• 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.

• Journal of Plant Biology
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• v.34 no.1
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• pp.37-43
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• 1991

Mesophyll protoplasts of Nicoliana labacum ($NR^{-}/SR^{+}$) and N glulinosa were electrofused with AC field of 0.5 MHz and 1 kV DC pulse for 2 ms. Fused protoplasts were selected and cultured to the green cell clusters in $MSNO_3$ medium containing 1.2 mg!ml streptomycin sulfate. Four plant lines regenerated from selected colonies showed both parental morphological characteristics of leaf and flower and these plant lines were confirmed as somatic hybrids based on electrophoretic patterns of leaf peroxidase. In XhoI restriction patterns of chloroplast DNA, these hybrid plant lines expressed both parent common restriction sites and parent specific sites. One of these hybrid lines exhibited interspecific pattern of both parental chloroplast genomes. indicating nine both parent common sites, one N labacum specific site and two N glutinosa specific sites. sites.

• Molecules and Cells
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• v.37 no.5
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• pp.372-382
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• 2014

In this study, the chloroplast (cp) genome sequences from three early diverged leptosporangiate ferns were completed and analyzed in order to understand the evolution of the genome of the fern lineages. The complete cp genome sequence of Osmunda cinnamomea (Osmundales) was 142,812 base pairs (bp). The cp genome structure was similar to that of eusporangiate ferns. The gene/intron losses that frequently occurred in the cp genome of leptosporangiate ferns were not found in the cp genome of O. cinnamomea. In addition, putative RNA editing sites in the cp genome were rare in O. cinnamomea, even though the sites were frequently predicted to be present in leptosporangiate ferns. The complete cp genome sequence of Diplopterygium glaucum (Gleicheniales) was 151,007 bp and has a 9.7 kb inversion between the trnL-CAA and trnV-GCA genes when compared to O. cinnamomea. Several repeated sequences were detected around the inversion break points. The complete cp genome sequence of Lygodium japonicum (Schizaeales) was 157,142 bp and a deletion of the rpoC1 intron was detected. This intron loss was shared by all of the studied species of the genus Lygodium. The GC contents and the effective numbers of codons (ENCs) in ferns varied significantly when compared to seed plants. The ENC values of the early diverged leptosporangiate ferns showed intermediate levels between eusporangiate and core leptosporangiate ferns. However, our phylogenetic tree based on all of the cp gene sequences clearly indicated that the cp genome similarity between O. cinnamomea (Osmundales) and eusporangiate ferns are symplesiomorphies, rather than synapomorphies. Therefore, our data is in agreement with the view that Osmundales is a distinct early diverged lineage in the leptosporangiate ferns.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.04a
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• pp.71-71
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• 2019

The genus Cenchrus (Poaceae), containing ca. 23 species, is distributed throughout Australia, Africa, Indian sub-continent, and America. In Korea, Cenchrus longispinus (Hack.) Fernald, especially introduced to Daecheong Island in 1999, is one of the most hazardous invasive plant which causes serious environmental threats, biodiversity damages and physically negative impact on humans and animals. Based on the next-generation sequencing (NGS) technology, we characterized the chloroplast (cp) genome sequences of C. longispinus which contains a large single copy (LSC; 80,223 bp), a small single copy (SSC; 12,449 bp), separated by a pair of inverted repeats (IRs; 22,236 bp). Additionally, we analyzed the cp genome sequences of Cenchrus echinatus L. which contains a large single copy (LSC; 80,220 bp), a small single copy (SSC; 12,439 bp), separated by a pair of inverted repeats (IRs; 22,236 bp). These cp genomes consist of 75 unique genes, 4 rRNA coding genes, 33 tRNA coding genes and 21 duplicated in the IR regions, of which the gene content and organization are similar to the other Poaceae cp genomes. We selected 40 potential regions in cp genomes of two Cenchrus species and one Korean Pennisetum species to develop new single nucleotide polymorphism (SNP) markers for identifying C. longispinus based on amplification-refractory mutation system (ARMS) technique. The markers, inferred from SNP in matK and ndhF genes, show effectiveness to recognize C. longispinus from C. echinatus and Korean native species Pennisetum alopecuroides (L.) Spreng.

• Korean Journal of Plant Taxonomy
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• v.44 no.1
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• pp.39-50
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• 2014

Phylogenetic positions of Sedirea and Neofinetia were addressed using the chloroplast matK and the nuclear ITS sequences. We also evaluate the usefulness of the makers for the identification of species and localities. Sedirea and Neofinetia form an independent monophyletic genus, respectively, in both matK and nuclear ITS trees. The sister genus of the Neofinetia was Vanda in both trees. In addition, our trees support the separate recognition of the Neofinetia from Vanda rather than the inclusion of Neofinetia into Vanda. The sister group of the Sedirea was (Dimorphorchis(Pteroceras(Saccolabiun+Phalaeonopsis))) clade. The Dimorphorchis was one of the most probable sister genus to the Sedirea. The sister group relationship between Sedirea and Aerides was suggested by their similar morphology, but not supported in molecular trees. The identification of species and localities of Neofinetia was possible using our two molecular markers. However, several pseudo-gene sequences are discovered from the public data base. In addition, the horizontal gene transfer of chloroplast genomes is frequent events in orchid hybrids. Therefore, we need a careful evaluation for the data prior to systematic use. Generation of sequence data from multiple accessions of a species may helpful to reduce these types of error.

• Journal of Plant Biotechnology
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• v.38 no.1
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• pp.42-48
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• 2011

Chloroplast genetic engineering of higher plants offers several unique advantages compared with nuclear genome transformation, such as high levels of transgene expression, a lack of position effect due to site-specific transgene integration by homologous recombination, multigene engineering in a single transformation event and reducing risks of gene flow via pollen due to maternal inheritance. We established a reproducible chloroplast transformation system of potato using a tobacco specific plastid transformation vector, pCtVG (trnI-Prrn-aadA-mgfp-TpsbA-trnA). Stable transgene integration into chloroplast genomes and the homoplasmic state of the transgenome were confirmed by PCR and Southern blot analyses. Northern, immunoblot analysis, and GFP fluorescence imaging revealed high expression and accumulation of GFP in the plastids of potato leaves. This system would provide new opportunities for genetic improvement and mass production of value added foreign proteins in this crop.

• Journal of Plant Biotechnology
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• v.49 no.1
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• pp.30-38
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• 2022

Solanum brevicaule is one of the tuber-bearing wild Solanum species. Because of its resistance to several important pathogens infecting potatoes during cultivation, it can be used for potato breeding. However, the fact that S. brevicaule used in this study has an EBN value of two causes the sexual reproduction barriers between the species and cultivated potatoes. In this study, specific markers for discriminating S. brevicaule from other Solanum species were developed on the basis of the results of sequence alignments with the whole chloroplast genomes of S. brevicaule and seven other Solanum species. The chloroplast genome of S. brevicaule was completed by next-generation sequencing technology described in other recent studies. The total sequence length of the chloroplast genome of S. brevicaule is 155,531 bp. Its structure and gene composition are similar to those of other Solanum species. Phylogenetic analysis revealed that S. brevicaule was closely grouped with other Solanum species. BLASTN search showed that its genome sequence had 99.99% and 99.89% identity with those of S. spegazzinii (MH021562) and S. kurtzianum (MH021495), respectively. Sequence alignment identified 27 SNPs that were specific to S. brevicaule. Thus, three PCR-based CAPS markers specific to S. brevicaule were developed on the basis of these SNPs. This study will facilitate in further studies on evolutionary and breeding aspects in Solanum species.

• Korean Journal of Plant Resources
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• v.36 no.4
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• pp.290-298
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• 2023

Several members of the genus Broussonetia are woody plants with high-quality cellulose fibers and are used to make a traditional type of Korean paper known as Hanji. Three of these species, Broussonetia kazinoki, Broussonetia monoica, and Broussonetia papyrifera, are found in the Korean Peninsula. Because it is challenging to distinguish different Broussonetia species based on morphology alone, we have developed a set of insertion/deletion (InDel) markers for genetic identification of these species. From twenty-two Broussonetia samples collected throughout Korea, we selected six for next-generation sequencing analysis. InDel marker candidates were identified by comparing this sequence information with the B. kazinoki chloroplast genome sequence. The marker candidates were used to screen the genomes of the twenty-two Broussonetia plants, and five useful chloroplast-based InDel markers were identified. Detailed genotyping using these five markers showed that the twenty-two plants of the genus Broussonetia could be clustered into five groups, verifying that the markers developed here can be used for breeding, identification, and analysis of species in the genus Broussonetia.

• Horticultural Science & Technology
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• v.33 no.3
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• pp.390-395
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• 2015

The synthesis of chimeras is a breeding approach for horticultural crops. In our breeding program, a new diploid citrus chimera, named 'Hongrou Taoye' (Citrus sinensis [L.] Osbeck + Citrus unshiu Marc.), was found arising at the junction where a 'Taoye' sweet orange (C. sinensis) scion was grafted onto Satsuma mandarin (C. unshiu). As an artificial chimera, its fruit traits derived from the L1 cell layer, with juice color and carotenoid complement, in which ${\beta}$-cryptoxanthin accumulated predominantly, similar to those of Satsuma mandarin. By contrast, traits originating from the L2/L3 cell layer, including pollen, seed, and rind aroma characteristics, were the same as those of 'Taoye' sweet orange (the scion). SSR and cpSSR analyses showed that both nuclear and chloroplast genomes of the chimera were a combination of both donor parents. 'Hongrou Taoye' thus combined the valuable traits of both donor plants, and therefore has good potential in citrus fresh market.

• Journal of Plant Biotechnology
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• v.2 no.2
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• pp.73-78
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• 2000

The plastid genomes of several plants contain ndh genes homologues of genes encoding subunits of the mitochondrial complex I. We sequenced the part of lupin ndhB, ndhD and ndhF genes in order to compare the structure of these genes with those of Nicotiana tabaum, Arabidopsis thaliana, Zea mays and Oryza sativa with the idea to detect the presence of stretches with identical aminoacid composition. We were only able to find one or two stretches of this kind of about 16 aminoacid- long in the analyzed fragments of the ndh genes. The total number of such stretches was different in particular gene products: for ndhc 1, ndhB 9, ndhD 3 and ndhF 6. We have also examined the transcription pattern of ndhC, ndhK and ndhJ genes during lupin development. We show that the greatest amount of ndhC, ndhK and ndhJ transcripts are observed in 7- to 14 day- old lupin seedlings. We also studied the level of transcription of those genes in plants growing at low temperature. All the data confirmed that the abundance of transcription of ndhC, ndhK, and ndhJ genes increased under chill conditions. It has to be noted that the level of transcription of the ndhC gene was higher than the other genes probably due to higher stability of this transcript.