Background : Cirsium plants have been used for perennial edible plants and grow wild in the mountainous regions of Korea, including Cirsium setidens and C. pendulum. In particular, C. setidens is commonly called 'gondre', and it has been used medicinally for diseases such as hematuria, hepatitis and hypertension. Hairy roots cultivation can be used as a method for increasing production through mass culture of medicinal plants, and elicitors such as methyl jasmonate (MeJa) can be treated to increase the content of certain useful ingredients. In this study, the hairy root was derived from the leaf tissues of C. setidens and C. pendulum, and the HPLC pattern was compared by MeJa treatment. Methods and Results : Agrobacterium rhizogenes R1000 was used to induce hairy roots in 1/2× MS medium. In addition, the hairy roots was treated with MeJa for different time (0, 1, 2, 5, 10, 24, 48, 72 h) and with various concentrations (0, 10, 50, 100, 200, 300 μM). The HPLC pattern changes were analyzed by on-line HPLC-ABTS. Four new peaks were observed in both Cirsium setidens and C. pendulum hairy roots, all of which showed antioxidant activity. In the case of C. pendulum, chlorogenic acid content was about 4 times higher than that of leaves. These peaks, including chlorogenic acid, were all affected by MeJa treatment. Conclusion : Four peaks were detected in the hairy roots of C. setidens and C. pendulum not in the leaves, and they were confirmed to be affected by the treatment of MeJa. It is necessary to clarify the structure through the subsequent compound separation.

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.

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.

The pollen grain is a unique tricellular structure suitable for the delivery of the sperm cells to the ovule. All nutrients required for microspore and pollen cell growth are derived by passage through the anther locule and secretion by the tapetum lining. During later stages the tapetum degenerates but contributes to produce pigments, waxes, lipids and proteins which form the pollen coat and function in signaling between male (pollen) and female (pistil) tissues. The development of both normal pollen and tapetum is necessary for the fertilization processes in rice and would be exploited for the induction of male-sterility which is very useful to improve economic value of crops. We aredeveloping new approaches using a conditional male-sterility for the F1 hybrid seed production in rice. The conventional three parental systems for F1 hybrid seed production requirethe following three lines: male-sterile line, maintainer line, and restorer line. In this system, a critical requirement is to maintain the male-sterile inbred lines. Here we suggest molecular approaches, in which the engineered male-sterile plants are generated by regulating endogenous hormonal balance through the loss-of-function of genes. We can expect the male-sterility can be restored by exogenous applications of hormones such as gibberellin or jasmonic acid. Based on two parental systems, we will address the answer onfollowing question: how can we maintain a male-sterile line producing 100% male-sterile progenies without a maintainer? This work was supported by grants from Crop Functional Genomics Center of the 21C Frontier Program (CG1517), RepublicKorea.

Nitrogen (N) fertilization is essential for alleviating nutrient deficiencies of the world’s population by increasing rice production, one of the most important food crops of our time. Here we established an in vivo hydroponics rice seedling culture system to investigate the physio-biochemical and molecular responses of various rice genotypes to low nitrogen application. Yoshida’s nutrient solution (YS) was used to grow rice seedlings, and at three-week-old the seedlings manifested highly stable and reproducible symptoms, such as reduced shoot growth and length. Out of 12 genetically selected or tested genotypes, almost all (11 genotypes) showed varied degrees of growth reduction response to applied nitrogen (4 and 40 ppm N for treatment and control, respectively), but SR19663-B-B-34-3-3-3-1 showed similar growth as the control though its leaf width was smaller than the control. The leaves of a 11 representative low nitrogen-responsive genotype as BG90-2 were sampled for revealing the protein profiles between low and normal (control) nitrogen application by using two-dimensional gel electrophoresis (2-DGE) followed by staining of separated proteins with silver. Fifty differentially expressed silver stained protein spots were excised from 2-D gels and 41 proteins identified using high-throughput mass spectrometry (MS) using matrix-assisted laser desorption/ionization-time of flight-MS and nano electrospray ionization liquid chromatography tandem MS. These proteins could be assigned as major (energy metabolism, photosynthesis and oxidative stress) and minor functional categories, revealing many novel low N-responsive proteins, including those having energy/photosynthesis, and defense/stress, and iron homeostasis-related functions.