We established pretreatment method of solidified cement ion-exchange resin samples generated before 2003 in nuclear power plants for measurement of non-volatile radionuclide activity. A microwave digestion system (MDS) with mixed acid (HCl-HNO3-HF-H2O2) was used to dissolve cement and to desorb non-volatile elements such as Ce, Co, Cs, Fe, Nb, Ni, Re, Sr and U from mixed ion-exchange resin. The content of Ce, Co, Fe, Nb, Ni, Re, Sr, U and Cs after pretreatment of cement plus mixed ion-exchange resin was measured by ICP-AES and ICP-MS, respectively. As iron and strontium are also present in cement, their content after dissolving a certain amount of cement was measured by ICP-AES. All elements except Nb were quantitatively recovered. Especially since the Nb recovery was low at 72.0±2.5%, the MDS following addition of the mixed acid to the resin was operated once more for desorbing Nb from it. Finally the recovery of Nb was over 95%. This sample pretreatment method will be applied to solidified cement ion-exchange resin samples generated in nuclear power plants for assessment of radionuclide inventory.

This study evaluates the gastroprotective effect of cabbage extract with sulforaphane content of 5.19 mg/L and Smethylmethionine content of 469.28 μg/L. In vitro, the lipopolysaccharide (LPS)-treated group had an increased NO activity compared to the normal group, and the concentration of NO was reduced when the cabbage extract was treated in the dose manner. The level of IL-6 induced by LPS was dose-dependently reduced when the extract was treated. The cabbage extract concentration was orally administered in rats at 5.75 mg/kg, 11.5 mg/kg, and 23 mg/ kg, and the inhibitory effect on gastric damage by HCl-ethanol was observed. Histological analysis exhibited mucosal erosion in the gastritis model compared to the normal group, while the ameliorating effect of the generated erosion was observed in the cabbage-treated group. The histamine concentration was significantly increased in the gastritis-induced animal model, and the histamine concentration was decreased in the 23 mg/L-treated group of cabbage extract. In conclusion, the results of this study suggest that cabbage extract not only down-regulates cytokines in vitro, but is also directly involved in histamine secretion in an animal model of gastritis; therefore, cabbage extract can help inhibit gastrointestinal disorders by improving the protective barrier.

애무늬고리장님노린재(Apolygus spinolae)는 포도원에서 생장 초기부터 신초를 흡즙 가해하여 잎의 기형화, 화진형상, 열매의 소립화, 기형, 열과 등의 피해를 일으킨다. 2017년과 2018년도에 경기도 화성 포도원의 캠벨얼리 품종을 대상으로 가온 하우스, 무가온 하우스, 비가림 재배, 노지 재배의 재배 양식과 생육단계별 애무늬고리장님 노린재의 발생 양상을 조사하였다. 재배양식별 피해는 1월부터 가온을 시작하여 7월 초순에 수확하는 조기가온 하우스와 3월부터 보온을 시작하여 8월 중순에 수확하는 무가온 하우스에서는 피해가 5% 미만으로 미미하였다. 그러나 비가림 및 노지재배에서는 잎과 열매의 피해가 관찰이 되었고, 특히 무농약 노지재배에서는 2017년도 27.6%, 2018년도 36%의 피해가지율을 나타내었다. 노지재배에서 피해 발생은 캠벨얼리 발아기 (BBCH(생물계절코드) 10)부터 이전 해에 포도나무 눈의 겉 인편 사이에 산란된 월동알이 부화하여 6 ~ 7엽(BBCH 16 ~ 17) 전개기까지 어린잎과 열매를 흡즙하여 피해를 주는 것으로 조사되었다. 애무늬고리장님노린재 온도 적산 모델에 따른 노지재배과원의 최적 방제시기는 월동알 부화 시기에는 2017년 4월 26일 (BBCH 11), 2018년 4월 30일(BBCH 11)이였고, 1세대 성충 출현시기에는 2017년 6월 5일 (BBCH 71), 2018년 6월 8일 (BBCH 74)로 예측되었다. 즉, 월동알 부화시기는 캠벨얼리 전엽 초기(BBCH 11), 1세대 성충 출현 시기는 과립 착과 초기(BBCH 71)로 예측되었다.

Genetically modified (GM) crops have been developed worldwide through the recombinant DNA technology and commercialized by global agricultural companies. Until now, GM crops have not been cultivated commercially in Korea. Commercialization of GM crops requires a compulsory assessment of environmental risk associated with the release of GM crops. This study was conducted to evaluate the frequency of pollen mediated gene flow from Bt transgenic rice (Agb0101) to japonica non-GM rice (Nakdongbyeo), indica non-GM rice (IR36), and weedy rice (R55). A total of 729,917, 596,318 and 230,635 seeds were collected from Nakdongbyeo, IR36, and R55, respectively, which were planted around Agb0101. Selection of the hybrids was determined by repeated spraying of herbicide and Cry1Ac1 immunostrip assay. Finally, the hybrids were confirmed by PCR analysis using specific primer. The hybrids were found in all non-GM rice and out-crossing ranged from 0.0005% at IR36 to 0.0027% at Nakdongbyeo. All of hybrids were located within 1.2 m distance from the Agb0101 rice plot. The meteorological elements including rainfall and temperature during rice flowering time were found to be important factors to determine rice out-crossing rate. Consideration should be taken for many factors like the meteorological elements of field and physiological condition of crop to set up the safety management guideline to prevention of GM crops gene flow.

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.

Map-based cloning is a basic method for identifying the mutated gene in plants. We selected the gametophytic mutant, named as AP-26-09, in activation-tagging pool. Mutant plant showed various kinds of pollen phenotype, such as the different number of nucleus or abnormal shapes. For the map-based gene cloning, we conducted phenotypic analysis of F2 mapping population through the screening of DAPI-stained pollen using fluorescence microscopy. Genomic DNA of F2 plants is prepared from leaves of approximately 1000 plants. In order to define chromosomal region where mutation is located, we designed SSLP markers and performed PCR amplification. In this study, we characterized gametophytic mutant and determined the chromosomal location using map-based approach.

We recently reported rice promoters that are active in late stages of pollen development. However, rice promoters that allow manipulation of gene expression at earlier stages of pollen development are still very limited to date. In this study, we have chosen 10 putative microspore promoters, OsMSP1 through OsMSP10, based on publicly available transcriptomic datasets in rice (Oryza sativa L.). Sequence analysis of these promoter regions revealed some cis regulatory elements involved in pollen-specific expression. We also examined promoter activities using the promoter-GUS reporter constructs in both transgenic rice and Arabidopsis. In rice, all of the 10 promoters directed GUS signals from the microspore stage throughout the all stages of pollen development. In addition, while GUS signals from 4 promoters, OsMSP2, OsMSP7, OsMSP9 and OsMSP10, seem to be expressed preferentially during pollen development, those from other six promoters were observed in vegetative tissues such as leaves, stems, and roots of seedlings. Similarly, in Arabidopsis, all of the 10 promoters directed GUS signals during pollen development. In detail, 8 promoters, OsMSP1 ~ OsMSP8 directed GUS signals from the microspore stage, whereas 2 promoters, OsMSP9 and OsMSP10, exhibited GUS signals from tricellular stage. Furthermore, seven promoters, except for OsMSP1, OsMSP2 and OsMSP10, showed GUS signals in shoot apical region or root tissues of seedlings. Furthermore, we verified microspore activity of four promoters, OsMSP1, OsMSP2, OsMSP3 and OsMSP6, by complementation analysis of the sidecar pollen (scp) mutant which displays microspore-specific defects. Currently, further analyses are underway for GUS expression of T2 generation in transgenic rice and scp complementation with remaining promoters.

To identify genes that play critical roles during male gametogenesis in Arabidopsis, we have isolated several pollen morphological mutants from a mutagenized seed pool generated with a T-DNA activation vector. In this study, we have focused on a mutant plant producing ~50% abnormal pollen grains including high levels of collapsed pollen at maturity. The pollen developmental analysis showed that the mutant pollen phenotype was first observed at tricellular stage. Interestingly, the mutation was only maintained as a heterozygote due to the severely reduced genetic transmission through both sexes. TAIL PCR analysis led to the identification of the responsible gene which encodes a conserved oligomeric golgi complex component-related protein (COGCC). RT-PCR analysis showed predominant expression of the gene in reproductive organs including developing spores. The gene identity was confirmed by the result that mutant plants harboring a T-DNA containing corresponding wild type gene produced less level of mutant pollen grains. Furthermore, confocal laser scanning microscopy using mature pollen expressing COGCC-RFP driven under the native promoter showed small punctate signals, which are likely to be from the Golgi complex. Further experiments for co-localization of the COGCC-RFP with the Golgi markers are underway.

In the course of map-based cloning, mutant genes are identified through linkage to specific region on genetic map. Here, we demonstrated gametophytic mutant line, named as AP-28-23, in which mutant gene was mapped on chromosome 2. Based on phenotypic analysis of mature pollen, mutant phenotype of AP-28-23 was classified into three classes, wild-type showing 2-4%, moderate 35-53% and severe type 97-100% on aberrant pollen frequencies, respectively. The severe type is completely sterilized with 100% unfertilized ovules. We also revealed that the transmission was reduced through male gametophyte in the AP-28-23 line. The transmission efficiency (TE) through the male gametophyte is only 0.67%, whereas in the female gametophyte is 89.87%.

Tissue-specific promoters are a very useful tool for manipulating gene expression in a target tissue or organ; however, their range of applications in other plant species has not been determined, to date. In this study, we identified two late pollen-specific rice promoters (ProOsLPS10 and ProOsLPS11) via meta-anatomical expression analysis. We then investigated the expression of both promoters in transgenic rice (a homologous system) and Arabidopsis (a heterologous system) using ProOsLPS10 or ProOsLPS11::GFP-GUS constructs. As predicted by microarray data, both promoters triggered strong GUS expression during the late stages of pollen development in rice, with no GUS signals detected in the examined microspores and sporophytic tissues. Interestingly, these promoters exhibited different GUS expression patterns in Arabidopsis. While in Arabidopsis, the OsLPS10 promoter conferred GUS expression at the uni- and bi-cellular macrospore stages, as well as at the shoot apical region during the seedling stage, the OsLPS11 promoter was not active in the pollen at any stage, or in the examined sporophytic tissues. Furthermore, by performing a complementation analysis using a sidecar pollen (scp) mutant that displays developmental defects at the microspore stage, we found evidence that OsLPS10, which can be an applied promoter expressed in Arabidopsis, is useful for directing gene expression in the early stages of pollen development. Our results indicate that the OsLPS10 and OsLPS11 promoters can drive the expression of target genes during the late stages of pollen development in rice, but not in Arabidopsis. Our results also emphasize the necessity of confirming the applicability of an established promoter to heterologous systems.

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.

본 연구는 Bacillus thuringiensis 유래의 살충성 mCry1Ac 유전자를 무선발 형질전환 방법으로 일미 벼에 도입하여 개 발된 마커프리 형질전환 Bt 벼 2계통의 일반성분 및 주요성분 (무기질, 아미노산) 함량을 확인하여 모본벼 및 다른 일반품 종과 함량차이를 비교 분석 함으로서 형질전환 벼의 영양성 분 동등성 여부를 확인하고자 수행되었다. 영양성분 분석결과 GM 벼 현미의 일반성분 조성 중 식이섬유 함량과 일부 무기 질 함량이 모본 벼인 일미와 비교하여 다소 유의적 차이가 있 었지만 일반품종에서 나타나는 함량범위 안에 포함되는 수치 이며, 아미노산 성분과 대부분의 일반성분 및 무기질의 함량 은 전반적으로 모본과 유의적 차이가 없었다. 따라서 형질전 환 벼에서 관찰된 일부 성분 차이는 Bt 유전자의 도입 효과가 아닌 재배 환경 및 토양성분의 차이에서 기인된 것으로 형질 전환에 의한 비의도적 영양성분 변화는 없는 것으로 판단된다.