Secretome analysis of the basidiomycete <Emphasis Type="Italic">Phanerochaete chrysosporium</Emphasis> grown on ammonia-treated lignocellulosic biomass from birch wood     

Kiyoshi Sakuragi  Chiaki Hori  Kiyohiko Igarashi  Masahiro Samejima 《Journal of Wood Science》2018,64(6):845-853

Ammonia pretreatment is a promising technique for enhancing enzymatic saccharification of lignocellulosic biomass. However, an enzymatic cocktail suitable for the breakdown of pretreated biomass samples is still being developed. The basidiomycete Phanerochaete chrysosporium is a well-studied fungus with regard to bioconversion of lignocellulosic biomass. In the present work, we analyzed proteins secreted by P. chrysosporium grown on untreated and ammonia-treated birch wood meal. Fungal growth, xylanase activity, and extracellular protease activity increased in the media containing the ammonia-treated biomass; however, cellulase production decreased compared to that observed in the untreated biomass. Secreted extracellular proteins were separated by two-dimensional electrophoresis and identified by liquid chromatography ion–trap mass spectrometry. Fifty-five spots corresponding to secreted proteins were chosen for further analysis. In the culture with ammonia-treated biomass, the relative concentration of a xylanase belonging to glycoside hydrolase (GH) family 11 increased, while acetyl xylan esterases belonging to carbohydrate esterase family 1 decreased. Moreover, GH family 10 xylanases were promoted proteolysis in the culture of ammonia-treated biomass, leading to the loss of family 1 carbohydrate-binding modules. These results indicated that P. chrysosporium produced enzymes related to the recognition of structural changes on xylan with de-acetylation and introduction of nitrogen by ammonia pretreatment of birch wood meal.  相似文献   

Role of prostatic fluid in cooled canine epididymal sperm          下载免费PDF全文

T Hori  T Masuda  M Kobayashi  E Kawakami 《Reproduction in domestic animals》2017,52(4):655-660

In this study, sperms collected from the right and left cauda epididymis were grouped into having canine prostatic fluid (PF) sensitization or not diluted with egg yolk Tris–fructose citrate extender, and stored at 4°C. The necessity of canine PF in cooled preservation was determined by elucidating the sperm quality after the storage. As a result, while there was no difference among all groups up to 48 hr of storage, after storage for 96 hr and more, a significantly lower sperm motility was observed in the group without being sensitized to PF than the groups with being sensitized to PF (p < .05, p < .01). Although sperm abnormality increased in all groups with increased storage time, the group without being sensitized to PF showed significantly higher sperm abnormality than did the groups with being sensitized to PF after storage for 24 hr and more (p < .01). From these findings, we concluded that PF was necessary for the cooled preservation of the canine sperm because these sperms were protected from any effects of low temperatures by being sensitized to PF.  相似文献   

Overwintering of <Emphasis Type="Italic">Taphrina wiesneri</Emphasis> within cherry shoots monitored with species-specific PCR     

Masabumi Komatsu  Machiko Taniguchi  Norihisa Matsushita  Yukiko Takahashi  Taizo Hogetsu 《Journal of General Plant Pathology》2010,76(6):363-369

Taphrina wiesneri, the pathogen of witches’ broom of cherry, is highly pathogenic to Cerasus × yedoensis, the most widely planted ornamental cherry species in Japan. For adequate control of this disease, it is necessary to understand the life history of T. wiesneri. However, sites inhabited by T. wiesneri within infected trees are little understood, except during flowering and leafing periods in spring. Therefore, we attempted to detect the location of T. wiesneri in shoots of witches’ broom before flowering and leafing in spring using PCR with a T. wiesneri-specific primer pair that was designed from 69 sequences in rDNA-internal transcribed spacer region of 32 Taphrina species. DNA extracted from symptomatic and asymptomatic C. × yedoensis sampled before leafing was amplified by PCR. T. wiesneri was detected in every bud and 5-mm stem segment of symptomatic shoots, except for one stem segment, and locally inside buds and the inner bark of stem segments. These results indicate that T. wiesneri overwinters inside symptomatic shoots. Fungal hyphae were observed with an epifluorescence microscope in intercellular spaces of young leaves in symptomatic buds but not in asymptomatic ones in thin sections stained with fluorescein-isothiocyanate-conjugated concanavalin A. This observation supports the results of PCR detection.  相似文献   

Comparison of fertility on intrauterine insemination between cryopreserved ejaculated and cauda epididymal sperm in dogs     

Hori T  Matsuda Y  Kobayashi M  Kawakami E  Tsutsui T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2011,73(12):1685-1688

The fertility was compared between ejaculated and cauda epididymal sperm sensitized with prostatic fluid in dog after freeze-thawing using the fertility of ova from the contralateral ovary after injection (2 × 10(8) sperm) into dog uterus on the unilateral ovariectomized side, on the basis of the presence or absence of conception. No significant difference was observed in sperm quality after freeze-thawing between the two groups and conception rates were equivalent and low. Therefore, to achieve a high fertility by intrauterine insemination of canine frozen-thawed ejaculated and cauda epididymal sperm, intrauterine insemination on both sides is recommended, rather than insemination with a lot of sperm of the uterine horn on one side.  相似文献   

In vitro development and postvitrification survival of cloned feline embryos derived from preadipocytes     

Tomii R  Ogawa B  Imai N  Handa Y  Sasayama N  Shirasu A  Nagashima H 《The Journal of reproduction and development》2011,57(2):273-279

The aim of the present study was to optimize the conditions for in vitro development and postvitrification survival of somatic cell cloned feline embryos. To determine the effects of cell cycle synchronization of the nuclear donor cells, we cultured preadipocytes under serum starvation or conventional conditions. After two days in serum starvation culture, the proportion of synchronized donor cells at the G0/G1 phase was 91.6%. This was significantly higher than the proportion of non-synchronized cells in the proliferative phase (72.6%, P<0.05). The in vitro development of somatic cell nuclear transfer (SCNT) embryos reconstructed using donor cells treated under serum starvation conditions (normal cleavage rate of 65.7%, 46/70, and blastocyst formation rate of 20.0%, 14/70) was comparable to that of the serum supplemented group (52.5%, 31/59, and 20.3%, 12/59). Use of in vitro or in vivo matured oocytes as recipient cytoplasts equally supported development of the SCNT embryos to the blastocyst stage (11.9%, 5/42, vs. 9.5%, 2/21). SCNT-derived blastocysts were vitrified using the original minimum volume cooling (MVC) or the modified (stepwise) MVC method. Although none (n=10) of the SCNT blastocysts survived following vitrification by the original MVC method, the stepwise MVC method resulted in 100% survival after rewarming (n=11). In conclusion, we demonstrated that feline somatic cell cloned embryos with a high developmental ability can be produced irrespective of cell cycle synchronization of donor cells using either in vivo or in vitro matured oocytes. Furthermore, by utilizing a stepwise vitrification method, we showed that it is possible to cryopreserve cloned feline blastocysts.  相似文献   

Establishment of a quantitative ELISA for the measurement of allergen-specific IgE in dogs using anti-IgE antibody cross-reactive to mouse and dog IgE     

Okayama T  Matsuno Y  Yasuda N  Tsukui T  Suzuta Y  Koyanagi M  Sakaguchi M  Ishii Y  Olivry T  Masuda K 《Veterinary immunology and immunopathology》2011,139(2-4):99-106

As IgE plays a pivotal role in type I hypersensitivity-mediated allergic diseases, it is valuable to measure absolute quantity of serum antigen-specific IgE for clinical and research purposes. Here we describe a novel ELISA system that enables quantification of antigen-specific IgE in ng/ml in dogs. A newly developed monoclonal antibody (CRE-DM) was shown to recognize canine and mouse IgE equally in a dose dependent manner, but it did not recognize canine IgG. The reactivity of CRE-DM to canine IgE was also confirmed by an inhibition ELISA using canine IgE as an inhibitor and the maximum inhibition rate was 91.3%. In order to know whether canine IgE specific to an allergen could be quantitatively measured with an ELISA using CRE-DM, we established a quantitative ELISA that could measure canine IgE recognizing Cry j 1, one of the major allergens of Japanese cedar pollen. In this ELISA, a standard curve was created by using concentration-predetermined Cry j 1-specific monoclonal mouse IgE. According to the standard curve, the concentration of Cry j 1-specific IgE in dogs that were experimentally sensitized to Japanese cedar pollen could be calculated and determined in ng/ml. The specificity of the Cry j 1-specific IgE ELISA using CRE-DM was also confirmed by inhibition ELISA using canine IgE as an inhibitor and the inhibition rate was 97.0%. Reproducibility of the ELISA in three independent assays was determined using groups of pooled canine sera whose Cry j 1-IgE titers ranged from 155.9 to 888.2 ng/ml. Intra- and inter-assay reproducibility was determined with coefficient of variation ranging between 3.1-5.2% and 2.2-8.0%, respectively. These results demonstrated that the ELISA utilizing CRE-DM was a specific, reliable and robust new laboratory test that could quantify absolute amount of antigen-specific IgE in canine serum. The ELISA will serve as a useful tool in the clinics to evaluate the change of serum IgE titers during anti-allergic treatments as well as during seasonal fluctuation of allergen exposure.  相似文献   

Rediscovery of Indian dwarf wheat (Triticum aestivum L. ssp. sphaerococcum (Perc.) MK.) an ancient crop of the Indian subcontinent     

Naoki Mori  Shoji Ohta  Hajime Chiba  Toshiya Takagi  Yukiko Niimi  Vasant Shinde  Mukund D. Kajale  Toshiki Osada 《Genetic Resources and Crop Evolution》2013,60(6):1771-1775

Indian dwarf wheat (Triticum aestivum L. ssp. sphaerococcum (Perc.) Mac Key, synonym: T. sphaerococcum Perc.) is endemic to southern Pakistan and northwestern India. It was one of the main winter crops grown by ancient Indian cultures. However, it disappeared from the record during the early twentieth century, especially after the Green Revolution brought modern wheat varieties into India and Pakistan. Whether or not Indian dwarf wheat is presently cultivated has been unclear. Here we report on the rediscovery of the cultivation of this wheat in northern Karnataka and southern Maharashtra in India. Molecular genetic analysis of the chloroplast DNA of the two specimens collected at location 3 revealed that both samples have a unique haplotype that is specific to Indian dwarf wheat. We found this wheat at three locations in 2010, but at only one of the three locations in 2011. Therefore, the future survival of this subspecies is uncertain. Further ethnobotanical research is urgently needed to conserve this unique genetic resource for the future.  相似文献   

Changes in proline-14C metabolism in barley seedlings germinating at low temperature     

Nobuyuki Shiomi  Shiro Hori 《Soil Science and Plant Nutrition》2013,59(1):121-129

Changes in the metabolic pattern of proline-¹⁴C were examined in barley seedlings germinated at the low temperature of 2°C (LT) and compared with those germinated at 25°C (HT). In the LT shoots. proline-¹⁴C incorporation was higher in the cationic fraction and lower in the acid-neutral fraction than that of HT, respectively. More proline-¹⁴C of LT was converted to other amino acids, especially to acidic amino acids in free amino acid state, than was the HT proline-¹⁴C which had a comparatively wide distribution.

In the LT protein fractions. more proline-¹⁴C was incorporated into the cytoplasmic protein than into the cell wall protein. On the contrary, the radioactivity of the lIT cell wall increased more distinctively. proline-¹⁴C in the two proteins of LT was a little less converted to other amino acids than in those of HT. A little higher radioactivity was found in the aspartate and glutamate of the LT protein hydrolysates. The hydroxyproline which is closely related with proline had a little lower level of radioactivity in the LT cell wall.  相似文献   

A comparison of physical properties, oxalate-oxalic acid soluble substances, protein content, and in vitro protein digestibility of the blue-green algaNostoc commune Vauch. from the Philippines and Japan     

Maria Portia P. Briones  Kohji Hori  Milagrosa R. Martinez-Goss  Genji Ishibashi  Takuo Okita 《Plant foods for human nutrition (Dordrecht, Netherlands)》1997,50(4):287-294

The physical properties, amount of oxalate-oxalic acid soluble substances (OOSS), protein and in vitro protein digestibility ofNostoc commune from the Philippines and Japan were determined. The samples were the spherical type from the Philippines (PS), the dry field discoid type from the Philippines (PD), and the dry field discoid type from Japan (JD). The discoid types, PD and JD, regardless of origin showed higher settling volume in water (SV) and bulk density than PS. The OOSS values obtained for all samples were directly related to SV and bulk density. The WHC level was highest in PS. This desirable characteristic makes PS a technologically potential sample for large-scale food production applications in the future. Protein and in vitro protein digestibility varied significantly among all samples. In general, the algal samples examined posses desirable physical properties which are expected to be related to physiological functions.  相似文献   

Structural changes in lignin of tropical woods during digestion by termite, <Emphasis Type="Italic">Cryptotermes brevis</Emphasis>     

Kyoko S. Katsumata  Zhenfu Jin  Keko Hori  Kenji Iiyama 《Journal of Wood Science》2007,53(5):419-426

Wood samples of apitong (Dipterocarpus grandiflorua) and ilang-ilang (Ilang-Ilang C. dadloyi) and feces of termites [Cryptotermes brevis (Walker)] fed on these woods were collected from University of the Philippines, Los Baňos. Lignin of each sample was isolated by Björkman’s procedure. There was no significant difference in ¹H nuclear magnetic resonance (NMR) spectra or in the methoxyl content between Björkman lignins from original woods and termite feces. Differences were detected in the contents of aliphatic and unconjugated phenolic hydroxyl groups, suggesting minor structural changes of lignin during digestion by termites. In addition, the ratio of syringyl to guaiacyl nuclei of Björkman lignin from termite feces determined by ¹H NMR spectra was higher than those from the original woods. The molar ratio of syringyl to guaiacyl nuclei of termite feces was higher than those from the original woods as determined by alkaline nitrobenzene oxidation. These results suggest that the structural changes of lignin in the termite gut are due to the insignificant formation of C-C linkages in guaiacyl nuclei. It was concluded that there were minor changes in the structural features of lignin under mostly anaerobic conditions, in contrast to the significant changes that occur through biological modification under aerobic conditions.  相似文献