• Journal of Life Science
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• v.7 no.4
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• pp.270-275
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• 1997

In order to produce betacyanins from sugar beet(Beta vulagris L.) in vitro, callus induction, shoot formation, root formation, betacyanin contents and protein contents determined from callus which was induced cotyledons of sugar beet seedling under an addition of NAA and BAP on the MS medium. The results were summarized as follows; The combination 3.0mg/l NAA and 1.0mg/l BAP treatment showed the most high callus induction rate, betacyanin and protein contents. The combination NAA and BAP treatments were not shoot formation, but BAP treatments showed high root formation rate. But high concentrations of BAP have not shown root formation.

• Journal of Life Science
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• v.11 no.3
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• pp.254-258
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• 2001

The biochemical change during regeneration of perilla callus were investigated by comparing total protein and lipid contents, protein band pattern in SDS-PAGE, and fatty acid composition in the calli cultured for various period(0, 1, 3, 5 and 6 weeks) Calli were induced from cotyledon and hypocotyl explants of peplants of perilla on perilla on MS medium containing BA(0.5 mg/L) and NAA(0.5mg/L). The protein contents reached the peak at 3 weeks after induction of calli, and then was decreased. Total lipid contents was decreased as the culture period increased. The band pattern of polypeptides showed that 30KD and 45KD polypeptides and 22KD and 45KD polypetides were major proteins in the cotyledon and hypocotyl explants, respectively. However increase of culture period only 30KD protein was highly accumulated.

• Journal of Life Science
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• v.9 no.1
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• pp.29-34
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• 1999

Cotyledon and hypocotyl explants of perilla were cultured on MS medium containing a combined concentration of BA(0.5, 1.0 and 1.5mg/$\ell$) and NAA(0.1, 0.5 and 2.0mg/$\ell$) in order to regenerate the explant and induce the callus. The best regeneration of the explant and induction of the callus were observed in a combined concenteration of 0.5mg/$\ell$ of BA and 0.5mg/$\ell$ NAA both in cotyledon and hypocotyl explants. In cotyledon explants, rooting was achieved upon transferring shoots to MS medium containing 0.5mg/$\ell$ of BA and 0.1mg/$\ell$ of NAA. We also investigated the change of protein and RNA content on developmental stage of callus and plant regeneration of perilla. Protein content was increased but RNA content was decreased as the culture period increases. The banding pattern of polypeptide revealed that both 30KD and 45KD polypeptides were obvious in cotyledon obtained from pre-culture explants, but only 30KD polypeptide was further getting obvious as the culture period increases.

• BMB Reports
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• v.30 no.1
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• pp.21-25
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• 1997

The growth. freezing resistance and electrophoretic protein patterns of carrot callus cultures were investigated following treatment with NaCl for various' intervals at 20$^{\circ}C$. Following 7 day exposure to 250 mM NaCl. freezing tolerance increased, which was measured by 2.3.5-triphenyl tetrazolium chloride (TTC) assay and fresh weight was reduced compared to control cells. Changes of electrophoretic patterns of total and boiling stable proteins were investigated using one or two dimensional gel system. Several proteins with molecular weight of 43 and 21 kDa increased by NaCl treatment. The most prominent change was detected in 21 kDa protein. The steady state level of this protein increased in NaCl treated cells, but decreased in control cells. Twenty one kDa protein was detected only in the NaCl treated cell when boiling stable protein was analyzed. The isoelectric point of 21 kDa protein was identified as 5.7. The timing of increase of 21 kDa protein was correlated to freezing resistance which implied the role of this protein in the induction of freezing resistance of the cell.

• Korean Journal of Breeding Science
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• v.49 no.3
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• pp.170-179
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• 2017

Plant molecular farming has attracted a lot of attention lately in the field of mass production of industrially valuable materials by extending application of the plant as a kind of factory concept. Among them, protein expression system using rice(Oryza sativa L.) callus is a technology capable of mass culture and industrialization because of a high expression rate of a target protein. This study was carried out to develop an Agrobacterium-mediated transformation system to increase the utilization of rice callus. The transformation efficiency was improved by using the hand when seeds were de-husked for callus induction. Furthermore, we were possible induction of callus from 6 years old seed smoothly. Selection of the callus contained the target gene was required a cultivation period of at least 3 weeks, and the most efficient selection period was after 6 weeks of culture including one passage. This selection was confirmed that the gene was stably inserted into the genomic DNA of the plant cell by the southern blot analysis and progeny test. Such an efficient selection system of rice callus that can be cultured in the long term will be contribute to the industrialization of useful recombinant proteins using rice.

• Journal of Plant Biotechnology
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• v.35 no.2
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• pp.133-140
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• 2008

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 $GeneFishig^{TM}$ 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.

• Journal of Plant Biotechnology
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• v.30 no.4
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• pp.315-321
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• 2003

Freezing and drought tolerances in plants are very important for survival in the desert. In an effort to reduce desertifcation in Gobi, a molecular breeding of Artemisia adamsii using the CLP (chitinase like protein, antifreeze protein) and Dhn5 (dehydrin5) genes from barley is performed by introducing them into Artemisia adamsii via Agrobacteria. We had found an optimal combinatorial concentration of hormones at 0.05mg/L of NAA and 0.5mg/L of BA for growth of callus in Artemisia adamsii. In addition, the higher rate of callus induction using hypocotyl as explant was observed comparing to explants of stem and leaf. There were some variations in the level of the proteins expressed among the transgenic lines such that the lines of CLP(CS1-5, 1-7,4-4) and Dhn5(DS2-2, 2-3) lines produce the protein to higher levels. The transgenic lines showing a higher level of Dhn5 exhibited better growth than nontransgenic callus in presence of 10 and 20％ PEG. In case of the CLP tansgenic lines, both CS1-5 and CS1-7 showed a higher level of freezing tolerance determined by ion leakage test.

• The Plant Pathology Journal
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• v.15 no.1
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• pp.38-43
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• 1999

Mutants of the monopartite geminivirus beet curly top virus (BCTV) have been screened for infectivity, systemic movement, replication and symptom development in Arabidopsis thaliana. As known by coding for coat protein, R1 mutant was not infectious and did not move systemically. R2, R3 and L2/L3 mutants produced milder symptoms compared to wild type BCTV but the infectivity was reduced by 40% to 60%. R2 ORF is thought to be involved in the regulation of ssDNA and dsDNA accumulation because only dsDNA was accumulated on R2-infected organs. Disruption of ORF L4 resulted in reduced infections, but the viral DNA was accumulated in infected organs from roots to shoot tips as much as wild type BCTV on Sei-O. In addition, 4 mutants did not produce callus-like tissues on infected organs, suggesting that L4 ORF may play a role in the induction of host cell divisions by virus infection. This result was supported by the patterns of mRNA expression and promoter analysis of the cell cycle marker gene, cycl, on Arabidopsis. cycl mRNA was accumulated on symptomatic organs by wild type BCTV infections but not by L4 mutant. We conclude that the BCTV L4 ORF is essential for symptom developments, specially callus-like formation on infected organs.

• KSBB Journal
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• v.6 no.3
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• pp.289-297
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• 1991

The insecticidal protein gene in the pKL-20-1 clone derived from Bacillus thuringiensis serovar. kurstaki plasmid was subcloned in the plant shuttle vector, pGA643. The 7.3 kb fragment was cloned in the BglII and Hpal sites of pGA643 vector and expressed in E. coli S17-1, which produced insecticidal proteins killing Bombyx mori larvae. The clone was named pHL-20. The protoplast formation, calli induction and plantlet regeneration of Nicotiana tabacum was carried out. A tremendous number of mesophyll protoplasts of N. tabacum were formed, up to 7$\times$105 protoplast per ml, for 20 hours in darkness in the enzyme solution of 0.5% cellulase and 0.1% macerosin, pH 5.8. The viabilities of the protoplasts were maintained above 80% for 6 days in the media containing 2mg/1 of NAA and 1mg/1 of kinetin. Calli were induced from the protoplasts and leaves of the N. tabacum on MS medium containing 0.5mg/1 BAP. Under the culture conditions the protoplasts underwent repeated cell division into calli. Plantlets were regenerated from callus cultures derived from protoplast and leaves. Shoots were induced in a medium containing 1mg/1 of BAP.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.2
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• pp.154-159
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• 2006

Interleukin-18 (IL-18), otherwise known as interferon-gamma-inducing factor (IGIF), is one of several well characterized and important cytokines that contribute to host defenses. The complementary DNA (cDNA) of mature human interleukin-18 gene (hIL-18) was fused with the signal peptide of the rice amylase 1A gene (Ramy1A) and introduced into the plant expression vector under the control of a duplicated CaMV 35S promoter. The recombinant plasmid was transformed into tobacco (Nicotiana tabacum L. cv Havana) using the Agrobacterium-mediated transformation method. The integration of the hlL-18 gene into the genome of transgenic tobacco plants was confirmed by polymerase chain reaction (PCR) amplification and its expression was observed in the suspension cells that were derived from the transgenic plant callus by using Northern blot analysis. The hlL-18 protein was detected in the extracts of the transgenic callus and in the medium of the transgenic tobacco suspension culture by using immunoblot analysis. Based upon enzyme-linked immunosorbant assay (ELISA) results, the expression level of the hlL-18 protein approximated $166{\mu}g/L$ in the suspension culture medium. Bioassay results from the induction of $interferon-{\gamma}$ from a KG-1 cell line indicated that the hlL-18 secreted into the suspension culture medium was bioactive.