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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
> Journal Vol & Issue
Journal of Plant Biotechnology
Journal Basic Information
Journal DOI :
The Korean Society of Plant Biotechnology
Editor in Chief :
Volume & Issues
Volume 38, Issue 4 - Dec 2011
Volume 38, Issue 3 - Sep 2011
Volume 38, Issue 2 - Jun 2011
Volume 38, Issue 1 - Mar 2011
Selecting the target year
Development of salt-tolerant transgenic chrysanthemum (Dendranthema grandiflorum) lines and bio-assay with a change of cell specificity
Kang, Chan-Ho ; Yun, Seung-Jung ; Han, Bum-So ; Lee, Gong-Joon ; Choi, Kyu-Hwan ; Park, Jong-Suk ; Shin, Yong-Kyu ;
Journal of Plant Biotechnology, volume 38, issue 1, 2011, Pages 1~8
DOI : 10.5010/JPB.2011.38.1.001
Recently the increasing of vinyl and green houses and development of reclaimed land including Saemangeum induced the need for breeding salt-tolerant crops which can survive and grow in high salinity soil. So we try to develop salt-tolerant transgenic chrysanthemum (Dendranthema grandiflorum.) lines by using anti-porter gene TANHX and HVNHX. Through marker selection and plant regeneration step, we could get 284 putative transgenic chrysanthemum lines. On selected putative transgenic plants, 40 candidates were used for genetic analysis and 30 lines could be made up of target size band on PCR, so about 75% of marker selected lines were decided as real transgenic lines. Selected 284 transgenic lines were also used for salt-tolerance test as a range of NaCl 0.2 ~ 1.2% (300 mM). As a result of salt-tolerance test, 15 selected transgenic lines could live and grow on the continuous supply of 0.8% (200 mM) NaCl solution and another 7 lines were could survive under 1.2% (300 mM) NaCl solution. This salt-tolerant transgenic lines under salt stress also lead a cell alternation especially a guard cell. A stressed guard cell be swelled and grow larger in proportion to NaCl concentration. TTC test for cell viability on transgenic chrysanthemum lines pointed out that more strong salt-tolerant lines can be live more than another under same salt stress. The numerical value of strong salt-tolerant 7 transgenic lines were 0.206 ~ 0.331 under 1.2% NaCl stress, and then it`s value is more larger than middle salinity lines` 0.114 ~ 0.193 and non-transgenic`s 0.046. And the proline contents as indicated stress compound also pointed out that HVNHX introduced salt-tolerant transgenic lines were less stressed than other under same salt stress. The contents of strong salt-tolerant transgenic lines were 2.255 ~ 2.638 mg/kg and it is much higher than that of middle salinity lines` 1.496 ~ 2.125.
Prediction and discrimination of taxonomic relationship within Orostachys species using FT-IR spectroscopy combined by multivariate analysis
Kwon, Yong-Kook ; Kim, Suk-Weon ; Seo, Jung-Min ; Woo, Tae-Ha ; Liu, Jang-Ryol ;
Journal of Plant Biotechnology, volume 38, issue 1, 2011, Pages 9~14
DOI : 10.5010/JPB.2011.38.1.009
To determine whether pattern recognition based on metabolite fingerprinting for whole cell extracts can be used to discriminate cultivars metabolically, leaves of nine commercial Orostachys plants were subjected to Fourier transform infrared spectroscopy (FT-IR). FT-IR spectral data from leaves were analyzed by principal component analysis (PCA) and Partial least square discriminant analysis (PLS-DA). The dendrogram based on hierarchical clustering analysis of these PLS-DA data separated the nine Orostachys species into five major groups. The first group consisted of O. iwarenge `Yimge`, `Jeju`, `Jeongsun` and O. margaritifolius `Jinju` whereas in the second group, `Sacheon` was clustered with `Busan,` both of which belong to O. malacophylla species. However, `Samchuk`, belong to O. malacophylla was not clustered with the other O. malacophylla species. In addition, O. minuta and O. japonica were separated to the other Orostachys plants. Thus we suggested that the hierarchical dendrogram based on PLS-DA of FT-IR spectral data from leaves represented the most probable chemotaxonomical relationship between commercial Orostachys plants. Furthermore these metabolic discrimination systems could be applied for reestablishment of precise taxonomic classification of commercial Orostachys plants.
Analysis of right border flanking sequence in transgenic chinese cabbage harboring integrated T-DNA
Ahn, Hong-Il ; Shin, Kong-Sik ; Woo, Hee-Jong ; Lee, Ki-Jong ; Kim, Hyo-Sung ; Park, Yong-Hwan ; Suh, Seok-Cheol ; Cho, Hyun-Suk ; Kweon, Soon-Jong ;
Journal of Plant Biotechnology, volume 38, issue 1, 2011, Pages 15~21
DOI : 10.5010/JPB.2011.38.1.015
We developed 14 transgenic lines of Chinese cabbage (Brassica rapa) harboring the T-DNA border sequences and CryIAc1 transgene of the binary vector 416 using Agrobacterium tumefaciens-mediated DNA transfer. Six lines had single copy cryIAc1 gene and four of them contained no vector backbone DNA. Of the left border (LB) flanking sequences six nucleotides were deleted in transgenic lines 416-2 and 416-3, eleven nucleotides in line 416-9, and 65 nucleotides including the whole LB sequences in line 416-17, respectively. And we defined 499 bp of genomic DNA (gDNA) of transformed Chinese cabbage, and blast results showed 96% homology with Brassica oleracea sequences. PCR with specific primer for the right border (RB) franking sequence revealed 834 bp of PCR product sequence, and it was consisted of 3` end of cryIAc1, nosterminal region and 52 bp of Chinese cabbage genomic DNA near RB. RB sequences were not found and the 58 nucleotides including 21 bp of nos-terminator 3` end were deleted. Also, there were deletion of 10 bp of the known genomic sequences and insertion of 65 bp undefined genomic sequences of Chinese cabbage in the integration site. These results demonstrate that the integration of T-DNA can be accompanied by unusual deletions and insertions both in transgenic and genomic sequences.
Stress-induced biphasic ethylene and ROS biosynthesis are synergistically interacted in cell damage
Ji, Na-Ri ; Park, Ky-Young ;
Journal of Plant Biotechnology, volume 38, issue 1, 2011, Pages 22~29
DOI : 10.5010/JPB.2011.38.1.022
Although reactive oxygen species (ROS) are inevitable by-products of many redox reactions in eukaryotic cells, they play a crucial role as signaling molecules in many cellular processes for development and defense response to abiotic stresses. The biphasic ROS production which was peaked twice in a first transient phase and a second massive phase was occurred after treatment of abiotic stress such as oxidative stress, high salinity. This biphasic generation of ROS was followed by the biphasic production of stress hormone, ethylene. The mechanism of interactions between ROS and ethylene biosynthesis is studied in tobacco (Nicotiana tabaccum L.) plants under the abiotic stresses. The stress-induced ethylene production was significantly inhibited in RbohD-AS and RbohF-AS, in which antisense expression of NADPH oxidase genes was performed. The accumulation of ROS, which was determined by DAB and DCFH-DA staining, was significantly decreased after abiotic stresses in transgenic plants. The suppression of signaling with ethylene and ROS induced more tolerance in response to abiotic stress. The transgenic plants were more tolerant in MS medium supplemented with salinity stress in contrast with wild-type. Stress-induced cell damage determined by DNA fragmentation was decreased at phase II in those transgenic plants. Therefore, the first burst of ROS is more responsible for making a role as a signaling molecule during stress-induced response. These results suggested that ethylene and ROS act in a positive feedback cycle that results in mutual enhancement of ethylene and ROS production during stress-induced cell death.
Influence of medium addition and agitation on the production of embryos in isolated microspore culture of hot pepper (Capsicum annuum L.)
An, Dong-Ju ; Park, Eun-Joon ; Kim, Moon-Za ;
Journal of Plant Biotechnology, volume 38, issue 1, 2011, Pages 30~41
DOI : 10.5010/JPB.2011.38.1.030
The influences of the agitation as well as the addition of medium during culture on the production of embryos were invested in isolated microspore culture of hot pepper (Capsicum annuum L.). When the culture medium was added during initial liquid culture step of liquid-double layer culture, the embryo yield and quality greatly increased. The most effective time point for medium addition was 5 days after the culture commenced. On the other hand, the effect of medium addition at later double layer culture step in liquid-double layer culture on the embryo production was less compared to that of medium addition during the initial liquid culture step. Agitating the culture for 1 week during later double layer culture step in liquid-double layer culture effectively increased the production of normal cotyledonary embryos. In the case of liquid culture, agitating the culture for 1 week from 7 days after the culture commenced was also effective for embryo development. However, when the total agitation time was longer (2 to 3 weeks) during liquid-double layer culture or liquid culture, the embryos developed abnormally in both cases. The normal cotyledonary embryos obtained in this study successfully developed to plants when transferred to regeneration media. These regenerated plants were either diploid or haploid, and there was a difference in the number of chloroplasts between guard cells of diploid and haploid. These results can be used as an important data for developing an efficient microspore culture system with high quality embryo production in hot pepper.
Production of stable chloroplast-transformed plants in potato (Solanum tuberosum L.)
Min, Sung-Ran ; Jeong, Won-Joong ; Park, Ji-Hyun ; Lyu, Jae-Il ; Lee, Jeong-Hee ; Oh, Kwang-Hoon ; Chung, Hwa-Jee ; Liu, Jang-R. ;
Journal of Plant Biotechnology, volume 38, issue 1, 2011, Pages 42~48
DOI : 10.5010/JPB.2011.38.1.042
Chloroplast genetic engineering of higher plants offers several unique advantages compared with nuclear genome transformation, such as high levels of transgene expression, a lack of position effect due to site-specific transgene integration by homologous recombination, multigene engineering in a single transformation event and reducing risks of gene flow via pollen due to maternal inheritance. We established a reproducible chloroplast transformation system of potato using a tobacco specific plastid transformation vector, pCtVG (trnI-Prrn-aadA-mgfp-TpsbA-trnA). Stable transgene integration into chloroplast genomes and the homoplasmic state of the transgenome were confirmed by PCR and Southern blot analyses. Northern, immunoblot analysis, and GFP fluorescence imaging revealed high expression and accumulation of GFP in the plastids of potato leaves. This system would provide new opportunities for genetic improvement and mass production of value added foreign proteins in this crop.
Basic seed production using aeroponic system in double cropping
Cho, Il-Chan ; Chang, Dong-Chil ; Kim, Hyun-Jun ; Cho, Hyun-Mook ;
Journal of Plant Biotechnology, volume 38, issue 1, 2011, Pages 49~53
DOI : 10.5010/JPB.2011.38.1.049
The present study was performed to select suitable potato (Solanum tuberosum L.) varieties for double cropping in seed production using aeroponic system, the nutrient solution was sprayed for 20 seconds in every 3 minutes, and potatoes were planted into styrofoam at a distance of 25 cm between plants. Transplantation was completed in September 9 and potatoes were harvested in November 12. The 10 trial cultivars, `Atlantic`, `Dejima`, `Haryong`, `Hongyoung`, `Jayoung`, `Jopung`, `Jowon`, `Namsuh`, `Seohong`, `Superior` were examined for growth and yield. The shoot growth of all cultivars at the early stage was better than `Superior`, however, no difference was observed at the late stage of plant growth. In addition, `Superior` showed a excellent root growth during the early plant growth stage, however, increased root growth was only found in `Jayoung`, `Seohong`, during the late plant growth stage. The stolon weight was greater in the order of `Seohong`>`Superior`>`Jayoung`>`Hongyoung`. Particularly, the number of tuber in `Jowon` was 22.4/plant that was over twice than `Superior`(11.3/plant). As results of the current study, therefore, we concluded that four cultivars `Seohong`, `Jayoung`, `Hongyoung`, `Jowon`were the most suitable cultivars for basic seed production using aeroponic system in Autumn potato.
Enhanced tolerance through increasing polyamine contents in transgenic tobacco plants with antisense expression of ACC oxidase gene
Wi, Soo-Jin ; Park, Ky-Young ;
Journal of Plant Biotechnology, volume 38, issue 1, 2011, Pages 54~61
DOI : 10.5010/JPB.2011.38.1.054
Antisense construct of cDNA for senescencerelated ACC oxidase (CAO) cDNA isolated from carnation flowers were introduced into tobacco by Agrobacteriummediated transformation. The decreasing expression of NtACO and the reduction of ethylene production were observed in these transgenic lines. In contrast, the SAMDC transcripts and spermidine content were increased. The findings that higher content of spermidine in the ethylene suppressed transgenic plants compared with wild-type should be directly resulted in the enhancement of SAMDC activity followed by the increased accumulation of SAMDC transcript. To investigate the pathogenic response in these transgenic plants, wild-type and transgenic plants were inoculated with Phytophthora parasitica pv. nicotianae. Transgenic plants suppressing ethylene production showed the increased resistance against fungal pathogen, comparing with wild-type plant. PR-protein genes expression in CAO-AS-2 and CAOAS-4 were also higher at the normal growth condition and pathogenic response than in wild-type plants. The results of higher spermidine content and SAMDC activity in transgenic plants, CAO-AS-2 and CAO-AS-4, support the possibility that an increase in spermidine content might induce the higher transcripts of PR-protein genes. This results agreed with the phenomena that spermidine promoted the expression of PR1a and a SAMDC inhibitor, MGBG, decreased the expression of PR1a in leaf discs. These results suggest that the resistance against fungal pathogen in transgenic tobacco impaired in ethylene production might be caused by increasing in polyamine, especially spermidine, biosynthesis.
AtCBP63, a Arabidopsis Calmodulin-binding Protein 63, Enhances Disease Resistance Against Soft Rot Disease in Potato
Chun, Hyun-Jin ; Park, Hyeong-Cheol ; Goo, Young-Min ; Kim, Tae-Won ; Cho, Kwang-Soo ; Cho, Hyeon-Seol ; Yun, Dae-Jin ; Chung, Woo-Sik ; Lee, Shin-Woo ;
Journal of Plant Biotechnology, volume 38, issue 1, 2011, Pages 62~68
DOI : 10.5010/JPB.2011.38.1.062
Calmodulin (CaM), a
binding protein in eukaryotes, mediates cellular
signals in response to a variety of biotic and abiotic external stimuli. The
-bound CaM transduces signals by modulating the activities of numerous CaM-binding proteins. As a CaM binding protein, AtCBP63 (
kD) has been known to be positively involved in plant defense signaling pathway. To investigate the pathogen resistance function of AtCBP63 in potato, we constructed transgenic potato (Solanum tuberosum L.) plants constitutively overexpressing AtCBP63 under the control of cauliflower mosaic virus (CaMV) 35S promoter. The overexpression of the AtCBP63 in potato plants resulted in the high level induction of pathogenesis-related (PR) genes such as PR-2, PR-3 and PR-5. In addition, the AtCBP63 transgenic potato showed significantly enhanced resistance against a pathogen causing bacterial soft rot, Erwinia carotovora ssp. Carotovora (ECC). These results suggest that a CaM binding protein from Arabidopsis, AtCBP63, plays a positive role in pathogen resistance in potato.
Myo-inositol increases the plating efficiency of protoplast derived from cotyledon of cabbage (Brassica oleracea var. capitata)
Jie, Eun-Yee ; Kim, Suk-Weon ; Jang, Hye-Rim ; In, Dong-Su ; Liu, Jang-Ryol ;
Journal of Plant Biotechnology, volume 38, issue 1, 2011, Pages 69~76
DOI : 10.5010/JPB.2011.38.1.069
This study describes the effect of myo-inositol on sustained cell division and plant regeneration from cotyledon-derived protoplast of cabbage (Brassica oleracea var. capitata). Freshly isolated protoplasts were cultured in modified Murashige and Skoog (MS) medium removed ammonia ions and containing
sucrose and several concentrations of myo-inositol (2, 4, 6, 8, 10% (w/v)) as an osmotic stabilizer. After 3 weeks of culture in the dark at
, the plating efficiency of cabbage protoplasts reached to
when cultured in modified MS medium supplemented with
sucrose and 8% (w/v) of myo-inositol at a density of
protoplasts/ml. Rapidly growing cell colonies after 3 weeks of culture were transferred to the same culture medium removed osmoticum. To induce shoot regeneration from calluses, calluses with about 2 mm in diameter were transferred to the MS medium containing
NAA. After further three weeks of incubation onto the medium in the light, green shoots were formed on the surface of calluses at a frequency of 30%. Upon transfer to half-strength MS basal medium, roots were formed onto the bottom of regenerated shoots without auxin treatments. These regenerated plantlets were successfully acclimatized to soil transfer, grown to normal mature plants. The cabbage protoplast culture system established in this study could be applied for production of somatic hybrids or cybrids by asymmetric protoplast fusion and mass proliferation of elite somatic clones of cabbage.
Isolation and characterization of Bradh1 gene encoding alcohol dehydrogenase from Chinese cabbage (Brassica rapa)
Abdula, Sailila E. ; Lee, Hye-Jung ; Melgar, Reneeliza J. ; Sun, Mingmao ; Kang, Kwon-Kyoo ; Cho, Yong-Gu ;
Journal of Plant Biotechnology, volume 38, issue 1, 2011, Pages 77~86
DOI : 10.5010/JPB.2011.38.1.077
Alcohol dehydrogenase (E.C.220.127.116.11) is an enzyme present in higher plants involved in the anaerobic fermentation pathway that catalyzes the reduction of pyruvate to ethanol, resulting in continuous
regeneration. It also plays an important role in many plant developments including tolerance to anoxia condition. Here, a cDNA clone encoding alcohol dehydrogenase (ADH) was isolated from Chinese cabbage (Brassica rapa) seedlings. The gene named Bradh1 had a total length of 1,326 bp that contains a single open reading frame of 1,140 bp. The predicted protein consists of 379 amino acid residues with a calculated molecular mass of 41.17 kDa. Expression pattern analysis revealed a tissue-specific expressing gene in different tissues and strongly expressed in the shoot, roots and seeds of Chinese cabbage. Agrobacterium transformation of full-length cDNA Bradh1 into rice Gopumbyeo showed high efficiency. Furthermore, induction of ADH in transgenic rice enhanced tolerance to anaerobiosis stresses and elevated mRNA transcripts. The overexpression of Bradh1 in rice increases germination under anaerobiosis stresses, implying the possibility of developing new varieties suited for direct seeding or flood-prone rice field.
Action mechanism of upstream open reading frame from S-adenosylmethionine decarboxylase gene as a in vivo translational inhibitor
Choi, Yu-Jin ; Park, Ky-Young ;
Journal of Plant Biotechnology, volume 38, issue 1, 2011, Pages 87~93
DOI : 10.5010/JPB.2011.38.1.087
S-Adenosylmethionine decarboxylase (SAMDC; EC 18.104.22.168), a key enzyme for polyamines biosynthesis, was tightly regulated for homeostatic levels. Carnation SAMDC gene (CSDC9) has an small upstream open reading frame (uORF) of 54 amino acids in 5`-leader sequence. To explore the functional mechanism of uORFs in controlling translation, we used a GUS reporter gene driven with the 35S promoter and uORF region of SAMDC gene for making transgenic tobacco plants. In our experiment, there were a translational inhibition of its downstream GUS ORF by SAMDC uORF sequence or SAMDC uORF protein. Expecially, translational inhibition was most effective in point-mutated construct, in which the start codon was changed. Therefore, this results suggested the ribosomal stalling might be involved in this translational inhibitory process. The frame shift in amino acid sequence of SAMDC uORF with start codon and stop codon resulted in a moderate increasing in GUS activity, suggesting the native amino acid sequence was important for a function as a translational inhibitor. Also, we showed that the production of GUS protein was significantly inhibited in the presence of the small uORF using histochemical analysis of GUS expression in seedlings and tobacco flowers. Importantly, the small uORF sequence induced a real peptide of 5.7 kDa, which was provided the presence of SAMDC uORF peptide band using an in vitro transcription/translation system. The peptide product of uORF might interact with other components of translational machinery as well as polyamines, which was resulted from that polyamine treatment was inhibited GUS protein band in SDS-PAGE experiment.
A transcription factor "OsNAC075" is essential for salt resistance in rice (Oryza sativa L.)
Jung, Yu-Jin ; Lee, Myung-Chul ; Kang, Kwon-Kyoo ;
Journal of Plant Biotechnology, volume 38, issue 1, 2011, Pages 94~104
DOI : 10.5010/JPB.2011.38.1.094
Salt stress is a major environmental factor influencing plant growth and development. To identify salt tolerance determinants, we systematically screened salt sensitive rice mutants by use of the Activator/Dissociation (Ac/Ds) transposon tagging system. In this study, we focused on the salt sensitive mutant line, designated SSM-1. A gene encoding a NAC transcription factor homologue was disrupted by the insertion of a Ds transposon into SSM-1 line. The OsNAC075 gene (EU541472) has 7 exons and encodes a protein (486-aa) containing the NAC domain in its N-terminal region. Sequence comparison showed that the OsNAC075 protein had a strikingly conserved region at the N-terminus, which is considered as the characteristic of the NAC protein family. OsNAC075 protein was orthologous to Arabidopsis thaliana ANAC075. Phylogenetic analysis confirmed OsNAC075 belonged to the OsNAC3 subfamily, which plays an important role in response to stress stimuli. RT-PCR analysis showed that the expression of OsNAC075 gene was rapidly and strongly induced by stresses such as NaCl, ABA and low temperature (
). Our data suggest that OsNAC075 holds promising utility in improving salt tolerance in rice.