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.

Radiation technologies have been widely used in mutation breeding of crops. Although gamma-radiation has been predominantly applied in radiation breeding, ion beam irradiation is recently emerging as a methodology highly effective in induction of mutations that are useful for plant breeding. Therefore, we investigated the biological effects caused by irradiation of two types of ion beams, which were proton and ion beams, respectively, in pepper. In the evaluation of survival rate, LD50 values were between 300 and 350 Gy, 30 and 40 Gy in irradiation of proton and carbon beams, respectively, while LD50 for gamma-ray was determined to be between 50 and 100 Gy. Growth traits including shoot length, root length, and root width were also examined in pepper seedlings according to does of proton and carbon ion beams to estimate biological effectiveness of each radiation. The result was applied in the construction of pepper mutant population which will be used to develop pepper breeding materials containing novel characteristics.

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.

Perilla frutescens (L.) is an annual herbaceous and ornamental plant in the Lamiaceae family. Perilla frutescens (L.) Britt.cv.Chookyoupjaso were irradiated using a 200 Gy gamma ray in 1995. By HPLC analysis, this new cultivar significantly induced isoegomaketone content compared with ‘Chookyoupjaso’ control. The phenotypical difference was the changed leaf color of the ‘Atom-Ketone’ from violet to green. The yield potential of this cultivar (106 kg/10a) was 1.83 folds higher than that of ‘Chookyoupjaso’ (57.65 kg/10a). The methanol extracts of ‘Atom-Ketone’ inhibit nitric oxide (NO) production in LPS-stimulated RAW 264.7 cells. This extract was further partitioned using ethyl acetate (EtOAc), butanol (BuOH), and water. The EtOAc fraction (EF-Atom-Ketone) was evaluated for antiinflammatory activities. These results indicated that the EF-Atom-Ketone reduced NO production by inhibiting inducible nitric oxide synthase (iNOS) expression. The EF-Atom-Ketone treatment also significantly diminished expression of MCP-1 and IL-6. Therefore, ‘Atom-Ketone’ reveals the potential therapeutic use of bioactive

Perilla frutescens (L.) is an edible plant, not only used as s food ingredient, but also in skin cream, soaps, and medicinal preparetions. ‘Atom-Ros’, a perilla (Perilla frutescenc (L.) Britt. cv. Chookyoupjaso was developed in 1995 by 200 Gy gamma irradiation-mutagenesis. This new cultivar has high rosmarinic acid content more than two fold compare with ‘Chookyoupjaso’ control. The observed phenotypical difference was changed leaf color of the ‘Atom-Ketone’ from violet to green. The yield potential of this cultivar (123.5 kg/10a) was 2.14 fold higher than that of ‘Chookyoupjaso’ (57.65 kg/10a). The methanol extracts of ‘Atom-Ros’ were tested for inhibition of nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophase cells. Atom-Ros showed significant inhibition of NO production. This rosmarinic acid extracted from ‘Atom-Ros’ has a good potential to be developed as an antioxidant agent.

This study was conducted to determine the optimal dose of gamma-ray for mutation breeding in sorghum (Sorghum bicolor M.). Gamma-rays irradiated to dry seeds with various doses (0 to 1000 Gy) at Korea Atomic Energy Research Institute. Lethal dosage (LD50) was approximately 256 Gy. Significant decreases in growth characteristics (plant height, tiller number and fresh weight) were observed by dose of increased over 300 Gy. Reduction doses (RD50) was approximately 363 Gy in plant height. We also conducted comet assay to observe nucleus DNA damage due to gamma irradiation. In comet assay, a clear difference was identified over 100 Gy treatments. With increasing doses of gamma-ray in the range of 100 to 400 Gy, the rate of head DNA was decreased significantly from 93.52% to 67.57%. The most of the Sorghum cells were severely damaged in the integrity of DNA by gamma-ray. These data provide valuable information when the optimal dose should be chosen for purpose of mutation breeding program of Sorghum.

In this study, the concentrations of isoflavones, anthocyanins and total phenol content (TPC) in 19 soybean mutant lines changed seed coat color from yellow to black or brown were determined. Among 19 soybean mutant lines, 5 soybean mutant lines with black pigment were detected 3 anthocyanins (delphinidin-3-O-β-D-glucoside, D3G; cyaniding 3-O-β -D-glucoside, C3G; petunidin 3-O-β-D-glucoside, Pt3G). The highest concentration of anthocyanins among 5 soybean mutant lines was D-16 (1280.0 ± 19.4 mg/100g seed coat) derived from cv. Danbaek. Although isoflavone contents of all soybean mutant lines showed lower levels compared to original cultivars, glycitein was detected only 5 soybean mutant lines (DP-37-2, DP-38, DP-39, DP-40, and DP-41 derived from cv. Daepung). In TPC of 19 soybean mutant lines, DP-10 was increase levels compared to original cultivar, while DP-37-2, DP-40, and DP-41 were decrease levels of TPC. Using reduction of DPPH, we measured the free radical scavenging activity (FRSA) among 19 soybean mutant lines. Five black and 4 brown soybean mutants showed significant increase in FRSA. On the basis of these results, it was concluded that gamma irradiation may change the isoflavone, anthocyanin, and total phenol contents of soybean. These mutant lines using in this study can be useful for the breeding of soybean varieties altering the nutritional values.

This study was carried out to determine the effects of different harvesting stage on nutritive value and the quality of ensiled kenaf after fermentation among six kenaf cultivars. Six kenaf cultivars including two different maturity groups, mid-late maturing (Auxu, Jangdae and Jinju) and early-maturing (Baekma, Jeokbong and C14), were planted on May 14, 2013. Four harvesting times were made at intervals of 20 days from 15 July to 16 September, 2013. In all cultivars, the CP (crude protein) contents were decreased by a delayed harvest; the CP contents of kenaf silage were ranged from 92 to 184 g kg-1. Interestingly, there were no significant difference of NDF (neutral detergent fiber) and ADF (acid detergent fiber) content between the cultivars, however NDF and ADF content of kenaf silage were significantly increased by a late harvest. The silages of all cultivars displayed a low pH ranges less than 4.0, which is sufficient for stable storage. The lactic acid contents in Auxu were from 2.57 to 3.21%, which is higher than other cultivars. The harvesting stages did not affect to the concentrations of butyric acid and acetic acid in all cultivars. These results indicate that the harvesting stage is more important for the quality of kenaf silage than cultivar differences. And kenaf silage could be also used as fodder for ruminants.

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.

KAS360-22 종자에 1998년 250 Gy의 감마선을 조사하였다. 그 후대에서 미이라병에 저항성을 가지는 계통을 선발･육성하여, ‘원율’로 명명하고 2009년에 최종적으로 품종보호권을 출원 신청하여 2012년 등록 완료하였다. 원품종 KAS360-22의 종피색이 노란색인데 반하여 신품종 ‘원율’은 갈색을 나타냈다. 100립중은 재래종 KAS360-22이 17.2 g, ‘원율’은 27.5 g으로 종자무게는 증가하였으며, ‘일품검정콩’(27.4g)과는 별다른 차이를 보이지 않았다. 신품종 ‘원율’의 10a 당 수량은 미이라병 발병 조건에서 233.3 kg으로 재래종 KAS360-22 (24.9 kg)에 비교하여 10.6배 가량 높았고, 대조구 ‘일품검정콩’(121.4 kg)에 비교하여도 1.9배 가량 높아 밥밑용 콩으로 안정적인 수량 확보가 가능할 것으로 사료된다.

Copy number variations (CNVs) are considered major sources of genetic variation, and CNVs may influence phenotypic variation and gene expression. To detect CNVs, rice seeds were exposed with 100～400 Gy of gamma-ray (GA, 60Co), cosmic-ray (CR) by Russia ISS, and 30 and 40 Gy of ion beam (IB, 220 Mev carbon ion). After the exposed rice seeds were cultured in 1/2 MS medium for 14 days, they were used for array-based Comparative genomic hybridization (CGH) analysis using Agilent’s RICE CGH array. As a results, the highest number of CNVs (Gain 808 and Loss 24,080) were detected in the CR treatment, whereas GA100 (100 Gy of GA) was identified the least CNVs. Compared individual chromosome, the chromosome 8 and 11 were identified the highest CNVs, the chromosome 3 had the least CNVs. Most of identified CNVs existed in the range of 10～500kb. In particular, the same CNV locations among different types of ionizing radiation were observed in chromosome 12, and these CNVs contained the commonly 5 amplified genes, containing retrotransposon protein, NADH-ubiquinone oxidoreductase chain 3, heavy metal transport/detoxification protein domain containing protein, and 2 unknown proteins. Other studies were reported that Ty1 (Long Terminal Repeat-retrotransposon family 1) transcription and retrotransposition were induced by different environmental stresses such as ionizing radiation, UV-light exposure, DNA damage and nutrient starvation in Saccharomyces cerevisiae. Our results also show that retrotransposon protein (LOC_Os12g34016) was specifically amplified by different types of ionizing radiation.

VitE (tocotrienols and tocopherols) are micronutrients with antioxidant properties synthesized by photosynthetic bacteria and plants that play important roles in animal and human nutrition. A new mutant line, T1001-1, was isolated from in vitro mutagenized population by ionizing radiation and shown to have increased VitE contents. The total VitE content was 26% increased in the T1001-1 mutant seeds compare with cv. Dongan (wild-type). In addition, we showed that the mutant confers retarded seedling growth during the early seedling growth stage in rice. To study the molecular mechanism of VitE biosynthesis, we used the rice microarray to identify genes that are upor down-regulated in T1001-1 mutant. In addition, we identified differentially regulated pathway using MapMan analysis, which provides deep insight into changes in transcript and metabolites. Our results enhanced the transcription of genes involved in starch and lipid metabolism in T1001-1 mutant. To identify the molecular mechanisms of the events involving transcription factors in tocopherol accumulation, we compared the expression patterns of transcription factors. The AP2-EREBP, WRKY, C2H2 transcription factor were up-regulated, whereas the MYB family was down-regulated in T1001-1 mutant. Our results demonstrate change of important transcript in high level of VitE accumulating rice mutant.

To determine the expression levels of genes related to the salt stress response in rice, gene expression profiles were investigated through microarray analysis using the rice mutant line Till-II-877. There were no significant changes in physiological response under salt stress of the mutant increased less than that in the WT. The intensity of gene expression was analyzed and compared between the wild type and mutant lines using a microarray. Among the most significantly affected pathways, α-linolenic acid metabolism and linoleic acid metabolism (in lipid metabolism), fructose and mannose metabolism and glycolysis-gluconeogenesis (in carbohydrate metabolism), cysteine and methionine metabolism (in amino acid metabolism), and carbon fixation (in the energy metabolism of photosynthetic organisms) showed changes in gene expression levels under salt stress. These results further our understanding of the effects of salt stress in rice and may aid in the development of salt-tolerant rice cultivars.

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.

Mutant lines induced by ethyl methane sulfonate (EMS) have been used for crop improvement and functional genomics. Since pepper is very recalcitrant to be transformed, EMS mutagenesis could be an alternative method to generate useful mutant lines and to characterize the function of genes. We have developed mutant lines consisting of about 3,938 M2 mutant lines using Korea local landrace, C. annuum ‘Yuwolcho’. Yuwolcho has suitable traits for mutagenesis such as early flowering and maturation, large number of seeds per fruit, and susceptibility to various diseases. Up to now, 917 M2 mutant lines were evaluated to confirm the effect of EMS. M2 mutant lines have shown variations in plant stature (small size, dwarfism, and early death), leaf development (light color, variegation and morphological change) and flower (inflorescence, morphological change) and fruit (size and color). We observed the largest morphological variation in leaf development. Most of these mutant phenotypes were inherited recessively. In addition, we are applying cel1-based TILLING to identify useful mutant lines. We will apply cel1-based TILLING to identify useful mutant lines. We are expecting that these mutant lines will be very useful to study the function of genes in C. annuum.

기내배양한 양란 심비디움 원괴체에 전자빔(2 MeV/n, 0.5 mA)을 10 Gy～300 Gy로 조사하여 생장 변화와 유전적 변이의 다양성을 검토하였다. 전자빔을 조사 후 배양 10주째에 선량의 증가에 따라 원괴체로부터 신초유도가 감소하였고, 200 Gy 이상의 선량을 조사한 처리구에서는 신초분화가 크게 억제되었다. 원괴체 18개체를 ISSR 분석한 결과 대조구는 12.5% 다형성을 나타내었고, 전자빔 조사구에서는 42.2%의 다형성을 나타내었다. RAPD 분석한 결과 대조구에서 13%의 다형성을 나타내었고, 전자빔 조사구에서는 43.8%의 다형성을 나타내었다. 따라서 RAPD 및 ISSR 분석 모두에서 전자빔 조사시 대조구보다 3.5배 정도 다형성이 증가한 것으로 나타났다. 유전적 유사도 지수(GSM)는 전자빔 처리가 대조구보다 낮게 나타나 전자빔 처리에 의하여 개체들 간의 유전적 다양성이 높아진 것으로 보인다. 군집분석 결과 RAPD와 ISSR 분석 모두 대조구와 전자빔 처리구가 분리되어 그룹을 이루었다. 이와 같이 전자빔 조사로 기내 배양 양란 심비디움의 유전적 다형성이 증대됨을 확인하였다.

Ionizing radiation is known to cause chromosomal alterations such as inversions and deletions and affects gene expression within the plant genome. To monitor the genome-wide transcriptome changes by ionizing radiation, we used rice Affimetrix GeneChip microarray to identify genes that are up- or down regulated by gamma-ray (200 Gy, 60Co source), cosmic-ray and ion beam (40 Gy, 220 MeV carbon ion). The overall expression patterns between gamma-ray and ion beam were similar but cosmic-ray was regulated differently. Combined results from all 3 radiations identified 27 up-regulated genes and 188 down regulated genes. These results mean the induction of similar mechanism changes in treatments of gamma ray and ion beam. However the different expression in treatment of cosmic-ray might be due to the other environmental conditions. Among the commonly up- or down- regulated genes, we chose highly up- or down- regulated several genes and confirmed its regulation in response to ionizing radiation exposure by RT-PCR analysis. Moreover, we showed that specific co-expression networks of candidate radio marker genes by ARACNE algorithm. Our results present profiles of gene expression related to different ionizing radiation and marker gene to predict sensitivity to ionizing radiation, such as GS (glutelin subunit) and FBX322.

A dietary deficiency of tryptophan can cause pellagra and lead to low levels of serotonin that is associated with depression, aggression, anxiety and overeating in humans. Thus, enhancement of tryptophan content in rice has great potential benefit for human and animal diets. In this study, a total of 1,350 rice mutant population was used to identify single nucleotide polymorphisms (SNPs) in Oryza sativa anthranilate synthase alpha1(OASA1) gene that was associated with negative feedback in tryptophan biosynthesis. For high-throughput TILLING analysis, 5 fluorescence-labeled primer sets were designed to cover exon regions of OASA1 locus and PCR amplifications were analyzed using ABI3130xl DNA sequencer. Through the screening of 1,350 mutant lines, nine mutant lines produced one or two cleaved fragments in the PCR products of OASA1 locus. The full sequencing of nine mutant lines revealed that total 31 SNPs were located in the regions of OASA1. In particular, three mutant lines contained SNPs in coding regions that resulted in an amino acid change. The tryptophan contents of the three mutant lines were 2.2- to 2.3-fold higher than the wild type. These high-tryptophan mutant lines will be used rice breeding programs and contribute directly to enhancing human nutrition.

Kenaf (Hibiscus cannabinus) is an annual herbaceous plant of the family Malvaceae that has been planted in Africa for more than 4000 years and used as source of fiber, energy and feed stock. Also, kenaf seeds are good source for edible oil used for first class cooking oil and margarine production. The seeds can be used for lubrication, soap, paint and varnishes. This study was carried out to evaluate fatty acids variation among sixteen kenaf germplasm and gamma-ray induced mutants derived from Jinju and Auxu. Linoleic, oleic, and palmitic acid were the predominant fatty acids in all kenaf seed oils. The sixteen accessions showed a wide range of fatty acid compositions, spanning from 28.94 to 43.36% saturated, 56.64 to 71.05% total unsaturated, 15.52 to 46.85% monounsaturated, and 13.56 to 48.97% polyunsaturated fatty acids. The mutant lines derived from Jinju, significantly surpassed parental mean for all the palmitic and oleic acid. Also, the mutant lines derived from Auxu showed broad ranges of variation in oleic and linoleic acid and narrow ranges of variation in stearic and palmitic acid. The relative amount of monounsaturated fatty acids (MUFA) were increased at all the gamma-ray induced mutants. These results will provide a valuable information to assist parental selection of kenaf breeding.

The protein in soybean seeds accounts for approximately 40% of the dry seed weight. Two major storage proteins, 7S and 11S, constitute 70-80% of the total storage proteins in the seeds. In this study, the variation of total soluble protein extracts from 1152 soybean landraces that have been collected from South Korea were studied using high-throughput screening method with HT Protein Express Labchip (Caliper Life Sciences, Inc.). Seven distinct protein band patterns - four protein sub-units of 11S and three sub-units of 7S, were taken into account and their presence or absence were analyzed. Among the 1152 landraces, 525 genotypes were identified as lacking lipoxygenase, 255 lacking α1 subunit, 680 lacking α subunit, 169 lacking β subunit, 140 lacking acidic, 114 lacking Kunitz Trypsin Inhibitor (KTi) and 199 lacking basic protein patterns. The high-throughput protein analysis is helpful in screening a large number of populations with less time and minimum labor. The selected genotypes with low amounts or lacking of anti-nutritional factors such as trypsin inhibitor, lipoxygenase and α subunit would be used for future breeding purpose of quality improvement in soybean protein.