Soybean seed is a good source of plant protein in human consumables such as baby formula and protein concentrate. The seeds contain an abundance of storage proteins, namely β-conglycin and glycinin that account for ~ 70-80% of the total seed protein content. Proteome profiling has been proved to be an efficient way that can help us to investigate the seed storage proteins. In the present study, the seeds were removed from the pods and the cotylendonary tissues were separated from the testa for proteome analysis in order to investigate the seed storage proteins. A systematic proteome profiling was conducted through one-dimensional gel electrophoresis followed by MALDI-TOF-TOF mass spectrometry in the seeds (cotyledonary tissue) of soybean genotypes. Two dimensional gels stained with CBB, a total of 10 proteins were identified and analyzed using MASCOT search engine according to the similarity of sequences with previously characterized proteins along with the UniProt database. A total of ten proteins such as glycinin Gy4 precursor, glycinin G3 precursor, glycinin G1 precursor, glycinin chain A2B1a precursor, glycinin chain A2B1a precursor were identified in our investigation. However, the glycinin subunit may be considered to play important roles in soybean breeding and biochemical characterization. In addition, the improved technique will be useful to dissect the genetic control of glycinin expression in soybean.

Field emergence of Azuki bean is poor due to hard seed coat as compared to other legumes. In this study, an attempt was made to develop prediction method with regression analysis based on various seed vigor tests in laboratory for field emergence of azuki bean. Azuki bean seeds artificially aged to provide various levels of seed quality were evaluated by the standard germination test (SGT), cold germination test (CT), cool germination test (CGT), complex stressing vigor test (CSVT), tetrazolium(TZ) vigor test and electroconductivity test. The SGT was suitable for predicting the field emergence in the unaged high vigor seeds. The abnormal seedling percentage and shoot length in the CGT were highly correlated with field emergence of moderate vigor seeds artificially aged for 2 days. Electroconductivity, seed viability in the CSVT, and vigor and predicted germinability in the tetrazolium vigor test were also useful for predicting field emergence. Percent of ungerminated seed in the CSVT was correlated with field emergence in the low vigor seeds artificially aged for 4 days. In a stepwise multiple regression analysis, seed viability in the SGT, normal seedling percentage and dry matter weight in the CGT accounted for 86.9% of the predicted value of field emergence in azuki bean.

Soybean (Glycine max (L.) Merr.) is an important crop for protein, oil, carbohydrates, isoflavones, and many other nutrients to humans and animals. But, antinutritional factors in the raw mature soybean are exist. Raffinose and stachyose are main antinutritional factors in soybean seed. Both raffinose and stachyose are carbohydrates, belonging to the raffinose family of oligosaccharides (RFOs). RFOs are not readily digested in humans and cause flatulence or diarrhea. The objective of this research is to obtain the information on raffinose and stachyose content according to genotype and environment. A total of twenty two soybean genotypes (11 cultivars, 3 germplasms and 8 breeding lines) were selected. Each genotype was grown in the field for two years with two replications and harvested in bulk at natural maturity for two years. Content of raffinose and stachyose was detected by HPLC. The raffinose content (g/kg) of 22 genotypes was 2.68±0.21 - 5.87±2.43 in year 1 and was 3.24±0.37 - 9.05±0.16 in year 2. The stachyose content (g/kg) was 4.23±0.98 - 27.68±9.90 at year 1 and was 5.11±1.09 - 25.32±0.35 in year 2. Genotype and environment have highly significant effects on raffinose and stachyose content. Three genotypes (Da-7, 116-13, and RS-78) have low stachyose content at 5% significant level in two years. A positive correlation (R2=0.1985*) between raffinose and stachyose was observed in year 2. These informations are valuable in soybean genetics and breeding program related with raffinose and stachyose content.

Sorghum seed is traditionally used as secondary food sources in addition to rice in Korea. While the hypoglycemia regulating phytochemicals have been found in sorghum seed, peptides related with hypoglycemia never been studied before. To obtain the peptide characteristics and the specifically high-expressed peptides in hypoglycemic sorghum seed, peptide profiles of seven hypoglycemic and five nonhypoglycemic sorghum lines bred in RDA were determined using surface-enhanced laser desorption/ionization time-offlight mass spectrometry (SELDI-TOF MS). The twelve sorghum lines exhibited 104 peptides on CM10 protein chip array (weak cation exchange) and 95 peptides on Q10 (weak cation exchange) in the molecular mass range from 2,000 to 20,000 Da. Heat map via supervised hierarchical clustering of the significantly different peptides (p < 0.01) in peak intensity among the 12 lines effectively revealed the specifically upregulated peptides in each line and distinguished between 7 hypoglycemic and 5 non-hypoglycemic lines. Through the comparison with hypoglycemic and non-hypoglycemic lines, 10 peptides including 2231.6, 2845.4, 2907.9, 3063.5, 3132.6, 3520.8, 4078.8, 5066.2, 5296.5, 5375.5 Da were specifically high-expressed in hypoglycemic lines at p < 0.00001. This study characterized seed peptides of 12 sorghums and found ten peptides highly expressed for hypoglycemic sorghum lines, which could be used as peptide biomarkers for identification of hypoglycemic sorghum.

Sorghum seed is traditionally used as health supplements and the secondary food mixed with rice in Korea. While the research of reserve protein in sorghum seed have been carried out in many countries used as major food, much less is known about reserve proteins of Korean local sorghum seeds. To obtain protein characteristics in 20 Korean local sorghum seed, quantitative content of reserve protein was determined after fractionation by modified ‘Osbone’ method and α-kafirin of prolamin was determined by SDSPAGE. Mean albumin, globulin, prolamin and glutelin contents based on total seed protein content of 20 Korean local sorghum seed were 6.2%, 0.9%, 57.9% and 35.1%, respectively. Sorghum cultivar with high prolamin were ‘Whin-susu’, ‘Whin-Chalsusu’, ‘Whanggeum-Chalsusu’, and ‘Daepungshushu’. Sorghum cultivar with high α-kafirin were ‘Whin-susu’, ‘Geumsan-Chalsusu’, ‘Whin-Chalsusu’, and ‘Jangmok-susu’. Among the 20 varieties, ‘Whin-susu’ and ‘Whin-Chalsusu’ were selected as high α -kafirin and prolamin sorghum cultivar, which showed 64.5 and 71.9% of prolamin contents, respectively.

The purpose of this study was to develop the cost-effective and efficiency seed longevity prediction method of rice (Oryza sativa L.) germplasm for viability monitoring. To find an optimum predicting method for rice seed longevity at genebank, an accelerated ageing (AA) test, a controlled deterioration (CD) test and a dry-heat treatment (DHT) were conducted to the four groups of rice germplasm based on ecotype, such as Indica, Japonica, Javanica and Tongil type. Among the three artificial aging treatments, the dry-heat treatment of 36 hours at 90°C is suggested as a routine predictive test method of rice germplasm longevity at a genebank. The distribution of germination rate on 3,066 accessions which conserved 26.5 years at 4°C showed similar trend with the result of distribution by dry-heat treatment at 90°C on 36 hours using 106 accessions of rice selected samples which composed four ecotype groups. The results show that the dry-heat treatment affect not only predicting the rice seed longevity but also determining effective interval for monitoring germination of rice germplasm in genebanks.

The purpose of this study was to know the seed longevity of rice (Oryza sativa L.) germplasm for effective viability monitoring. The longevity was determined via germination tests of 3,066 accessions of rice germplasm from the National Agrobiodiversity Center, Rural Development Administration, Korea. The rice germplasm accessions have been conserved at a mid-term storage (4°Csealed metal can on either sides for 25~26.5 years. The final germination percentages of 3,066 rice germplasm accessions of 6.5±1.0% seed moisture content with 94% initial germination stored at 4°C for 26.5 years declined to 47% while at -18°C for 25 years maintained high germinability as 93%. Germination time courses, which represent the average performance of rice ecotypes stored at 4°C and 30% RH, were fitted regression equation, to calculate the time at which germination characteristically declined to 50% (P50). These P50 values of Indica, Japonica, Javanica and Tongil type in rice were 39.9, 22.9, 25.4 and 31.8 years, respectively. The rice germplasm stored at 4°C could be clustered in 4 groups using quartile of final germination after 26.5 years storage. The seed longevity (P50) of each group was estimated by regression equation of changed germination percentages according to storage periods. The P50 values of group Ⅰ, group Ⅱ, group Ⅲ and group Ⅳ were 21.1, 23.6, 30.0 and 75.7 years.

Seed spray는 종자, 비료 및 기타 재료들을 섞어 살포하는 방법으로 주로 대면적의 경관을 조성할 때 주로 사용되는 기법이다. 살포된 종자는 온도조건에 따라 경관형성 소요시간은 달라지나 시공시 식물생장과 온도간의 관계를 고려되지 않는다. 본 연구에서는 수레국화 종자의 효율적인 사용을 위하여 수레국화종자발아에 필요한 적정온도와 발아소요시간을 설정하는데 있다. 시험방법은 2013년 3월부터 4월까지 국립원예특작과학원에서 진행되었다. 시험재료는 수레국화이며 생장상을 이용하여 온도처리를 하였다. 시험방법은 페트리디쉬와 필터페이퍼에 100립씩 4반복으로 치상 후 10ml씩 1차 증류수를 관수하여 7수준의 온도를 처리하였다. 처리온도는 5, 10, 15, 20, 25, 30℃이다. 시험 중 광원은 4~4.5 μmoles/m2/s를 유지하였다. 조사방법은 실내시험에서는 페트리 디쉬에 치상 후 2일 간격으로 발아율을 조사하였다. Parabolic 함수를 이용하여 발아율 50% 기준으로 온도와 발아소요시간의 관계 나타낸 결과 수레국화 종자의 발아 최저온도는 0℃, 최적온도는 12.3℃, 최고온도는 24.5℃로 나타났다. 그리고 최적온도에 따른 발아율 50%까지 발아소요시간은 6.6일이었다.

Arabidopsis E3 SUMO ligase controls vegetative growth and development including responses to nutrient deficiency and environment stresses. Here, we analyzed seed proteins of its mutant siz1-2. Proteomic analysis showed that the amount of three major nutrient reservoir proteins were decreased in siz1-2 mutants. However, quantitative real-time RT-PCR showed that their transcript levels were significantly high in siz1-2 mutants compared to wild-type plants, which means that these proteins are stabilized by E3 SUMO ligase. In addition, yeast two hybrid assay showed that they interact with E3 SUMO ligase, suggesting that they must be sumoylated by E3 SUMO ligase. Furthermore, tthe analysis of amino acid composition by HPLC showed that the contents of amino acids were a bit high in siz1-2 mutants. Our data indicate that AtSIZ1 plays an important function for accumulation of seed storage proteins through its ligase activity

It is well known that Dharial (Bangladesh origin and weedy rice line) has longer seed longevity than indica and japonica rice varieties. To study the genetic basis of seed longevity of Dharial, we developed 240 BC3F7 backcross recombinant inbred lines derived from the crosses between Dharial (a donor parent) and two korea rice accessions (recurrent parents) including Ilmi and Gopum, respectively. Among these lines, we selected two introgression lines with longer seed longevity and named them Ilmi-NIL and Gopum-NIL. Also, we developed an EMS-induced mutant line from Dharial which has shortened seed longevity, and named it Dharial-EMS. We performed re-sequencing of four rice accessions that are Dharial, Dharial-EMS, Ilmi-NIL, and Gopum-NIL. A total of 706×106 raw reads were generated which provided sequence data over 46x rice genome coverage per each accession. We did genome-wide variation analysis comparing produced re-sequencing data and the re-sequencing data of Ilmi from NABIC database with the Nipponbare reference sequence. By graphical analysis of SNP distribution in rice genome of the five accessions, we could select candidate chromosomal segments introgressed from Dharial in Ilmi-NIL and Gopum-NIL. The introgressed chromosomal segments were in seven regions in Ilmi-NIL and eight regions in Gopum-NIL, and four common introgressed regions between Ilmi-NIL and Gopum-NIL were identified. 2,758 SNPs between Dharial and Dharial-EMS were found in the introgressed regions. Also, we detected 450 genes including at least one SNP among these SNPs. This result will facilitate identification of genes and development of molecular markers for improvement of seed longevity.

Spatial- and temporal-specific expression patterns are primarily regulated at the transcriptional level by the promoter. Therefore, it is important to determine the binding motifs of transcription factors to understand the networks associated with embryogenesis. Here, we used a protein-binding microarray (PBM) to determine the binding motif of OsSMF1, which is a basic leucine zipper transcription factor that is involved in the regulation of rice seed maturation. OsSMF1 (previously called RISBZ1) is known to interact with GCN4 motifs (TGA(G/C)TCA) to regulate seed storage proteins (SSPs). In addition, OsSMF1 (also known as OsbZIP58) functions as a key regulator of starch synthesis in the rice seed. Quadruple 9-mer-based PBM (Q9-PBM) and electrophoretic mobility shift assay (EMSA) experiments revealed that OsSMF1 binds to the ACGT (CCACGT(C/G)), GCN4 (TGA(G/C)TCA), and GCN4-like (GGATGAC) motifs with Kd values of 0.3353 μM, 0.6458 μM, and 1.117 μM, respectively. We also identified 60 putative OsSMF1 target genes using a combination of data from expression microarrays and RiceArrayNet (RAN) analysis. Of these OsSMF1 target genes, 20, 22, and 17 genes contained ACGT, GCN4, and GCN4-like motifs within the 2-kb promoter region, respectively. In addition to known target genes, we also identified 35 potential OsSMF1 target genes that have not been previously described in immature seeds. We also confirmed that OsSMF1 directly regulates Os03g0168500 (thioredoxin-related protein), RPBF, NAC6, and two hypothetical proteins (Os12g0621600 and Os11g0582400) in vivo. This study suggests that OsSMF1 functions in a wide range of seed development processes with specific binding affinities for three DNA binding motifs

Rice (Oryza sativa L.) is important crop as a staple carbohydrate sources and was regarded as domesticated in Asia region. The seed dormancy is one of the domesticated traits, and this character allows that plants survive in various environment. The degree of dormancy have been targeted for controling in breeding program while weak dormancy cause pre-harvest sprouting contrary to the nonuniform seed germination by strong dormancy. In this study, we surveyed the variation of germiability in diverse rice genetic resources including japonica and indica ecotype. Overall, the degree of seed dormancy of rice germplasm was distinguished into four groups and admixed types; first group(G-1) included accessions revealing high germiability, the accessions of second group(G-2) acquired the high germiability by after-ripening process, third group(G-3)’s accessions showed high germiability after dormancy breaking process and the accessions of last group(G-4) maintained the low germiability in spite of the dormancy breaking process. Among the 51 japonica accessions G-1, G-2, G-3 and G-4 included 15, 15, 11 and 10 accessions, respectively, and among 40 indica accessions, G-1, G-2, G-3 and G-4 groups included 15, 15, 9 and 1 accessions, respectively. Based on these primary test, we plan to detail examine the seed germiability and survey the genetic factors affecting the degree of germiability in representative accessions of each group

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.

In southern region of Korea, double cropping system including soybean as a second crop is widely adopted because prolonged high temperature in October enables soybean maturation even in late autumn. Global warming during decades derives transition of soybean planting dates as late as late-July. Delayed planting beyond the critical time moves reproductive stage to less favorable weather conditions like low temperature and short photoperiod, which can affect soybean yield and seed composition. Therefore selection of appropriate cultivar for changed climate and cultural practice is required. Effects of different eco-type and planting date on soybean yield was investigated for three sowing dates of 20 June, 5 July and 20 July with ten cultivars. Strong interaction between cultivar and sowing date was detected in number of branch, node and pod. No significant interaction was observed in 100-grain weight and yield, and this might be resulted from similar yield reduction pattern. Daepung, Taekwang and Uram recorded the highest yield when planted late, proving that these three cultivars are the most adaptable to delayed planting. Delayed planting results in higher sugar content, especially stachyose and raffinose which have detrimental effect on human health by causing flatulence and diarrhea. Protein content was decreased and oil content was increased in July 20 planting. Decrease in protein content was the largest in Hwangeumol and maintained high in Saedanbaek. This information will provide a guideline when select appropriate soybean cultivar in delayed planting to maintain yield and seed quality.

Seed viability testing provides valuable information for assessing seed lot germinability. However, most testing methods require destruction of seed prior to test. Because the dissected seeds for viability test cannot be used further evaluation, the nondestructively X-ray photography technique that can be applied for the evaluation of seed quality has been developed. In order to know the validity and accuracy of X-ray photography technique in seed evaluation test that conducted to remove the abnormal seed from a seed lot, we have compared the results from tetrazolium viability test, germination test and X-ray contrast method in cotton. Metallic salts treatment increased the efficiency of X-ray photographic method by enhancing the penetration of X-ray in abnormal or damaged seeds rather than normal seeds that have strong and well-organized tissues in seed. Cotton seeds presoaked for 16 hr in distilled water followed by soaking into metallic salt solution (5% NaI in water) for 60 min were easily classified seeds into dead seed and viable seed based on the radiography images obtained by X-ray radiation. We concluded that soft X-ray photography was reliable to find out the various defective characters due to heat and mechanical damage of seeds.

본 연구는 참깨 유전자원 핵심집단과 주요 품종 및 우량계통의 세사민과 세사몰린, 지방 함량변이를 분석하여 리그난 함량이 높은 참깨 품종육성을 위한 기초자료로 활용하고자 하였다. 본 시험에 이용된 참깨 유전자원 143점의 리그난 함량 범위는 2.33 mg/g에서 12.17 mg/g으로 나타났다. 그 중 터키에서 수집된 IT184615은 리그난 함량이 12.17 mg/g으로 가장 높아 리그난 함량 개량을 위한 유전자원으로 활용성이 높은 것으로 판단되었다. 참깨 유전자원을 14개의 수집원산지별, 4개의 종피색(백, 황, 갈, 흑)별, 4개의 SSR마커그룹으로 분류하고 리그난 함량과의 유의성을 검정한 결과 세사민, 세사몰린 함량은 수집원산지, 종피색 간에 유의한 차가 인정되었다. 러시아(10.0 mg/g), 네팔(9.08 mg/g)에서 수집된 유전자원은 다른 국가에서 수집된 유전자원보다 평균 리그난 함량이 높았다. 세사민 함량은 세사몰린 함량과 정의 상관관계가 있어 리그난 함량이 높은 참깨 품종 육성을 위해서는 두 성분을 동시에 높이는 방향으로 선발되어야 할 것으로 판단된다. 참깨 주요 품종과 우량계통의 리그난, 지방 함량의 지역간, 연차간 변이를 분석하여 품질특성에 미치는 환경의 영향을 분석한 결과 리그난과 지방 함량은 지역간, 연차간에 고도로 유의적인 차이가 인정되었고 지역간 변이가 계통 간 변이보다 커서 유전변이보다 환경변이의 변이의 폭이 넓었다. 따라서 리그난, 지방 함량이 높은 참깨 품종 육성을 위해서는 함량이 높은 유전자원을 육종재료로 활용함과 동시에 리그난 축적에 관계되는 재배환경에 관한 연구가 지속적으로 이루어져야 할 것이다.