Recently, in Korea various kinds of genetically modified (GM) crops have been imported and used as a raw material to manufacture foods and livestock feeds, but the different social concerns about the benefits and the potential risks of GM crops are being shown with a different reaction from the public. Thus a persistent management is required for the safe utilization of genetically modified organism (GMO). PCR analysis of transgene into crop is generally performed for the efficient post management of GMOs. The most important prerequisite for the application of nucleic acid detections is to decide the effective DNA-extraction methods. Particularly, in the case of processed feeds, the nucleic acids of which may be damaged by heating, high pressure, pH treatments, fermentation, etc. in processing, DNA must be extracted with high sensitivity from the samples to perform the PCR successfully. In this study, seven of DNA-extraction methods used commercially and non-commercially were compared with respect to the yields and quality of DNA extracted from livestock feeds and those crop materials. Amounts of genomic DNA obtained from the extraction methods varies according to feed configurations and crop materials. The DNA yield and uniformity of samples extracted with PG, CTAB, and QF method is greater than that obtained from other extraction methods. In the DNA integrity of the selected extraction methods, PCR analysis showed distinct amplifications and similar patterns in detecting crop endogenous genes and GMO genes. These results would be applicable for the selection of an adequate DNA-extraction method in extracting processed feeds and/or crop materials.

Rice is the most important staple food and the consumption is increasing in Europe and America. It is expected that the rice price will be increased due to the unstable supply and demand because of abiotic stress. Experts expect that the rice supply of Korea will be deficient in 2050 for the reason of global warming. Conventional breeding methods need to be supplemented with recent biotechnology to meet the problem such as growing population and climate change. Using rice biotechnology, a number of agronomically important traits and nutritional value have been improved. In this paper, the current situation of rice cultivation and production situation and supply and consumption in the major country were investigated to prepare future food security. Also the research trends of rice biotechnology and kinds of transformed rice as well as safety issues were reviewed by considering the scientific, social and economic values of rice.

본 연구의 목적은 우리나라에서 개발된 병저항성 GM벼의알레르기 유발성을 평가하고자 하였다. 벼 유래의 Cholinekinase 1 유전자(OsCK1)를 과발현 하도록 형질전환한 벼를이용하여 알레르기 유발인자와 상동성이 있는지 확인한 결과,연속되는 80개 이상의 아미노산 절편에서 35% 이상의 상동성을 보이는 서열은 없었으며 8개씩의 인접 아미노산과 일치하는 서열도 확인되지 않았다. 또한 GM벼와 non-GM 벼 사이의 단백질 분포가 일치하는 것을 확인하였으며, 인공 위액에서 빠르게 분해됨을 확인하였다. 발현단백질의 당화 발생 여부를 확인한 결과, OsCK1단백질에 대한 당화는 발생하지 않는 것을 알 수 있었으며 열안정성 검사에서OsCK1 단백질을100oC에서 가열하였을 때 30분 이후부터는 약하게 분해됨을확인하였다. 본 연구의 결과를 통해 병저항성 GM벼는 알레르기 유발과는 상관관계가 없음을 확인하였다.

농업적, 환경적, 경제적 및 사회적인 이익으로 농업생명공학에 의한 유전자변형(GM) 작물의 재배는 점차 증가되고 있다.국내에서도 주요 작물을 대상으로 유용 GM작물이 개발되고있으며, 최근 Choline kinase 유전자(OsCK1)가 도입된 병저항성 형질전환벼가 개발되었다. GMO의 안전성과 관련하여, 표시제의 시행 또는 사후 이력추적을 위해서 검정법이 필수적으로 요구되고 있다. 본 연구에서 각 134, 306, 243bp의 PCR증폭산물을 갖는 유전자 특이, 구조 특이 및 이벤트 특이primer를 병저항성(OsCK1) GM벼의 검출에 사용하였고, 다른어떤 작물에서도 반응산물을 나타내지 않았다. 이벤트 특이primer CKRB32-1/02-2를 사용한 정성 duplex PCR을 통해서OsCK1 GM벼에 대한 검출한계(LOD)가 0.05%임이 확인되었다. Real-time PCR을 이용한 정량검정을 위해서 벼 내재유전자 염기와 OsCK1 GM벼의 5’-인접염기를 갖는 pSPSCKR을표준물질로 제조하였고, 10 copies 범위까지 정량검출이 가능한 것으로 나타났다. 따라서 도출된 real-time PCR 방법의 정확성 및 정밀성을 확인하고자 0.5, 1, 3, 5 및 10%로 GM시료에 대하여 정량 분석하였으며, 표준편차 및 상대표준변이가 20% 내로 확인되었다. 이상의 결과로, 개발된 이벤트 특이정성 및 정량 PCR 방법이 OsCK1 GM벼의 사후 GMO 모니터링 및 이력추적에 효과적으로 적용 가능할 것으로 판단된다.

The increase in carbon stock and sustainability of crop production are the main challenges in agricultural fields relevant to climate change. Methane is the most important greenhouse gas emitted from paddy fields. This study was conducted to investigate the effects of tillage and cultivation methods on methane emissions in rice production in 2014 and 2015. Different combinations of tillage and cultivation were implemented, including conventional tillage-transplanting (T-T), tillage-wet hill seeding (T-W), minimum tillage-dry seeding (MT-D), and no-tillage-dry seeding (NT-D). The amount of methane emitted was the highest in T-T treatment. In MT-D and NT-D treatments, methane emissions were significantly decreased by 77%, compared with that in T-T treatment. Conversely, the soil total carbon (STC) content was higher in MT-D and NT-D plots than in tillage plots. In both years, methane emissions were highly correlated with the dry weight of rice (R 2 = 0.62~0.96), although the cumulative emissions during the rice growing period was higher in 2014 than in 2015. T-T treatment showed the highest R 2 (0.93) among the four treatments. Rice grain yields did not significantly differ with the tillage and cultivation methods used. These results suggest that NT-D practice in rice production could reduce the methane emissions and increase the STC content without loss in grain yield.

A variety of genetically modified (GM) crops have been developed in Korea. In these crops, the resveratrol-enriched transgenic rice plant (Agb0102) has moved ahead to generate the dossier for regulatory review process required for commercialization of GM crop. The resveratrol-enriched transgenic rice plant could be released to farmers for cultivation after national regulators have determined that it is safe for the environment and human health. Here, we developed a PCR-based DNA marker based on flanking sequences of transgene for the discrimination of resveratrol-enriched transgenic rice plant. This DNA markers will be useful for identifying of resveratrol-enriched transgenic rice plant, and can also be used to estimate transgene movement occurred by pollen transfer or seed distribution. Moreover, it is helpful for prompt screening of a homozygote-transgenic progeny in the breeding program.

Natural and artificially induced mutants have provided valuable resources for plant genetic studies and crop improvement. Some variations induced in the process of plant transformation have often been observed in regenerated plants. In this study, we investigated the insertion number of transgene and the flanking sequences of T-DNA in tall-induced line BP23, which was unexpectedly gained in the process of transformation of insect-resistant rice with cryBP1 gene, and also analyzed the whole-genome sequencing by using the NGS technologies to gain a better understanding of the sequence and structural changes between tall line or natural cultivar and rice reference. than others, was confirmed with two copies of foreign gene insertion, which was inserted in one genomic site facing each other between the position 2,430,152~2,430,151 of rice chromosome 12 without any deletion of genomic sequences. Sequencing analysis also revealed that 18bp-unknown sequences were added in the 5′ insertion site of T-DNA. This position in rice genome was confirmed with none of expressed gene sites. By the NGS analysis, we detected 86560 SNPs and 1091/1472 large insertion/deletion (indel) sites (100bp) between BP23 and rice reference, and 84743 SNPs and 1094/1451 large indels between natural cultivar Nagdong and rice reference. The possible mechanisms for the gene mutation, the developmental and tissue expression of the taller height in BP23 line may need to be scrutinized a few more.

The selectable marker-free rice plants containing mcry1Ac insecticidal gene isolated from Bacillus thuringiensis (Bt) were generated using a non-selection approach by Agrobacterium tumefaciens-mediated transformation. The nutritional composition of two lines of transgenic rice plants (RTB5 and RTB11) was compared with that of its non-transgenic counterpart. The results showed that, except for small differences in dietary fiber and some minerals, there was no significant difference between transgenic rice and conventional counterpart variety with respect to their nutrient composition. Most of measured levels of nutrients were within the range of values reported for other commercial cultivars, showing substantial equivalency. Therefore, the insertion of transgenes did not affect the nutritional composition of transgenic RTB5 and RTB11 rice grains.

The β-carotene biofortified transgenic soybean was developed recently through Agrobacterium -mediated transformation using the recombinant PAC (Phytoene synthase-2A-Carotene desaturase) gene in Korean soybean (Glycine max L. cv. Kwangan). GM crops prior to use as food or release into the environment required risk assessments to environment and human health in Korea. Generally, transgenic plants containing a copy of T-DNA were used for stable expression of desirable trait gene in risk assessments. Also, information about integration site of T-DNA can be used to test the hypothesis that the inserted DNA does not trigger production of unintended transgenic proteins, or disrupt plant genes, which may cause the transgenic crop to be harmful. As these reasons, we selected four transgenic soybean lines expressing carotenoid biosynthesis genes with a copy of T-DNA by using Southern blot analysis, and analyzed the integration sites of their T-DNA by using flanking sequence analysis. The results showed that, T-DNA of three transgenic soybean lines (7-1-1-1, 9-1-2, 10-10-1) was inserted within intergenic region of the soybean chromosome, while T-DNA of a transgenic soybean line (10-19-1) located exon region of chromosome 13. This data of integration site and flanking sequences is useful for the biosafety assessment and for the identification of the β-carotene biofortified transgenic soybean.

A variety of genetically modified (GM) crops have been developed in Korea. In these crops, the resveratrol-enriched transgenic rice plant has moved ahead to generate the dossier for regulatory review process required for commercialization of GM crop. The resveratrol-enriched transgenic rice plant could be released to farmers for cultivation after national regulators have determined that it is safe for the environment and human health. Here we developed a PCR-based DNA marker based on flanking sequences of transgene for the discrimination of zygosity in resveratrol-enriched transgenic rice plant. This DNA marker will be useful for identifying of resveratrol-enriched transgenic rice plant, and can also be use to estimate transgene movement occurred by pollen transfer or seed distribution.

It is necessary to carry out a risk assessment to determine the consequences of releasing a particular plant species containing specific transgenes before transgenic plants can be grown under filed conditions. Gene flow from transgenic plants to wild closely related species has raised concern recently. Since transgenic crops were released in 1996, the global area of transgenic crops has been increasing rapidly. The transgene introgression from transgenic crops to their wild relatives is unavoidable in some species. Transgene introgression is of concern because the crop–wild plant hybrids might be conferred with a selection advantage to increase their performance, which could result in negative ecological consequences to natural ecosystems. The genus Brassica has 159 species, including a number of wild species that are of great importance to the economy. Most transgenic Brassica gene flow research has focused on the most successful cross between transgenic oilseed rape Brassica napus and its wild relatives Brassica rapa, a widely distributed weed in the farming system in Europe and America, since the hybridization can spontaneously happen and the generations can backcross to B. rapa easily in the wild conditions. In this study, we aimed to characterize transgene introgression, segregation, and expression in backcrossed generations between tramsgenic B. napus and B. rapa. These results will contribute to the environmental risk assessment and assist in biosafety management.

The β-carotene biofortified transgenic soybean was developed recently through Agrobacterium-mediated transformation using the recombinant PAC (Phytoene synthase-2A-Carotene desaturase) gene in Korean soybean (Glycine max L. cv. Kwangan). GM crops prior to use as food or release into the environment required risk assessments to environment and human health in Korea. Generally, transgenic plants containing a copy of T-DNA were used for stable expression of desirable trait gene in risk assessments. Also, information about integration site of T-DNA can be used to test the hypothesis that the inserted DNA does not trigger production of unintended transgenic proteins, or disrupt plant genes, which may cause the transgenic crop to be harmful. As these reasons, we selected four transgenic soybean lines expressing carotenoid biosynthesis genes with a copy of T-DNA by using Southern blot analysis, and analyzed the integration sites of their T-DNA by using flanking sequence analysis. The results showed that, T-DNA of three transgenic soybean lines (7-1-1-1, 9-1-2, 10-10-1) was inserted within intergenic region of the soybean chromosome, while T-DNA of a transgenic soybean line (10-19-1) located exon region of chromosome 13. This data of integration site and flanking sequences is useful for the biosafety assessment and for the identification of the β-carotene biofortified transgenic soybean.

The cultivation of genetically modified (GM) crops has increased due to their economic and agronomic advantages. Before commercialization of GM crops, however, we must assess the potential risks of GM crops on human health and environment. The aim of this study was to investigate the possible impact of Bt rice on the soil microbial community. Microbial communities were isolated from the rhizosphere soil cultivated with Bt rice and Nakdong, parental cultivar and were subjected to be analyzed using both culture-dependent and molecular methods. The total counts of bacteria, fungi, and actinomycetes in the rhizosphere of transgenic and conventional rice were not significantly different. Denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA genes revealed that the bacterial community structures during cultural periods were very similar each other. Analysis of dominant isolates in the rhizosphere cultivated with Bt and Nakdong rice showed that the dominant isolates from the soil of Bt rice and Nakdong belonged to the Proteobacteria, Cloroflexi, Actinobacteria, Firmicutes, and Acidobacteria. These results indicate that the Bt rice has no significant impact on the soil microbial communities during cultivation period. Further study remains to be investigated whether the residue of Bt rice effect on the soil environment.

A Transgenic Kimch cabbage has been developed harboring T-DNAs expressing delta-endotoxin insecticidal protein, herbicide (basta) resistant protein, and antisense transcript of AsMADS2 gene. Three transgenic lines, #24, #45, and #51, originated from the same T0 plant were analyzed in terms of molecular characterization, phenotype, and agronomic traits. Flanking sequence analysis confirmed that T-DNA, with 7132 bp intact structure, was inserted onto the pseudochromosome A10 of B. rapa and all the genes in T-DNA were functionally active. Three of GM cabbage showed 69.2～75.3% of plant height and 81.8～89.7% of diameter to those of the isogenic variety ‘Nowon’, respectively. Curving upward leaf lamina attitude was observed on GM cabbage, while straight or slight concave on non-GM cabbage. In addition, an average range of 86～91.5% of head height and 87.4～94.8% of head diameter were observed on GM cabbage to those of the isogenic variety ‘Nowon’, respectively Moreover, curled inwards or slight overlap of head-forming leaf overlap at terminal region was observed on GM cabbage, but curled outwards or erect on non-GM cabbage. AsMADS2, a transcription factor reported to be involved in early flowering, was stably expressed to RNA in the GM cabbage, but it was not shown the significant influences to flowering time.