• Korean Journal of Plant Taxonomy
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• v.53 no.1
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• pp.47-53
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• 2023

We have determined the complete chloroplast genome of Erigeron Canadensis isolated in Korea. The circular chloroplast genome of E. canadensis is 152,767 bp long and has four subregions: 84,317 bp of large single-copy and 18,446 bp of small single-copy regions are separated by 25,004 bp of inverted repeat regions including 133 genes (88 protein-coding genes, eight rRNAs, and 37 tRNAs). The chloroplast genome isolated in Korea differs from the Chinese isolate by 103 single-nucleotide polymorphisms (SNPs) and 47 insertions and deletion (INDEL) regions, suggesting different invasion sources of E. canadensis in Korea and China. A nucleotide diversity analysis revealed that the trend of the nucleotide diversity of E. canadensis followed that of 11 Erigeron chloroplasts, except for three peaks. The phylogenetic tree showed that our E. canadensis chloroplast is clustered with E. canadensis reported from China. Erigeron canadensis can be a good target when attempting to understand genetic diversity of invasive species.

• Journal of Photoscience
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• v.9 no.2
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• pp.55-58
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• 2002

The reaction center Dl protein of photosystem II is the target of photodamage by excess illumination. The Dl protein is damaged by reactive oxygen species generated by photochemical reactions and then degraded by specific proteolytic enzymes. We found that the Dl protein also cross-links with the surrounding polypeptides, such as D2 and CP43 in isolated thylakoids or photosystem II-enriched membranes from spinach under the illumination with strong visible light. The cross-linking was observed in spinach leaf discs as well when they were illuminated at higher temperature (40°C). It was also shown that the cross-linked products are digested efficiently by a protease(s) in the stroma. Thus the cross-linking/digestion processes of the Dl protein seem to comprise a new pathway in the turnover of the photodamaged Dl protein. It should be noted, however, that the cross-linked products of the Dl protein and CP43 induced by endogenous cationic radicals in the donor-side photoinhibition are resistant to proteolytic digestion. Accumulation of these cross-linked products in the thylakoids may lead to the decay of the function of chloroplasts and finally to the death of plant cells. Thus, we suggest that the quality control of photosystem II, especially removal of the cross-linked products of the Dl protein, is crucial for the survival of chloroplasts under the light stress.

• Journal of Plant Biology
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• v.34 no.4
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• pp.267-273
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• 1991

Polyamine levels in the male and female cells as well as DNA methyltransferase activity in the female cells during gametogenesis of Chlamydomonas reinhardtii indicated that both spermidine and spermine levels were decreased while DNA methyltransferase activity was markedly increased about 12 hours after the onset of gametogenesis. In vitro, putrescine and spermine at 1 mM inhibited methylation of chloroplasts DNA isolated from vegetative female cells by 35% and 65%, respectively. Spermine was found to be more inhibitory than putrescine at all concentrations tested. The pattern of the inhibition by polyamines appeared different from that caused by cations. The results obtained in this work suggest that the polyamine inhibition of DNA methylation is due to an action of polyamines on the enzyme involved instead of on the DNA itself.

• Journal of Plant Biology
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• v.29 no.3
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• pp.157-165
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• 1986

The onset of photophosphorylation at the various stages of greening showed different patterns with varying concentrations of Pi. With further greening, ATP formation occurred at the lower concentration of Pi (48 hrs; 0.05 mM). At early stages of greening, more Pi was required for photophosphorylation (6 hrs; 5.0 mM). The addition of cell-free extracts of etiolated barley seedlings resulted in the competitive inhibition of photophophorylation. The apparent inhibition by cell-free extracts were gradually decreased during greening of etiolated barley seedlings. We found that the inhibitors of photophosphorylation in cell-free extracts were some organic phosphates and most of them was P-choline. P-choline inhibited photophosphorylation competitively with Pi and its content was decreased considerably in greening. It is likely that P-choline partly delay the photophorylation in early stages of greening.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.29-29
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• 2003

Incompatible plant-pathogen interactions result in the rapid cell death response known as hypersensitive response (HR) and activation of host defense-related genes. To understand the molecular and cellular mechanism controlling defense response better, several approaches including isolation and characterization of novel genes, promoter analysis of those genes, protein-protein interaction analysis and reverse genetic approach etc. By using the yeast two-hybrid system a clone named Tsipl, Tsil -interacting protein 1, was isolated whose translation product apparently interacted with Tsil, an EREBP/AP2 type DNA binding protein. RNA gel blot analysis showed that the expression of Tsipl was increased by treatment with NaCl, ethylene, salicylic acid, or gibberellic acid. Transient expression analysis using a Tsipl::smGFP fusion gene in Arabidopsis protoplasts indicated that the Tsipl protein was targeted to the outer surface of chloroplasts. The targeted Tsipl::smGFP proteins were diffused to the cytoplasm of protoplasts in the presence of salicylic acid (SA) The PEG-mediated co-transfection analysis showed that Tsipl could interact with Tsil in the nucleus. These results suggest that Tsipl-Tsil interaction might serve to regulate defense-related gene expression. Basically the useful promoters are valuable tools for effective control of gene expression related to various developmental and environmental condition.(중략)

• Proceedings of the Botanical Society of Korea Conference
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• 1999.07a
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• pp.57-62
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• 1999

Light-regulated translation of chloroplast mRNAs requires nuclear-encoded trans-acting factors that interact with the 5' untranslated region (UTR) of these mRNAs. A set of four proteins (60, 55, 47, and 38 kDa) that bind to the 5'-UTR of the psbA mRNA had been identified in C. reinhardtii. 47 kDa protein (RB47) was found to encode a chloroplast poly (A)-binding protein (cPABP) that specifically binds to the 5'-UTR of the psbA mRNA, and essential for translation of this mRNA, cDNA encoding 60 kDa protein (RB60) was isolated, and the amino acid sequence of the encoded protein was highly homologous to plants and mammalian protein disulfide isomerases (PDI), normally found in the endoplasmic reticulum (ER). Immunoblot analysis of C. reinhardtii proteins showed that anti-PDI recognized a distinct protein of 56 kDa in whole cell extract, whereas anti-rRB60 detected a 60 kDa protein. The ER-PDI was not retained on heparin-agarose resin whereas RB60 was retained. In vitro translation products of the RB60 cDNA can be transported into C. reinhardtii chloroplast in vitro. Immunoblot analysis of isolated pea chloroplasts indicated that higher plant also possess a RB60 homolog. In vitro RNA-binding studies showed that RB60 modulates the binding of cPABP to the 5'-UTR of the psbA mRNA by reversibly changing the redox status of cPABP using redox potential or ADP-dependent phosphorylation. Site-directed mutagenesis of -CGHC- catalytic site in thioredoxin-like domain of RB60 is an unique PDI located in the chloroplast of C. reinhardtii, and suggest that the chloroplast PDI may have evolved to utilize the redox-regulated thioredoxin like domain as a mechanism for regulating the light-activated translation of the psbA mRNA.

• Proceedings of the Botanical Society of Korea Conference
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• 1985.08b
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• pp.7-18
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• 1985

Light-regulated translation of chloroplast mRNAs requires nuclear-encoded trans-acting factors that interact with the 5' untranslated region (UTR) of these mRNAs. A set of four proteins (60, 55, 47, and 38 kDa) that bind to the 5'-UTR of the psbA mRNA had been identified in C. reinhardtii. 47 kDa protein (RB47) was found to encode a chloroplast poly (A)-binding protein (cPABP) that specifically binds to the 5'-UTR of the psbA mRNA, and essential for translation of this mRNA, cDNA encoding 60 kDa protein (RB60) was isolated, and the amino acid sequence of the encoded protein was highly homologous to plants and mammalian protein disulfide isomerases (PDI), normally found in the endoplasmic reticulum (ER). Immunoblot analysis of C. reinhardtii proteins showed that anti-PDI recognized a distinct protein of 56 kDa in whole cell extract, whereas anti-rRB60 detected a 60 kDa protein. The ER-PDI was not retained on heparin-agarose resin whereas RB60 was retained. In vitro translation products of the RB60 cDNA can be transported into C. reinhardtii chloroplast in vitro. Immunoblot analysis of isolated pea chloroplasts indicated that higher plant also possess a RB60 homolog. In vitro RNA-binding studies showed that RB60 modulates the binding of cPABP to the 5'-UTR of the psbA mRNA by reversibly changing the redox status of cPABP using redox potential or ADP-dependent phosphorylation. Site-directed mutagenesis of -CGHC- catalytic site in thioredoxin-like domain of RB60 is an unique PDI located in the chloroplast of C. reinhardtii, and suggest that the chloroplast PDI may have evolved to utilize the redox-regulated thioredoxin like domain as a mechanism for regulating the light-activated translation of the psbA mRNA.

• Applied Biological Chemistry
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• v.40 no.6
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• pp.501-507
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• 1997

Chloroplasts isolated from Stevia rebaudiana Bertoni leaves contained an enzyme activity which catalyzed hydroxylation of ent-kaurenoic acid (ent-kaur-16-en-19-oic acid; ent-KA) to steviol (ent-13-hydroxy kaur-16-en-19-oic acid), the diterpenoid carboxylic alcohol which is the aglycone of sweet stevioside-related glycosides. $[^(14)C]-methylated$ ent-KA was used to localize ent-KA hydroxylase. $[^(14)C]-methyl-KA$ was most actively was transformed into methyl-steviol in chloroplast. The enzymatic activity was found in stroma fraction but not in thylakoid membrane in Stevia rebaudiana Bertoni. However, ent-KA 13-hydroxylase activity was not detected in stroma fraction of either Spinacia oleracea and Solidago altissima. The reaction products using $[^(14)C]-methyl-KA$ were purified and identified on TLC autoradiogram. The hydroxylation of ent-KA from stromal protein to form steviol required NADPH and oxygen. FAD and riboflavin stimulated the enzyme activity 1.5-and 1.7-fold, respectively. It also turned out that the activity of this enzyme using methyl-KA as a substrate was 16.7% that of ent-KA. The purified ent-KA 13-hydroxylase did not act on t-cinnamic acid, 4-hydroxyphenyl acetic acid, choline and resorcinol, known as monooxygenase and hydroxylase substrates.

• Journal of Plant Biology
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• v.37 no.2
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• pp.231-236
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• 1994

The effect of $Cd^{2+}$ on the electron transport rate of PSI and PSII was investigated in isolated spinach chloroplasts. In photosystem II, the rate of electron transport was decreased as the concentration of $Cd^{2+}$ was increased from 1 to $100\;\mu\textrm{M}$. The inhibitory effect of $Cd^{2+}$ was reduced when diphenylcarbazide was added to the reaction medium, indicating that $Cd^{2+}$ affects primarily psn oxygen evolving complexes of thylakoid membrane. The inhibitory effect of $Cd^{2+}$ was reduced when $Mn^{2+}\;and\;Ca^{2+}$ were added to the reaction medium, but the inhibitory effect was not fully relieved. Although the activity of psn was decreased significantly by the treatment of $50\;\mu\textrm{M}\;Cd^{2+}$, Fv/Fm was decreased slightly. However, the treatment of $100\;\mu\textrm{M}\;Cd^{2+}$ resulted in the marked decrease of Fv/Fm. In photosystem I, the rate of electron transport decreased as the concentration of $Cd^{2+}$ was increased from 0.2 to 3.2 mM. The inhibitory effect of $Cd^{2+}$ was decreased when the chloroplast treated with $Cd^{2+}$ was washed by centrifugation.gation.

• BMB Reports
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• v.34 no.4
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• pp.316-321
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• 2001

Dehydroascorbate (DHA) reductase (DHAR, EC 1.8.5.1) catalyzes the reduction of DHA to reduced ascorbate (AsA) using glutathione (GSH) as the electron donor in order to maintain an appropriate level of ascorbate in plant cells. To analyze the physiological role of DHAR in environmental stress adaptation, we developed transgenic tobacco (Nicotiana tabacum cv. Xanthi) plants that express a human DHAR gene isolated from the human fetal liver cDNA library in the chloroplasts. We also investigated the DHAR activity, levels of ascorbate, and GSH. Two transgenic plants were successfully developed by Agrobacterium-mediated transformation and were confirmed by PCR and Southern blot analysis. DHAR activity and AsA content in mature leaves of transgenic plants were approximately 1.41 and 1.95 times higher than in the non-transgenic (NT) plants, respectively In addition, the content of oxidized glutathione (GSSG) in transgenic plants was approximately 2.95 times higher than in the NT plants. The ratios of AsA to DHA and GSSG to GSH were changed by overexpression of DHAR, as expected, even though the total content of ascorbate and glutathione was not significantly changed. When tobacco leaf discs were subjected to methyl viologen at $5\;{\mu}M$, $T_0$ transgenic plants showed about a 50% reduction in membrane damage compared to the NT plants.