• Korean Journal of Agricultural Science
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• v.43 no.2
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• pp.205-214
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• 2016

A drought-tolerant transgenic rice (Agb0103) was developed using a pepper methionine sulfoxide reductase (CaMsrB2) under the control of rice Rab21 promoter with a selection marker, the phosphinothricin acetyltransferase (PAT) gene. Commercialization of genetically modified (GM) crops will require the evaluation of risks associated with the release of GM crops. With the potential problems associated to GM crops safety testing, the investigation of their effects on non-target organisms is necessary for environmental risk research. This study was carried out to assess acute toxicity of a GM crop using the water flea (Daphnia magna) for non-target organism risk evaluation. The effect of acute toxicity on Daphnia magna of Agb0103 rice and a non-GM rice, Ilmibyeo, were investigated at different concentrations (0, 625, 1,250, 2,500, 5,000, and 10,000 mg/L). The Agb0103 rice used for the test was confirmed to express the CaMsrB2/PAT gene by the PCR and ELISA. Daphnia magna feeding tests showed no significant differences in cumulative immobility or abnormal response with either Agb0103 rice or non-GM rice. The 48hr-EC50 values showed no difference between Agb0103 rice (2243 mg/L) and non-GM rice (2694 mg/L). These results suggest that there is no significant difference in toxicity to Daphnia magna between Agb0103 rice and its non-GM counterpart.

• Korean Journal of Environmental Agriculture
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• v.33 no.2
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• pp.121-128
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• 2014

BACKGROUND: The disease resistant (OsCK1) rice was generated by inserting choline kinase (CK1) and phosphinothricin acetyltransferase (PAT) genes isolated from Oryza sativa and Streptomyces hygroscopicus into the genome of the rice, Nakdongbyeo. With the potential problems of safeties, the evaluations on non-target organisms are essentially required for the environmental risk assessment of genetically modified (GM) crops. In the present study, we conducted the evaluation of acute toxicity on Daphnia magna that commonly used as a model organism in ecotoxicological studies for non-target organism evaluation. METHODS AND RESULTS: Effect of acute toxicity to Daphnia magna by each concentration were investigated in the disease resistant (OsCK1) rice and non-genetically modified (non-GM) rice, Nakdongbyeo, as concentration (0, 1,000, 1,800, 3,240, 5,830, 10,500 and 20,000 mg/L). The OsCK1 rice used for the test was confirmed to express the OsCK1/PAT gene by the PCR(Polymerase chain reaction) and western blot analysis. Feeding test showed that no significant differences in cumulative immobility and abnormal response of Daphnia magna fed on OsCK1 rice or non-GM rice. The 48hr-$EC_{50}$ values showed no difference between OsCK1 rice (3,147.18 mg/L) and non-GM rice (3,596.27 mg/L). CONCLUSION: This result suggested that there was no significant difference in toxicity to Daphnia magna between OsCK1 rice and non-GM counterpart.

• Korean Journal of Plant Resources
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• v.17 no.1
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• pp.28-33
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• 2004

This study was conducted to simple transformants selection by herbicide Basta treatment after transformation with Agrobacteium tumefaciens MP90/PAT in hybrid poplar(Populus alba ${\times}$ P. glandulosa No. 3). In preliminary study, we determined that the lethal concentration of herbicide Basta was 1.0mg/L in callus culture, adventitious bud formation and axillary bud elongation experiment. By the treatment of 1.0mg/L Basta, we could be selected the transformed shoots effectively from the various cultures. In addition, the treatment was useful on selection of transformants which are growing in soil pot. Finally, we also confirmed the transformation by PAT assay, Above results show that the herbicide Basta treatment and PAT assay can be a very simple and effective method for the identification of PAT gene transformed hybrid poplar.

• Korean Journal of Plant Tissue Culture
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• v.28 no.4
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• pp.197-200
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• 2001

Agrobacterium tumefaciens MP90 harboring PAT (phosphinothricin acetyltransferase) and NPTII-GUS gene were used for the genetic transformation of lettuce (Lactuca Sativa L.). Shoot regeneration from cotyledon explants were obtained from the MS medium supplemented with 0.1 mg.L$^{-1}$ NAA, 1.0 mg.L$^{-1}$ 2ip, 50 mg.L$^{-1}$ kanamycin and 500 mg.L$^{-1}$ carbenicillin after cocultivation with A. tumefaciens for 2 days. Kanamycin resistance test of transgenic plants indicated that the NPTII gene was integrated into the lettuce genome and was stably expressed. PCR and northern blot analysis indicated that bialaphos resistance gene (PAT) was stably integrated into the lettuce genome. The transgenic plant sprayed with Basta (1500x) remained healthy with continuous growth, while the control group exhibited fatality.

• Journal of The Korean Society of Clinical Toxicology
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• v.17 no.1
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• pp.7-13
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• 2019

Purpose: Owing to the increased agricultural use of the herbicide glufosinate ammonium (GLA), the incidence of GLA poisoning has recently increased. Therefore, we investigated the possible predictive factors associated with severe complications following GLA poisoning. Methods: A retrospective analysis of medical records was conducted based on 76 patients who had visited our regional emergency medical center with GLA poisoning from 2006 to 2017. Severe complications were defined as respiratory failure requiring intubation, systolic blood pressure less than 90 mmHg, Glasgow Coma Scale (GCS) less than 8, and presence of seizure. Results: Age, ingested amount and ingested amount per weight were significantly greater in the severe group (p<0.001). PSS grade 2 or higher was more common in the severe group (p<0.001), and In addition, the APACHE II score was significantly higher in the severe group (p<0.001), as were the SOFA scores (p=0.002). Serum ammonia levels were significantly higher in the severe group (p=0.007), while MDRD-GFR was smaller in the severe group (p=0.002). The spot urine protein levels were significantly higher in the severe group (p=0.005), as was the urine protein to creatinine ratio (p=0.001). Upon multivariate analysis, the amount ingested per weight and PSS grade 2 or higher were identified as significant predictors. Conclusion: Our study showed that MDRD-GFR was significantly lower in the severe group after GLA poisoning. PSS grade 2 or higher and ingested amount per weight may be useful to evaluate the severity of complications after GLA poisoning.

• Korean Journal of Agricultural Science
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• v.47 no.4
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• pp.791-802
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• 2020

Theoredoxin (TRX) transgenic soybeans were developed using the human Theoredoxin gene under the control of the ��-conglycinin promoter with a selection marker, the phosphinothricin acetyltransferase (PAT) gene. This study was done to assess the acute toxicity of a genetically modified (GM) soybean using the fresh water planktonic crustacean Daphnia magna. The acute toxicity effect of the TRX soybean and non-GM soybean (Gwangan) on D. magna was investigated at different concentrations (0, 156, 313, 625, 1,250, 2,500, and 5,000 mg·L-1). The TRX soybean used for the test was confirmed to express the TRX/PAT genes by PCR and enzyme-linked immunosorbent assay (ELISA). D. magna feeding tests showed no significant differences in the cumulative immobility or an abnormal response with either the TRX soybean or non-GM soybean. The feeding study showed a similar abnormal response and cumulative immobility of the D. magna between the TRX soybean and Gwangan treatments. Additionally, the 48 h-EC50 values for the TRX and Gwangan soybeans were 755.6 and 778 mg·L-1, respectively. The soybean NOEC (no observed effect concentration) value for D. magna was suggested to be 156 mg·L-1. These results suggest that there is no significant difference in toxicity to Daphnia magna between the TRX soybean and its non-GM counterpart.

• Korean Journal of Agricultural Science
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• v.47 no.4
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• pp.815-827
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• 2020

The epidermal growth factor (EGF) transgenic soybean was developed and biosynthesis of human epidermal growth factor (hEGF) in soybean seeds was confirmed. Also, EGF transgenic soybean were found to contain a herbicide resistance selectable marker by introduction of phosphinothricin acetyltransferase (PAT) gene from the Streptomyces hygroscopicus. For biosafety assessment, the EGF transgenic soybean expressing the EGF biosynthesis gene EGF and herbicide resistant gene PAT was tested to determine effects on survival of Cyprinus carpio, commonly used as a model organism in ecotoxicological studies. C. carpio was fed 100% ground soybean suspension, EGF soybean or non-genetically modified (GM) counterpart soybean (Gwangan). Gene expression of EGF soybean was confirmed by PCR and ELISA to have EGF/PAT. Feeding test showed that no significant differences in cumulative immobility or abnormal response between C. carpio samples fed on EGF soybean and non-GM counterpart soybean. The 48 h-EC50 values of the EGF and non-GM soybean were 1,688 mg·L-1 (95% confidence limits: 1,585 - 1,798 mg·L-1) and 1,575 mg·L-1 (95% confidence limits: 1,433 - 1,731 mg·L-1), respectively. The soybean NOEC (no observed effect concentration) value for C. carpio was suggested to be 625 mg·L-1. We concluded that there was no significant difference in toxicity for non-target organisms (C. carpio) between the EGF soybean and non-GM counterparts.

• Journal of Plant Biotechnology
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• v.47 no.4
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• pp.316-323
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• 2020

This study aimed to evaluate the potential allergenicity and oral toxicity of the phosphinothricin acetyltransferase (PAT) protein expressed in Zoysia japonica, a herbicide-tolerant genetically modified (GM) zoysiagrass. In silico analysis of PAT showed no similarities with any known allergenic or toxic proteins, with <35% amino acid sequence homology with known allergens across a length of 80 amino acids and no continuous eight amino acid identity with known allergens. The PAT protein expressed in Z. japonica degraded very rapidly in the simulated gastric fluid in the presence of pepsin, and, no glycosylation of PAT was observed. The oral toxicity test revealed no mortality or toxic effect in mice following PAT administration at 4,000 mg/kg body weight. Our findings indicate that the PAT protein expressed in Zoysia japonica does not exhibit allergenic or toxic properties.

• FLOWER RESEARCH JOURNAL
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• v.18 no.1
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• pp.1-8
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• 2010

Sensitivities of PLBs of four Phalaenopsis cultivars, P. 'Taisuco Windian', P. 'Nancy Amour', P. 'Pink Twilight' and P. 'Taipei Gold' to kanamycin, spectinomycin and hygromycin at different concentrations (0, 25, 50, 100, 200, and $400mg{\cdot}L^{-1}$) were examined. Hygromycin was favorable for selecting the transformants in the genetic transformation of Phalaenopsis as PLBs of four cultivars were all dead at even $25mg{\cdot}L^{-1}$ hygromycin. Responses of PLBs of P. 'Maki Watanabe' and P. 'Brother Lawrence' to DL-phosphinothricin (PPT) were determined at different concentrations (0, 0.1. 0.25, 0.5, 1.0, 2.0, 2.5, and $5.0mg{\cdot}L^{-1}$) and $0.5mg{\cdot}L^{-1}$ PPT was thought to be suitable for selecting the transformants of Phalaenopsis. The optimum conditions for Agrobacterium cocultivation with Phalaenopsis PLBs were examined using a two-step cocultivation method in Dtps. 'City Girl' and A. tumefaciens LBA4404. In the first infection period in a 1 : 10 suspension of Agrobacterium to a VW medium, 1 hr infection showed the highest PLB survival ratio. And then, PLBs were cocultivated with a bacterial strain and a 3-day cocultivation period was better for Phalaenopsis PLBs than a prolonged period. Agrobacterium tumefaciens strains LBA4404 (pTOK233) and EHA105 (pGA643) were used to compare their efficiency on the genetic transformation of Phalaenopsis PLBs. The PLBs infected with EHA105 survived more than those infected with LBA4404 after two days in a dark condition and two weeks in light condition on a selective medium. About 1,000 PLBs for each of P. 'Maki Watanabe' and P. 'Brother Lawrence', and each bacterial strain of AGL1 (pCAMBIA3301) and LBA4404 (pTOK233) were used for the regeneration of transgenic plants. The bacterial strain AGL1 had a higher genetic transformation efficiency than LBA4404, with no significant difference between cultivars. In this study, 11 hygromycin-resistant plantlets and 32 PPT-resistant plantlets were produced, but these putative transgenic plantlets need further examinations.

• Journal of The Korean Society of Clinical Toxicology
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• v.14 no.2
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• pp.107-114
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• 2016

Purpose: The incidence of glufosinate poisoning is gradually increasing, and it can be fatal if severe poisoning occurs. However, factors useful for predicting the post-discharge neurological prognosis of patients who have ingested glufosinate have yet to be identified. Our objective was to evaluate the utility of the acute physiology and chronic health evaluation (APACHE) II score measured in the emergency department for predicting the neurological prognosis. Methods: From April 2012 to August 2014, we conducted a retrospective study of patients who had ingested glufosinate. The outcome of the patients at discharge was defined by the Cerebral Performance Category Score (CPC). The patients were divided into a good prognosis group (CPC 1, 2) and a poor prognosis group (CPC 3, 4, 5), after which the APACHE II scores were compared. The Hosmer-Lemeshow test and the area under the receiver operating characteristic (ROC) curve from patients determined calibration and discrimination. Results: A total of 76 patients were enrolled (good prognosis group: 67 vs poor prognosis group: 9). The cut-off value for the APACHE II score was 12 and the area under the curve value was 0.891. The Hosmer and Lemeshow C statistic x2 was 7.414 (p=0.387), indicating good calibration for APACHE II. Conclusion: The APACHE II score is useful at predicting the neurological prognosis of patients who have ingested glufosinate.