A Monitoring System for Green Fluorescence Protein Gene-transformed Fusarium oxysporum in Melon Seedlings     

Teruo NONOMURA  Yoshinori MATSUDA  Mikako TAKASUGI  Takashi OOTANI  Tomoya HASEGAWA  Kazutomo MIYAJIMA  Tetsuya HATASA  Hideyoshi TOYODA 《Journal of General Plant Pathology》2001,67(4):273-280

Using melon seedlings at the cotyledon stage and genetically marked fungi, a system for monitoring pathogenic and nonpathogenic Fusarium oxysporum was devised in the present study. Protoplasts were prepared from three formae speciales (melonis, radicis-lycopersici and fragariae )of F. oxysporum and transformed with a synthetic gene for green fluorescence protein. Transformants were primarily isolated in the presence of hygromycin B and then screened by the emission of bright green fluorescence. Roots of melon seedlings were inoculated with fluorescing microconidia of these fungi, and fungal infection behavior was traced. Using fluorescence microscopy, we directly observed not only the fungus at the root surface, but also the mycelia elongating in the trachea of roots. Both pathogenic and nonpathogenic fungi germinated and hyphae elongated superficially on the surface of root. Only pathogenic fungi caused root necrosis at the inoculation site. Hyphae grew within the stem to induce constriction or cracking of lower hypocotyls, then causing wilting of the seedlings. Infection behavior of genetically marked pathogenic and nonpathogenic F. oxysporum could be successfully monitored after inoculation of cotyledons of seedlings. Received 6 June 2001/ Accepted in revised form 3 August 2001  相似文献   

Effect of trimetoquinol on skeletal muscle contraction in rabbits. Comparison with epinephrine and isoproterenol     

K Yamada  M Iizuka 《Nippon juigaku zasshi. The Japanese journal of veterinary science》1973,35(3):183-191

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Evaluation of duodenal perfusion by contrast‐enhanced ultrasonography in dogs with chronic inflammatory enteropathy and intestinal lymphoma     

Khoirun Nisa  Sue Yee Lim  Masayoshi Shinohara  Tatsuyuki Osuga  Nozomu Yokoyama  Masahiro Tamura  Noriyuki Nagata  Kazuyoshi Sasaoka  Angkhana Dermlim  Rommaneeya Leela‐Arporn  Tomoya Morita  Noboru Sasaki  Keitaro Morishita  Kensuke Nakamura  Hiroshi Ohta  Mitsuyoshi Takiguchi 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2019,33(2):559-568

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Correction to: Dispersion and degradation of environmental DNA from caged fish in a marine environment     

Murakami  Hiroaki  Yoon  Seokjin  Kasai  Akihide  Minamoto  Toshifumi  Yamamoto  Satoshi  Sakata  Masayuki K.  Horiuchi  Tomoya  Sawada  Hideki  Kondoh  Michio  Yamashita  Yoh  Masuda  Reiji 《Fisheries Science》2019,85(6):1109-1109

Fisheries Science - In the original publication the text in right column of page 330, the sequences of primers were incorrectly published as.  相似文献   

Isolation and enzymatic formation of lignans ofDaphne genkwa andDaphne odora     

Tomoya Okunishi  Toshiaki Umezawa  Mikio Shimada 《Journal of Wood Science》2001,47(5):383-388

Four lignans — pinoresinol, lariciresinol, secoisolariciresinol, matairesinol — were isolated from each ofDaphne odora andDaphne genkwa (Thymelaeaceae). Matairesinol isolated from both plants was optically pure (>99% e.e.) and dextrorotatory. Pinoresinol and lariciresinol isolated from the plants were not optically pure, and their enantiomeric compositions ranged from 88% to 95% e.e. in favor of (–)-enantiomers. As for secoisolariciresinol, the one fromD. odora was optically pure [(+)-enantiomer, >99% e.e.], and that fromD. genkwa was 97% e.e. in favor of the (+)-enantiomer. Lignan-synthesizing enzyme activity was detected from a Thymelaeaceae plant for the first time; cell-free extracts fromD. genkwa catalyzed the formation of (–)-lariciresinol (23% e.e.) from racemic (±)-pinoresinols. The stereochemistry of the enzymatic reaction is discussed in relation to the stereochemical features of the isolated lignans.Parts of this report were presented at the 42nd Lignin Symposium, Sapporo, October 1997; 48th Annual Meeting of the Japan Wood Research Society, Shizuoka, April 1998; and 49th Annual Meeting of the Japan Wood Research Society, Tokyo, April 1999  相似文献   

Characterization of the reproductive cycle in female bharals (Pseudois nayaur) based on the changes in serum progesterone concentrations and parturition season     

Kusuda S  Nagami H  Ueda K  Nishikaku T  Nakagawa D  Takida T  Kurita D  Uemichi K  Fukai M  Kubota H  Ueda K  Ooe T  Okuda K  Kusunoki H  Doi O 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2006,68(8):847-851

The purpose of present study was to determine annual changes in serum progesterone (P(4)) concentrations and to clarify basic reproductive characteristics, such as breeding season, estrous cycle, and puberty in female bharals (Pseudois nayaur). Blood was collected from 9 female bharals once or twice weekly for approximately one year. Serum P(4) concentrations were determined by radioimmunoassay. Serum P(4) concentrations showed remarkable and cyclic changes between November/December (winter) and May/June (late spring). The mean estrous cycle was 24.9 +/- 0.5 days. Chasing insistently to other females and discharge of mucus from the vulva were observed around the time when the serum P(4) concentrations began to increase. The chasing behavior and discharge of mucus were considered to be external indicators of estrus in female bharals. Serum P(4) concentrations of a pregnant female had non-cyclic changes, and the values remained high. In this study, all 37 deliveries were between April and September, and about 70% of these were concentrated in May and June. The conception month determined on the day of birth was between October and April for all animals, and the most common month was in December (54%). This month corresponded to an early stage of the period when the serum P(4) concentrations changed cyclically. These results indicate that many female bharals become pregnant at the beginning of the breeding seasons and, if they do not become pregnant, the estrous cycle, about 25 days in length, is repeated.  相似文献   

Analyses of leaves from open field-grown transgenic poplars overexpressing xyloglucanase     

Tomomi Kaku  Kei’ichi Baba  Toru Taniguchi  Manabu Kurita  Ken-ichi Konagaya  Katsuaki Ishii  Teiji Kondo  Satoshi Serada  Haruka Iizuka  Rumi Kaida  Teruaki Taji  Yoichi Sakata  Takahisa Hayashi 《Journal of Wood Science》2012,58(4):281-289

The transgenic expression of Aspergillus xyloglucanase cDNA (AaXEG2) with 35S promoter in the leaves of open field-grown poplars was studied. The level of xyloglucan in the transgenic poplars was decreased to 15–16% in the non-fertile soil (forest-field soil) and to 21–22% in the fertile soil (farming-field soil) compared with that of the wild-type poplars. The leaves exhibited a smaller surface area with more rounded teeth than those of the wild-type plants, similar to the sun leaf variety that was grown in the incubation room and subsequently greenhoused. The majority of total veins with water-conducting vascular bundles were shorter in the leaves of the transgenic poplars than those of the wild type. This decrease in vein length may result from a decrease in xyloglucan during leaf development, from which large numbers of proteins were markedly downregulated in the leaves of the transgenic plants via proteomic analysis. It seems likely that the leaves of the transgenic poplars came to relax the edges of their tooth rather than extend their veins as a result of the loosening of the xyloglucan cellulose networks in the leaves.  相似文献   

Clinical and clinico-pathologic characteristics of Shiba dogs with a deficiency of lysosomal acid beta-galactosidase: a canine model of human GM1 gangliosidosis     

Yamato O  Masuoka Y  Yonemura M  Hatakeyama A  Satoh H  Kobayashi A  Nakayama M  Asano T  Shoda T  Yamasaki M  Ochiai K  Umemura T  Maede Y 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2003,65(2):213-217

The present study was conducted to determine the clinical and clinico-pathologic characteristics of Shiba dogs with GM1 gangliosidosis, which is due to an autosomal recessively inherited deficiency of lysosomal acid beta-galactosidase activity. Clinical and clinico-pathological features were investigated in 10 homozygous Shiba dogs with GM1 gangliosidosis. The age at onset was 5 to 6 months and the dogs manifested progressive neurologic signs including loss of balance, intermittent lameness, ataxia, dysmetria and intention tremor of the head. The dogs were unable to stand by 10 months of age due to a progression of ataxia and spasticity in all limbs. Corneal clouding, a visual defect, generalized muscle rigospasticity, emotional disorder and a tendency to be lethargic were observed at 9 to 12 months. The dogs became lethargic from 13 months of age. The survival period seemed to be 14 to 15 months. As a clinico-pathologic feature, lymphocytes with abnormally large vacuoles were observed in peripheral blood (30 to 50% of total lymphocytes) through the lifetime of the dogs. The clinical and clinico-pathologic characteristics of this animal model are useful for not only the development and testing of potential methods of therapy, but also the diagnosis of affected homozygous Shiba dogs in veterinary clinics.  相似文献   

Increased concentration of GM1-ganglioside in cerebrospinal fluid in dogs with GM1- and GM2-gangliosidoses and its clinical application for diagnosis.     

Hiroyuki Satoh  Osamu Yamato  Tomoya Asano  Masahiro Yamasaki  Yoshimitsu Maede 《Journal of veterinary diagnostic investigation》2004,16(3):223-226

GM1- and GM2-gangliosidoses are lethal lysosomal diseases that are caused by a defect of acid hydrolases, resulting in the intralysosomal accumulation of the specific physiological substrates, GM1- and GM2-gangliosides, respectively. In the present study a method for the diagnosis of canine GM1-gangliosidosis was established using canine cerebrospinal fluid (CSF). The concentration of GM1-ganglioside in CSF was determined by thin-layer chromatography-enzyme immunostaining using biotin-conjugated cholera toxin B, which specifically binds with GM1-ganglioside. The concentration of CSF GM1-ganglioside was increased in Shiba dogs with GM1-gangliosidosis, and the increased level was approximately proportional to the age of the dogs. The concentration was high in the affected dog even at 5 months of age, when Shiba dogs with GM1-gangliosidosis first manifest neurologic signs. In addition, the concentration of CSF GM1-ganglioside in a dog with the GM2-gangliosidosis 0 variant (Sandhoff disease) was also 7 times the normal level. From these results it was concluded that this laboratory technique enables a definitive and early diagnosis of canine GM1-gangliosidosis even if tissues and organs cannot be obtained. However, because GM1-ganglioside can also be elevated in cases of GM2-gangliosidosis, it is necessary to assay for specific enzyme deficiencies to definitively separate GM1- from GM2-gangliosidosis.  相似文献   

Comparison of polymerase chain reaction-restriction fragment length polymorphism assay and enzyme assay for diagnosis of G(M1)-gangliosidosis in Shiba dogs.     

Osamu Yamato  Asogi Kobayashi  Hiroyuki Satoh  Daiji Endoh  Toru Shoda  Yukiko Masuoka  Ayano Hatakeyama  Eun-Og Jo  Tomoya Asano  Madoka Yonemura  Masahiro Yamasaki  Yoshimitsu Maede 《Journal of veterinary diagnostic investigation》2004,16(4):299-304

In the present study, diagnostic methods for canine G(M1)-gangliosidosis were examined by comparing a DNA mutation assay with an enzyme assay. Sixty-two Shiba dogs of a pedigree with G(M1)-gangliosidosis were differentiated into 3 genotypes, i.e., normal, heterozygous, and homozygous affected dogs, using a DNA mutation assay, which consists of polymerase chain reaction amplification and the determination of restriction fragment length polymorphisms. The beta-galactosidase activity in leukocytes, umbilical cords, and plasma was measured using 4-methylumbelliferyl beta-D-galactoside and p-nitrophenyl beta-D-galactoside as artificial substrates and compared among the 3 genotypes. The results showed that it was possible to identify homozygous dogs with the enzyme assay using leukocytes and umbilical cords. When using leukocytes, heterozygous carriers could be differentiated from normal dogs in many cases. However, the use of the DNA mutation assay is essential for a complete determination of heterozygous carriers because of the overlap in the distribution of enzyme activity between these 2 groups. When umbilical cords were used, heterozygous carriers could not be differentiated from normal dogs because of no significant difference in enzyme activity between these 2 groups. The beta-galactosidase activity in plasma was not applicable to the diagnosis and genotyping of G(M1)-gangliosidosis in Shiba dogs.  相似文献