadmium and lead for plants can be increased by the YCF1 gene expression. Moreover, the recent studies have shown that YCF1 gene transgenic plants increase the accumulation of cadmium and lead into plant vacuoles. The effect of plant growth regulator such as 2,4-dichlorophenoxyacetic acid (2,4-D),
-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and thidiazuron (TDZ) were studied to evaluate the propagation of plants through internode explants. The efficient induction of multiple adventitious shoots and callus were observed on a medium supplemented with 0.1 mg/L TDZ + 0.2 mg/L BA. To induce shoot elongation and rooting, regenerated shoots were transferred into basal MS medium without any plant growth regulator. Successful Agrobacterium tumefaciens mediated transformation was obtained by 20 min vacuum-infiltration with
acetosyringone on the optimal multiple shoot induction medium with 30 mg/L hygromycin and 300 mg/L cefotaxime. To confirm the integration and expression of transgene, Polymerase Chain Reaction (PCR) and Reverse Transcriptase PCR (RT-PCR) were performed with specific primers. The frequency of transformation was approximately 18.94%. This study can be used to genetic engineering of phytoremediator.
Enhanced Tolerance to Oxidative Stress of Transgenic Potato (cv. Superior) Plants Expressing Both SOD and APX in Chloroplasts
Tang, Li ; Kwon, Suk-Yoon ; Kim, Myoung-Duck ; Kim, Jin-Seog ; Kwak, Sang-Soo ; Lee, Haeng-Soon ;
Journal of Plant Biotechnology, volume 34, issue 4, 2007, Pages 299~305
DOI : 10.5010/JPB.2007.34.4.299
Oxidative stress is a major damaging factor for plants exposed to environmental stresses. Previously, we have generated transgenic potato (cv. Superior) plants expressing both CuZnSOD and APX genes in chloroplast under the control of an oxidative stress-inducible SWPA2 promoter (referred to as SSA plants) and selected the transgenic potato plant lines with tolerance against methyl viologen (MV)-mediated oxidative stress. When leaf discs of SSA plants were subjected to
methyl viologen (MV), they showed approximately 40% less damage than non-transgenic (NT) plants. SSA plantlets were treated with
MV stress, SSA plants also exhibited reduced damage in root growth. When 350 MV was sprayed onto the whole plants, SSA plants showed a significant reduction in visible damage, which was approximately 75% less damage than leaves of NT plants. These plants will be used for further analysis of tolerance to environmental stresses, such as high temperature and salt stress. These results suggest that transgenic potato (cv. Superior) plants would be a useful plant crop for commercial cultivation under unfavorable growth conditions.
Genetic Transformation of Chlamydomonas reinhardtii with the RNAi Suppressor p19 Gene of Tombus Virus
Jeong, Won-Joong ; Liu, Jang-Ryol ; Cerutti, Heriberto ;
Journal of Plant Biotechnology, volume 34, issue 4, 2007, Pages 307~312
DOI : 10.5010/JPB.2007.34.4.307
Chlamydomonas reinhardtii was transformed with the coding sequence of the Tombus virus gene p19 to determine whether the gene functions as an RNAi suppressor in C. reinhardtii. Transformants were confirmed to have 1 to several copies of p19 gene in their chromosomes. When an RNAi strain of C. reinhardtii generated by transforming the inverted repeat (IR) sequence homologous to the 3'UTR region of the MAA7 gene was re-transformed with the gene p19, MAA7 transcript levels of transformants fluctuated and proliferation of trans-formants on the medium containing 5-FI was suppressed. Overall results suggest that p19-mediated silencing suppression works at a low level in C. reinhardtii because of difference in codon usage resulting in weak P19 expression unless p19-mediated silencing suppression in C. reinhardtii works in a different manner from higher plants.
Biological Effect and Chemical Composition Variation During Self-Fermentation of Stored Needle Extracts from Pinus densiflora Siebold & Zucc.
Paudyal, Dilli P. ; Park, Ga-Young ; Hwang, In-Deok ; Kim, Dong-Woon ; Cheong, Hyeon-Sook ;
Journal of Plant Biotechnology, volume 34, issue 4, 2007, Pages 313~322
DOI : 10.5010/JPB.2007.34.4.313
Extract of Japanese red pine needles has been used in Asia pacific regions since long periods believing its valuable properties as tonic and ability of curing diseases of unidentified symptoms. Some selective compounds present in the extract and their effects were analyzed. Carbohydrates and vitamin c were identified using HPLC; terpenoid compounds by GC-MS; anti-bacterial analysis by paper discs, plates count and gastrointestinal motility by whole cell patch clamp. The extract is a mixture of compounds therefore its diverse effect was expected. Self-fermentation in extract proceeds after spontaneous appearance of yeast strains without inoculation. Effects and composition of the extract vary with varying period of self-fermentation. Extract inhibits the growth of bacteria dose dependently exhibiting its antibacterial properties however effectiveness increases with increase in fermentation period. The extract also can modulate gastrointestinal motility in murine small intestine by modulating pace maker currents in ICC mediated through ATP sensitive potassium channel.
Gene Silencing Induced by Cytosine Methylation in Transgenic Tomato
Jung, Seo-Hee ; Min, Sung-Ran ; Lee, Soo-Young ; Park, Ji-Young ; Davarpanah, S Javad ; Chung, Hwa-Jee ; Jeon, Jae-Heung ; Liu, Jang-Ryol ; Jeong, Won-Joong ;
Journal of Plant Biotechnology, volume 34, issue 4, 2007, Pages 323~329
DOI : 10.5010/JPB.2007.34.4.323
Transgene expression was analyzed in tomato plants. Four lines of neomycin phosphotransferase II gene (NPTII) and the trehalose biosynthetic fusion gene (TPSP) transformed
plants showed kanamycin resistance on selection medium. However, the analysis of phenotype (kanamycin resistance) and mRNA expression in
plants indicated that the expression of the NPTII and TPSP transgenes was down-regulated to an undetectable level in two independent lines 1 and 11. Southern analysis demonstrated that the lines 1 and 11 had multicopies of the transgenes, whereas the typical transgenic lines 2 and 10 had 1 or 2 copies. DNA methylation analysis using methylation sensitive enzyme detected accumulated CpG DNA methylation on TPSP coding region and CaMV35S promoter region in the line 11, but not the typical transgenic line 2. These results suggest that multicopy transgene in plants is attributed to down-regulation of the transgene expression via transcriptional gene silencing.
Microtuberization and Morphological Development by Culture Condition In Vitro Node Culture of Potato
Hwang, Hye-Yeon ; Lee, Young-Bok ;
Journal of Plant Biotechnology, volume 34, issue 4, 2007, Pages 331~338
DOI : 10.5010/JPB.2007.34.4.331
One-node stem pieces ca. 1 cm in length containing a axillary bud and a fully expanded leaf were obtained from it in vitro plants of potato (Solanum tuberosum L.). Leaves were removed and the nodes were cultured on the MS medium to investigate the effects of temperature, day length, sucrose, and CCC in microtuber formation and development. The fresh weight of microtubers after 80 days increased significantly at 8% sucrose and
. The tuberization and development were reduced at
except short-day treatment of 8 hours at 8% sucrose. The fresh weight and diameter were increased on the culture medium added CCC 500 mg/L. The potato tuberization was promoted under short daylength, and it showed great effect by treatment with the CCC. Though the tuberization was promoted at low temperature of
in a histologic change of an axillary bud part cell of a potato, the cells were able to observe the swelling growth. Swelling growth of tissue was stimulated in the darkness and was more remarkable by addition of CCC. In particular, in the visual ratio of cell division for each position in the tissue, the cortex part showed larger ratio of cell expansion than that of the pith part. The effect of CCC was identified at 8% sucrose in the darkness. The effect of CCC was not showed in sucrose 3% under long daylength of 16 hours. As a result, the fact of a substance with AGPase important for starch composition was certified by the result with the inclose of AGPase activity on high concentration of sucrose, CCC, and dark treatment by which tuber formation and development are promoted.
Effect of Boric Acid on In Vitro Pollen Germination in Transgenic Plants Expressing Monoclonal Antibodies
Ahn, Mi-Hyun ; Lee, Kyung-Jin ; Ko, Ki-Sung ;
Journal of Plant Biotechnology, volume 34, issue 4, 2007, Pages 339~346
DOI : 10.5010/JPB.2007.34.4.339
Pollen germination viability is an essential factor to produce seeds from pollination and fertilization, which are required to maintain plant generation. In this study, we tried to identify the effect of boric acid on pollen germination and tube grouch in non-transgenic and transgenic plants expressing monoclonal antibodies (anti-colorectal cancer mAb CO17-1A, anti-breast cancer mAb BR55, and anti-rabies virus mAb57). The pollen of non-transgenic plant was treated with different concentration of boric acid (0, 5, 10, 15, 20,
) in germination buffer to investigate its effect on in vitro pollen germination. At
of boric acid, tile pollen germination rate was the highest (49.5%) compared to other concentrations. In general, the germination rate significantly increased 3-10 folds in boric acid (
) treated group in non-transgenic and transgenic plants. Also, the pollen tube length increased in boric acid (
) treated groups. In the treated group, the pollen tube length increased until 3 h boric acid treatment and decreased after the 3 h, indicating that the 3 h is the most appropriate incubation time period. Western blot analysis showed that the mAb transgene expression was more stable in leaf than pollen in transgenic plants. This study suggested that
of boric acid is ideal concentration to induce in vitro pollen germination of transgenic plants expressing therapeutic monoclonal antibodies, indicating stable pollination and fertilization in transgenic plants.
Molecular Characterization and Event-Specific Marker Development of Insect Resistant Chinese Cabbage for Environmental Risk Assessment
Lim, Sun-Hyung ; Kim, Na-Young ; Lee, Si-Myung ; Woo, Hee-Jong ; Shin, Kong-Sik ; Jin, Yong-Moon ; Cho, Hyun-Suk ;
Journal of Plant Biotechnology, volume 34, issue 4, 2007, Pages 347~354
DOI : 10.5010/JPB.2007.34.4.347
Commercialization of genetically modified (GM) plants will be required the assessment of risks associated with the release of GM plants that should include a detailed risk assessment of their impacts in human health and the environment. Prior to GM plant release, applicants should provide the information on GM crops for approval. We carried out this study to provide the molecular data for risk assessment of the GM Chinese cabbage plants with insect-resistance gene, modified CryIAc, which we obtained by Agrobacterium-transformation. From the molecular analysis with GM Chinese cabbage, we confirmed the transgene copy number and stability, the expression of the transgene, and integration region sequences between the transgene and the Chinese cabbage genome. Based on the unique integration DNA sequences, we designed specific primer set to detect GM Chinese cabbage and set up the GM cabbage detection method by qualitative PCR analysis. Qualitative analysis with GM Chinese cabbage progenies analysis was revealed the same as the result of herbicide treatment. Our results provided the molecular data for risk assessment analysis of GM Chinese cabbage and demonstrated that the primer set proposed could be useful to detect GM Chinese cabbage.
Purification and Partial Characterization of a Peroxidase from Perilla Callus
Hur, Yeon-Jae ; Lee, Han-Gil ; Hu, Gaosheng ; Chung, Won-Bok ; Jeong, Soon-Jae ; Yi, Young-Byong ; Nam, Jae-Sung ; Chung, Young-Soo ; Lee, Jai-Heon ; Kim, Doh-Hoon ;
Journal of Plant Biotechnology, volume 34, issue 4, 2007, Pages 355~361
DOI : 10.5010/JPB.2007.34.4.355
Cotyledons of perilla6 were cultured on MS medium containing 0.5 mg/l NAA and 0.5 mg/l BA for 7 weeks. The activity of perilla peroxidase was observed to increase following culture stages as assessed by peroxidase assay. A peroxidase (POD) was purified from perilla tissue cultured on MS medium for 7 weeks. The peroxidase was purified using ion exchange and gel nitration chromatography. The perilla peroxidase had a molecular mass of 30 kDa by SDS-PAGE. We showed that the N-terminal amino acid sequence of this protein shared 67% identity with the tea peroxidase. As indicated by SDS-PAGE, the banding pattern of the 30 kDa polypeptide present in total soluble protein from perilla tissue was increased following culture stages. Immunoblot analysis indicated that perilla peroxidase protein appeared after 3 weeks of perilla tissue culture, and continued to increase with extended duration of tissue culture for at least 7 weeks.
Influence of donor plant growth condition, microspore isolation method, culture medium, and light culture on the production of embryos in microspore culture of hot pepper (Capsicum annuum L.)
Lee, Jong-Suk ; Park, Eun-Joon ; Kim, Moon-Za ;
Journal of Plant Biotechnology, volume 34, issue 4, 2007, Pages 363~373
DOI : 10.5010/JPB.2007.34.4.363
To establish an efficient and reliable microspore culture system for pepper (Capsicum annuum L.), the effect of light intensity used for donor plant's growth, microspore isolation methods, the composition of culture medium, and culture period in light on the production of embryos were investigated. The viability of microspores taken from the plants grown under the light intensity of 10,000 lux was almost same as that from the lower (5,500 lux) light intensity, and the embryo induction and development were a bit higher when donor plants were grown under the lower light intensity. This result implies that lower light intensity does not interfere with the embryo induction and development. However, it was very difficult to prepare microspores for culture since only a small number of flower buds could be harvested from plants grown under the light intensity of 5,500 lux. Microspore isolation methods greatly affected microspores viability; that is, when microspores were isolated by blending rather than maceration, the greater number of viable microspores were easily generated (about 13 times). Among media used for microspores culture in this study, MN medium was most efficient for embryo induction and development. Total number of embryos and the number of cotyledonary embryos were highest when microspores were cultured in dark for 4 weeks, and then in light for one week. These results will be provide valuable information to set up efficient microspore culture system of hot pepper with a high frequency of embryo production, which are applicable to gene transformation and mutagenesis.
Expression of CP4 5-Enol-Pyruvylshikimate-3- Phosphate Synthase Transgene in Inbred Line of Korean Domestic Maize (Zea may L.)
Cho, Mi-Ae ; Kwon, Suk-Yoon ; Kim, Jin-Seog ; Lee, Byoung-Kyu ; Moon, Choo-Yeun ; Choi, Pil-Son ;
Journal of Plant Biotechnology, volume 34, issue 4, 2007, Pages 375~380
DOI : 10.5010/JPB.2007.34.4.375
This study was conducted to develop herbicide-resistance domestic maize plants by introducing the CP4 5-enol-pyruvylshikimate-3-phosphate synthase (CP4 EPSPS) gene using Agrobacterium tumefaciens-mediated immature embryo transformation. Immature embryos of five genotypes (HW1, KL103, HW3, HW4, HW7) were co-cultivated with strains Agrobacterium tumefaciens (strain C58C1) containing the binary vector (pCAMBIA2300) carrying Ubiquitin promoter-CP4 EPSPS gene and Cauliflower mosaic virus 35S (CaMV35S) promoter-nptll gene conferring resistance to paromomycin as a selective agent. The presence and expression of CP4 EPSPS transgene were confirmed by PCR, RT-PCR and Northern blot analysis, respectively. Also, the resistance to glyphosate in the transgenic maize (
) was analyzed by shikimate accumulation assay. The frequency (%) of paromomycin-resistance callus was 0.37, 0.03, 2.20, 2.37, and 0.81% in pure lines HW1, KL103, HW3, HW4 and HW7, respectively. EPSP transgene sequences were amplified in putative transgenic plants that regenerated from paromomycin-resistance calli of two inbred lines (HW3, HW4). Of them, RT-PCR and Northern blot analyses revealed that the transgene was only expressed in two transgenic events (M266, M104) of HW4 inbred line, and a mild glyphosate resistance of transgenic event (M266) was confirmed by the lower shikimate accumulation in leaf segments. These results demonstrate that transgenic maize with herbicide-resistance traits in Korean genotype can be genetically obtained.