• Korean Journal of Medicinal Crop Science
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• v.9 no.1
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• pp.8-14
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• 2001

The purpose of this research was to investigate the effects of extracts from cultured hairy roots of R. undulatum on human kidney epithelial cells. The cytotoxicity was measured by colorimetric assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), neutral red (NR) and sulforhodamine protein B (SRB) with human kidney epithelial cell lines A498. MTT, NR and SRB quantities decreased propotionally in cultured A498 cells treated with the water or chloroform extracts of cultured hairy roots at increasing concentrations. These results suggest that extracts of cultured hairy roots are cytotoxic on human epithelial cells. The cytotoxicity of chloroform fraction was stronger than that of water fraction. The values of $MTT_{50},\;NR_{50}\;SRB_{50}$ of the extracts of chloroform fraction and those of water fraction were measured to be $289.3{\mu}g/ml,\;302.7{\mu}g/ml,\;433.8{\mu}g/ml\;and\;475.8{\mu}g/ml,\;428.3{\mu}g/ml,\;549.5{\mu}g/ml$, in A498 cell line.

• Journal of Plant Biotechnology
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• v.46 no.3
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• pp.205-216
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• 2019

Since the RNA interference (RNAi) had been discovered in many organisms, small non-coding RNA-mediated gene silencing technology, including RNAi have been widely applied to analysis of gene function, as well as crop improvement. Despite the usefulness of RNAi technology, RNAi transgenic crops have various potential environmental risks, including off-target and non-target effects. In this study, we developed methods that can be effectively applied to environmental risk assessment of RNAi transgenic crops and verified these methods in 35S::dsRNAi_eGFP rice transgenic plant we generated. Off-target genes, which can be non-specifically suppressed by the expression of dsRNAi_eGFP, were predicted by using the published web tool, pssRNAit, and verified by comparing their expressions between wild-type (WT) and 35S::dsRNAi_eGFP transgenic rice. Also, we verified the non-target effects of the 35S:: dsRNAi_eGFP plant by evaluating horizontal and vertical transfer of small interfering RNAs (siRNAs) produced in the 35S::dsRNAi_eGFP plant into neighboring WT rice and rhizosphere microorganisms, respectively. Our results suggested that the methods we developed, could be widely applied to various RNAi transgenic crops for their environmental risk assessment.

• Journal of Plant Biotechnology
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• v.44 no.2
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• pp.149-155
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• 2017

Bacterial blight in rice caused by Xanthomonas oryzae pv. oryzae (Xoo) greatly reduces the growth and productivity of this important food crop. Therefore, we sought to increase the resistance of rice to bacterial blight by using a NAC (NAM, ATAF, and CUC) transcription factor, one of the plant-specific transcription factors that is known to be involved in biotic/abiotic stress resistance. By isolating the OsNAC58 gene from rice and analyzing its biological functions related to Xoo resistance, phylogenetic analysis showed that OsNAC58 belongs to group III. To investigate the biological relationship between bacterial leaf blight (BLB) and OsNAC58 in rice, we constructed a vector for overexpression in rice and generated transgenic rice. The expression analysis resulting from use of RT-PCR showed that OsNAC58-overexpressed transgenic rice exhibited higher levels of OsNAC58 expression than wild types. Further, subcellular localization analysis using rice protoplasts showed that the 35S/OsNAC58-SmGFP fusion protein was localized in the nuclei. Thirteen OsNAC58-overexpressed transgenic rice lines, with high expression levels of OsNAC58, showed more resistant to Xoo than did the wild types. Together, these results suggest that the OsNAC58 gene of rice regulates the rice disease resistance mechanism in the nucleus upon invasion of the rice bacterial blight pathogen Xoo.

• Journal of Plant Biotechnology
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• v.32 no.1
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• pp.37-44
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• 2005

Microspores were isolated from pepper (Capsicum annuum L.) anthers by using a micro-blender and cultured in modified NLN medium at $25^{\circ}C$. The influence of pretreatment period at $32^{\circ}C$, adding the 2-hydroxynicotinic acid to a pretreatment medium, and co-pretreatment anthers with microscopes on the induction of embryo were examined. Globular and torpedo embryos were observed from 3 weeks after culture. Embryo development was not synchronized within culture. After 4 weeks in culture, in addition to globular and torpedo embryos, cotyledonary embryos were observed. Normal cotylodonary embryos developed into plantlets when transferred to a solid hormone free B5 medium containing $2\%$ sucrose. Embryo yields were significantly higher after 1- and 2-day pretreatment at $32^{\circ}C$. However the development of embryo ceased at the globular or heart stage. In contrast, embryo yields were lower after 3- to 6-day pretreatment at $32^{\circ}C$ and embryo developed at the cotyledonary stage. After adding the 2-hydroxynicotinic acid to anther pretreatment solution, embryo yields were slightly increased. However most embryos occurred were at the globular or heart stage. Co-pretreatment of microspores with anthers was deleterious for embryo induction and development. AS far as we know, this is the first report of success in obtaining high frequency of embryogenesis and plantlets formation from isolated microspores of pepper. Although the culture conditions have to be optimized further, this promising microspore culture system can be used for genetic transformation, selection for dominant and recessive traits as well as for the production of homozygous doubled haploid plants.

• Journal of the Institute of Electronics Engineers of Korea SP
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• v.43 no.6 s.312
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• pp.28-35
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• 2006

Statistical models of shape variability based on active shape models (ASMs) have been successfully utilized to perform segmentation and recognition tasks in two-dimensional (2D) images. Three-dimensional (3D) model-based approaches are more promising than 2D approaches since they can bring in more realistic shape constraints for recognizing and delineating the object boundary. For 3D model-based approaches, however, building the 3D shape model from a training set of segmented instances of an object is a major challenge and currently it remains an open problem in building the 3D shape model, one essential step is to generate a point distribution model (PDM). Corresponding landmarks must be selected in all1 training shapes for generating PDM, and manual determination of landmark correspondences is very time-consuming, tedious, and error-prone. In this paper, we propose a novel automatic method for generating 3D statistical shape models. Given a set of training 3D shapes, we generate a 3D model by 1) building the mean shape fro]n the distance transform of the training shapes, 2) utilizing a tetrahedron method for automatically selecting landmarks on the mean shape, and 3) subsequently propagating these landmarks to each training shape via a distance labeling method. In this paper, we investigate the accuracy and compactness of the 3D model for the human liver built from 50 segmented individual CT data sets. The proposed method is very general without such assumptions and can be applied to other data sets.

• Horticultural Science & Technology
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• v.31 no.2
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• pp.246-254
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• 2013

Five isolates of Tomato yellow leaf curl virus (TYLCV) collected from various regions of Korea were amplified using PCR and determined the sequences of full-length genome, respectively. The PCR-amplified DNA of each TYLCV isolate was introduced into a binary vector to construct infectious clone containing 1.9 copies of the corresponding viral genome. Various cultivars and breeding lines of tomato were inoculated with Agrobacterium tumefaciens harboring infectious clone of each TYLCV isolate to assess resistance against TYLCV. Susceptible cultivar 'Super-sunread' revealed typical yellowing and narrowing of the upper leaves. In contrast, breeding linesTY12, GC9, GC171, and GC173, which contained the TY-1 and/or TY-3 genes that confer resistance against TYLCV in nature, were completely symptomless, suggesting that the lines were resistant to challenging TYLCV isolates. Symptoms of TYLCV in susceptible tomato cultivars are significantly different from those of TYLCV in the resistant tomato cultivars at 30 days after agroinfiltration. Although genomic DNAs of TYLCV were detected from the breeding lines TY12, GC9, GC171, and GC173 using real-time PCR analysis with specific primers, levels of TYLCV DNA accumulation in the resistant breeding lines were much lower than those of TYLCV DNA accumulation in susceptible tomato cultivars. Similar symptom severity and levels of TYLCV DNA accumulation were observed from TYLCV infections mediated by Bemisia tabaci in the resistant and susceptible tomato cultivars. Concentration of agrobacterium did not affect the response of tomato cultivars against TYLCV inoculation. Taken together, these results suggest that TYLCV inoculation via agroinfiltration is as effective as inoculation through Bemisia tabaci and is useful for breeding programs of TYLCV-resistant tomato.

• Journal of Plant Biotechnology
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• v.40 no.1
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• pp.18-26
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• 2013

Genetically modified (GM) rice Agb0101, which expresses the insecticidal toxin modified cry1Ac (mcry1Ac1) gene, was developed by the Rural Development Administration in Korea. To monitor the probable release of Agb0101 in the future, it is necessary to develop a reliable detection method. Here, we developed the PCR detection method for monitoring and tracing of GM rice. The primer pair (RBEgh-1/-2) from a starch branching enzyme (RBE4) gene was designed as an endogenous reference, giving rise to an expected PCR amplicon of 101 bp. For the qualitative PCR detection, construct- and event-specific primers were designed on the basis of integration sequence of T-DNA. Event-specific PCRs amplified specifically 5'- or 3'-junction region spanning the native genome DNA and the integrated gene construct, while none of amplified product was shown on crops, rice varieties, and other insect-resistant transgenic rice lines. The event-specific real-time PCR method was performed using TaqMan probe and plasmid pRBECrR containing both rice endogenous gene RBE4 sequence and 5'-junction sequence as the reference molecule. The absolute limit of quantification (LOQ) of real-time PCR was established with around 10 copies for one plasmid molecule pRBECrR. Thereafter, the different amounts of transgenic rice (1, 3, 5, and 10%, respectively) were quantified by using the established real-time PCR method, with a range below 19.55% of the accuracy expressed as bias, 0.06-0.40 of standard deviation (SD) and 3.80-7.01% of relative standard deviations (RSD), respectively. These results indicate that the qualitative and quantitative PCR methods could be used effectively to detect the event Agb0101 in monitoring and traceability.

• Journal of The Korean Society of Grassland and Forage Science
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• v.29 no.1
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• pp.1-6
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• 2009

In order to optimize tissue culture conditions for genetic transformation of zoysiagrass (Zoysia japonica Stued.), the effect of plant growth regulators and culture medium supplements on embryogenic callus induction from mature seeds of a cultivar 'Zenith' were investigated. The optimal concentration and treatment period of NaOCl is 30% (v/v) for 60 minutes. Cultivation of mature seed on the callus Induction medium containing 3 mg/L 2,4-D and 3 mg/L dicamba showed 17.5% of embryogenic callus formation frequency. Supplementation of 1 g/L casein hydrolysate and 500 mg/L L-proline improved frequency of embryogenic callus induction. Audition of the medium with 5 mg/L $AgNO_3$ and 20 mg/L cysteine enhanced frequencies of embryogenic callus induction. Efficient callus induction system established in this study will be useful for molecular breeding of Boysiagrass through genetic transformation.