Molecular analysis of transgenic rice overexpressing UDP-glycosyltransferase from Brassica rapa

Me-Sun Kim,Hye-Jung Lee,Joonki Kim,Dal-A Yu,Sanguk Byeon,Kwon-Kyoo Kang,Illsup Nou,Yong-Gu Cho 한국육종학회 2014 한국육종학회 심포지엄 Vol.2014 No.07

Secondary plant metabolites undergo several modification reactions, including glycosylation and physiological functions. Glycosylation, which is mediated by UDP-glycosyltransferase (UGT), plays a role in the storage of secondary metabolites and in defending plants against stress. In this study, a UDP-glucosyltransferase cDNA was isolated from Brassica rapa hereinafter referred to as BrUGT. It has a full-length cDNA of 1,236 bp that contains a single open reading frame of 834 bp which encodes a polypeptide of 277 amino acid residues with a calculated mass of 31.19 kDa. BLASTX analysis hits a catalytic domain of glycos_transf_1 super family (c112012) that belongs to the glycosyltransferases group 1 with tetratricopeptide (TPR) regions. UGT gene expression analysis showed high mRNA transcripts in pistil, followed by petal, seed and calyx of flower in Brassica rapa. Furthermore, we constructed a recombinant pFLCIII vector carrying the BrUGT gene under the control of ubiquitin promoter and NOS terminator and transformed into rice using Agrobacterium tumefaciens. The UGT overexpressing rice lines were then characterized at the physiological and molecular levels. To further understand the biological function of BrUGT, transcriptional profiling of the gene in transgenic rice lines under cold, salt, PEG, H2O2, ABA and drought stress condition is underway.

Overexpression of CIPK 15 improved tolerance to pre-harvest sprouting in rice

Dal-A Yu,Hye-Jung Lee,Joonki Kim,Me-Sun Kim,Marjohn Nino,Sothea Ouk,Seong-Dong Kim,Ill-sup Nou,Yong-Gu Cho 한국육종학회 2015 한국육종학회 심포지엄 Vol.2015 No.07

Since global climate changes drastically, pre-harvest sprouting (PHS) is expected to pose serious problems in rice production. CBL-interacting serine/threonine protein kinases (CIPKs) have been implicated to play important role in regulating various abiotic stresses such as cold, salinity and drought. In this study, to understand the function of this gene under pre-harvest sprouting in rice, a cDNA clone encoding CBL-interacting protein kinase 15 was isolated from rice flowers. We constructed a recombinant vector carrying the CIPK15 under the control of the CaMV 35S promoter and Tnos terminator and transformed into rice using Agrobacterium tumefaciens. Insertion of the gene was verified in transformants using HPT resistance test and genomic PCR. Transcriptional profiling using tissues of wild type, Gopum, revealed expression of the gene in whole plant tissues with level of expression highest in the seeds suggesting possible role in dormancy. Comparative expression analysis of the gene in transgenic and wild type through semi-quantitative RT-PCR and real-time PCR showed higher expression in transgenic rice lines. Moreover, screening in the mist chamber showed overexpression lines that were resistant to the PHS. This result suggests the involvement of CIPK15 in the regulation of pre-harvest sprouting.

Changes in the Electric Parameters of Mesoporous Silicon under Adsorption of Plant Viruses

Yu.A. Vashpanov,Kae-Dal Kwack,Han-Sub Yoon,Jung-Young Son 한국물리학회 2009 THE JOURNAL OF THE KOREAN PHYSICAL SOCIETY Vol.54 No.6

Changes in the electric parameters of porous silicon under adsorption of nematode-transmitted polyhedral-type plant viruses, such as tomato ringspot virus, grapevine virus A and grapevine fan leaf virus, are investigated at room temperature. The pore sizes of the porous silicon are bigger than the characteristic sizes of the plant viruses, which are considered as having a spherical shape. Plant viruses adsorbed on the surface of porous silicon change the voltage-current and the voltage-capacitance characteristics. The changes in these characteristics depend on the nature of the adsorbed virus and are mostly caused by modifications in the characteristics of the potential barriers formed between nanowires which are thought to be composed of mesoporous silicon. The adsorbed biological particles (plant viruses) on the voltage-current characteristic redistribute the potential barriers in accordance with the applied voltage and the nature of the virus.

Overexpression of CBL-interacting protein kinase 15 improves tolerance to pre-harvest sprouting in rice (Oryza sativa L.)

Dal-A Yu,Hye-Jung Lee,Joonki Kim,Me-Sun Kim,Marjohn Nino,Sanguk Byeon,Jeongmi Kim,Illsup Nou,Kwon-Kyoo Kang,Yong-Gu Cho 한국육종학회 2014 한국육종학회 심포지엄 Vol.2014 No.07

Since global climate changes drastically, pre-harvest sprouting (PHS) is expected to pose serious problems in rice production. CBL-interacting serine/threonine protein kinases (CIPKs) have been implicated to play important role in regulating various abiotic stresses such as cold, salinity and drought. In this study, to understand the function of this gene under pre-harvest sprouting in rice, a cDNA clone encoding CBL-interacting protein kinase 15 (CIPK15) was isolated from rice flowers. This gene is 2,818 bp long with 1,332 bp coding region that encodes a polypeptide of 443 amino acids. We constructed a recombinant vector carrying the OsCIPK15 under the control of the CaMV 35S promoter and Tnos terminator and transformed into rice using Agrobacterium tumefaciens. Insertion of the gene was verified in transformants using HPT resistance test and genomic PCR. Transcriptional profiling using tissues of wild type, Gopum, revealed expression of the gene in whole plant tissues with level of expression highest in the seeds suggesting possible role in dormancy. Comparative expression analysis of the gene in transgenic and wild type through semi-quantitative RT-PCR and real-time PCR showed higher expression in transgenic rice lines. Moreover, screening in the mist chamber showed overexpression lines that were resistant to the PHS. This result suggests the involvement of OsCIPK15 in the regulation of pre-harvest sprouting.

Modification of Starch Composition Using Downregulation of GBSS1 gene in Japonica Rice

Hye-Jung Lee,Moo-Geun Jee,Dal-A Yu,Me-Sun Kim,Franz Nogoy,Mingmao Sun,Sailila E. Abdula,Kwon-Kyoo Kang,Illsup Nou,Yong-Gu Cho 한국육종학회 2014 한국육종학회 심포지엄 Vol.2014 No.07

There is a great consideration on rice eating quality aside from improving its tolerance to various stresses. High yielding and pest and disease tolerant rice is highly desirable but it is more commercially important if it also has a high eating quality. There are various factors contributing to the good eating quality of rice. This study focuses on modifying the expression of GBSS1 genes which are responsible for amylopectin and amylose synthesis in rice by using RNAi and antisense techniques. We have developed 40 transgenic plants with RNAi-GBSS1 gene and 60 transgenic lines with antisense-GBSS1 gene. The transgenic plants show diverse amylose contents in rice seed. We selected candidate lines according to PCR, RNA expression and amylose contents. A semi-quantitative RT-PCR was carried out to measure the expression level of GBSS1 gene at several time points after the flowering of transgenic plants. The expression level of GBSS1 gene in rice grains decreases over time and the mRNA expression among the transgenic plants were lower compare to its wild type. In the SEM analysis, the starch granule of wild type Gopumbyeo has very large structures accompanied with small ones around the area. However, the starch structures in transgenic plants were smaller and more uniform in size and shape throughout the viewing area

Effects of the Anti-sepsis Drug, (S)-1-(α-Naphthylmethyl)-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline (CKD-712), on Mortality, Inflammation, and Organ Injuries in Rodent Sepsis Models

Jae-Hwan Park,Dal-hyun Kim,In-Chang Hwang,Nina Ha,Sera Lee,Jung Min Kim,Sung Sook Lee,Hosung Yu,In-Taek Lim,Jun-A You 대한약학회 2011 Archives of Pharmacal Research Vol.34 No.3

CKD-712 is a 1-naphthyl analog of higenamine that has been reported to have potent antiinflammatory and thus anti-sepsis effects. The purpose of this study was to investigate the potential of CKD-712 as a medicine for sepsis and to confirm its protective effects on organs in animal sepsis models. Pretreatment with CKD-712 dose-dependently increased survival rate in a lipopolysaccharide-induced sepsis model in mice. Body temperature decrease, an important pre-symptom of septic death, was also prevented by CKD-712. CKD-712 still significantly increased survival rate even when administered one and four hours after lipopolysaccharide injection. Therapeutic efficacy of CKD-712 was also confirmed against sepsis following zymosan-induced endotoxemia and in cecal ligation and puncture surgery in mice. In a disseminated intravascular coagulation model in rats, CKD-712 showed organ-protective effect by reducing serum glutamate-oxaloacetate transaminase, glutamate-pyruvate transferase, blood urea nitrogen, and creatinine levels. CKD-712 also prevented histological damage to the lung and liver. In this same model, CKD-712 showed anti-inflammatory effects through decreases in tumor necrosis factor-α and interleukin-6 in the blood and reduced translocation of nuclear factor-κB to the nuclei of lung cells. CKD-712 administration also diminished infiltration of leukocytes into the lung and liver. Taken together, these results show that CKD-712 has excellent potential as an effective medicine for sepsis.

Biological function of Brassica rapa cysteine protease in transgenic rice and Xanthomonas oryzae pv. oryzae pathosystem

Marjohn Nino,Sailila E. Abdula,Hye-Jung Lee,Dal-A Yu,Mi-Sun Kim,Sanguk Byeon,Franz Nogoy,Kwon-Kyoo Kang,Illsup Nou,Yong-Gu Cho 한국육종학회 2013 한국육종학회 심포지엄 Vol.2013 No.07

Bacterial blight is a serious problem of rice in irrigated and rainfed lowlands. It is caused by Xanthomonas oryzae pv. oryzae (Xoo) which is represented by many pathotypes, making it difficult to control. Plant proteases are important players in immunity acting either in the execution of attack, in signaling cascade or in perception of invader. This study demonstrates the response of cysteine protease (CP) upon interaction with the pathogen. The cysteine protease encoding full-length cDNA was identified and characterized using web-based tools. Conserved domain of the gene revealed its affinity to Peptidase_CIA family. The full-length cDNA of CP in Brassica rapa was then cloned and overexpressed in rice. Insertion of gene was verified in the transformants through PCR assay. Spatiotemporal expression of the gene was performed in transgenic rice. To evaluate the resistance of CP-overexpression lines to Xoo, transgenic plants were inoculated with two races of Xoo. In planta analysis of enzymatic activity of CP was also performed before and after infection by the pathogen.

Classification of Korean rice varieties based on growth characteristics

Me-Sun Kim,Hye-Jung Lee,Dal-A Yu,Joonki Kim,Franz Nogoy,Eun-Ju Jeong,Jang-Hwan You,Yong-Gu Cho 한국육종학회 2015 한국육종학회 심포지엄 Vol.2015 No.07

The International Union for the Protection of New Varieties of Plants (UPOV) promotes an effective system of plant variety protection and encourages the development of new varieties of plants. International convention was initiated to standardized the system efforts and strengthen the policy. This study was conducted to establish a database for rice identification using morphological characters which include number of tillers and panicle per plant, spikelets per panicle, yield, plant maturity, height, leaf pigments, flag leaf angles, and rice bran. The whole rice population was grouped into three based on leaf angles, majority members of which retained the flag leaf angle-character until maturity stage. Most rice accessions did not exhibit anthocyanin pigments on the leaves particularly on the first leaf, leaf blade, leaf sheath and auricle, except for varieties classified as black rice. In the case of grain, many accessions produced secondary branching, and showed no awn. For agronomic traits, productive tiller and panicle per plant were higher in early flowering varieties, while spikelets per panicle and ripened grain were higher in late flowering varieties, and yield was higher in medium flowering varieties. All data were then pooled for cluster analysis which revealed three major independent clusters and four minor clusters.

Modification of starch composition using RNAi targeting of SSS1 gene in rice

Hye-Jung Lee,Moo-Geun Jee,Dal-A Yu,Me-Sun Kim,Joonki Kim,Seon-Kyeong Song,Kwon-Kyoo Kang,Yong-Gu Cho 한국육종학회 2015 한국육종학회 심포지엄 Vol.2015 No.07

An increasing preference for good eating quality of rice among consumers has become one of the important considerations in rice breeding. Amylose content is a leading factor affecting eating quality of rice. Amylose composition is determined by the relative activity of soluble starch synthase (SSS) and granule-bound starch synthase (GBSS). This study focused on modifying the expression of SSSI gene which is responsible for amylopectin and amylose synthesis in rice by using RNA interference (RNAi) technology. The transgenic rice plants showed various amylose content in rice grains. Favorable rice lines were selected according to genomic PCR, transgene expression and amylose contents analysis. A semi-quantitative RT-PCR was carried out to determine the expression level of SSSI gene after flowering of transgenic rice and wild type. Down-regulation of SSSI gene in transgenic plants was evident in the decreasing expression in rice grains. Accordingly, scanning electron microscopy (SEM) analysis revealed uniform size with smooth curves starch granules in down-regulation rice lines, in contrast with the non-uniform granules in wild type. Results indicated that RNAi-SSSI transgenic lines produced low amylose contents that fell between glutinous and non-glutinous rice. This study showed that down-regulation of endogenous SSSI may improve the eating quality in rice.

Overexpression of OsMLD encoding MYB-like DNA binding domain increases tolerance to salt stress in rice (Oryza sativa L.)

Hye-Jung Lee,Sailila E Abdula,Moo-Geun Jee,Dae-Won Jang,Dal-A Yu,Sung-Han Park,Ung-Han Yoon,Tae-Ho Kim,Yong-Gu Cho 한국육종학회 2012 한국육종학회 심포지엄 Vol.2012 No.07

MYB-like domain (MLD) gene is a transcription factor that plays a diverse role in plant development and in response to abiotic stresses. In this study, we isolated and developed CaMV35S::OsMLD rice lines and determined its expression pattern under abiotic stresses. It has Myb_CC_LHEQLE superfamily similar to most transcription factor genes but with a very unique binding domain of SHLQKYR in the C-terminal region. Overexpressing rice lines showed enhanced tolerance to salinity with elevated mRNA transcript. Additionally, mRNA transcripts were up-regulated by ABA, H2O2 and dehydration stresses. Further investigation in the enhanced tolerance to salinity showed an increased accumulation of proline and a decreased in malondialdehyde contents indicating that OsMLD gene may be involved in the regulation of proline and osmolytes during abiotic stresses. These results showed that OsMLD gene could be used in the development of rice intended for soil with salinity-related problem.