• The Plant Pathology Journal
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• v.16 no.3
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• pp.151-155
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• 2000

A cDNA (Oslls1) encoding Lls1-homologue of maize was isolated from cDNA library of rice (Oryza sativa cv. Ilpum). The 2,138 bp of full length Oslls1 clone contains an open reading frame of 1,623 nucleotides encoding 575 amino acid residues. The deduced amino acid sequence of Oslls1 has a high level of homology with chlorophyll a oxygenases of Arabidopsis thaliana (67%) and Marchantia polymorpha (65%). Southern blot analysis of genomic DNA indicates the existence of a small gene family for Oslls1 in the rice genome. The expression of Oslls1 mRNA was induced in leaves and germinating seeds. Treatment of $H_2O$$_2$significantly down-regulated Oslls1 expression. The expression of Oslls1 mRNA was consititutively down-regulated in the blm, a rice mutant exhibiting spontaneous necrotic lesions. These results suggest that this Oslls1 gene may be involved incell death mechanisms in the blm mutant of rice.

• Journal of Plant Biotechnology
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• v.37 no.1
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• pp.89-95
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• 2010

Dihydroflavonol 4-reductase (DFR) is a key enzyme of the flavonoid biosynthesis pathway which catalyses the NADPH-dependent reduction of 2R,3R-trans-dihydroflavonols to leucoanthocyanidins. In this study we describe cloning and expression of the genes encoding the flavonoid-biosynthetic enzyme DFR in Gypsophila paniculata L. Inspection of the 1279 bp long sequence revealed an open reading frame 1063 bp, including a 36 bp 5' leader region and 181 bp 3' untranslated region. Comparison of the coding region of this DFR cDNA sequence including the sequences of Arabidopsis thaliana, Citrus sinensis, Dianthus caryophyllus, Ipomoea batatas, Matthiola incana, Nierembergia sp, Petunia hybrida, Solanum tuberosum, Vitis vinifera reveals an identity higher than 69% at the nucleotide level. The function of this nucleotide sequences was verified by comparison with amino acid sequences of the amino-terminus and tryptic peptides from purified plant enzyme, by northern blotting with mRNA from wild type and mutant plants, by in vitro expression yielding and enzymatically active reductase, as indicated by the small leucopelargonidin peak. Genomic southern blot analysis showed the presence of only one gene for DFR in Gypsophila paniculata.

• Horticultural Science & Technology
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• v.29 no.2
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• pp.135-143
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• 2011

In the Doritaenopsis hybrid, like most of the orchid species and hybrids, temperature is crucial for the vegetative-to-reproductive transition, and low temperature is required for bud differentiation. To understand the molecular mechanism of this process, an orchid GIGANTEA (GI) gene, DhGI1, was isolated and characterized by using the rapid amplification of cDNA ends (RACE) PCR technique. Sequence analysis showed that the full-length cDNA is 4,022 bp with a major open reading frame of 3,483 bp, and the amino acid sequence showed high similarity to GI proteins in Zea mays, Oryza sativa, Arabidopsis thaliana and other plants. Semi-quantitative RT-PCR revealed that DhGI1 was expressed throughout development and could be detected in roots, stems, leaves, peduncles and flower buds. The expression level of DhGI1 was higher when the plants were flowering at low temperature (22/$18^{\circ}C$ day/night) than the other growth stages. Further analysis indicated that the accumulation of DhGI1 transcripts was significantly increased at low temperature, and concomitantly, initiation of the peduncle was observed. However, DhGI1 levels were low under high temperature (30/$25^{\circ}C$) conditions, and flower initiation was inhibited. These results indicate that the expression of DhGI1 is regulated by low temperature and that DhGI1 may play an important role in inflorescence initiation in this Doritaenopsis hybrid at low temperatures.

• Korean Journal of Plant Resources
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• v.16 no.3
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• pp.257-263
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• 2003

The NDK1 is an ubiquitous enzyme that transfer phosphate groups from triphosphate nucleoside diphosphates(NDPs) in eukaryotes and prokaryotes. We isolated and characterized a cDNA encoding a nucleoside diphosphate kinase 1(CNDK 1) in Codonopsis lanceolata. The CNDK 1 is 444bp long and open reading frame of 447bp with a deduced amino acid of 148 residue. The CNDK 1 has an ATP binding site in 12­16 residue and phosphohistidine intermediate in 115 residue of amino acid sequence. Although several NDK 1 genes have been cloned in plants, but little is known about the functional significance of this enzyme during plant growth and development. The CNDK 1 shows the identities to Arabidopsis thaliana (71％), Oryza sativa(75％), Glycine max (79％), Brassica rapa (77％), Mesembryanthemum crystallinum (85 ％), Spinacia oleracea (83％), Pisum sativum (82％). The CNDK 1 of C. laceolata have a closer relationship of Glycine max and Pisum sativum at the phylogenic analysis.

• Plant Biotechnology Reports
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• v.2 no.1
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• pp.41-46
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• 2008

Salinity stress is a major limiting factor in cereal productivity. Many studies report improvements in salt tolerance using model plants, such as Arabidopsis thaliana or standard varieties of rice, e.g., the japonica rice cultivar Nipponbare. However, there are few reports on the enhancement of salt tolerance in local rice cultivars. In this work, we used the indica rice (Oryza sativa) cultivar BR5, which is a local cultivar in Bangladesh. To improve salt tolerance in BR5, we introduced the Escherichia coli catalase gene, katE. We integrated the katE gene into BR5 plants using an Agrobacterium tumefaciens-mediated method. The introduced katE gene was actively expressed in the transgenic BR5 rice plants, and catalase activity in $T_1$ and $T_2$ transgenic rice was approximately 150% higher than in nontransgenic plants. Under NaCl stress conditions, the transgenic rice plants exhibited high tolerance compared with nontransgenic rice plants. $T_2$ transgenic plants survived in a 200 mM NaCl solution for 2 weeks, whereas nontransgenic plants were scorched after 4 days soaking in the same NaCl solution. Our results indicate that the katE gene can confer salt tolerance to BR5 rice plants. Enhancement of salt tolerance in a local rice cultivar, such as BR5, will provide a powerful and useful tool for overcoming food shortage problems.

• Molecules and Cells
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• v.43 no.1
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• pp.48-57
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• 2020

The microalga Chlamydomonas reinhardtii accumulates triacylglycerols (TAGs) in lipid droplets under stress conditions, such as nitrogen starvation. TAG biosynthesis occurs mainly at the endoplasmic reticulum (ER) and requires fatty acid (FA) substrates supplied from chloroplasts. How FAs are transferred from chloroplast to ER in microalgae was unknown. We previously reported that an Arabidopsis thaliana ATP-binding cassette (ABC) transporter, AtABCA9, facilitates FA transport at the ER during seed development. Here we identified a gene homologous to AtABCA9 in the C. reinhardtii genome, which we named CrABCA2. Under nitrogen deprivation conditions, CrABCA2 expression was upregulated, and the CrABCA2 protein level also increased. CrABCA2 knockdown lines accumulated less TAGs and CrABCA2 overexpression lines accumulated more TAGs than their untransformed parental lines. Transmission electron microscopy showed that CrABCA2 was localized in swollen ER. These results suggest that CrABCA2 transports substrates for TAG biosynthesis to the ER during nitrogen starvation. Our study provides a potential tool for increasing lipid production in microalgae.

• Korean Journal of Environmental Biology
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• v.39 no.3
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• pp.354-361
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• 2021

Understanding the phytotoxic mechanism of methyl bromide (MB), an essential fumigant during the quarantine and pre-shipment process, is urgently needed to ensure its proper use and reduce international economic losses. In a previous study, two main MB-induced toxic mechanisms such as reactive oxygen species (ROS) and auxin distribution were selected by analyzing transcriptomic analysis. In the study, a 3-week-old A. thaliana was supplied with 1 mM ROS scavengers [N-acetyl-L-cysteine (NAC) or L-glutathione (GSH)] and 1µM indole-3-acetic acid(IAA) three times every 12 h, and visual and gene expression assessments were performed to evaluate the reduction in phytotoxicity by supplements. Phytotoxic effects on the MB-4h exposed group were decreased with GSH application compared to the other single supplements and a combination of supplements at 7 days post fumigation. Among these supplements, GSH at a concentration of 1, 2, and 5mM was suppled to A. thaliana with MB-fumigation. During a long-term observation of 2 weeks after the fumigation, 5 mM GSH application was the most effective in minimizing MB-induced phytotoxic effects with up-regulation of HSP70 expression and increase in main stem length. These results indicated that ROS was a main key factor of MB-induced phytotoxicity and that GSH can be used as a supplement to reduce the phytotoxicity of MB.

• BMB Reports
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• v.40 no.2
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• pp.247-260
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• 2007

Cinnamate 4-hydroxylase (C4H) is a key enzyme of phenylpropanoid pathway, which synthesizes numerous secondary metabolites to participate in development and adaption. Two C4H isoforms, the 2192-bp BnC4H-1 and 2108-bp BnC4H-2, were cloned from oilseed rape (Brassica napus). They both have two introns and a 1518-bp open reading frame encoding a 505-amino-acid polypeptide. BnC4H-1 is 57.73 kDa with an isoelectric point of 9.11, while 57.75 kDa and 9.13 for BnC4H-2. They share only 80.6% identities on nucleotide level but 96.6% identities and 98.4% positives on protein level. Showing highest homologies to Arabidopsis thaliana C4H, they possess a conserved p450 domain and all P450-featured motifs, and are identical to typical C4Hs at substrate-recognition sites and active site residues. They are most probably associated with endoplasmic reticulum by one or both of the N- and C-terminal transmembrane helices. Phosphorylation may be a necessary post-translational modification. Their secondary structures are dominated by alpha helices and random coils. Most helices locate in the central region, while extended strands mainly distribute before and after this region. Southern blot indicated about 9 or more C4H paralogs in B. napus. In hypocotyl, cotyledon, stem, flower, bud, young- and middle-stage seed, they are co-dominantly expressed. In root and old seed, BnC4H-2 is dominant over BnC4H-1, with a reverse trend in leaf and pericarp. Paralogous C4H numbers in Brassicaceae genomes and possible roles of conserved motifs in 5' UTR and the 2nd intron are discussed.

• Journal of Plant Biotechnology
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• v.45 no.3
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• pp.155-170
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• 2018

In this paper, mechanisms of gene editing technologies including ZFN, TALENS and CRISPR were briefly discussed with mutual advantages and disadvantages. Classification criteria of gene edited, site-directed mutagenesis (SDN) crops for regulatory purpose were also discussed. The number of studies using CRISPR technology was high and studies conducted on Arabidopsis thaliana and rice were highest, followed by tobacco, tomato, wheat, and corn. It has been applied to a variety of plants such as other grain crops, flower crops, vegetable crops, and fruit trees. The number of studies focused on practical application or commercialization in the future were also increasing yearly, and the scope of studies also expanded to include research on metabolic engineering for mass production of useful proteins or substances, development of disease resistant crops against viruses, bacteria, and fungi, abiotic environmental stressresistant crops, and increased yields. In addition to this, it was revealed that application range is becoming more diversified, including the development of parthenocarpic tomatoes, hybrid rice lines using male sterility and increased shattering resistance Brassica napus. It was also revealed that the number of CRISPR gene edited crops permitted by the USDA(APHIS) increases yearly, to be released in the international seed market soon.

• Molecules and Cells
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• v.38 no.3
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• pp.259-266
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• 2015

The regulation of flowering time has crucial implications for plant fitness. MicroRNA156 (miR156) represses the floral transition in Arabidopsis thaliana, but the mechanisms regulating its transcription remain unclear. Here, we show that two AGAMOUS-like proteins, AGL15 and AGL18, act as positive regulators of the expression of MIR156. Small RNA northern blot analysis revealed a significant decrease in the levels of mature miR156 in agl15 agl18 double mutants, but not in the single mutants, suggesting that AGL15 and AGL18 co-regulate miR156 expression. Histochemical analysis further indicated that the double mutants showed a reduction in MIR156 promoter strength. The double mutants also showed reduced abundance of pri-miR156a and pri-miR156c, two of the primary transcripts from MIR156 genes. Electrophoretic mobility shift assays demonstrated that AGL15 directly associated with the CArG motifs in the MIR156a/c promoters. AGL18 did not show binding affinity to the CArG motifs, but pull-down and yeast two-hybrid assays showed that AGL18 forms a heterodimer with AGL15. GFP reporter assays and bimolecular fluorescence complementation (BiFC) showed that AGL15 and AGL18 co-localize in the nucleus and confirmed their in vivo interaction. Overexpression of miR156 did not affect the levels of AGL15 and AGL18 transcripts. Taking these data together, we present a model for the transcriptional regulation of MIR156. In this model, AGL15 and AGL18 may form a complex along with other proteins, and bind to the CArG motifs of the promoters of MIR156 to activate the MIR156 expression.