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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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Journal of Plant Biotechnology
Journal Basic Information
Journal DOI :
The Korean Society of Plant Biotechnology
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Volume & Issues
Volume 35, Issue 4 - Dec 2008
Volume 35, Issue 3 - Sep 2008
Volume 35, Issue 2 - Jun 2008
Volume 35, Issue 1 - Mar 2008
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Molecular cloning and expression of glyceraldehyde-3-phosphate dehydrogenase gene under environmental stresses in sweetpotato
Kim, Young-Hwa ; Song, Young-Sun ; Huh, Gyung-Hye ;
Journal of Plant Biotechnology, volume 35, issue 2, 2008, Pages 95~100
DOI : 10.5010/JPB.2008.35.2.095
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a main enzyme in the glycolytic pathway, is involved in cellular energy production and regarded as a housekeeping gene. Previously, cytosolic GAPDH was selected as the most significantly abundant gene in EST library of sweetpotato suspension cells. In this study, a full-length of cDNA clone (IbGAPDH) encoding GAPDH was isolated from suspension-cultured cells of sweetpotato (Ipomoea babatas), and its expression was investigated with a view to understanding the physiological function of GAPDH in relation to environmental stresses. IbGAPDH encoded a 36.9 kDa polypeptide consisting of 337 amino acids. When the deduced amino acid of IbGAPDH was compared with other higher plants, IbGAPDH showed high homology with cytosolic GAPDH. The mRNA level of IbGAPDH significantly increased under environmental stresses, such as
, MV and cold treatments. Among them, the transcript level of IbGAPDH gene was the highest under cold stress. Further investigation of the transcription level under
was performed with different tissues of sweetpotato. The transcription of IbGAPDH was increased by cold stress with tissue-specificity, moreover, showed different patterns according to temperature.
Analysis of junction between T-DNA and plant genome in insect resistance GM Chinese cabbage
Lim, Sun-Hyung ; Park, Seung-Hye ; Kim, Jung-Hwan ; Kim, Na-Young ; Won, So-Youn ; Lee, Si-Myung ; Shin, Kong-Sik ; Woo, Hee-Jong ; Kim, Dong-Hern ; Cho, Hyun-Suk ;
Journal of Plant Biotechnology, volume 35, issue 2, 2008, Pages 101~108
DOI : 10.5010/JPB.2008.35.2.101
The Agrobacterium-mediated transformation has been successfully used method to introduce foreign genes into some monocotyledonous as well as a large number of dicotyledonous plants genome, We developed transgenic Chinese cabbage plants with insect-resistance gene, modified CryIAc, by Agrobacterium-transformation and confirmed transgene copy number by Southern blot analysis. We confirmed that twenty-nine out of 46 transgenic Chinese cabbage plants have single copy of CryIAc. To obtain the sequences information on the transferred DNA (T-DNA) integration into plant genome, we analyzed left border (LB) flanking sequences by genome walking (GW) PCR method. Out of 46 transgenic Chinese cabbage plants examined, 37 carried the vector backbone sequences. This result indicates that the transfer of the vector backbone from the binary vectors resulted mainly from inefficient termination of LB site. Analysis of T-DNA LB flanking region of 9 transgenic Chinese cabbage plants without vector backbone revealed that all LB ends were not conserved and nucleotides up to 36bp from the LB cleavage site were deleted.
GFP expression in the microspore-derived early embryo through co-culturing with Agrobacterium
Jung, Min ; In, Dong-Su ; Kim, Bong-Kyu ; Jang, In-Chang ; Park, Eun-Joon ; Kim, Moon-Za ; Harn, Chee-Hark ;
Journal of Plant Biotechnology, volume 35, issue 2, 2008, Pages 109~114
DOI : 10.5010/JPB.2008.35.2.109
The aim of this research is to establish the conditions for Agrobacterium-mediated genetic transformation using microspore. The embryo induction from the microspore was examined under several Kanamycin concentration in media, and the induction rate decreased about 4, 8, 10 times when the Kanamycin concentration increased 10, 50, 100 mg/L, respectively. This indicates that the transformation rate would be much lower if the Kanamycin was used for selection marker. In order to apply the GFP gene as a reporter gene for Agrobacterium-mediated genetic transformation, GFP expression from the microspore-mediated embryos was observed using GFP filter under microscope. The GFP expression occurred when the microspore cultured toward the embryo development for 12, 24 and 48 days. The microspore formed a cluster by microspore division from 12 days culture and continuously became a bigger mass. We obtained a total of 8 GFP-expressing embryos suggesting that the transformation of microspore occurred. However, those young embryos were not fully developed. Further study pertinent to culture conditions is required to fulfill the Agrobacterium-mediated genetic transformation using microspore.
Transgenic lettuce (Lactuca sativa L.) with increased vitamin C levels using GalUR gene
Lim, Mi-Young ; Cho, Yi-Nam ; Chae, Won-Ki ; Park, Young-Soo ; Min, Byung-Whan ; Harn, Chee-Hark ;
Journal of Plant Biotechnology, volume 35, issue 2, 2008, Pages 115~120
DOI : 10.5010/JPB.2008.35.2.115
L-Ascorbic acid (vitamin C) in vegetables is an essential component of human nutrition. The objective is to transform lettuce (Lactuca sativa L.) with GalUR gene that is involved in the vitamin C biosynthesis. The cotyledons of Hwoahong (Nongwoo Bio Co.) were used to induce the callus and shoot under the selection media with MS + 30 g/L Sucrose + 0.5 mg/L BAP + 0.1 mg/L NAA + 100 mg/L kanamycin + 200 mg/L lilacillin, pH 5.2. The shoot was developed from the cut side of the explants after 3 weeks on the selection media. We successfully transformed the lettuce with GaIUR gene and analyzed the levels of vitamin C. We found that some of the lettuce transgenic lines contained higher levels of vitamin C compared with the normal one (non-transformed). Especially, some of
lettuces inserted by GalUR showed about
times higher content of vitamin C compared to the non-transformed lettuce. This data support the previously work performed with GLOase transgenic
lettuces from which several times higher content of vitamin C were identified. The
lettuces with high content of vitamin C have been selected for further analysis.
Regeneration of symmetric protoplast fusion between cabbage (Brassica oleracea L.) and radish (Raphanus sativus L.)
In, Dong-Su ; Song, Min-Jung ; Jang, In-Chang ; Min, Byung-Whan ; Nahm, Seok-Hyeon ; Shin, Jong-Sub ; Lee, See-Woo ; Harn, Chee-Hark ;
Journal of Plant Biotechnology, volume 35, issue 2, 2008, Pages 121~126
DOI : 10.5010/JPB.2008.35.2.121
Protoplasts from cabbage and radish were isolated and fused symmetrically by PEG treatment. The PEG treated mixture of high concentrated protoplasts produced lots of micro-calli after
weeks. The microcalli developed to normal calli and shoots were regenerated from the calli. A total of 218 shoots were regenerated, but none of them contained the NWB-CMS specific DNA marker, indicating that the transfer of the radish NWB-CMS character into cabbage did not occur. However, ISSR analysis revealed that the cell fusion between protoplasts from radish and cabbage was occurred (3 out of 208 plantlet). The fused regenerants possessed the characteristics of source plants used for protoplast fusion. After vernalization, three regenerants were flowered with white petal color as seen in radish. Only three seeds were able to obtain from one regenerant by backcrossing with the cabbage pollen.
Plant let growth, leaf stomata, and photosynthesis of grape rootstock '5BB' as affected by inoculum density in bioreactor cultures
Choi, Eun-Jung ; Hahn, Eun-Joo ; Paek, Kee-Yoeup ;
Journal of Plant Biotechnology, volume 35, issue 2, 2008, Pages 127~132
DOI : 10.5010/JPB.2008.35.2.127
In bioreactor cultures of plants, inoculum density is an important factor affecting growth and proliferation of the plantlets. To maximize shoot growth and proliferation of grape rootstock '5BB' in bioreactors, inoculum density varied at 15, 30, 45 and 60 single nodes in a 3-liter scale balloon type bioreactor, respectively and cultured for 40 days. Results suggested that the growth and the photosynthesis of the plantlet were greatly affected by inoculum density in the bioreactor. The inoculum density of 45 nodes resulted in the greatest growth (910.4 mg/shoot FW, 764.4 mg/root FW) followed by 30 nodes.
assimilation rate, stomatal conductance, transpiration rate of the plantlet were also highest at the inoculum density of 45 nodes. Significant reduces in shoot and root growth (426.5 mg/shoot FW, 248.4 mg/root FW) were observed at the inoculum density of 60 nodes. When the inoculum density decreased by 15 nodes, plantlets were malformed due to hyperhydricity, resulting in the highest transpiration rate and the lowest
assimilation rate. The plantlets stressed by the inoculum density at 15 nodes and 60 nodes showed larger number and irregular shape of stomata compared to the plantlets inoculated with 45 nodes.
Effects of 5-azacytidine, a DNA methylation inhibitor, on embryogenic callus formation and shoot regeneration from rice mature seeds
Lee, Yeon-Hee ; Lee, Jung-Sook ; Kim, Soo-Yun ; Sohn, Seong-Han ; Kim, Dool-Yi ; Yoon, In-Sun ; Kweon, Soon-Jong ; Suh, Seok-Chul ;
Journal of Plant Biotechnology, volume 35, issue 2, 2008, Pages 133~140
DOI : 10.5010/JPB.2008.35.2.133
The modification of DNA and histone plays an important role for gene expression in plant development. The objective of this research is to observe the effects of methylation on the gene expression during dedifferentiation from rice mature seeds to callus and differentiation from callus to shoots. The embryogenic callus with ability to shoot regeneration was not induced on the N6A medium supplemented with 5-azacytidine and abnormal callus with brown color was formed. When the normal rice callus was placed on the regeneration MSRA medium supplemented with 5-azacytidine, the shoot regeneration was inhibited. The results showed that 5-azacytidine, DNA demethylating agent, had negative effects on normal embryogenic callus formation and shoot regeneration. This suggested that DNA methylation of some genes was required for normal cell dedifferentiation and differentiation in tissue culture. The microarray and
DEG screening were used to observe the gene transcript profile in callus induction and regeneration on N6A (N6 medium + 5-azaC) and MSRA (MS regeneration medium + 5-azaC). Subsets of genes were up-regulated or down-regulated in response to 5-azaC treatments. The genes related with epigenetic regulation, electron transport, nucleic acid metabolism and response to stress were up and down regulated. The different expression of some genes (germin like protein etc.) during callus induction and shoot regeneration was confirmed using RT-PCR and northern blot analysis.
Cyanobacterial bioreporters for detection of heavy metals, herbicide, and antibiotics
Kim, Soo-Youn ; Jeong, Won-Joong ; Suh, Kye-Hong ; Liu, Jang-Ryol ; Park, Youn-Il ;
Journal of Plant Biotechnology, volume 35, issue 2, 2008, Pages 141~145
DOI : 10.5010/JPB.2008.35.2.141
In this study, glucose-inducible intergenic sequences were used to generate bioreporters of the cyanobacterium Synechocystis sp. PCC 6803 that could monitor environmental pollutants. Luciferase genes LuxAB from the marine bacterium Vibrio fischeri under the control of glucose-inducible intergenic seqeucens of eight genes (atpI, ndbA, ctaD1, tkt, pgi, pdh, ppc, and cydA) were successfully expressed in the cyano-bacterial transformants, showing 5-25 fold increases in biolumeniscence upon exposure to glucose. In addition, glucose-inducible cyanobacterial bioreporters were very sensitive to various chemicals such as heavy metals (
), electron transport inhibitors (DCMU, DBMIB,
), and antibiotics (chloramphenicol and rifampicin). These glucose-inducible cyanobacterial bioreporters would be useful to develop biosensors for rapid screening of environmental samples.
Agrobacterium-mediated transformation of Bacillus thuringiensis cry1Ac gene in chrysanthemum (Dendranthema grandiflorum Kitamura) 'Linneker Salmon'
Han, Bong-Hee ; Lee, Su-Young ; Lim, Jin-Hee ;
Journal of Plant Biotechnology, volume 35, issue 2, 2008, Pages 147~153
DOI : 10.5010/JPB.2008.35.2.147
Cry1Ac gene was introduced into chrysanthemum (Dendranthema grandiflorum Kitamura) 'Linneker Salmon' through Agrobacterium-mediated gene transformation to develop new lines showing resistance to tobacco cutworm (Spodoptera litura). Cry1Ac gene was transferred into chrysanthemum by Agrobacterium C58C1 containing pCAMBIA2301. After infection of Agrobacterium C58C1 with leaf segments, the segments were cultured on regeneration medium (MS + 1.0 mg/L BA + 0.5 mg/L IAA) containing 10 mg/L kanamycin for the first selection, on the same medium containing 20 mg/L kanamycin for the second selection, and on rooting medium (MS basal medium) containing 20 mg/L kanamycin for the third selection. Until the third selection, sixty nine plantlets (1.6%) were survived and rooted. Thirty six ones (0.8%) among them were confirmed as putative transformants with nptll gene by nptll primer PCR, and 35 (0.8%) of 36 ones as transformants with nptll gene and cry1Ac gene by Southern analysis. The gene transformation efficiency of cry1Ac gene was favorable with 0.8%. The resistance of tobacco cutworm (Spodoptera litura) in chrysan-themum transformant introduced cry1Ac gene was tested in green house. Three transformants were confirmed to have resistance to tobacco cutworm.
Induction and growth of adventitious roots and bioreactor culture in Codonopsis lanceolata
Ahn, Chang-Ho ; Bae, Kee-Hwa ; Yi, Jae-Seon ; Choi, Yong-Eui ;
Journal of Plant Biotechnology, volume 35, issue 2, 2008, Pages 155~161
DOI : 10.5010/JPB.2008.35.2.155
This paper reported the establishment of mass production system of adventitious roots of Codonopsis lanceolata through shake flask and bioreactor culture. Induction of adventitious roots was started from the explants of leaf, stem and root on 1/2 strength Murashing and Skoog (MS) solid medium. Stem segments were the best explants for induction of adventitious roots compared to leaf and root segments. Among the different auxins tested (NAA, IBA and IAA), number of adventitious root per explant was highest on solid medium with 1.0 mg/L IBA, and produced
roots per explant. However, growth of adventitious roots was fast in the presence of IBA at low concentration (0.1 mg/L). In shake flask culture, maximum production of adventitious roots (fresh weight) was obtained in half-strength MS medium compared to full-strength and one-third MS medium. When the adventitious roots produced in shake-flask culture were transferred to 5 L air-lift bioreactor, 16 times of fresh weight increase was gained after one month of culture. These results indicate that this protocol for the production of C. lanceolata adventitious roots can be applied to large scale culture for practical application.
The identification of optimum condition for direct regeneration in black raspberry
Ran, Choi-Heh ; Park, Pill-Jae ; Lee, Hee-Kwon ; Joong, Yun-Song ; Lee, In-Sok ;
Journal of Plant Biotechnology, volume 35, issue 2, 2008, Pages 163~167
DOI : 10.5010/JPB.2008.35.2.163
Adventitious buds appeared within 2 weeks on the base of the petiole explants and increased for two months. A maximum of regeneration (15.6%) was obtained on the medium containing
TDZ in combination with
IBA. To know which explants are the best for the induction of regeneration, three explants such as leaf, petiole and leaf-petiole were used. Among the explant types, the leaf-petiole explant was significantly more effective than leaf and petiole for promoting adventitious shoots, with leaf-petiole inducing at the highest regeneration frequency (33.7%). The regeneration frequency of adventitious shoots in the leaf-petiole explants was significantly affected by leaf size and the position of explants. The leaf-petiole smaller than 5 mm leaf in width was induced at the highest regeneration frequency (68.9%). The smaller leaf sizes, the greater regeneration frequency. Also when the leaves are nearer to the shoot tip, the regeneration frequency is higher. When the rooted micro-shoots were transferred to the soil after growing for 6 weeks in the media, the survival rate was 90%.