본 연구는 환자용 균형영양식으로 제조하여 감마선 조사처리를 통해 살균한 모델식품의 유전독성 여부를 평가하기 위해 수행되었다. 4 kGy 이상의 흡수선량으로 감마선 조사처리한 시료에서 생균수가 1 log CFU/g의 검출한계 이하로 나타났기 때문에 살균을 위한 조사처리 선량으로 4 kGy를 설정하였다. 복귀돌연변이 유발성 평가 결과, 모델식품의 열수 추출물과 메탄올 추출물은 처리한 용량과 대사활성계존재 여부와는 상관없이 음성대조구(멸균증류수 또는 DMSO)와 유사한 수준의 복귀돌연변이 발생빈도를 나타내었다. 염색체 이상 시험에서 실험군들의 정상 염색체의 수는 음성대조군과 유사하였으며, 용량 및 대사활성계에 대한 비의존성을 나타내어 염색체 이상 유발능은 없는 것으로 판단하였다. 또한 최고 2,000 mg/kg 체중의 농도로 모델식품을 경구 투여한 마우스의 골수세포에서도 소핵 다염성적 혈구의 발생빈도는 음성대조군에 비해 유의적인 차이를 나타내지 않았다. 따라서 4 kGy의 흡수선량으로 감마선 조사처리한 환자용 균형영양식은 본 실험 조건하에서 유전독성을 나타내지 않는 것으로 사료된다.

This study examined radiation damage and the optimal gamma-ray dose for mutation breeding in oat (Avena sativa L. cv. Samhan). The seed germination rate decreased as the dose increased over 500 Gy. The median lethal dose (LD50) was approximately 392 Gy. The median reduction dose (RD50) for plant height, tiller number, root length, and flash weight was 411, 403, 394, and 411 Gy, respectively. The optimal dose of gamma irradiation for inducing oat mutation appears to be in the range 300-400 Gy. We performed the comet assay to observe nuclear DNA damage induced by gamma-ray irradiation. This assay showed a clear difference with gamma-ray treatments. DNA damage increased temporarily 7 days after treatment depending on the dose, while no significant difference was identified in response to 300 Gy 30 days after the gamma-ray treatments. The growth characteristics of the M2 generation decreased as the dose increased over 400 Gy.

본 연구에서는 감마선 조사가 떫은감 과육의 세포벽변화, 수렴성 및 경도변화에 미치는 영향을 살펴보았다. 감마선 조사 결과 떫은감 과육 중의 가용성 탄닌 함량의 변화는 대조구의 181.04±1.72 μg/g에 비해 처리 방사선의 양이 5～20 kGy로 증가할수록 146.31 μg/g에서 128.09 μg/g으로 과육내의 가용성 탄닌의 양이 감소하는 것이 확인되었다. 감 과육의 경도는 방사선을 조사하지 않은 대조구에 비해 방사선 조사한 감과육의 경도가 감소하였다. 세포벽 성분의 변화는 알콜불용성 물질의 경우 대조구의 39.3 mg/g에 비해 방사선 처리의 경우 37.2 mg/g로 낮아졌다. 수용성 물질의 함량은 대조구의 경우 11.4 mg/g이었으나, 방사선 조사의 경우 13.9 mg/g로 높아졌으며, 세포벽 성분의 함량은 대조구의 경우 26.6 mg/g였으나, 방사선 조사의 경우 23.1 mg/g로 낮아져, 떫은감 과육이 방사선조사에 의해 숙성이 일어나 세포벽성분의 함량이 감소하고 수용성물질의 함량이 증가한 것으로 나타났다. 세포벽 구성다당류 중 lignin의 함량과 pectin질의 함량은 대조구의 0.82 mg/g, 3.56 mg/g에 비교해 방사선 조사의 경우 0.77 mg/g, 3.14 mg/g로 감소하였다. 산가용성 hemicellulose의 함량은 lignin과 pectin처럼 방사선 조사에 의해 감소하는 경향을 나타내었으나, 알칼리 가용성 hemicellulose와 cellulose는 방사선조사에 의해 증가하는 양상을 나타내었다. 감과육에 존재하는 세포벽 분해 및 연화효소인 polygalacturonase, pectinesterase와 β-galactosidase의 활성의 경우 방사선 조사에 의해 모든 효소활성이 증가하였다. 방사선 처리감의 관능검사 결과 수렴성은 대조구의 경우 매우 떫은 것으로 나타났으나 방사선처리 후에는 거의 떫지 않게 느껴졌고, 과육의 경도에 대한 관능검사 결과 대조구는 딱딱한 것으로 나타났으나 방사선처리 후에는 약간 물렁해 지는 것으로 나타나 방사선처리에 의해 탄닌의 중합이 일어나며 과육의 숙성도 동시에 발생하였다. 따라서 감마선 처리는 떫은 감의 수렴성 제거와 숙성 기간을 단축시킬 수 있는 새로운 방법으로 활용이 가능할 것으로 판단되었다.

In recent years, the efficiency and accuracy of QTL analysis for identification of useful traits have been increased by high-throughput genotyping. In a previous study, the genome variation of significant DNA polymorphism was observed in early maturing type rice mutant (EMT) by comparing with that of wild type (WT). For detection of major QTL for flowering time, we constructed a linkage map of 36 InDel- and 6 SNP- markers. In the linkage analysis of F2 plants derived from the cross “WT x EMT”, we have detected one potential QTL region on chromosome 6 by M6-3 marker. Also, the Hd1, which contained the target fragment of M6-3 marker, exhibited the relatively high nonsynonymous substitutions in genes located on chromosomal region from M6-2 to M6-4. To evaluate the reliable allele segregation related to expected Mendelian ratio between M6-3 and its flanking markers, M6-3 marker developed in Hd1 gene exhibited the 1:2:1 ratio as clear monogenic segregation in heterozygous F3 plant. Additionally, we further analyzed the different transcript regulations of OsGI and Hd3a gene related to Hd1 involved in photoperiodic flowering pathway. Although the mRNA levels of Hd1 had no difference between WT and EMT, the Hd3a as downstream effector of Hd1 significantly upregulated in EMT, suggesting that Hd1 gene may become nonfunctional.

Rose (Rosa Hybrida Hort.) are of a high symbolic value and a great cultural importance in different societies. They are widely used as garden ornamental plants and as cut flowers. For the induction of mutation, gamma-rays are widely used as a mutagen. This study was carried out to establish a system for mutation breeding by irradiation of gamma-ray in rose. The rooted cuttings of five cultivar roses (Lovelydia, Vital, Aqua, Yellowbabe and Haetsal) are grown by in a greenhouse. They were two difference treatment (Before rooting gamma-ray irradiation, After rooting gamma-ray irradiation) were exposed to dose of 70 Gy using a 60Co gamma-irradiator (150 TBq of capacity ; ACEL, Canada) at the Korea Atomic Energy Research Institute. The irradiated plants were planted in a greenhouse, and investigated survival rate, mutation rate, flower buds number, and shoot length were planted after 80days. The two treatments of and growth characters was significantly reduced to 20% to 40% compared with the control. In addition, survival rate and mutation rate were ‘after rooting γ-ray irradiation (37.4~67.3% and 0.5~5.6%)’ higher than ‘before rooting γ-ray irradiation (18.3~50.8% and 0.3~3.4%)’. Mutation types were solid type, chimeric and mosaic petal mutants with various colors were induced from five rose. These results indicate that efficiency of mutation induction in rose by gamma-ray irradiation on petal colors and petal shapes in two difference treatment with rooted cutting system.

Gamma irradiation has been used as a tool for plant mutation breeding to select new cultivar with improved characteristics. Generally, the irradiation of seeds with high doses of gamma rays disturbs the synthesis of protein, hormone balance, and enzyme activity. And also, high dose of gamma rays to reduce plant height, number of tiller, and root length, although the effect of gamma-irradiated plants may depend on the species and cultivar or stress conditions. Biological effects of radiations can be divided into two types according to dose range and periods of exposure. Acute irradiation represents exposure to high-dose of irradiation over short period time, whereas the chronic irradiation is comprised of exposure to low doses of radiation over extended period of time. To compare the effects of acute and chronic exposure to ionizing radiation on two wheat cultivars (K4191 and Geumgangmil), we measured their germination rate, seedling height, and root length. In order to understand the influence of antioxidant-related genes and DNA repair-related genes, we used qRT-PCR methods to identify their expression levels. To study the behavior of a radiation-induced free radical, gamma-irradiated seeds were used for ESR spectroscopy. Plant growth pattern was showed positive correlation with ESR results. This study indicates that low level chronic radiation exposure is even more serious effects than short doses of high level radiation according to different wheat cultivar.

Chronic gamma irradiation can be used an alternative mutation breeding methods for induction of many useful mutants. Seedlings of purple-colored wheat plants were irradiated with wide range doses of chronic gamma-rays (20, 25, 30, 40, 50, 70, 100, 125, 150, 200, 250, 300 Gy) during 6 weeks at gamma-phytotron in the Korea Atomic Energy Research Institute, respectively. To identify the biological responses purple-colored wheat, we examined the plant height, chlorophyll, carotenoid and total anthocyanin contents in leaf. Plant growth, chlorophyll and carotenoid contents in leaf were decreased when the dose rate increased. Anthocyanin contents were increased with the increase of the radiation dose until 50 Gy treatment. To confirm the real contents of anthocyanin, we also investigated cyanidin-3-glucoside in purple-colored wheat leaf by using UPLC analysis. These results indicate that anthocyanin accumuation was observed under chronic gamma irradiation.

본 연구에서는 후춧가루의 위생화를 위해 감마선, 전자선 및 X-선을 조사한 후 이에 따른 미생물학적 및 관능적 품질변화를 평가하였다. 후춧가루에 감마선, 전자선 및 X-선을 2, 4, 6, 8 및 10 kGy로 각각 조사한 후 미생물 변화와 관능품질 변화를 평가한 결과, 미생물의 경우 선종에 관계없이 흡수선량이 증가할수록 미생물 제어 효과가 증가하는 것으로 나타났으며, 선종별 D10값의 경우 2.24-2.75 kGy범위를 보였으며 방사선 선종에 따른 유의차가 없는 것으로 나타났다. 색도의 경우 명도, 적색도 및 황색도 모두 방사선 선종에 따른 변화가 없는 것으로 판단하였다. 관능적 품질 역시 방사선 선종에 관계없이 흡수선량에 따라 품질저하가 발생되는 것으로 나타났다. 이상의 결과를 종합하여 볼 때 방사선 선종에 따른 후춧가루의 품질변화 차이가 없는 것으로 판단하였으며, 이상의 결과는 X-선의 상업적 이용을 위한 기초 연구 자료로 활용될 수 있을 것으로 사료되었다.

국화 6 품종을 대상으로 감마선(35 Gy) 조사에 의해 유기 된 화색변이체를 선발하고, 식물체 재생을 위한 배지내 식물생 장조절제 적정농도를 구명하여 신품종 육성 가능성을 검토하고 자 수행하였다. 고유품종과 다른 꽃잎의 화색변이율은 ‘SP-410’ 이 15.7%로 가장 높았으며, ‘예스스타’가 10.6%, ‘무지개’가 10.3%순 이었다. ‘무지개’는 이중색(황색/적색, Y7D/R40A) 에서 적색(R42B)으로, ‘예스라인’은 자주색(RP74C)에서 진자 주색(RP63B)으로, ‘예스스타’는 황색/적색(Y4A/RP59A) 이 중색에서 적자주색(RP64A)으로, ‘SP-410’은 백색(W155C)에서 황색(Y2C)으로, ‘예스투게더’의 화심색은 백색(W155C)에서 연보라색(P76B)로, ‘예스우리’의 화심색은 연녹색(YG149A)에 서 짙은 녹색(GY1C)으로 화색이 변한 변이체를 선발하였다. 화색변이체 꽃잎의 신초형성을 위한 배지내 식물생장조정제 의 최적조합은 BA 1.0 mg/L와 NAA 0.5 mg/L, BA 2.0 mg/L 와 NAA 0.5 mg/L의 2조합이였으며, 신초형성율은 품종간에 차이가 있어 ‘예스라인’이 91.6%로 가장 높았다. 감마선 조 사에 의한 화색 변이체의 꽃잎배양 유래 재생식물체는 영양 번식으로 유지되었으며, 3년동안 재배한 결과 화색과 꽃잎의 형태적 특성 차이가 없었다.

Rice is one of the most important food crops in the world, and has been used as model monocots for genetic studies, because of its relatively small genome size. We have previously reported the different functions of several RING (Really Interesting New Gene) proteins to respond the various abiotic stresses. In order to study a regulation of RING proteins in rice under ionizing irradiation such as gamma ray (GA), we have identified the expression patterns of these genes by RT-PCR. We found Gamma-ray induced RING finger protein (OsGRP) gene, which were associated with cytosol by subcellular localization analysis. in vitro ubiquitination assay revealed that OsGRP possess E3 ligase activity. Also, we demonstrate that C196A point mutation in the RING finger domain of OsGRP can have a critical effect to the breakdown of structural integrity in RING constructs. To identify the interaction partners for OsGRP in protein-protein interactions, we found the seven genes interacted with OsGRP by Yeast Two Hybrid method. To examine the GA-influence of interaction partners by RT-PCR, two genes were specifically down-regulated in rice during GA treatment. These interaction partners were identified the reliable interactions and subcellular localizations via BiFC method. Interestingly, five genes associated with plastid, while two down-regulated genes associated with cytosol and plastid. These results of OsGRP based on genetic approach might provide a clue to understanding the GA responsive mechanism in rice.

Brachypodium has been focused as new model plant for grass species. Like small size, small room requirement, and fast growth, Brachypodium shows numerous advantages as a model plant. Brachypodium is a typical grass at the genome level, which also exhibits an overall similarity of gene content and gene families when compared with rice (Oryza sativa) and sorghum (Sorghum bicolor) genomes. Brachypodium is an excellent material for structural and functional genomic studies in grass species. Targeting-Induced Local Lesions IN Genomes (TILLING) is a high-throughput technique and an approach for reverse genetics study. Moreover, it has been wildly utilized to find induced mutation. Bradi3g45515 is orthologue of the cellulose synthase-like HvCslF8 in barley. For TILLING library construction, 384 M2 Brachypodium mutants induced by chronic-gamma irradiation were used. Single nucleotide polymorphysm (SNP) and small deletion in Bradi3g45515 were searched through TILLING analysis. Template DNA for PCR reaction were prepared according to two dimensional pooling (eightfold) strategy. Heteroduplex DNA was digested by SURVEYOR nuclease (TRANSGENOMIC) and the DNA fragment was detected using polyacrylamide gel electrophoresis. Positive signal appeared at polyacrylamide gel from more than 4 lines and their Bradi3g45515 region were sequenced. SNP(s) were identified in 509-2 and 677-3 mutant line. Cellulose content and/or cell wall materials content will be measured using these mutants.

Soluble sugar content in soybean seed is an important quality attribute for soyfood and feed. Usually, soluble sugars comprise 6 to 17% of total dry wt. in mature soybean seeds. In this study, 414 soybean mutant lines induced by gamma-ray were screened by colormetric assay, FACE (Fluorophore-assisted carbohydrate electrophoresis), and GC-MS to identify the change of soluble sugar contents. Among 414 soybean mutant lines, 12 mutant lines derived from three different soybean cultivars (Hwanggum, Paldal, and Bangsa) showed higher level of soluble sugar content compared to their original cultivars. However, 5 mutant lines derived from soybean landrace KAS 636-15 showed lower level in the colormetric assay. In FACE, 17 soybean mutant lines selected by colormetric assay also showed different band intensity compared with their original cultivars. However, there were no different soluble sugar patterns between soybean original cultivars and mutant lines. Finally, the variations of soluble sugar content in 17 soybean mutant lines were confirmed by using GC-MS. These mutant lines will be used for genetic study to find mutations of genes related soluble sugar biosynthesis.

The effects of gamma irradiation at medium dose levels on commercial starches (corn starch/CS, sweet potatostarch/SS, and potato starch/PS) were investigated in terms of its color and lipid oxidation. The CS, SS, and PS samples were irradiated at 0, 1.5, 3, 4.5 and 6 kGy by a Co-60 gamma irradiator and used for measuring Hunter's colors and TBA value. Irradiation doses applied did not cause apparent changes in Hunter color parameters between the control and irradiated groups. But, the decrease in lightness (L value) for CS and the increase in both redness (a value) for SS and yellowness (b value) for PS were observed, respectively, thereby resulting in slight increase in overall color difference (ΔE) upon irradiation. Browning intensity observed with the naked eye for gelatinized starches by irradiation doses was also gradually increased along with irradiation dose, which was more remarkable in SS. The thiobarbituric acid (TBA) values of samples showed a dose-dependant increase with ≥ 0.9964 of R2 (p<0.05).

식물유전자원의 소재를 확대하고자 동진 1호 벼의 건조 종자에 감마선(60Co, 300 Gy)을 조사하여 선발한 조기출수 계통의 유전적 변이성을 분석하였다. 선발한 조기출수 계통의 M7(2010년)과 M8(2011년) 세대에서 평균 출수기는 대조품종인 동진 1호(중만생종) 보다 각각 11일(γ-2 계통), 10일(γ-5 계통), 6일(γ-1 계통), 5일(γ-3 계통), 4일(γ-4 계통)이 빠르게 나타났다. ISSR 분석 결과, 선발계통의 유전적 다형성은 γ-2 계통 5.9%, γ-1 계통 7.5%, γ-4 계통 15.1%, γ-3 계통 15.3%, X-1 계통 19.4%, γ-5 계통 23.4%로 대조품종의 4.3% 보다 높게 나타나 방사선 조사로 DNA 수준에서 변이가 증가되었음을 확인하였다. 엽록체 DNA(rps16-trnK 영역) 분석 결과, 염기 길이는 대조 품종과 같은 664 bp인 반면, 총 5개 염기서열 영역에서 치환 변이가 발생하였고 그 중 6 bp 및 587 bp 영역의 변이는 선발 계통에서만 나타났다.