• Korean Journal of Breeding Science
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• v.41 no.3
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• pp.261-270
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• 2009

Oxidative stress is a major damaging factor for plants exposed to environmental stresses. In order to develop transgenic rice plants with enhanced tolerance to various environmental stresses, PsAPX1, the gene of ascorbate peroxidase isolated from Pisum sativum was expressed in chloroplast under the control of an oxidative stress inducible sweet potato peroxidase2 (SWPA2) promoter (referred to as PsAPX1 plants). PsAPX1 transgenic plants showed enhanced tolerance to various environmental stresses, such as 170 mM NaCl, UV-B, ozone, 20% PEG, and drought in compared with non-transgenic (NT) plants. These results suggest that chloroplast-targeted over-expression of PsAPX1 gene could be very useful strategy for developing transgenic rice plants with increased tolerance to environmental stresses.

• Food Science and Preservation
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• v.30 no.3
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• pp.483-491
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• 2023

This study aimed to compare the bioactive compounds in Centella asiatica (C. asiatica) cultivated in a smart farm and a field and their effects on human keratinocyte cells. C. asiatica was collected in Jeju-do, Korea, and cultured in a smart farm and a field. The main bioactive compounds in the two differentially cultured C. asiatica were identified, and their activation in keratinocytes were assessed. Amplification and sequencing of the internal transcribed spacer (ITS) DNA in the nucleus and psbA-H DNA in the chloroplast were performed for species analysis. A comparison of DNA of plants reported in the NCBI GenBank was performed. The ITS DNA and psbA-H DNA sequences of C. asiatica cultivated in a smart farm and a field were consistent with No. MH768338.1 and No. JQ425422.1, respectively. Analysis of the triterpenes was performed using high performance liquid chromatography (HPLC) and as a result, C. asiatica cultured in a smart farm had more triterpenes than those cultured in a field. The effects of C. asiatica grown in a smart farm on cell proliferation and scratch recovery in HaCaT cells were greater than those grown in a field. These results suggest that C. asiatica cultivated in a smart farm can be effectively utilized as a health functional food.

• Journal of Korean Society of Forest Science
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• v.104 no.4
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• pp.549-557
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• 2015

Novel cpSSR primers were developed based on the sequence information of the Pinus koraiensis chloroplast genome. A total of 30 cpSSR loci were detected in the chloroplast genome, and a total of 30 primer sets flanking those loci were designed. All primer sets were successfully amplified for chloroplast DNA in P. koraiensis. The cross-species transferability of the 30 primer sets was considerably high in P. pumila (100%) and P. paviflora (97%) belonging to the same Subgenus (Strobus) of P. koraiensis. Meanwhile, the transferability was relatively low (73%) in P. densiflora and P. sylvestris belonging to Subgenus Pinus. A total of 13 cpSSR loci out of the 30 loci were polymorphic in the Mt. Jumbong population of P. koraiensis. The mean of haploid diversity(H) was 0.512. The number of haplotypes(N) and the haplotype diversity($H_e$) were 25 and 0.992, respectively. Of the 25 haplotypes, 22 were unique in the analyzed population. The unique haplotypes differentiated 22 individuals (79%) from the total of 28 individuals. In conclusion, the novel cpSSR primers developed in this study would be applicable to other Pinus species, especially the subgenus Strobus, and provide a high level of polymorphism for the study of genetic variation of P. koraiensis.

• Journal of Plant Biotechnology
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• v.50
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• pp.34-44
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• 2023

The tetraploid Solanum hjertingii, a wild tuber-bearing species from Mexico is a relative of potato, S. tuberosum. The species has been identified as a potential source of resistance to blackening for potato breeding. It does not exhibit enzymatic browning nor blackspot which are physiological disorders. However, due to their sexual incompatibility, somatic hybridization between S. hjertingii and S. tuberosum must be used to introduce various traits from this wild species into potato. After somatic hybridization, molecular markers are essential for selecting fusion products. In this study, the chloroplast genome of S. hjertingii was sequenced by next-generation sequencing technology and compared with those of other Solanum species to develop specific markers for S. hjertingii. The chloroplast genome has a total sequence length of 155,545 bp, and its size, gene content, order and orientation are similar to those of the other Solanum species. Phylogenic analysis including 15 other Solanaceae species grouped S. hjertingii with S. demissum, S. hougasii, and S. stoloniferum. After detailed comparisons of the chloroplast genome sequence with eight other Solanum species, we identified one InDel and seven SNPs specific to S. hjertingii. Based on these, five PCR-based markers were developed for discriminating S. hjertingii from other Solanum species. The results obtained in this study will aid in exploring the evolutionary aspects of Solanum species and accelerating breeding using S. hjertingii.

• Journal of Life Science
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• v.17 no.6 s.86
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• pp.772-777
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• 2007

A rice OsLeuD gene for small subunit of 3-isopropylmalate isomerase (IPMI) (EC 4.2.1.33) has been isolated. OsLeuD gene is located on 109.3 cM of chromosome 2. OsLeuD gene was expressed abundantly in metabolically active organs including leaves and developing seeds, indicating that OsLeuD gene expression is developmentally regulated. The cDNA of OsLeuD gene was coded for 257 amino acids which showed 58% and 48% homology to small subunits of IPMI in OsLeuD genes of cyanobacteria and green sulfur bacteria, respectively. The molecular character of OsLeuD is closely related to those of photosynthetic bacteria rather than those of eukaryotes including fungi and yeast. This suggests that OsLeuD gene in chromosomal genome of plants may possibly be originated from chloroplast genome.

• Journal of Life Science
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• v.24 no.10
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• pp.1055-1062
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• 2014

We investigated the expression of cucumber genes involved in lipid mobilization and metabolism during cotyledon development to compare gene activity and to study the direction of carbon (acetyl unit) transport between glyoxysomes and mitochondria. The core metabolic pathway involving 10 genes was examined in four intracellular compartments: glyoxysomes (peroxisomes), mitochondria, chloroplasts, and cytosol. Additionally, we tested the early germination response of dark-grown seedlings and the immediate light response for a further 3 days. According to the reverse transcription polymerase chain reaction (RT-PCR), 3-L-ketoacyl-CoA thiolase 2 (Thio2), isocitrate lyase (ICL), and malate synthase (MS), the genes involved in storage lipid mobilization showed a similar and consistent pattern of gene expression in seedling development. Furthermore, coordinate expression of the A BOUT DE SOUFFLE (BOU) gene with ICL and MS during seedling emergence pointed to a possible secondary route of acetyl unit (acetyl-CoA) transport between peroxisomes and mitochondria in cucumber. The expression of the BOU gene was light dependent, as shown by BOU activity in Arabidopsis, suggesting that the dark condition also results in weak membrane biogenesis. In addition, several genes were active throughout the development of the green cotyledon, even during senescence. In conclusion, this study summarizes oil-seed germination and gene expression during cucumber cotyledon development and proposes an additional route for acetyl unit transport.

• Journal of Plant Biotechnology
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• v.44 no.2
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• pp.135-141
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• 2017

Cudrania tricuspidata Bureau is a widely used medicinal perennial woody plant. For conservation and germplasm utilization of the plant, it is imperative to obtaining information regarding the genetic diversity of the plant populations. Although C. tricuspidata is an important medicinal plant registered in South Korea, no molecular markers are currently available to distinguish Korean-specific ecotypes from other ecotypes of different countries. In this study, we developed single nucleotide polymorphism (SNP) markers derived from chloroplast genomic sequences to identify distinct Korean-specific ecotypes of C. tricuspidata via the amplification refractory mutation system (ARMS)-PCR analyses. Molecular authentication of twelve C. tricuspidata ecotypes from different regions was performed, using DNA sequences in the trnL-F chloroplast intergenic region. The SNP markers developed in this study are useful for rapidly identifying specific C. tricuspidata ecotypes from different regions.

• Korean Journal of Plant Taxonomy
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• v.44 no.1
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• pp.51-58
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• 2014

This study aims to define the phylogenetic relationship within Korean section sabina and find molecular markers which resolve the phylogenetic relationship in genus Juniperus and section sabina. cpDNA matK and psbA-trnH were used as molecular markers. The combined analyses of two genes suggested that section sabina was a clade supported by 100% BP. The relationships of [J. chinensis var. sargentii+J. davurica] clade and [J. chinensis var. chinensis+J. chinensis var. procumbens+J. chinensis var. horizontalis] clade were supported by 91% BP and 100% BP, respectively. Thus, the classification of Korean section sabina would be appropriate at follows, (1) J. chinensis var. sargentii+J. davurica, and (2) J. chinensis var. chinensis+J. chinensis var. procumbens According to the results of separate analyses, matK seems to work better resolving power to clarify the phylogenetic ambiguity in Juniperus and section sabina than psbA-trnH.

• Korean Journal of Plant Tissue Culture
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• v.24 no.2
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• pp.93-97
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• 1997

For the molecular genetic study of cold tolerance mechanism in plants, a cDNA encoding fatty acid desaturase (fad3), converting linoleic acid (18:2, $\omega$-6) to linolenic acid (18:3, $\omega$-3), was isolated from $\lambda$ZAPII Arabidopsis thaliana cDNA expression library by plaque hybridization using fad3 cDNA probe derived from Brassica napus. A 1.8 kb-EcoRI fragment from a lambda clone showing a strong positive hybridization signal was subcloned into pGEM7 and analyzed for its nucleotide sequence. From deduced amino acid sequences, the fad3 gene was revealed to have an open reading frame(ORF) consisting of 386 amino acids with a molecular mass of 44,075 Da. The fad3 gene was compared to chloroplast $\omega$-3 fatty acid desaturase (fad7) and endoplasmic reticulum Δ12 fatty acid desaturase (fad2) to show 70% and 58% amino acid sequence homology, respectively, Especially, amino acids of internal (82 to 151) and carboxy terminal (276 to 333) regions were highly conserved, implying their requisite role for enzymatic functioning of fatty acid desaturases. IPTG-induced fad3 cDNA expression in E. coli cells was suggested to be toxic to bacterial growth.

• Journal of Life Science
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• v.32 no.12
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• pp.956-961
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• 2022

Plant Chloroplast have several advantages as an expression platform of biopharmaceuticals over conventional expression platforms such as mammalian cells, yeast and bacteria. First, plants do not serve as a host for mammalian infectious virus and have endotoxin like bacteria which can cause anaphylactic shock. In addition, high copy number of chloroplast genome allows for chloroplast transformants to reach the high level of expression of heterologous genes. Moreover, the integration of transgenes into specific region of chloroplast genomes makes chloroplast transformants unaffected by positional effect which can be frequently observed from nuclear transformants, resulting in loss of transgene expressions. Antimicrobial peptides (AMPs) are a kind of innate immunity which is found from bacteria to humans. Unlike conventional antibiotics, very less dosage of AMPs can have catastrophic effect on bacterial survival. Further, the repeated use of AMPs does not trigger the development of bacterial resistance. Moricin, one of the AMPs, was isolated from Bombyx mori, a silkworm moth. The C-terminal of moricin consists largely of basic amino acids, and the N-terminal has an α-helix structure. Moricin was chosen and expressed in a SUMO/SUMOase without leaving any unwanted amino acids which could potentially affect the anti-bacterial activity of the moricin. The transformation vector used in this study has already been created in this lab for the expression in both prokaryotic systems such as E. coli and chloroplast. The expressed moricin was purified using Ni columns and SUMOase, and the antibacterial activity of the purified moricin was confirmed using an agar diffusion assay.