dlm mutants controlling disease lesion mimic leaf trait may be useful in basic research of disease hypersensitive response and programmed cell death in soybean. The study on genetic relationship between dlm trait and other morphological C trait, position of dlm allele on classical linkage group, and a molecular marker linked to dlm allele was little reported. Two populations [T173 (ffDlmDlm) x T363 (FFdlmdlm), T363 (dlmdlmY9Y9) x T135 (DlmDlmy9y9)] were made to find independent assortment or linkage between dlm locus and f locus or between dlm locus and y9 locus. The segregation ratios of 3 : 1 were observed in the F2 population and the Chi-square values suggested that the disease lesion mimic leaf, fasciation stem, and chlorophyll-deficient leaf traits were controlled by a single recessive gene. Segregation ratios of 78 Dlm_F_: 19 Dlm_ff: 17 dlmdlmF_ : 3 dlmdlmff based on F2 phenotype showed that dlm allele was inherited independently with the f allele controlling fasciation stem trait in soybean. Also, segregation ratios of from 149 Dlm_Y9_: 41 Dlm_y9y9: 38 dlmdlmY9_ : 5 dlmdlmy9y9 based on F2 phenotype confirmed that dlm allele was inherited independently with the y9 allele controlling chlorophyll- deficient leaf trait in soybean. From these results, dlm allele would not be located on linkage group 11 (molecular linkage group: D1b+W) and linkage group 14 (molecular linkage group: E) in soybean. This results will helpful to attempt to position the Dlm locus on the soybean molecular linkage map.

RAPD analysis showed that all the OTUs of 'Sandolbae' were the same species because amplified band patterns of all samples generated by each of 5 random primers were identical. Even though there were different environmental conditions, all the "Chuiangne" trees from three different places were the same species, and also all the "Cheongshilli" trees were the same species too. No genetic variations were detected between native Korean pears grown in the habitats and in the research field. Because 212 polymorphic bands were generated by 9 primers selected through primer screening, they were possible to analyze genetic relationship among naturally growing three native Korean pears and nine cultivars of Pyrus pyrifolia and P. communis. Based on the RAPD analysis, three main groups were formed. The first group represented the Six P. pyrifoia cultivars, the second group was the three native Korean pears, and the last group was the three P. communis cultivars. Genetic distance between 'Wonwhang' and 'Chojuro' was closer than other cultivars in group 1 since dissimilarity index value between these two cultivars was 50.82. However, genetic distance between 'Niitaka' and 'Chojuro' was the most distant compared to the others in group 1. In group 2, 'Sandlobae' was genetically closer to 'Chuiangne' than 'Cheongshilli' because dissimilarity index value between 'Sandlobae' and 'Chuiangne' was smaller, 50.82, than the value between 'Sandlobae' and 'Cheongshilli', 63.636. In group 3, 'Old Home' was genetically closer to 'Bartlett' than 'Kaiser Alexander(or Bosc)'. Group 3 composed of P. communis cultivars was genetically further than other two groups, P. pyrifolia cultivars and native Korean pears.

GE interaction is the expression of differential genotypic adaptation across environments. GE interactions through different stability parameters and performance of the traits of genotypes were studied. The traits were days to maturity, pod length, number of pods/ plant, 100-seed weight and seed yield/plant in ten soybean genotypes across five environments. Significant differences were observed for genotypes, environments and GE interactions. Stability analysis after Eberhart and Russell's model suggested that the genotypes used in this study were all more or less responsive to environmental changes. Most of the genotypes perform better in Env.3. Based on phenotypic indices(Pi), regression (S2di) genotype Garurab was found fairly stable for days to maturity. BS-23 and G-2120 may be considered as stable genotype for pod length. All the genotypes except G-2120 showed that the genotypes were relatively unstable under environmental fluctuation for the number of pod/plant. Genotype BS-23 was found most stable among all the genotypes for 100-seed weight. BS-3 and Gaurab was the most stable and desirable genotypes for seed yield in soybean.

The removal of nitrogen compounds from a wastewater is essential and it is often accomplished by biological process. An aerobic nitrate-removing bacterium was isolated from a municipal sewage treatment plant and soil. On the basis of its morphological, cultural and physiological characteristics and 16S rRNA sequencing data, this strain was identified as Pseudomonas fluorescens, and named as P. fluorescens K4. The optimal conditions of the initial pH and temperature of media for its growth were 7.0~8.0 and 30℃, respectively. P. fluorescens K4 was able to remove 99.9% of nitrate after 24 h in a culture. The strain could grow with a nitrate concentration up to 800 mg/l and was able to remove 99.9% of nitrate after 104 h of incubation. The optimal electron donor was sodium citrate for a nitrate removal. The strain K4 showed a capability of a complete nitrate removal when the initial C/N ratio was 1.0. An effect of the initial seed concentration was observed for a cell of 10% (v/v) for a nitrate removal. Especially P. fluorescens K4 could completely remove 200 mg/l ammonium for 3 days.

A phosphate starvation-induced acid phosphatase cDNA was cloned from the rice, Oryza sativa. The cDNA encoding O. sativa acid phosphatase (OsACP1) has 1100 bp with an open reading frame of 274 amino acid residues. The deduced amino acid sequence of OsACP1 cDNA showed 53% identity to tomato acid phosphatase and 46-50% identity to several other plant phosphatases. OsACP1 expression was up-regulated in the rice plant and in cell culture in the absence of phosphate (Pi). The induced expression of OsACP1 was a specific response to Pi starvation, and was not affected by the deprivation of other nutrients. OsACP1 expression was responsive to the level of Pi supply, with transcripts of OsACP1 being abundant in Pi-deprived root. The OsACP1 cDNA was expressed as a 30 kDa polypeptide in baculovirus-infected insect Sf9 cells. In addition, the OsACP1 gene was introduced into Arabidopsis via Agrobacterium mediated transformation. Functional expression of the OsACP1 gene in the transgenic Arabidopsis lines was confirmed by Northern blot and Western blot analyses, as well as phosphatase activity assays. These results suggest that the OsACP1 gene can be used to develop new transgenic dicotyledonous plants able to adapt to Pi-deficient conditions.

We isolated low temperature inducible genes using suppression subtractive hybridization (SSH) method and were able to obtain to cloneMLT107 gene encoding peroxiredoxin and aminotransferase. The full-length cDNA of MLT107 is 1,049 bp with an open reading frame (ORF) consisting of 261 amino acid (aa). Genomic southern blot confirmed that mungbean genome has two copies of MLT107 gene. Northern blot analysis was also carried out for the gene expression during ABA, NaCl, drought, wounding and H2O2 stresses. The expression of MLT107 gene significantly decreased by ABA, NaCl and drought stress, but wounding and H2O2 stress significantly induced MLT107 gene expression. Especially, H2O2 strongly induced the MLT107 gene expression. The expression of MLT107gene during low temperature stress started to increase in 3 h after treatment, and than slightly decreased and again increased at 24 h. Using GFP fusion vector, smGFP-MLT107 was targeted both to mitochondria and chloroplast. However, it was mostly targeted to mitochondria and partially targeted to chloroplast. For the functional analysis of MLT107, MLT107 recombinant protein was heterologously expressed in E.coli. The MLT107 recombinant cells showed enhanced antioxidant activity compared to that of vector control cells.

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.

We isolated wound-inducible genes using suppression subtractive hybridization (SSH) method and were able to obtain to clone w123 gene encoding dnaJ like protein. The full-length cDNA of w123 is 689 bp with an open reading frame (ORF) consisting of 163 amino acid (aa). Genomic southern blot confirmed that soybean genome has two copies of w123 gene. Northern blot analysis was also carried out for the gene expression during heat, NaCl, drought, wounding stresses. The expression of w123 gene specifically induced by heat, NaCl, wounding and drought stress. Using GFP fusion vector, w123-smGFP was targeted both to nucleus. For the functional analysis of w123, His-tagged w123 recombinant protein was heterologously expressed in E. coli. The w123 recombinant cells showed enhanced heat tolerance compared to that of vector control cells. We suggest that dnaJ-like w123 protein function as molecular chaperone in the nucleus of the plant cell during various stresses.

In this study, a 141 BC3F4 lines from across between the O. sativa cv. Milyang23 as there current parent, and O. glaberrima as the donor parent was used to identify favorable QTL alleles from O. glaberrima for yield and yield components. To detect the introgressions, 198 microsatellite markers of known chromosomal position were used for the parental survey. Of the 178 markers, 128 (64.6%) showed polymorphism. Among them, 115 SSR markers were used to construct a genetic linkage map with average interval length of 12.7 cM based on the previous rice molecular genetic map. The mean number of O. glaberrima segments in the population was 1.84 ranging from 0 to 7. The average length of the segments was 16.6 cM and ranged from 0.5 to 232.5 cM. This population consisting of 141 lines was used to evaluate for six traits of agronomic importance and genotypes were determined for 141 BC3F5 using SSR markers. A total of 22 QTLs for 6 traits were detected on chromosomes 1, 2, 3, 4, 5, 6, 7 and 9. Phenotypic variance associated with each QTL ranged 9.5% ~ 58.2%. For 26 of the QTLs identified in this study, the O. glaberrima alleles contributed a desirable agronomic effect despite the over all undesirable characteristics of the wild phenotype. Favorable wild alleles were detected for culm length, panicle length, yield, panicles per plant and 1000-grain weight. When compared with previous studies involving interspecific crosses, it can be concluded that O. glaberrima is useful asa source of valuable alleles for rice improvement. There sults will be discussed.

ABSTRACT: Wide hybridization was used to broaden the gene pool of japonica rice. Different approaches involving direct crosses between four japonica cultivars and eight wild species in combination with anther culture, embryo rescue, and molecular markers were used to produce interspecific hybrids, advanced alien introgression lines, dihaploids, and characterization of alien introgression. Interspecific hybrids were produced between Jinmibyeo, Ilpumbyeo, and Hwaseongbyeo cultivars of rice (O. sativa, 2n=24 AA) and wild species, O. rufipogon (2n=24, AA), O. longistaminata (2n=24, AA), O. punctata (2n=24, BB), O. minuta (2n=48, BBCC), O. alta (2n=24, CCDD), including African rice species, O. glaberrima (2n=24, AA). Crosses involving species other than A genome were produced through embryo rescue and were sterile. Following backcrossing with the recurrent japonica parents, advanced progenies have been produced for transfer of alien genes into japonica rice. As results of yield trials, three elite lines were generated from this cross; they are "Suweon 487" with resistance to black streak dwarf virus, "Suweon 497" with blast and bacterial blight resistance and "Suweon 506" with blast resistance, emonstrating that wild species genes have now become important component of japonica rice breeding.

Plant proteomic study requires a high-throughput separation to the detection and analysis of peptides and proteins by mass spectrometry (MS) to detect low abundant proteins. Together, efficient separation and MS can lead to the detection of thousands of plant proteins in a cell or tissue and help build proteome maps that can provide a global snap-shot of the cell or tissue status. Recently efficient separations based on the HPLC were introduced to allow deeper protein detection and improve throughput. For the HPLC based methods, Multidimensional Protein Identification Technology (MudPIT) and 1D-Gel-LC-MS/MS will be introduced. In MudPIT analysis, all proteins in a sample are digested into peptides before the separation step then the mixture of peptides are separated through the biphasic capillary column and sequentially eluted into the mass spectrometer and analyzed. 1D-Gel-LC-MS/MS separates protein samples in 1D-SDS-Gel then the proteins in each band were ingel-digested into peptides followed by peptides separation with Reverse Phase column and elution into the mass spectrometer. The main goal of this presentation is to introduce the recent protein separation and identification methods based on the HPLC coupled with MS analysis including conventional method of 2D-PAGE.

Organ abscission is a programmed cell separation process that results in the detachment of an entire organ from a plant. Our goal is to understand the signaling pathway that regulates this physiological process. The receptor-like protein kinase, HAESA (HAE), and its paralog, HAESA-like 2 (HSL2), are both expressed in the floral abscission zones in Arabidopsis thaliana. Loss-of-function analyses of either gene do not show any phenotypical change, but the hae hsl2 double mutant shows an abscission-defect phenotype. Examination of the abscission zone by light and scanning electron microscopy showed that the abscission zone in the hae hsl2 appears structurally normal. The force required to remove the petals in wild type and hae hsl2 flowers was measured using a petal breakstrength meter. The force required to remove petals from the hae hsl2flowers at all stages of development was similar to that of wild type flowers that have not yet begun to abscise their petals. Taken together, these data support the role of HAE and HSL2 in the activation of cell separation, rather than differentiation of the abscission zone. Ethylene is also known to promote abscission; therefore we tested the ethylene-induced triple response and the effect of exogenous treatment on floral organ in the hae hsl2, revealing that HAE and HSL2 act independently of ethylene. This implies that the HAE is critical for floral abscission in concert with the action of HSL2.

Echinacea purpurea is an important medicinal plant native of North America, which contains caffeic acid derivatives, alkamides, glycoproteins, and polysaccharides. For commercial roduction of bioactive compounds from Echinacea purpurea, adventitious roots were induced and proliferated in 5 and 20-liter bioreactors under various cultural conditions (Medium composition, inoculum density, air volume, light, temperature et al.). In a 5-liter bioreactor, more than 10-fold of biomass increment was achieved after 5 weeks of culture. Increases of root biomass and bioactive compounds were correlated with increased consumption of sucrose and macro/micro nutrients in the medium. Elicitations have been conducted with UV-C, NO, and SA to increase the accumulation of total polyphenolics, flavonoids and caffeic acid derivatives in the adventitious root cultures, which resulted in 1.2-1.6 times of increment. Based on the previous experimental results, 500-liter and 1000-liter scale bioreactors were applied for large scale cultures of Echinacea purpurea adventitious roots. Total root biomass of 3.62 kg and 5.05kg (dry weight) were achieved in a 500 L and a 1000 L bioreactor, respectively. The accumulation of 22.55 mg/g DW cichoric acid, 4.92 mg/g DW chlorogenic acid and 3.99 mg/g DW caftaric acids were also achieved from the adventitious root in a 1000-liter scale bioreactor.

Ornithine deacetylase (argE) gene as a negative selection marker gene was successfully co-transformed with SOD-APX or NDPK2 as a target gene to develop marker free transgenic rice. E.coli argE gene encodes an enzyme which activates N-acetyl phosphinothricin (N-AcPt) through deacetylation. The enzyme was reported that deacetylate N-AcPt, a non cyto-toxic compounds, to produce cyto-toxic PPT. So the argE gene can be used as a negative selection marker and N-AcPt can be used as a substrate of argE gene. Using former purification method (Kriete G., et al., 1996) was very expensive because the small amount of N-AcPt produced by the method. We developed a strategy to produce N-AcPt by means of chemical method. N-AcPt function was identified using hygromycin resistant T1 seeds on MS basal medium contained N-AcPt prior to utilization of argE gene as a negative selection marker in marker free transgenic rice. Negative selection effects were performed with T1 seeds containing argE gene under N-AcPt. This system provides a efficient negative selection effect from transgenic rice and that will be efficiently used for the production of marker gene free rice plants.

Aluminum (Al) toxicity in plants is one of the major limitations to crop growth on acid soils. The Al-induced change of H+-ATPase expression has been regarded as an important mechanism for Al tolerance in soybean. To investigate whether translational regulation of plasma membrane H+-ATPase is involved in the response to Al stress, we conducted western - blot of this protein. The results show that western - blot of plasma membrane H+-ATPase in the "Sowon" (Al tolernace) significantly increased in translational expression level, while citric acid (50 μM) with Al (50 μM) treatment has not effected. In contrast, Al sensitive cultivar "Poongsannamool" inhibited expression level of plasma membrane H+-ATPase with Al treatment. Two - dimensional gel analysis were performed to determine the protein induction patterns of control and Al (50 μM, 24 h) treated soybean. There are many changes of plasma membrane proteins in both cultivars under Al stress. Especially "Sowon" was significantly enhanced the expression of the plasma membrane H+-ATPase in Al treatment. But protein expression of "Poongsannamool" was less than "Sowon". These results suggest that the regulatory role of plasma membrane H+-ATPase may involved the tolerance mechanism in soybean roots. At the present, transcriptional level of H+-ATPase is under investigation.

The powdery mildews are obligate biotrophic fungi and are one of the most economically important groups of plant pathogens (Agrios, 1997). As a family, the powdery mildews infect a broad range of plant species including barley, wheat, pea, apple, sugar beet, and grape (Braun, 1987). Powdery mildew pathogens uptake nutrients by forming a feeding structure, the haustorium, within 12‐18 hours after infection (hai) in their respective host plants. Because of the initial stages of these plant pathogen invasion are mostly confined to a limited number of host cells, it’s often too late to find out till the infections widely spread out. To identify the earliest and often transient responses to pathogen attack, there is considerable interest in monitoring the subcellular events that occur specifically in living host cells. Recent improvements in live cell imaging using fluorescent‐tagged markers have expanded the scope of experiments that can be performed. Changes in the subcellular distribution of organelles as well as fluorescently tagged proteins can be monitored in real time in living tissues during pathogen attack, and the dynamic nature of such changes across space and over time can be determined. The application of these sensitive imaging methods has extended earlier observations made with Nomarski microscopy or inferred from static transmission electron micrographs about the focal accumulation of subcellular organelles at sites of pathogen attack. In addition, recent experiments have demonstrated the focused accumulation and interaction of specific plant proteins at penetration sites, opening a new window on early host responses and raising questions about the underlying plant processes that sense and direct this marshalling of host resources to block pathogen entry.

Magnaporthe oryzae is a causal pathogen of the rice blast, the most destructive disease of rice worldwide. This ascomycete fungus has been intensively studied as a model organism in plant-pathogen interactions. Recently, the genome sequence of M. oryzae 70-15 was published by International Rice Blast Consortium. This led us to investigate comparative and functional analysis of this pathogen at genome-wide level. As a first step, sequencing project of M. oryzae Korean strain KJ201 was launched to face practical interests on diversity of plant pathogens. We constructed a fosmid library with a copy number controllable pCC1 vector. Over 13,000 end reads from 6637 fosmid clones were generated and anchored on genome sequence of the strain 70-15. Currently, 9,793 end reads, in which 2.684, clones with both ends and 1,757 clones with on end matched, are aligned to the sequence of strain 70-15. In the next step, shotgun sequencing of fosmid clones was conducted for fine scale comparative analysis of chromosome 7 that is well defined in genome sequencing of 70-15. Totally 119 fosmid clones were pooled in region and analyzed. For reasonable intra-species comparison in genome of M. oryzae, two strains, KJ201 and Guy11, were chosen for whole genome shotgun sequencing analysis with GS454 FLX system. About 8X genome coverage sequences were obtained for each strain and being merged for comparative analysis. Genome information of other strains is being also available from China and Japanese research groups and could be joined together for extended comparative analysis and the results give more detail in genome diversity in species level.

A new garden pea cultivar, Daehyup 2, was developed at the Yeongnam Agricultural Research Institute (YARI) in 2006. It was selected from the cross between Olwandu and Sachulwandu in 1996. The preliminary, advanced, and regional yield trials for evaluation and selection of this cultivar were carried out from 2003 to 2006. It has white flower, yellow cotyledon, absent or very weakly curved pod shape and yellow seed color. It was resistant to virus and powdery mildew disease. The 100 green seed weight of Daehyup 2 was 65.5 g, which was 8.9 g heavier than check cultivar. The sucrose and total sugar contents of fresh pod of Daehyup 2 were 11.1% and 12.5%, respectively, which were 3.2% and 4.3% higher than those of Sachulwandu. The fresh pod yield of Daehyup 2 was similar to that of Sachulwandu with 12.16 ton per hectare in the regional yield trials carried out in three locations of Korea from 2005 to 2006.

A new black seed coat soybean variety, “Socheong” was developed at the Yeongnam Agricultural Research Institute (YARI) in 2006. The goal to breed the black seed coat soybean is to develop the cultivar with high yield, lodging tolerance, resistant to disease such as soybean mosaic virus (SMV), and bacterial pustule and seed size. Socheong was selected from the cross between Milyang 78, which was late maturing, susceptible to lodging and SMV and with large seed size and green cotyledon, and Peking which was tolerant to lodging and with small seed size. The preliminary, advanced and regional yield trials for evaluation and selection of this variety were carried out from 2002 to 2006. It has determinate growth habit, white flower, brown pubescence, brown pod color, black seed coat, green cotyledon, elongated flattened seed shape, oval leaf shape and small seed size (15.7 grams per 100 seeds), and it was 3 days later in maturity than the check cultivar Cheongjakong. Socheong was higher, in the seed quality of sucrose and total sugar contents (6.8 and 8.2%) and isoflavone contents (1,754 ㎍/g) than the check cultivar. Futhermore, it has good characteristics for mechanical harvest, such as lodging tolerance, pod shattering and stem diameter. It also has been identified to have resistance to soybean mosaic virus symptom which was the troublesome soybean diseases. The average yield of Socheong was 2.21 ton per hectare in the regional yield trials carried out in four locations of Korea among seven locations from 2004 to 2006, which was 5 percent lower than the check cultivar Cheongjakong.

A new black seed coat soybean cultivar, “Heugmi” was developed at the Yeongnam Agricultural Research Institute (YARI) in 2006. The goal to breed the black seed coat soybean is to develop the cultivar with high yield, lodging tolerance, resistant to disease such as soybean mosaic virus (SMV), and bacterial pustule and seed size. Heugmi was selected from the cross between Milyang 78, which was late maturing, susceptible to lodging and SMV and with large seed size and green cotyledon, and Milyang 68, which was middle maturing, resistant to lodging and SMV and with middle seed size and yellow cotyledon. The preliminary, advanced and regional yield trials for evaluation and selection of this cultivar were carried out from 2001 to 2006. It has determinate growth habit, purple flower, brown pubescence, black pod color, black seed coat, green cotyledon, spherical seed shape, oval leaf shape and middle seed size (24.8 grams per 100 seeds), and it was 2 days earlier in maturity than the check cultivar Cheongjakong. Heugmi was better in the seed quality of crude protein contents, sucrose and total sugar contents than the check cultivar. It also has been identified to have resistance to SMV which was the troublesome soybean diseases. The average yield of Heugmi was 2.53 ton per hectare in the regional yield trials carried out in seven locations of Korea from 2004 to 2006, which was 8 percent higher than the check cultivar Cheongjakong.