• 농업과학연구
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• 제43권2호
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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.

• 한국환경농학회지
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• 제33권2호
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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.

• 한국자원식물학회지
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• 제17권1호
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• pp.28-33
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• 2004

본 연구는 개량 포플러 현사시 3호를 재료로 제초제 저항성 유전자 PAT으로 형질전환시 킨 다음 제초제 Basta처리에 의한 효율적인 형질전환체 선발을 목적으로 시험되었다. Basta 처리에 따른 캘러스유도, 부정아유도 및 액아유도 시험을 통해 조직치사 농도는 1.0mg/L로 결정할 수 있었으며, 이 농도처리에 의해 형질전환체의 조기선발에 사용하였다. 이 농도처리로 비형질전환체는 모두 고사되었지만, 형질전환체는 정상적으로 반응을 보이며, 생육이 가능하였다 .한편, 형질전환체에 제초제를 직접 살포한 결과, 정상적 생육이 가능했고, PAT activity측정 결과에서도 양성 반응을 나타내어 현사시 3호 세포내에 PAT유전자가 정상 발현됨이 확인되었다. 이로써 현사시에 PAT유전자 도입 및 발현을 위하여 제초제 Basta 처 리와 PAT assay가 매우 효과적인 것으로 나타났다.

• 식물조직배양학회지
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• 제28권4호
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• pp.197-200
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• 2001

상추에 제초제 저항성을 도입하기 위해 상추 자엽 조직을 NPTII-GUS 및 제초제 저항성 유전자 (PAT)가 삽입된 A. tumefaciens MP 90과 2일간 공동배양한 다음 0.1 mg.L$^{-1}$ NAA, 1.0 mg.L$^{-1}$ 2ip, 50 mg.L$^{-1}$ kanamycin, 500 mg.L$^{-1}$ carbenicillin을 첨가한 MS배지에 배양하여 식물체를 재분화시켰다. 재분화 식물체는 kanamycin 내성 검정을 통해 NPTII 유전자의 도입과, PCR, Northern blot 분석을 통해 제초제 저항성 유전자가 식물체의 게놈상에 삽입된 형질전환체임이 확인되었다. 또한 1500배액의 농도로 Basta를 살포한 결과 살포 10일 후 대조 식물체는 완전히 고사하는 데 반해 형질전환체는 지속적인 생육을 보여 얻어진 형질전환 상추가 제초제 저항성 개체임이 확인되었다.

• 대한임상독성학회지
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• 제17권1호
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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.

• 농업과학연구
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• 제47권4호
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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.

• 농업과학연구
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• 제47권4호
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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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• 제47권4호
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• pp.316-323
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• 2020

본 연구는 제초제저항성 GM 들잔디에서 발현된 PAT 단백질의 잠재적인 알레르기성 및 독성을 평가하기 위해 수행되었다. PAT 단백질의 in silico 분석에서 PAT 단백질은 알려진 알레르겐 또는 독성 단백질과 유사성을 보이지 않았다. PAT 단백질은 80개의 아미노산으로 이루어진 절편에 걸쳐 기지의 알레르겐과 35% 미만의 아미노산 서열 상동성을 보였고, 기지의 알레르겐과 연속한 8개의 아미노산에서 일치하는 서열도 확인되지 않았다. 또한 제초제저항성 GM 들잔디에서 발현된 PAT 단백질은 펩신 존재 하의 인공위액에서 매우 빠르게 분해되었고, GM 들잔디에서 발현된 PAT 단백질은 번역 후 수식(glycosylation)도 일어나지 않았음이 확인되었다. 경구투여 독성 시험에서는 체중 1 kg 당 4,000 mg 을 투여한 실험구에서도 PAT 단백질 투여 후 마우스에서 사망이나 독성 영향은 관찰되지 않았다. 이들 결과는 제초제저항성 GM 들잔디에서 발현된 PAT 단백질이 잠재적 알레르겐이나 독소로 작용할 가능성이 없음을 시사한다.

• 화훼연구
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• 제18권1호
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• pp.1-8
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• 2010

호접란의 형질전환시스템을 확립하기 위한 제 연구를 수행하였다. 항생제 kanamycin, hygromycin 및 spectinomycin 농도(0, 25, 50, 100, 200, and $400mg{\cdot}L^{-1}$)가 품종별 PLB 생존율에 미치는 영향을 알아보기 위한 실험에서 hygromycin은 $25mg{\cdot}L^{-1}$에서 모든 품종이 괴사하였으므로 형질전환 개체의 선발 항생제로는 hygromycin이 유리할 것으로 보였다. P. 'Maki Watanabe'와 P. 'Brother Lawrence' 두 품종에서 형질전환체 선발을 위한 DL-Phosphinothricin (PPT)의 적정 농도는 $0.5mg{\cdot}L^{-1}$이었다. 형질전환시 가장 높은 효율을 얻기 위한 공동배양 일수를 결정하기 위한 실험은 Dtps. 'City Girl'과 A. tumefaciens LBA4404를 이용하여 2단계로 이루어졌다. 균주와 VW 배지의 1 : 10 현탁액에 균주와 PLB를 감염시킨 결과 1시간 처리구에서 PLB 생존이 가장 많았다. 그런 다음 공동배양한 결과 5일 배양에서 PLB 생존수가 가장 많았지만, 4일 이상의 공동배양할 경우 PLB 조직이 연화가 되고 약해져서 죽게 되었다. 따라서 오히려 3일 공동배양 기간이 적당한 것으로 판단되었다. 박테리아 균주의 종류가 호접란 PLB의 형질전환에 미치는 효율을 비교하기 위해 A. tumefaciens LBA4404(pTOK233)와 EHA105(pGA643)를 이용하였다. LBA4404 보다 EHA105로 감염시킨 PLB의 생존율이 더 높았다. A. tumefaciens LBA4404(pTOK233)와 AGL1(pCAMBIA3301)을 이용한 형질전환 실험에서 치상된 PLB가 초기에 백변하는 정도가 LBA4404를 이용한 경우 눈에 띄게 빠르게 나타났고 새로운 PLB가 유도되는 정도도 매우 낮았다(1% 미만). 반면에 AGL1을 이용한 경우 40% 정도의 새로운 PLB 및 유식물체 형성율을 나타내었다. 형질전환 실험에서 최종적으로 hygromycin 저항성 식물체 11개체와 PPT 저항성 식물체 32개체를 얻어냈으나 진정한 형질전환체인지는 차후에 더 검정이 되어야 할 것으로 보인다.

• 대한임상독성학회지
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• 제14권2호
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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.