• BMB Reports
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• v.37 no.5
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• pp.538-545
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• 2004

A new CRT binding factor (CBF) gene designated Cbcbf25 was cloned from Capsella bursa-pastoris, a wild grass, by the rapid amplification of cDNA ends (RACE). The full-length cDNA of Cbcbf25 was 898 bp with a 669 bp open reading frame (ORF) encoding a putative DRE/CRT (LTRE)-binding protein of 223 amino acids. The predicted CbCBF25 protein contained a potential nuclear localization signal (NLS) in its N-terminal region followed by an AP2 DNA-binding motif and a possible acidic activation domain in the C-terminal region. Bioinformatic analysis revealed that Cbcbf25 has a high level of similarity with other CBF genes like cbf1, cbf2, and cbf3 from Arabidopsis thaliana, and Bncbf5, Bncbf7, Bncbf16, and Bncbf17 from Brassica napus. A cold acclimation assay showed that Cbcbf25 was expressed immediately after cold triggering, but this expression was transient, suggesting that it concerns cold acclimation. Our study implies that Cbcbf25 is an analogue of other CBF genes and may participate in cold-response, by for example, controlling the expression of cold-regulated genes or increasing the freezing tolerance of plants.

• Journal of Plant Biotechnology
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• v.37 no.2
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• pp.166-173
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• 2010

The purpose of functional genome research is to identify biological function of useful gene and to give an agricultural value in plant biotechnology. Brassica rapa is an economic crop which recorded 1,000 billion won of domestic market and 100 million dollar of exports and it produces 2.5 million ton in 50,000 ha as a major ingredient of representative Korean food, Kimchi. Furthermore, it is very important crop economically and commercially because Korea is major seed exporter. The fact that Multinational Brassica Genome Project (MBGP) was launched and Arabidopsis thaliana, affiliated to same genus with B. rapa, has been fully sequenced activated functional genome research of B. rapa. Besides new technologies related to gene function analysis keep developing, many results are reporting every year by international research including Korea. This review paper introduces development of Chinese cabbage mutants which is a first step in functional genome research, variant phenotypes of mutants, flanking DNA analysis in B. rapa genome, gene identification, gene analysis using microarray, and representative researches.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 1995.06b
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• pp.27-53
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• 1995

In plant pathology there is an increasing necessity for improved cytological techniques as basis for the localization of cellular substances within the dynamic fine structure of the host-(plant)-pathogen-interaction. Low temperature (LT) preparation techniques (shock freezing, freeze substitution, LT embedding) are now successfully applied in plant pathology. They are regarded as important tools to stabilize the dynamic plant-pathogen-interaction as it exists under physiological conditions. - The main advantage of LT techniques versus conventional chemical fixation is seen in the maintenance of the hydration shell of molecules and macromolecular structures. This results in an improved fine structural preservation and in a superior retention of the antigenicity of proteins. - A well defined ultrastructure of small, fungal organisms and large biological samples such as plant material and as well as the plant-pathogen (fungus) infection sites are presented. The mesophyll tissue of Arabidopsis thaliana is characterized by homogeneously structured cytoplasm closely attached to the cell wall. From analyses of the compatible interaction between Erysiphe graminis f. sp. hordei on barley (Hordeum vulgare), various steps in the infection sequence can be identified. Infection sites of powdery mildew on primary leaves of barley are analysed with regard to the fine structural preservation of the haustoria. The presentation s focussed on the ultrastructure of the extrahaustorial matrix and the extrahaustorial membrane. - The integration of improved cellular preservation with a molecular analysis of the infected host cell is achieved by the application of secondary probing techniques, i.e. immunocytochemistry. Recent data on the characterization of freeze substituted powdery mildew and urst infected plant tissue by immunogold methodology are described with special emphasis on the localization of THRGP-like (threonine-hydrxyproline-rich glycoprotein) epitopes. Infection sites of powdery mildew on barley, stem rust as well as leaf rust (Puccinia recondita) on primary leaves of wheat were probed with a polyclonal antiserum to maize THRGP. Cross-reactivity with the anti-THRGP antiserum was observed over the extrahaustorial matrix of the both compatible and incompatible plant-pathogen interactions. The highly localized accumulation of THRGP-like epitopes at the extrahaustorial host-pathogen interface suggests the involvement of structural, interfacial proteins during the infection of monocotyledonous plants by obligate, biotrophic fungi.

• Korean Journal of Weed Science
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• v.31 no.3
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• pp.240-249
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• 2011

The potential of juglone a plant naphthoquinone as a natural herbicide on new target, 7-keto-8-amino pelargonic acid synthetase (KAPAS) in the early step of biotin biosynthesis pathway, was performed in vitro and in vivo. Juglone effectively inhibited KAPAS activities in vitro and the $IC_{50}$ was $9.5{\mu}M$. Foliar application of juglone showed very good herbicidal activity to the eight-tested weed species. Among them, Solanum nigrum was completely controlled at a concentration of $250{\mu}g\;mL^{-1}$ with main symptoms of desiccation or burndown. Digitaria sanguinalis and Aeschynomene indica were also sensitive to juglone treatment. All eight weed species were controlled by 90~100% at a concentration of $500{\mu}g\;mL^{-1}$. However, soil application of juglone to Digitaria sanguinalis did not show any herbicidal symptoms. Cellular leakage from cucumber leaf squares treated with juglone increased depending on the concentrations increased from 6.25 to $100{\mu}M$ after 24 hours incubation with or without light. However, chlorophyll loss in cucumber leaf squares was negligible. Biotin supplements significantly rescued the inhibition of germination rate of Arabidopsis thaliana seeds previously inhibited by the juglone. Our results suggest that the juglone is a possible environmental friendly herbicide candidate with a new target KAPAS inhibiting activity.

• Journal of Plant Biotechnology
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• v.33 no.2
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• pp.85-91
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• 2006

Flavanone 3$\beta$-hydroxylase (FHT) is an enzyme acting in the central part of the flavonoid biosynthesis pathway. FHT catalyses the hydroxylation of flavanone to dihydroflavonols in the anthocyanin pathway. In this paper we describe the cloning and expression of the genes encoding the flavonoid-biosynthetic enzyme FHT in Gypsophila paniculata L. A heterologous cDHA probe from Dianthus cavophyllus was used to isolate FHT-encoding cDHA clones from Gypsophila paniculata L.. Inspection of the 1471 bp long sequence revealed an open reading frame 1047 bp, including a 190 bp 5' leader region and 288 bp 3' untranslated region. Comparison of the coding region of this FHT cDHA sequence including the sequences of Arabidopsis thaliana, Citrus sinensis, Dianthus caryophyllus, Ipomoea batatas, Matthiola incana, Nierembergia sp, Petunia hybrida, Solanum tuberosum, Vitis vinifera reveals a identity higher than 69% at the nucleotide level. The function of this nucleotide sequences were verified by comparison with amino acid sequences of the amino-terminus and tryptic peptides from purified plant enzyme, by northern blotting with mRHA from wild type and mutant plants, by in vitro expression yielding and enzymatically active hydroxylase, as indicated by the small dihydrokaempferol peak. Genomic southern blot analysis showed the presence of only one gene for FHT in Gypsophila paniculata.

• Korean Journal of Breeding Science
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• v.42 no.5
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• pp.534-539
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• 2010

'Hongro' is early-mid maturing cultivar with good quality like 'Tsugaru' and it has not preharvest drop. The CAX1 gene was introduced into Korean apple cultivar 'Hongro' by Agrobacterium tumefaciens LBA4404 harboring pBI121 to obtain transgenic apple with enhanced Ca level. The CAX1 gene playing the role of $H^+/Ca^{2+}$ transporter from Arabidopsis thaliana increases Ca concentration in several plants. Regenerated transgenic lines were confirmed by polymerase chain reaction (PCR) analysis and Southern blot analysis of genomic DNA for the existence of CAX1 gene. Southern blot analysis of 'Hongro' transformants showed that two putative transgenic lines were integrated with CAX1 gene in genomic DNA. The CAX1 comparative expression levels of two transgenic lines were higher than that of non-transformant evaluated by comparative quantification analysis using a real-time PCR. These two lines were multiplied in vitro, and micro-grafted on apple rootstocks 'M.9' in the isolated greenhouse. Since two years after micro-grafting, the fruits came into bearing. Compared to Ca level of the non-transgenic 'Hongro', that of the CAX1 transgenic 'Hongro' in the flesh and leaves was higher.

• Journal of Plant Biotechnology
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• v.33 no.3
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• pp.153-160
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• 2006

Brassica rape is an important species used as a vegetable, oil, and fodder worldwide. It is related phylogenically to Arabidopsis thaliana, which has already been fully sequenced as a model plant. The 'Multinational Brassica Genome Project (MBGP)'was launched by the international Brassica community with the aim of sequencing the whole genome of B. rapa in 2003 on account of its value and the fact that it has the smallest genome among the diploid Brassica. The genome study was carried out not only to know the structure of genome but also to understand the function and the evolution of the genes comprehensively. There are two mapping populations, over 1,000 molecular markers and a genetic map, 2 BAC libraries, physical map, a 22 cDHA libraries as suitable genomic materials for examining the genome of B. rapa ssp. pekinensis Chinese cabbage. As the first step for whole genome analysis, 220,000 BAC-end sequences of the KBrH and KBrB BAC library are achieved by cooperation of six countries. The results of BAC-end sequence analysis will provide a clue in understanding the structure of the genome of Brassica rapa by analyzing the gene sequence, annotation and abundant repetitive DHA. The second stage involves sequencing of the genetically mapped seed BACs and identifying the overlapping BACs for complete genome sequencing. Currently, the second stage is comprises of process genetic anchoring using communal populations and maps to identify more than 1,000 seed BACs based on a BAC-to-BAC strategy. For the initial sequencing, 629 seed BACs corresponding to the minimum tiling path onto Arabidopsis genome were selected and fully sequenced. These BACs are now anchoring to the genetic map using the development of SSR markers. This information will be useful for identifying near BAC clones with the seed BAC on a genome map. From the BAC sequences, it is revealed that the Brassica rapa genome has extensive triplication of the DNA segment coupled with variable gene losses and rearrangements within the segments. This article introduces the current status and prospective of Korea Brassica Genome Project and the bioinformatics tools possessed in each national team. In the near future, data of the genome will contribute to improving Brassicas for their economic use as well as in understanding the evolutional process.

• Korean Journal of Weed Science
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• v.31 no.3
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• pp.250-259
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• 2011

This study was conducted to assess the possibility of 5,8-dihydroxy-1,4-naphthoquinone (DHNQ) as a environmental friendly herbicide candidate. Foliar application of DHNQ showed excellent herbicidal effect to the 3 grasses and 5 broad-leaved weeds. Among them, Digitaria sanguinalis and Solanum nigrum were completely controlled by $250{\mu}g\;mL^{-1}$ of DHNQ with main symptoms of desiccation or burndown within 24 hours. Aeschynomene indica was also sensitive to DHNQ treatment. All of the eight weed species were controlled by 90~100% at a concentration of $1000{\mu}g\;mL^{-1}$. However, soil application of DHNQ to Digitaria sanguinalis did not show any herbicidal symptoms. DHNQ strongly inhibited KAPAS activities in vitro and the $IC_{50}$ was $4.4{\mu}M$. Cellular leakage from cucumber leaf squares treated with DHNQ increased depending on the concentrations increased from 6.25 to $100{\mu}M$ after 24 hours incubation with or without light. However, chlorophyll loss in cucumber leaf squares was negligible. Biotin supplements significantly rescued the inhibition of germination rate of Arabidopsis thaliana seeds previously inhibited by the DHNQ. According to above results, DHNQ is a good natural herbicide candidate having a new target KAPAS, which is involved in biotin biosynthesis pathway, with environmental friendly.