Construction of artificial promoters highly responsive to iron deficiency     

Takanori Kobayashi  Yuko Nakayama  Michiko Takahashi  Haruhiko Inoue  Hiromi Nakanishi  Toshihiro Yoshihara 《Soil Science and Plant Nutrition》2013,59(7):1167-1175

Iron deficiency-responsive element 1 (IDE1) and IDE2 are cis-acting elements that are responsible for Fe-deficiency-inducible and root-specific expression of the barley (Hordeum vulgare L.) gene IDS2 (Fe-deficiency-specific clone no. 2). Using these cis-acting elements, we aimed to construct super-promoters that would induce prominent gene expression in the roots of Fe-deficient rice plants (Oryza sativa L.). Modules containing IDE1 and IDE2 of the IDS2 promoter were used as repeats or were linked to the Fe-deficiency-responsive promoter of barley IDS3, and were connected to known enhancer-like sequences. Five artificial promoters, as well as the native promoters of barley IDS2 or IDS3, were connected individually upstream of β-glucuronidase (GUS) and were introduced into rice. Transgenic rice plants were grown under control or Fe-deficient conditions, and GUS expression was analyzed. The artificial promoter that contained one module of IDE1 and IDE2 conferred strong Fe-deficiency-inducible GUS expression to the roots of rice plants. Each of the five artificial promoters induced a similar level of GUS expression in Fe-deficient roots, which did not exceed the GUS expression driven by the native IDS2 or IDS3 promoter. Artificial and native promoters induced GUS expression in response to Fe-deficiency in leaves, although the level of expression was lower than that in roots. Histochemical observations revealed that GUS expression driven by artificial and native promoters was spatially similar, and expression was dominant within vascular bundles and root exodermis. These findings suggest that there is coordinated expression of the genes that are involved in Fe-deficiency-induced Fe uptake in rice.  相似文献   

Influences of aluminum ions on the determination of zpc (zero point of charge) of variable charge soils     

Katsutoshi Sakurai  Akinori Nakayama  Tsutomu Watanabe  Kazutake Kyuma 《Soil Science and Plant Nutrition》2013,59(4):623-633

Influence of Al dissolution on soil ZPC (zero point of charge) measured by a potentio-metric titration (PT) method and a modified salt titration (STPT) method was examined using two strongly weathered soils from Thailand and two volcanic ash soils from Japan. The amount of dissolved Al ions increased with the increase in the concentration of a supporting electrolyte for the strongly weathered soils, while the increase was negligible for the volcanic ash soils.

ZPC value of the strongly weathered soils determined by the PT method was lower than that by the STPT method, due to the greater Al dissolution associated with the higher electrolyte concentration used in the PT method. Al ions adsorbed onto the soil surface would shift the ZPC to a higher pH value not as a result of the formation of hydroxy Al polymers, but due to the blocking of permanent negative charge sites, which could otherwise lower the ZPC. The σp value, as a measure of permanent charge or the amount of 11 or O11 adsorbed by a soil required to attain the ZPC, could be used to describe this phenomenon.

In the STPT method, the salt concentration was not high enough to causc a significant Al dissolution at the ZPC, which is considered to be a more suitable condition than in the PT method because the ZPC value can be evaluated at a low salt concentration as in the ease of field conditions for crop production. Thus, the STPT method is rccommendcd for the determination of the ZPC.  相似文献   

Comparative analysis of floral pigmentation between wild-type and white-flowered varieties of Cyclamen graecum     

Y. Akita  H. Ishizaka  M. Nakayama  A. Shimada  S. Kitamura  Y. Hase 《The Journal of Horticultural Science and Biotechnology》2013,88(5):437-443

Summary

The flower colour of Cyclamen graecum gra6 (wild-type) is pink-purple in the main part of the petal, referred to as the ‘slip’, and deep purple at the petal base, referred to as the ‘eye’. On the other hand, flowers of C. graecum gra50 (a white-flowered variant) exhibit a white colour in both the ‘slip’ and ‘eye’ regions. In this study, the relationship between floral pigmentation and the expression of several anthocyanin biosynthesis genes was investigated in C. graecum gra6 and gra50. The pigments in the ‘slip’ and ‘eye’ regions consist mainly of malvidin 3,5-diglucoside in gra6, suggesting that the difference between the colour of the ‘slip’ and ‘eye’ regions is related to the amount of anthocyanin present. White-flowered C. graecum gra50 possessed lower amounts of anthocyanins, but higher amounts of flavonols compared to gra6, suggesting a change in metabolism caused by a disruption of anthocyanin biosynthesis. Gene expression analysis demonstrated that expression of the dihydroflavonol 4-reductase gene 2 (CgraDFR2) was lower in gra50 compared with gra6, whereas expression of the three other key genes (dihydroflavonol 4-reductase gene 1, flavonoid 3’,5’-hydroxylase, and anthocyanidin synthase) did not differ greatly. These results suggest that the white-flowered variant (gra50) may result from a defect in expression of the CgraDFR2 gene.  相似文献   

Goat cathelicidin‐2 is secreted by blood leukocytes regardless of lipopolysaccharide stimulation          下载免费PDF全文

Supreena Srisaikham  Wisitiporn Suksombat  Yukinori Yoshimura  Naoki Isobe 《Animal Science Journal》2016,87(3):423-427

It has been reported that goat cathelicidin‐2, an antimicrobial peptide, localizes in leukocytes and is present in milk. Here, we examined whether cathelicidin‐2 is secreted by leukocytes. Different concentrations (10⁵–10⁸ cells/mL) of blood leukocytes were cultured for 0–48 h with or without lipopolysaccharide (LPS). After culture, the concentrations of cathelicidin‐2 in the conditioned media were measured. Blood was collected from male goats 0–24 h after the intravenous injection of Escherichia coli O111:B4 LPS. The plasma cathelicidin‐2 concentrations were determined and the blood leukocytes immunostained with anti‐cathelicidin‐2 antibody to calculate the proportion of cathelicidin‐2‐positive cells in the total leukocytes. When higher concentrations of leukocytes were cultured, the cathelicidin‐2 concentrations in the media increased significantly, whereas the addition of LPS to the media caused no further increase. The plasma cathelicidin‐2 concentrations did not increase with time after LPS infusion. The proportion of cathelicidin‐2‐positive cells in the total leukocytes was significantly reduced 1 h after LPS injection compared with that at 0 h, but increased again at 6 h and thereafter. These results suggest that cathlicidin‐2 is secreted by leukocytes even without LPS stimulation, whereas LPS may be required for cathelicidin‐2‐containing leukocytes to be recruited from the blood to tissues showing inflammation.  相似文献   

Partial cloning of CYP2C23a genes and hepatic protein expression in eight representative avian species          下载免费PDF全文

K. P. Watanabe  Y. K. Kawai  S. M. M. Nakayama  Y. Ikenaka  H. Mizukawa  N. Takaesu  M. Ito  S.‐I. Ikushiro  T. Sakaki  M. Ishizuka 《Journal of veterinary pharmacology and therapeutics》2015,38(2):190-195

Large interspecies differences in avian xenobiotic metabolism have been revealed by microsome‐based studies, but specific enzyme isoforms in different bird species have not yet been compared. We have previously shown that CYP2C23 genes are the most induced CYP isoforms in chicken liver. In this study, we collected partial CYP2C23a gene sequences from eight avian species (ostrich, blue‐eared pheasant, snowy owl, great‐horned owl, Chilean flamingo, peregrin falcon, Humboldt penguin, and black‐crowned night heron) selected to cover the whole avian lineage: Paleognathae, Galloanserae, and Neoaves. Genetic analysis showed that CYP2C23 genes of Galloanserae species (chicken and blue‐eared pheasant) had unique characteristics. We found some duplicated genes (CYP2C23a and CYP2C23b) and two missing amino acid residues in Galloanserae compared to the other two lineages. The genes have lower homology than in other avian lineages, which suggests Galloanserae‐specific rapid evolutionary changes. These genetic features suggested that the Galloanserae are not the most representative avian species, considering that the Neoaves comprise more than 95% of birds. Moreover, we succeeded in synthesizing an antipeptide polyclonal antibody against the region of CYP2C23 protein conserved in avians. However, comparative quantitation of CYP2C23 proteins in livers from six species showed that expression levels of these proteins differed no more than fourfold. Further study is needed to clarify the function of avian CYP2C23 proteins.  相似文献   

A NOVEL SUPPORT DEVICE FOR HEAD IMMOBILIZATION DURING RADIATION THERAPY THAT IS APPLICABLE TO BOTH CATS AND DOGS          下载免费PDF全文

Yuki Nemoto  Takuya Maruo  Yasuhiro Fukuyama  Shinpei Kawarai  Takuo Shida  Tomohiro Nakayama 《Veterinary radiology & ultrasound》2015,56(6):680-686

Repeatable head immobilization is important for minimizing positioning error during radiation therapy for veterinary patients with head neoplasms. The purpose of this retrospective cross‐sectional study was to describe a novel technique for head immobilization (Device II) and compare this technique with a previously described technique (Device I). Device II provided additional support by incorporating three teeth (vs. two teeth with Device I). Between 2011 and 2013, both devices were applied in clinically affected cats (Device I, n = 17; Device II, n = 11) and dogs (Device I, n = 85; Device II, n = 22) of various breeds and sizes. The following data were recorded for each included patient: variability in the angle of the skull (roll, yaw, and pitch), coordinates of the isocenter, and distance from the reference mark to the tumor. Devices I and II differed for skull angle variability during the treatment of dogs (roll, P = 0.0007; yaw, P = 0.0018; pitch, P = 0.0384) and for yaw of during the treatment of cats (P < 0.0001). In each case, Device II was superior to Device I. The distance from the reference mark to the center of the tumor was significantly decreased for Device II vs. Device I (dogs, P < 0.0001; cats, P = 0.0002). Device II also provided more accurate coordinates for the isocenter. Authors recommend the use of, Device II for future clinical patients.  相似文献   

Deficient proliferation and apoptosis in the granulosa and theca interna cells of the bovine cystic follicle   总被引：1，自引：0，他引：1  

Isobe N  Yoshimura Y 《The Journal of reproduction and development》2007,53(5):1119-1124

The aim of the present study was to examine the frequencies of cell proliferation and death of granulosa and theca interna layers during development of cystic follicles in order to understand the mechanisms of cystic follicle formation. Paraffin sections of cystic follicles were immunostained with antibodies against proliferating cell nuclear antigen (PCNA) and cleaved caspase-3 in order to observe proliferating and apoptotic cells, respectively. The concentrations of estradiol-17beta and progesterone in the follicular fluid of these follicles were measured by ELISA. The granulosa and theca interna layers contained both PCNA- and caspase-3-positive cells, although their numbers were limited. There was significant negative correlation between the estradiol-17beta and progesterone concentrations in the follicular fluid. Regression analysis revealed no significant correlation, except for that between the PCNA-positive cells in the theca interna and the caspase-3-positive cells in the granulosa layer. These results indicate that the granulosa and theca interna cells of the cystic follicle show weak proliferative activity and low apoptotic frequency; this implies that the cystic follicle grows slowly and then maintains a static condition without degeneration, which leads to long-term persistence of the follicle.  相似文献   

Studies of the constituents of Uruguayan propolis   总被引：4，自引：0，他引：4  

Kumazawa S  Hayashi K  Kajiya K  Ishii T  Hamasaka T  Nakayama T 《Journal of agricultural and food chemistry》2002,50(17):4777-4782

Eighteen flavonoids including two new compounds, four aromatic carboxylic acids, and eleven phenolic acid esters including one new compound were isolated and identified from the ethyl acetate soluble fraction of the 70% ethanol extract of Uruguayan propolis. The new compounds were elucidated as pinobanksin 3-(2-methyl)butyrate (1; recently reported in Usia, T.; Banskota, A. H.; Tezuka, Y.; Midorikawa, K.; Matsushige, K.; Kadota, S. J. Nat. Prod. 2002, 65, 673-676) pinobanksin 3-isobutyrate (2), and 2-methyl-2-butenyl ferulate (24). The constituents isolated from Uruguayan propolis in this study were similar to those of propolis of European and Chinese origin. Thus, it is suggested that the Uruguayan propolis has a plant origin similar to those of propolis from Europe and China.  相似文献   

Genetic Variation of Blast Resistance in Foxtail Millet (<Emphasis Type="Italic">Setaria italica</Emphasis> (L.) P. Beauv.) and its Geographic Distribution     

Hiroki Nakayama  Tsukasa Nagamine  Nagao Hayashi 《Genetic Resources and Crop Evolution》2005,52(7):863-868

To clarify the geographic variation and isolate specificity in blast resistance, 20 cultivars of foxtail millet (Setaria italica (L.) P. Beauv.) originating from Eurasia were examined for their resistance using 11 Japanese Setaria isolates of blast fungus. Cultivars from the countries east of Pakistan generally showed resistance to most of the 11 fungus isolates, whereas those from the countries west of Afghanistan indicated higher susceptibility. The origin of this geographical regularity was discussed in relation to the deficiency or specificity of relevant resistance gene(s). No identical reaction pattern was observed among cultivars, indicating their highly distinctive isolate specificity. The virulence of isolates was also diverse, since only 2 out of 10 virulent isolates were identical for their reaction patterns. Preliminary genetic analysis of blast resistance to four fungus isolates suggests that they are governed by more than two dominant genes.  相似文献   

A tetraploid Echinochloa with plagiotropic tillers: Its distribution and habitat in the northern part of the main island of Japan     

Kentaro Yasuda  Kaoru Mori  Yuichiro Nakayama 《Weed Biology and Management》2020,20(2):82-88

An annual Echinochloa with plagiotropic tillers (semi‐erect type) was found on a paddy levee in Miyagi prefecture in Japan. This semi‐erect type had tetraploid chromosome numbers (2n = 36) and would be expected to be E. oryzicola. However, its appearance was distinct from that of E. oryzicola. To collect seed samples and clarify its distribution and habitat, exploration and collection surveys were carried out in the northern part of Honshu, the main island of Japan in 2014, 2017, and 2018. Thirty‐two populations that were assumed to be the semi‐erect type were found and their mature seeds were collected. Based on the results of chromosome counts and growth form analyses, all of them were confirmed to be the semi‐erect type. The 32 populations were distributed in the prefectures of Aomori, Iwate, Akita, Miyagi, Yamagata, Fukushima, and Niigata, and located in paddy fields, abandoned paddy fields, paddy levees, roadsides, vacant land, and scree slopes. Our surveys indicated that the semi‐erect type grew in wet and dry disturbed habitats at an early stage of secondary succession including paddy fields in the northern part of the main island of Japan.  相似文献