금속염화물계 방사성 폐기물은 전해공정으로 이루어진 파이로프로세싱공정의 주요한 방사성 폐기물이 다. 이와 같은 폐기물은 탄산염이나 질산염과 달리 고온에서 분해되지 않고 바로 휘발되며, 기존의 규산 계 유리와 상용성이 낮아 처리가 쉽지 않다. 본 연구팀은 금속염화물계 폐기물을 고화처리하는 방법으로 탈염화처리법을 채택하였다. 본 연구에서는 그 후속적인 연구로서, 탈염화물질로 제안된 SAP (SiO2- Al2O3-P2O5)의 조성을 변화시켜 LiCl-KCl과의 반응성을 향상시키고 고화공정을 단순화시키고자 하였다. 기본물질계에 Fe2O3를 첨가할 경우 무게반응비 SAP/Salt를 3에서 2.25로 낮출수 있으며, Fe가 Al을 치환 하는 몰분율이 0.1이상이 될 경우에는 오히려 반응성이 점진적으로 감소하는 것으로 확인되었다. 또한 M-SAP에 B2O3를 첨가할 경우에는 유리매질을 사용하지 않고 monolithic form을 제조할 수 있었다. 침출 시험결과 U-SAP 1071이 가장 높은 내구성을 보여주었으며, 1 g의 금속폐기물을 처리시 약 3∼4 g의 고 화체가 발생되며, 이는 기존의 고화처리법보다 약 ⅓∼¼배정도 최종처분부피가 감소되는 효과를 얻을 수 있다. 이상의 실험결과로부터, 기존의 유리고화공정으로 처리가 어려운 휘발성 금속염화물계 폐기물 을 단 하나의 물질을 이용하여 처리할 수 있음을 확인하였으며, 이러한 처리방법은 고화처리시 발생되는 부피를 최소화활 수 있는 대안적인 고화처리방법이 될 것으로 판단된다.

The wild relatives of soybean [Glycine soja Sieb. and Zucc.] have curly/wavy nature whereas cultivated varieties are upright. Such morphological characteristics have agronomic importance too. To investigate the molecular mechanism of development contributing to coiled morphology, screening was carried out to look for Arabidopsis mutants in activation tagging lines obtained by activation T-DNA treatment that have curly/wavy morphology. A mutant named Coiled Branch 1 (cbr1), is found to have a wavy and curly morphology with coiling branches. Plasmid rescue and genomic southern blot analysis revealed the site of T-DNA insertion in the genome. RT-PCR was performed to monitor expression levels of the genes adjacent to the T-DNA integration sites, and showed the activation of an E3 ubiquitin ligase gene. Database search showed that the gene with the RING domain belongs to a family of E3 ubiquitin ligases. Complementation test by overexpression and RNA interference of the gene was also carried out. The complementation test results showed that the novel gene activation tagging affected the cbr1 mutant phenotypes. Ubiquitylation has been linked virtually to every cellular process including plant development. E3 ubiquitin ligase has been reported to recognize target proteins that are to be ubiquinated for further degradation by the proteasome complex. Further, more detailed studies are needed to identify the specific substrate(s) of the novel E3 ubiquitin ligase gene.

Arabidopsis E3 SUMO ligase SIZ1 (AtSIZ1) controls vegetative growth and development including responses to nutrient deficiency and environment stresses. Here, we analyzed the effect of AtSIZ1 on the stability and amount of seed proteins. Proteomic analysis showed that the amount of three major nutrient reservoir proteins, CRUCIFERIN (CRU) 1, 2 and 3, were decreased in siz1-2 mutants. However, quantitative real-time RT-PCR showed that transcript levels of CRU1, 2 and 3 genes were rather significantly higher in siz1-2 mutants than wild-type plants. Yeast two hybrid analysis revealed that AtSIZ1 interacts with CRU1, CRU2 and CRU3, strongly suggesting that CRU1, 2 and 3 proteins are sumoylated by AtSIZ1. In addition, the 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.

Soybean [Glycine max (L.) Merr.] seeds are abundant in high-quality proteins and fats. In addition, soybean seeds are also rich in secondary metabolites, such as isoflavones, lecithin, and saponins. Triterpene saponins are major components of these physiologically active metabolites in soybean seeds. Soybean saponins are classified as group A and DDMP saponins. Among them group A saponins are undesirable component of food products due to bitterness and astringency and also cause foaming in tofu production. Whereas, DDMP saponins and their derivatives are less bitter and astringent and beneficial to human health when consumed as regular diet. Therefore, reducing the group A saponins or increasing the DDMP saponins are required to improve the food quality. The present study focused to identify and characterize the gene which is encoding a protein responsible for biosynthesis of DDMP saponins. EMS mutant lines (sg-7-1 & sg-7-2) which lack DDMP saponins were developed. The breeding cross has been made with these two mutants with two cultivars, Pungsannamul and Wooram to study the segregation and genetic linkage analysis, respectively. The segregation analysis showed that the mutant phenotype is controlled by single recessive gene. TLC analysis for phenotyping F2 population of Wooram X sg-7-1 showed mutant, wild and heterozygous types. To surprise two more patterns were detected and they were named as strange type1 (ST1) and strange type2 (ST2). Further, SSR marker analysis will be carried out to locate the gene which encoding a protein responsible for biosynthesis of DDMP saponins.

Soybean germplasm have diverse accessions with great variation in their ability to survive and reproduce under salt stress conditions. In general, cultivated soybeans are more sensitive to salt stress than their wild relatives, however exceptions are found in both the groups. These variations in response to salt stress makes soybean germplasm an interesting collection of genetic resources to be explored for the identification of salt-tolerance genes, and their mechanism of action. Here, in this report we presented a data showing differential response of selected accessions of both cultivated and wild soybeans to salt stress. Two modes of salt treatment; gradual salt stress (GS) as well as salt shock (SS) were used in this study. The GS was found more effective in finding the difference in response of soybean accessions to salt stress. Various genetic marker based methods are in use to identify and isolate the potential genes contributing to the salt tolerance in soybean. Even then there is a paucity of knowledge on the key genes contributing to the salt tolerance in soybean. We expect that a recently developed functional screen based method, like yeast based functional screen, using cDNA library generated from different salt tolerant accessions of soybean could lead to identification of novel genes responsible for salt tolerance in soybean. Also, we propose for the use of RNA isolated from different stages of GS and SS for making cDNA library to be used for functional screening.

Soybean [Glycine max (L.) Merr.] have a variety of flower colors which are controlled by six different genes (W1,W2,W3,W4,Wm, and Wp). Among these genes, mutation in W3 gene causes near white flowers in the background of w4 genotype whereas the genotype W3w4 does purple throat flowers. Earlier studies showed that dihydroflavonol 4-reductase1 (DFR1) gene was closely linked to the flower color variants for W3 locus. In order to find out the W3 gene responsible for w3 phenotype, we first, studied the candidate gene Glyma14g07940 (DFR1) which is having 100% similarity with DFR probe sequence. Sequence analysis of DFR1 between W3 and w3 soybeans showed one base substitution in exon 6 of w3 mutant soybean resulting in one amino acid change in the amino acid sequence. However, comparison of amino acid sequences of DFR proteins from various crop plants showed that there is no functional change in the protein. Besides, the promoter analysis showed that, 311 bp of indel was traced in 5’-upstream promoter region of DFR1 gene in the w3 mutant. Here, we show that the near white or purple throat phenotypes in G. max is associated with existence or nonexistence of indel at 5’- upstream promoter region and low or high expression of DFR1, respectively. These results suggest that w3 phenotype may be caused by certain regulator of DFR1 gene located near or distant from DFR1 in G. max. In further study, we need to check the correlation between promoter indel with W3 expression level through GUS analysis.

Gene expression is regulated by DNA and histone methylation by DNA and histone methyltransferases, respectively. In animal system, DNA methyltransferase with CG methylation activity is modified by SUMO conjugation and then its activity was increased, which means that the activity of DNA methyltransferase is modulated by posttranslational modification. so Chromatin remodeling is a new concept for expression of controlling of gene function. We thus analyzed the effect of E3 SUMO ligase AtSIZ1 in CMT3 (chromometnylase 3)-mediated genome methylation by next-generation sequencing (NGS), methyl binding domain MeDIP-sequencing and gene analysis using siz1-2 and cmt3 mutants. we carried out CG-enrich analysis by MeDIP sequencing revealed that the methylation level of the genome including transposons was significantly low in siz1-2 mutants compared to wild-type. Result showed the genes regulated by methylation, that genes related of embryo and root development, cellulose metabolism, and post-translational modifications. All of our data indicate that the methyltransfearse activity of CMT3 may be able to be regulated by AtSIZ1 and thereby CMT3-mediated gene expression and plant development also can be controlled by E3 SUMO ligase activity. Besides, our data also suggest that ammonium (NH4+) can stimulate AtSIZ1- and CMT3- mediated DNA methylation.

The transition from vegetative growth to flowering is a major developmental switch in the plant cycle and the timing of flowering is very critical for reproduction of plant species. In transition to flowering in plants, Flowering locus C (FLC) is one of the crucial factors. Here, we showed How the stability and activity of FLC are regulated by sumoylation mechanism. By pull-down assay, we showed that FLC interact with E3 SUMO ligase in vitro and vivo. And we showed that FLC is sumoylated in vitro condition with AtSUMO1 protein. In transgenic plants with overexpression of FLC and inducible expression of AtSIZ1, sumo E3 ligase led to increase of FLC protein level and delayed the post-translation degradation of FLC indicating that Arabidopsis E3 sumo ligase AtSIZ1 stabilizes FLC. Also, the plants with overexpression of mutant FLC (K154R, a mutation of the sumoylation site on FLC) flowered considerably earlier than plants with overexpression of FLC but comparable with wild type indicating that sumoylation is a important part for function of FLC. Our data indicate that the sumoylation of FLC is critical for its role in the control of flowering time.

Flowering time is a important agronomic trait for grain production in rice. So the control of flowering time is a critical step. In Arabidopsis, expression of certain key flowering gene such as FLOWERING LOCUS C (FLC) is known to be epigenetically regulated by chromatin modification through Enhancer of Zeste[E(z)], a histone methyltransferase, that core component of repressive complex, polycomb repressive complex2(PRC2). However, the chromatin mechanism involved in the regulation of rice flowering genes is presently not well known. Here we show that predict coding region of a intronic LncRNA[termed rice COLDAIR(OsCOLDAIR)], which is expected to associate with a component of PRC2, is predicted at rice FLC gene. And additionally we suggest interaction of histone methyltransferase and E3 SUMO ligase that indicate possibility of interaction with rice E(z) gene and rice E3 SUMO ligase. Our study contribute to control of rice flowering time by observing two factor that can regulate expression of related of rice FLC gene.

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

Protein disulfide isomerase (PDI) is a chaperone protein that involves in oxidative protein folding by acting as catalysts and folding assistants in the endoplasmic reticulum (ER). Genome database showed that rice contains three PDI-like genes. But, their functions and subcellullar localization are not clearly identified. Here, we show possible functions of rice PDI (OsPDI) during seed development. Seeds of OsPDI T-DNA insertion mutants which were identified by genomic DNA PCR and western blot display chalky phenotype. Electron microscope analysis revealed that endosperms of the OsPDIL1-1Δ mutant show imperfect packing of round starch granules, causing floury-white color. Abnormal form of protein body I (PB-I) containing prolamin and thick aleurone layer were also observed in the OsPDIL1-1Δ mutants. Protein content per seed was significantly low in the OsPDIL1-1Δ mutant. However, free sugar content was high in the OsPDIL1-1Δ mutant seed. Northern and western blot analyses showed that during seed development, OsPDI protein is steadily accumulated in the seed until maturation while its transcript level was highest at 10 days after flowering and rapidly decreased to basal level. In addition, OsPDI strongly interacts with cysteine protease OsCP1 and chaperone BiP protein accumulates in OsPDIL1-1Δ mutant. Besides, proteomic analysis of the OsPDIL1-1Δ mutant seed showed that OsPDI is post-translationally regulated and its loss causes accumulation of many types of seed proteins. Our results indicate that OsPDI plays a critical role in seed development through its regulatory activity for various proteins.

Grain yield, one of the most important agronomic traits, is greatly affected by architecture in rice. Here, we show that an OsPrMC3, a rice PrMC3 orthologue with a lipase or esterase domain, involves in yielding by tillering. Phenotypic analysis of T-DNA insertion mutant revealed that it has high number of tillers than wild type although height and leaf width are shorter and narrower than wild type. Size and branch number of panicle were greatly reduced in the mutant, which resulted in significant decrease of seed number per panicle and dry weight of the seeds. OsPrMC3 is highly expressed in the leaf during the early stage of development. However, it is mainly expressed in mature seed and root after flowering although its expression is detected in all of the tissues. Our result indicates that OsPrMC3 involves in leaf growth and tillering during vegetative growth and also seed development after flowering, suggesting its crucial regulatory role in yielding

Post-translational covalent modifications by small molecules or peptides remodel target proteins. One such modification, made by ubiquitin or small ubiquitin-related modifier (SUMO), is a rapidly expanding field in cell signaling pathways. Ubiquitin attachment controls the turnover and degradation of target proteins while SUMO conjugation regulates their activity and function. Recent studies report many examples of cross-talk between ubiquitin and SUMO pathways, indicating that the boundary is no longer clear. Here, we review recent progress concerning how ubiquitin and SUMO participate in new regulatory roles in plant cell, and how ubiquitination and sumoylation control plant growth and development.

and distribution of seed storage proteins are responsible for the quality of soybean and seed development. Among storage proteins, lipoxygenase isoforms (Mw. ~97 kDa) play a major role in the distinct bean flavor during storage. In this study, we compared three soybean elite cultivars viz., JIMPUM, JINPUM2 and TANMI2 (lipoxygenase null mutants, originated from Japan) along with WILLIAMS 82 (control plant, USA) to determine the seed storage proteins by proteomic approach. Phenotype of the mature seeds showed the variation in seed coat, color and appearance. Total seed proteins of the above cultivars were subjected to two dimensional gel electrophoresis (2-DE). The resulted protein profiling showed the intensity of the different quantitative spots varied among the four cultivars. We are now investigating by using other proteomic tool and the resulted difference in proteins may helpful in quality improvement or the functional roles in the seed development.

The composition and distribution of seed storage proteins are important factors for eating quality such as grain flavor and quality in rice (OryzasativaL.) Rice protein disulfide isomerase (OsPDI) and binding protein (OsBIP) regulate synthesis, stability and sorting of storage proteins. We thus have tried to develop a marker protein for selection of rice cultivars which have different eating quality. Immunoblot analysis revealed that protein levels of OsPDI and OsBIP have no direct correlation with eating quality, suggesting that they may indirectly participate in control of eating quality through their-interacting partners or other regulatory mechanism.