Establisment of rice library is an essential approach for rice functional genomics study. Utilizaing maize transposable element Ac/Ds is a promising method to construct insertional mutagenesis library of rice. Ac/Ds tagging system has received extensive application in rice during the past several years. The maize Ds element is one of the main tagging vehicles used in rice. Narrow leaf mutant have short height, narrow leaf width and large angle. To compare with wild type and narrow leaf mutant in detail, we observed the leaves under microscope. In specific portion(large and small vein), no significantly reduce cell size and number of cell. Knock-out of the OsNLR(narrow leaf ribokinase) gene inhibits internodes, panicles, angle(between leaf and stem), leaf, seed. OsNLR was shown to specifically expressed on leaf. In real time PCR analysis with mature leaf of wild type and mutant, there might be a functional association between OsAGO7, NRL1, NAL1 and NAL7 in regulating leaf development. We tested on the experimental field using wild type and mutant plants. In agricutural traits that contain leaf and seed related traits(except angle) significantly reduce in mutant plants. These results demonstrate that OsNLR gene may be associated with leaf development.

We used an efficient system to create rice mutant by Ac/Ds transposon insertion mutagenesis, such as selected homozygous mutant in dwarf phenotypes. We reported here the identification of function of dwarf OsGASD gene(Oryza sativa Gibberellin Acid Sensitive Dwarf). OsGASD gene encodes a 344 amino acid polypeptide and no homology proteins in Gene Bank. The osgasd mutnat was sensitive to exogenous gibberellic acid(GA) level. We performed experiment to controlled expression the OsGASD gene, its role in plant development, a quantitative analysis of endogenous GA content and sensitivity to GA. The osgasd mutant includes smaller amount of active GAs than wild-type. osgasd mutant plant of GA biosynthesis pathway causes GA deficiency and dwarf plants, and endogenous GA suppliance can restore the wild type phenotype in this mutant. There result indicated that OsGASD gene regulated the elongation of shoot, stem and plant height. The increased expression of OsGASD gene dramatically induces expression of the factors associated with GA biosynthesis, whereas osgasd mutant suppression of the factors associated with GA biosynthesis, loading to dwarf phenotypes. That applied GA3 at the plant development stage to survey the response of OsGASD gene to GA3. We suggest that OsGASD gene is related to factors of GA biosynthesis pathway regulating rice internodes development.

R2R3 MYB transcription factors play regulatory roles in plant responses to various environmental stresses and nutrient deficiency. In this study, we isolated MYB-like gene respond to phosphorus deprivation in rice and designated OsMYB4P, an R2R3 MYB transcription factor, from rice under low-phosphate conditions. OsMYB4P is 993bp long and encodes a 330 amino acid polypeptide. OsMYB4P was localized in the nucleus and acted as a transcriptional activator. Transcriptional levels of OsMYB4P in cell suspension, shoots, and roots of rice increased under low phosphate conditions. Shoots and roots of OsMYB4P overexpressing plants grew well in high and low phosphate conditions. In addition, root system architecture was altered considerably as a result of OsMYB4P overexpression. Under both phosphate sufficient and deficient conditions, more Pi accumulated in shoots and roots of OsMYB4P overexpressing plants than in the wild type. Overexpression of OsMYB4P led to greater expression of Pi transporter-family proteins OsPT1, OsPT2, OsPT4, OsPT7, and OsPT8 in shoots, and to decreased or unchanged expression of these proteins in roots, with the exception of OsPT8. These results demonstrate that OsMYB4P may be associated with efficient utilization of Pi in rice.

The ubiquitin-26S proteasome system is important in the quality control of intracellular proteins. The ubiquitin-26S proteasome system includes the E1 (ubiquitin activating), E2 (ubiquitin conjugating) and E3 (ubiquitin ligase) enzymes. U-box proteins are a derived version of RING-finger domains, which have E3 enzyme activity. Here, we present the isolation of a novel U-box protein, OsUPS, from rice (Oryza sativa).The cDNA encoding the O.sativa U-box protein(OsUPS) comprises 1338bp, with an open reading frame of 445 amino acids. The open reading frame of the OsUPS protein is comprised of notable domains: a single ~70-amino acid domain and a GKL domain that contains conserved glycine, lysine/ arginine residues and leucine-rich feature. We found that full-length expression of OsUPS was up-regulated in both rice plants and cell culture in the absence of inorganic phosphate (Pi). A self-ubiquitination assay indicated that the bacterially expressed OsUPS protein had E3 ligase activity, and subcellular localization results showed that OsUPS was located in the chloroplast. Suppression of OsUPS resulted in servre signs of toxicity caused by the over-accumulation of Pi. These results support the notion that OsUPS plays an important role in the Pi signaling pathway through the ubiquitin-26S proteasome system.

Fibroin silk proteins from spider or silkworm are attractive biomaterials that are of particular biotechnological interest for industrial and medical purposes because of their unique physical and mechanical properties. In this study, we generated and characterized the transgenic rice plant expressing a spider silk protein. Spider silks have great potential as biomaterials with extraordinary properties. Here, we report the cloning and characterization of the major ampullate silk protein gene from the spider Araneus ventricosus. A cDNA encoding the partial major ampullate silk protein (AvMaSp) was cloned from A. ventricosus. An analysis of the cDNA sequence shows that AvMaSp consists of a 240 amino acid repetitive region and a 99 amino acid C-terminal non-repetitive domain. The peptide motifs that were found in the spider major ampullate silk proteins, (A)n, (GA)n, and (GGX)n, were conserved in the repetitive region of AvMaSp. Phylogenetic analysis further confirmed that AvMaSp belongs to the spider major ampullate spidroin family of proteins. Recombinant AvMaSp-R was degraded abruptly by trypsin. However, AvMaSp-R was stable at 100 °C for at least 30 min. Additionally, the AvMaSp-R was stable at pH values from 2 to 12 for at least 1 h. Taken together, our findings describe the molecular structure and biochemical properties of the A. ventricosus major ampullate silk protein and demonstrate its potential as a biomaterial.

The molecuar prossing of upsteam regulation of Pi response genes during Pi starvation remains inadequately understood. R2R3 MYB transcription factors play regulatory roles in plant responses to various environmental stresses and nutrient deficiency. In this study, we isolated and designated OsMYB4P, an R2R3 MYB transcription factor, from rice under low-phosphate conditions. OsMYB4P was localized in the nucleus and acted as a transcriptional activator. Transcriptional levels of OsMYB4P in cell suspension, shoots, and roots of rice increased under low phosphate conditions. To investigate the function of OsMYB4P in Pi-starvation signaling, we developed transgenic rice plants overexpressing OsMYB4P for analysis of Pi signaling and uptake. Shoots and roots of OsMYB4P overexpressing plants grew well in high and low phosphate conditions. In addition, root system architecture was altered considerably as a result of OsMYB4P overexpression. Under both phosphate sufficient and deficient conditions, more Pi accumulated in shoots and roots of OsMYB4P overexpressing plants than in the wild type. Overexpression of OsMYB4P led to greater expression of Pi transporter-family proteins OsPT1, OsPT2, OsPT4, OsPT7, and OsPT8 in shoots, and to decreased or unchanged expression of these proteins in roots, with the exception of OsPT8. These results demonstrate that OsMYB4P lead to Pi accumulation and acts as a Pi-dependent regulator in controlling the expression of Pi transporters.

We used an efficient system to create rice mutant by Ac/Ds transposon insertion mutagenesis, such as selected homozygous mutant in dwarf phenotypes. We reported here the identification of function of dwarf OsGASD gene(Oryza sativa Gibberellin Acid Sensitive Dwarf). OsGASD gene encodes a 344 amino acid polypeptide and no homology proteins in GeneBank. The osgasd mutnat was sensitive to exogenous GA level. We performed experiment to controlled expression the OsGASD gene, its role in plant development, a quantitative analysis of endogenous GA content and sensitivity to GA. The osgasd mutant includes smaller amount of active GAs than wild-type. osgasd mutant plant of GA biosynthesis pathway causes GA deficiency and dwarf plants, and endogenous GA suppliance can restore the wild type phenotype in this mutant. There sult indicated that OsGASD gene regulated the elongation of shoot, stem and plant height. The increased expression of OsGASD gene dramatically induces expression of the factors associated with GA biosynthesis such as CPS, KO, KAO, GA 20 ox and GA 2 ox, whereas osgasd mutant suppression of the factors associated with GA biosynthesis, loading to dwarf phenotypes. That applied GA3 at the plant development stage to survey the response of OsGASD gene to GA3. These results indicated that OsGASD gene is involved in GA biosynthesis factors, not only in the internodes, but also leaf length at the developing stage.

In this study, we generated and characterized the transgenic rice plant expressing a spider silk protein. Spider silks have great potential as biomaterials with extraordinary properties. We report the cloning and characterization of the major ampullate silk protein gene from the spider Araneus ventricosus. A cDNA encoding the partial major ampullate silk protein (AvMaSp) was cloned from A. ventricosus. An analysis of the cDNA sequence shows that AvMaSp consists of a 240 amino acid repetitive region and a 99 amino acid C-terminal non-repetitive domain. The peptide motifs that were found in the spider major ampullate silk proteins, (A)n, (GA)n, and (GGX)n, were conserved in the repetitive region of AvMaSp. Phylogenetic analysis further confirmed that AvMaSp belongs to the spider major ampullate spidroin family of proteins. The AvMaSp-R cDNA, which encodes the 240 amino acid repetitive domain, was expressed as a soluble 22 kDa polypeptide in baculovirus-infected insect cells. To produce transgenic rice plant with high contents of glycine and alanine, the prolamin promoter-driven AvDrag was introduced into rice plant via agrobacterium tumefaciens-mediated gene transformation. The introduction and copy number of the AvDrag gene in transgenic rice plants were determined by PCR and Southern blot analysis. AvDrag expression in transgenic rice seeds was examined by Northern blot and Western blot analysis. Immunofluorescence staining with the AvDrag antiserum revealed that the recombinant AvDrag protein were localized in transgenic rice seed. Furthermore, the amino acid content analysis showed that transgenic rice seeds were greatly increased in glycine and alanine as compared to controls

The molecular processing of upstream regulation of Pi response genes during Pi starvation remains inadequately understood. Several transcription factor have been studied that appear to regulate subsets of the responses to Pi stress either positively or negatively. MYB genes are responsive to one or multiple type of hormone and stress treatments. In this study, cDNA of the MYB have been cloned, and we generated Rice overexpressing plants for characterization of these genes. OsMYB gene function focused on phosphate conditions with rice and Arabidopsis transgenic plants. We selected 30 - T1 transgenic lines from T0 transgenic rices. those are shown high Pi content. The Pi contents of shoots part of transgenic plants were shown 10~20% increased Pi contents than WT, whereas roots have 30% increased Pi contents. As a result, OsMYB genes affect Pi uptake in plants. To investigate interactions between MYB proteins and phosphate signaling related genes. We demonstrate that Myb-binding sites (MBSs) exist in putative promoter of OsPT transporter by analysis of bioinformatics, and its bind specific MYB transcription factor. OsMYB expression is induced by low Pi and Pi deficiency, and its overexpression plants are shown morphological phenotype in Pi stress.

The utilization of several genetic resources is the most important for developing rice, such as high yield seeds or various stresses. We used an efficient system to create rice mutant by Ac/Ds transposon insertion mutagenesis, such as selected homozygous mutant in dwarf phenotypes. We reported here the identification of function of dwarf OsGASD gene (Oryzasativa Gibberellin Acid Sensitive Dwarf). OsGASD gene encodes a 344 amino acid polypeptide and nohomology proteins in GeneBank. The osgasd mutnat was sensitive to exogenous gibberellic acid (GA) level. We performed experiment to controlled expression the OsGASD gene, its role in plant development, a quantitative analysis of endogenous GA content and sensitivity to GA. The osgasd mutant includes smaller amount of active GAs than wild-type. osgasd mutant plant of GA biosynthesis pathway causes GA deficiency and dwarf plants ,and endogenous GA suppliance can restore the wildtype phenotype in this mutant. The result indicated that OsGASD gene regulated the elongation of shoot, stem and plant height. The increased expression of OsGASD gene dramatically induces expression of the factors associated with GA biosynthesis such as CPS, KO, KAO, GA20ox and GA2ox, whereas osgasd mutant suppression of the factors associated with GA biosynthesis, loading to dwarf phenotypes. That applied GA3 at the plant development stage to survey the response of OsGASD gene to GA3. We suggest that OsGASD gene is related to factors of GA biosynthesis pathway regulating rice internodes development.

In this study, we generated and characterized the transgenic rice plant expressing a spider silk protein. A cDNA coding for the C-terminus of spider silk protein (AvMaSp) was cloned from the spider Araneus ventricosus. Analysis of the cDNA sequence shows that the C-terminus of AvMaSp consists of 165 amino acids of are petitive region and 99 amino acids of a C-terminalnon-repetitive region. The peptide motifs found in spider silk proteins, GGX and An, were conserved in the repetitive region of AvDrag. The AvMaSp cDNA was expressed as a 28kDa polypeptide in baculovirus-infected insect cells. To produce transgenic rice plant with high contents of glycine and alanine, the prolamin promoter-driven AvMaSp was introduced into rice plant via Agrobacteriumtumefaciens-mediated gene transformation. Because of seeds pecific prolamin promoter, expression of AvMaSp protein has been achieved inriceseed. The introduction and copy number of the AvMaSp gene in transgenic rice plants were determined by PCR and Southern blot analysis. AvMaSp expression in transgenic rice seeds was examined by Northern blot and Western blot analysis. Immuno fluorescence staining with the AvMaSpantiserum revealed that the recombinant AvMaSp proteins were localized in transgenic rice seeds. Furthermore, the amino acid content analysis showed that transgenic rice seeds were greatly increased in glycine and alanine as compared to controls. The present study is the first to show the expression of spider silk protein in rice seed.

Farmers have use phosphate fertilizer to provide sufficient yields. However, overuse of phosphorus accumulate in soil and causes soil and water pollution. We evaluated the phosphate acquisition and growth characteristics of OsPT1 transgenic rice (OsPT1-OX, over-expressing the high affinity phosphate transporter 1) in high phosphate soils with different level of nitrogen fertilizer treatment to investigate removing ability of excessive phosphate from soil. OsPT1-OX had shorter culm length but more tillers than those of wild-type plants in each soil conditions. Phosphate content per dry weight of OsPT1-OX was 1.8 times higher than that of wild-type under control fertilizer treated conditions. Although the dry weight of OsPT1-OX was not different from that of wild-type plants, whole plant phosphate content was 1.7 times higher than that of wild-type plants under control fertilizer conditions. Tiller number and phosphate content per dry weight of wild-type plants increased following high levels of phosphate application but did not change by following additional nitrogen application. Tiller number and phosphate content per dry weight of OsPT1-OX did not change under the high phosphate condition, but increased following nitrogen application under similar conditions. Whole plant phosphate content was highest under high nitrogen and high phosphate application conditions. These results suggest that OsPT1-OX may reduce phosphate content in soils containing excess phosphate and may be further effective under high nitrogen condition.

The molecular processing of upstream regulation of Pi response genes during Pi starvation remains inadequately understood. several transcription factor have been studied that appear to regulate subsets of the responses to Pi stress either positively or negatively. MYB gene is responsive to one or multiple type of hormone and stress treatments. In this study, cDNA of the MYB have been cloned, and we generated Rice overexpressing plants for characterization of these genes .OsMYB gene’s functions focused on phosphate conditions with rice and Arabidopsis transgenic plants. We selected 30 - T1 transgenic lines from T0 transgenic rices. those are shown high Pi content. The Pi contents of shoots part of transgenic plants were shown 10~20% increased Pi contents than WT, whereas roots have 30% increased Pi contents. As a result, OsMYB genes affect Pi uptake in plants. To investigate interactions between MYB proteins and phosphate signaling related genes.

In this study, we generated and characterized the transgenic rice plant expressing a spider silk protein. A cDNA coding for the C-terminus of spider dragline silk protein (AvDrag) was cloned from the spider Araneus ventricosus. Analysis of the cDNA sequence shows that the C-terminus of AvDrag consists of 165 amino acids of are petitive region and 99 amino acids of a C-terminalnon-repetitive region. The peptide motifs found in spider drag line silk proteins, GGX and An, were conserved in the repetitive region of AvDrag. The AvDrag cDNA was expressed as a 28kDa polypeptide in baculovirus-infected insect cells. To produce transgenic rice plant with high contents of glycine and alanine, the prolamin promoter-driven AvDrag was introduced into rice plant via Agrobacteriumtumefaciens-mediated gene transformation. Because of seeds pecific prolamin promoter, expression of AvDrag protein has been achieved inriceseed. The introduction and copy number of the AvDrag gene in transgenic rice plants were determined by PCR and Southern blot analysis. AvDrag expression in transgenic rice seeds was examined by Northern blot and Western blot analysis. Immuno fluorescence staining with the AvDrag antiserum revealed that the recombinant AvDrag proteins were localized in transgenic rice seeds. Furthermore, the amino acid content analysis showed that transgenic rice seeds were greatly increased in glycine and alanine as compared to controls. The present study is the first to show the expression of spider silk protein in rice seed.

본 연구는 압화의 효과적인 누름건조법의 개발을 위해서 산과 들에서 직접 채집하거나 산지에서 구입한 야생화,절화,분화,관엽식물을 대상으로 계절별로 분류했다. 압화용 식물들은 수분을 효과적으로 제거하여 변색을 방지하고 꽃의 원형을 잘 유지하기 위해 전처리의 과정을 밟았다. 건조가 용이하거나, 건조 과정에서 화색이 쉽게 변하는 꽃들은 전처리를 하지 않는 것을 원칙으로 하였다. 수술의 수가 많은 경우는 핀셋으로 일부를 솎아주는 것이 좋았다. 꽃봉오리, 꽃, 화경(줄기), 화병, 그리고 가지는 부분적으로 나이프 처리를 해 주는 것이 건조에 효과적이었다. 압화용 식물의 잎은 기본적으로 잎의 뒷면과 엽병 부위를 #100수준의 샌드페이퍼에 살짝 눌러주는 것이 효과적이었다. 장미처럼 꽃잎이 여러 겹으로 포개져 있거나, 건조가 다소 까다로운 양란 등은 꽃잎을 분리시켜 건조한 다음 원래 형태로 복원시키는 꽃 조립 과정을 밟는 것이 유리했다. 그러나, 유스토마의 꽃은 효과적으로 건조시키는데 다리미 처리가 필요했다. 일반 전처리로 건조가 어려운 안스리움의 포엽 등은 열탕 처리가 필요했다. 주름이 지지 않고 원형에 가까운 꽃을 얻기 위해서는 스펀지를 깔아주는 것이 좋았다. 꽃잎이 얇은 경우는 2∼3㎏ 정도의 무게 처리로 투명화 현상이 나타나지 않았고,꽃잎 수가 많거나, 꽃잎이 두껍거나, 자방 부위가 두터운 경우는 7∼12㎏ 정도의 무게 처리로 수축 현상을 최소화시킬 수 있었다. 이상의 기본 처리 과정을 거친 후에도 쉽게 건조되지 않는 경우에는 건조 매트를 1회∼2회 교환해 주는 것이 필수적이었다.

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.

It is important to understand pedigree of rice cultivars commonly used for breeding. In this paper, pedigree tables for tracking the pedigree of 17 representative rice cultivars recommended by Rural development Adminstration (RDA) were completed and analyzed using two kinds of web database system; 'IRIS' and 'RRDB'. Seven cultivars, namely, 'Sangmibyeo', 'Ilpumbyeo', 'Saegewhabyeo', 'Surabyeo', 'Shindongjinbyeo', 'Ilmibyeo' and 'Jungwhabyeo' had 'Koshihikari' on the pedigree of their ancestor. Besides 'Koshihikari', the most feguently used ancestral germplasms among the high quality rice cultivars were 'Fujisaka 5', 'Kameno o' and 'Asahi', 'Fujisaka 5' was used as ancestral parent in 12 out of 17 cultivars. Interestingly, 'Kameno o' was used in pedigree of 16 out of 17 high quality varieties and 'Asahi' was used in the ancestral pedigree of all 17 varieties. 'Hwayeongbyeo' was used as one of parent in the breeding of 'Dongjin 1', 'Hwabongbyeo', 'Saegewhabyeo' and 'Junambyeo'. 'Ilpumbyeo' was used in the breeding pathway of 'Junambyeo' and 'Saegewhabyeo', 'Mangeumbyeo' itself was not enlisted as one of high quality rice cultivars, but was used as a breeding parent of three high quality varieties, namely, 'Saegewhabyeo', 'Hwabongbyeo' and 'Nampyeongbyeo'. Incorporated with evaluation data, pedigree will provide a valuable chance to genealogical tracking of agronomic traits such as disease resistance, grain quality and etc.