To determine the patterns and the levels of expression of the cauliflower mosaic virus (CaMV 35S) promoter and the rice actin 1 (Act1) promoter in rice, transgenic rice plants containing CaMV 35S-$\beta$-glucuronidase (GUS) and Act1-GUS constructs were generated and examined by fluorometric and histochemical analyses. The fluorometric analysis of stably transformed calluses showed that the activity of the rice Act1 promoter was stronger than that of the CaMV 35S promoter in rice cells. In a histochemcial study of the transgenic rices, it was shown that the GUS activity directed by the CaMV 35S promoter was localized mainly in parenchymal cells of vascular tissues of leaves and roots and mesophyll cells of leaves. These results are similar to those of potato, a dicot plant. In contrast, rice plant transformed with Act1-GUS fusion construct revealed strong GUS activity in parenchymal cells of vascular tissue, mesophyll cells, epidermal cells, bulliform cells, guard subsidiary cells of leaves and most cells of the root, suggesting that the rice Act1 promoter is more constitutive than the CaMV 35S promoter. It was also confirmed that in both types of transgenic rice little or no staining was localized in metaxylen tracheary elements of vascular tissue from leaves or roots. These results indicate that the rice Act1 promoter can be utilized more successfully for expression of a variety of foreign gene in rice than the CaMV 35S promoter.
Kim, Eun-Ji;Moon, Jungsun;Kang, Jung-Il;Lee, Young-Ki;Koh, Young-Sang;Yoo, Eun-Sook;Kang, Hee-Kyoung;Yim, Dongsool
Korean Journal of Pharmacognosy
/
v.44
no.3
/
pp.269-274
/
2013
In this study, we investigated the anticancer effect of rice bran extract in HL-60 human promyelocytic leukemia cells. The extract of rice bran inhibited the proliferation of HL-60 cells. When treated with the rice bran extract, we could observe the apoptotic characteristics such as apoptotic bodies and the increase of sub-G1 hypodiploid cell population, increase of Bax level, decrease of Bcl-2 expression, cleavage of procaspase-3, cleavage of procaspase-9 and cleavage of poly(ADP-ribose) polymerase(PARP) in HL-60 cells. Furthermore, the apoptosis induction of HL-60 cells treated with the rice bran extract was also accompanied by the inactivation of mitogen-activated protein kinases (MAPK) such as ERK1/2 MAPK and p38 MAPK. In addition, the rice bran extract induced the down-regulation of c-myc. These data suggested that the rice bran extract could induce the apoptosis via the inactivation of ERK1/2 MAPK and p38 MAPK, and the down-regulation of c-myc in HL-60 acute pomyelocytic leukemia cells. The results support that the rice bran extract might have potential for the treatment of acute promyelocytic leukemia.
The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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v.16
no.2
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pp.57-77
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2003
Objectives: The aim of this study was to investigate the depigmentation effects of Barley, Unpolished rice, Job's-tear. Metbods: We investigated that the extracts of Barley, Unpolished rice, Job's-tear inhibit activity of tyrosinase, the enzyme which convert ts 3-(3,4-dihydroxyphenyl) alanine to dopachrom in the biosynthetic process of melanin, the UV absorbance of the extracts in the UV-A region and UV-B region was measured by UV scanning, the effect of extracts on cell viability and melanin production in cultured B16 mouse melanoma cells were measured, and cytoprotective effects of extracts on PC12 cells injured by hydrogen peroxide measured by MTT assay. Results: The extracts of Barley, Unpolished rice, Job's-tear inhibited activity of tyrosinase in low density. The Barley, Job's-tear extracts not only showed inhibitory effects on melanin production in cultured B16 mouse melanoma cells, but also exhibited cytoprotective effects on PC12 cells injured by hydrogen peroxide in low density. Unpolished rice extract showed inhibitory effect on melanin production in cultured B16 mouse melanoma cells, but did not showed cytoprotective effect Barley, Unpolished rice, Job's-tear extracts did not showed an absorbance effect in the UV-A region and UV-8 region. Conclusions: These results suggest that Barley, Unpolished rice, Job's-tear inhibit melanin biosynthesis which is involved in hyperpigmentation and could be used as a whitening agent.
Black rice contains many biologically active compounds. The aim of this study was to investigate the protective effects of black rice extracts (whole grain extract, WGE and rice bran extract, RBE) on tert-butyl hydroperoxide (TBHP)-induced oxidative injury in HepG2 cells. Cellular reactive oxygen species (ROS), antioxidant enzyme activities, malondialdehyde (MDA) and glutathione (GSH) concentrations were evaluated as biomarkers of cellular oxidative status. Cells pretreated with 50 and $100{\mu}g/mL$ of WGE or RBE were more resistant to oxidative stress in a dose-dependent manner. The highest WGE and BRE concentrations enhanced GSH concentrations and modulated antioxidant enzyme activities (glutathione reductase, glutathione-S-transferase, catalase, and superoxide dismutase) compared to TBHP-treated cells. Cells treated with RBE showed higher protective effect compared to cells treated with WGE against oxidative insult. Black rice extracts attenuated oxidative insult by inhibiting cellular ROS and MDA increase and by modulating antioxidant enzyme activities in HepG2 cells.
Pigmented rice is mainly black, red, and dark purple, and contains a variety of flavones, tannin, polyphenols, sterols, tocopherols, ${\gamma}$-oryzanols, amino acids, and essential oils. The present study evaluated the cytotoxic effects of purple rice extracts (PREs) combined with chemotherapeutic drugs on human cancer cells and mechanisms of cell death. Methanolic (MeOH) and dichloromethane (DCM) extracts of three cultivars of purple rice in Thailand: Doisaket (DSK), Nan and Payao (PYO), were tested and compared with white rice (KK6). Cytotoxicity was determined by 3-(4, 5-dimethyl)-2, 5-diphenyltetrazolium bromide (MTT) assay in human hepatocellular carcinoma HepG2, prostate cancer LNCaP and murine normal fibroblast NIH3T3 cells. MeOH-PYO-PRE was the most cytotoxic and inhibited HepG2 cell growth more than that of LNCaP cells but was not toxic to NIH3T3 cells. When PREs were combined with paclitaxel or vinblastine, they showed additive cytotoxic effects on HepG2 and LNCaP cells, except for MeOH-PYO-PRE which showed synergistic effects on HepG2 cells when combined with vinblastine. MeOH-PYO-PRE plus vinblastine induced HepG2 cell apoptosis with loss of mitochondrial transmembrane potential (MTP) but no ROS production. MeOH-PYO-PRE-treated HepG2 cells underwent apoptosis via caspase-9 and-3 activation. The level of ${\gamma}$-oryzanol was highest in DCM-PYO-PRE (44.17 mg/g) whereas anthocyanin content was high in MeOH-PYO-PRE (5.80 mg/g). In conclusion, methanolic Payao purple rice extract was mostly toxic to human HepG2 cells and synergistically enhanced the cytotoxicity of vinblastine. Human HepG2 cell apoptosis induced by MeOH-PYO-PRE and vinblastine was mediated through a mitochondrial pathway.
The purpose of this study is to promote the substitution of rice flour for wheat flour in making bread and thus the consumption of rice by examining the effect of particle size of rice flour on leavening rice bread. For this purpose, several experiments were carried out. With regard to particle size distribution, 59.45% of wheat flour had passed 200 mesh and 3 kinds of rice flour milled to penetrate into 20, 35, and 45 mesh (S1, S2 and S3) had passed 21.88%, 33.1% and 36.38% of those for 200 mesh, respectively. To leaven the rice flour dough to bread, 25% of vital gluten was needed. To determine the optimal water quantity for rice bread dough, the hardness of wheat and rice flour dough was measured by rheometer. The appropriate water quantity for S1, S2 and S3 was set at 285 ml , 295 ml and 335${\sim}$340m1, respectively. The loaf volume index of the wheat flour bread was 6.24, while that of and rice flour bread S1, S2 and S3 was 5.38, 5.50 and 5.75, respectively. These results indicated that the loaf volume index of rice flour bread is lower than that of wheat flour bread, but that the volume of rice flour bread was increased with fuel particle size of rice flour. Scanning electron microscopy (SEM), image of the wheat bread tissue at a magnification of 35 times showed long, large, oval-shaped, air cells and thin cell membrane, as well as small air cells, whereas the images of rice flour showed angular, circular, air cells and rough and thick cell membrane. The size and number of air cells in the rice bread were larger in S2 and S3 with fuel particle flours than in S1. In particular, the bread made with S3 contained many air cells that were as large as those in the wheat bread were. In addition, when the inner cell wall was magnified 1500 times, almost no small air cell was observed in C and S3, whereas many fine air cells were observed in the cell wall of S1 and S2.
We evaluated a more practical and cost-effective immobilization carriers for ethanol production using the yeast Saccharomyces cerevisiae. Three candidate materials-rice hull, rice straw, and sawdust-were tested for their cell-adsorption capacity and operational durability. Derivatizations of rice hull, rice straw, and sawdust with the optimal concentration of 0.5 M of 2-(diethylamino)ethyl chloride hydrochloride (DEAE HCl) resulted in > 95% adsorption of the initial yeast cells at 2 hr for DEAE-rice hull and DEAE-sawdust and in only approximately 80% adsorption for DEAE-rice straw. In addition, DEAE-sawdust was found to be a more practical carrier for immobilizing yeast cells in terms of operational durability in shaking flask cultures with two different speeds of 60 and 150 rpm. Furthermore, the biosorption isotherms of DEAE-rice hull, -rice straw, and -sawdust for yeast cells revealed that the $Q_{max}$ of DEAE-sawdust (82.6 mg/g) was greater than that of DEAE-rice hull and DEAE-rice straw. During the 404-hr of continuous column reactor operation using yeast cells immobilized on DEAE-sawdust, no serious detachment of the yeast cells from the DEAE-sawdust was recorded. Ethanol yield of approximately 3.04 g/L was produced steadily, and glucose was completely converted to ethanol at a yield of 0.375 g-ethanol/g-glucose (73.4% of the theoretical value). Thus, sawdust is a promising practical immobilization carrier for ethanol production, with significance in the production of bioethanol as a biofuel.
Journal of the Korean Society of Food Science and Nutrition
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v.31
no.3
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pp.521-526
/
2002
A number of experimental and epidemiological studies have implicated that antiestrogenic effects of estrogen-like compounds in legumes and plant seeds are responsible for lowering breast cancer risk in human. However, few studies have been conducted to illustrate the possible chemopreventive effects of Korean traditional food materials. This study was performed to determine the cytotoxic and apoptotic effects of yellow soybeans, black soybeans and brown rice extracts on hormone-dependent and hormone-independent human breast cancer cells. Methanol-or acetone-soluble fractions of soybeans or brown rice were incubated with hormone-dependent cells (MCF-7) or hormone-independent cells (MDA-MB-231). Cell cytotoxicity was measured by MTT assay at 24, 48 and 72 hrs of incubation. Apoptotic effects of these extracts toward breast cancer cells were also determined at 48 hrs of incubation by measuring DNA fragmentation. Results indicated that the acetone-soluble fraction of brown rice exerted strongest cytotoxic effect on MCF-7 ceIls, although other fractions also reduced the number of viable MCF-7 cells after 48 hrs of incubation. Both acetone and methanol soluble fractions of all samples exerted a significant cytotoxicity towards MDA-MB-231 cells after 24 hrs of incubation, and acetone and methanol soluble fractions of brown rice were especially effective in these cells. At 48 hrs of incubation, methanol fractions of all three samples induced apopotosis of MDA-MB-231 cells. These results indicate methaol or acetone soluble fractions of yellow soybeans, black soybeans and brown rice induce cytotoxicity in both hormone-dependent and hormone-independent breast cancer cells. Therefore, possible mechanisms of cell cytotoxicity do not necessarily include antiestrogenic effects of soybean or brown rice extract. A possible anticarcinogenic effect of brown rice methanol-soluble fraction may mediated through their apoptotic effect. Further studies are requried to elucidate responsible compounds and mechanisms involved in observed anticarcinogenesis.
Proceedings of the Korean Society of Crop Science Conference
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2017.06a
/
pp.134-134
/
2017
Recently, host-induced gene silencing (HIGS) system has been successfully applied into development of resistant crops against insects, fungal and viral pathogens. To test HIGS-mediated resistance in rice against rice blast fungus, Magnaporthe oryzae, we first tested possibility of movement of small non-coding RNA from rice cells to rice blast fungus. The rice blast fungus expressing GFP transgene were inoculated to transgenic rice plants ectopically expressing dsRNAi construct targeting fungal GFP gene. Expression of dsRNAi construct for GFP gene in transgenic plants significantly suppressed GFP expression in infected fungal cells indicating that small RNAs generated in plant cells can move into infected fungal cells and efficiently suppress the expression of fungal GFP gene. Consistent with these results, expression of dsRNAi constructs against 3 fungal pathogenic genes of M. oryzae in transgenic rice specifically and efficiently suppressed not only the expression of fungal pathogenic genes, but also fungal infection. The conidia of M. oryzae applied on leaf sheath of transgenic rice expressing dsRNAs against 3 fungal pathogenic genes showed abnormal development of primary hyphae and malfunction of appressorium, which is consistent with the phenotypes of corresponding fungal knock-out mutants. Taken these results together, here, we suggest a novel strategy for development of antifungal crops by means of HIGS system.
This study was conducted to investigate the anatomical and ultrastructural responses of the oxyfluorfen-tolerant and -susceptible rice cultivars with barnyardgrass, a typical susceptible weed by oxyfluorfen and the herbicides having similar mode of action treatment. After the treatment of $10^{-5}M$ oxyfluorfen, the tolerant rice cultivars no showed the structural damage of leaf surface, but the susceptible rice cultivate was damaged in the wax and the susceptible barnyardgrass was even destroyed in the tissue irregularly. Also in the susceptible rice cultivars and barnyardgrass the thickness of leaf blade was greatly decreased. The anatomical change was not observed in the tolerant rice cultivars but epidermal cells, mesophyll cells and bundle sheath cells were badly broken in the susceptible rice cultivars and barnyardgrass and especially after 24 hours of the treatment the structure of susceptible rice cultivars was completely disintegrated. The irregularity of chloroplast shape and the distortion of chloroplast envelope were generally observed and the starch tended to decrease by oxyfluorfen treatment regardless of rice cultivars. Such a structural damage were appeared more badly in the susceptible rice cultivars and bamyardgrass than in the tolerant rice cultivars. By the treatment of diphenyl ether herbicides, the thickness of leaf blade greatly reduced in the order of oxyfluorfen > acifluorfen > bifenox > oxadiazon, and the susceptible rice cultivars showed more reduction than the tolerant rice cultivars. Especially, the susceptible rice cultivars showed that the leaf structure was badly broken down with damage epidermal cells and bundle sheath cells.
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