MATSUDA Yoshinori

    Department of Agricultural Science Professor
Last Updated :2024/03/02

Researcher Information

Degree

  • (BLANK)

URL

J-Global ID

Research Interests

  • 雑草防除   植物病虫害防除   Plant Protection   

Research Areas

  • Environmental science/Agricultural science / Agricultural environmental and information engineering
  • Environmental science/Agricultural science / Conservation science (plants)

Academic & Professional Experience

  • 2008 - Today  Kindai UniversityFaculty of Agriculture教授
  • 2002 - 2008  Kindai UniversityFaculty of Agriculture准教授
  • 1997 - 2002  Kindai University農学総合研究所講師
  • 1994 - 1997  Kindai University農学総合研究所助手
  • 1993 - 1994  ミネソタ大学米国農務省研究員

Education

  • 1989 - 1992  Kindai University  Graduate School, Division of Agriculture  作物保護
  • 1984 - 1986  Kindai University  農学研究科博士前期課程  作物保護
  • 1979 - 1983  Kindai University  Faculty of Agriculture  農学科

Association Memberships

  • JAPANESE SOCIETY OF AGRICULTURAL, BIOLOGICAL AND ENVIRONMENTAL ENGINEERS AND SCIENTISTS   THE INSTITUTE OF ELECTROSTATICS JAPAN   

Published Papers

  • Yoshinori Matsuda; Koji Kakutani; Hideyoshi Toyoda
    American Journal of Civil Engineering and Architecture Science and Education Publishing Co., Ltd. 12 (1) 1 - 7 2328-398X 2024/01 [Refereed]
  • Koji Kakutani; Yoshinori Matsuda; Yoshihiro Takikawa; Hideyoshi Toyoda
    American Journal of Public Health Research 11 (6) 211 - 218 2023/12 [Refereed]
  • Yoshinori Matsuda; Koji Kakutani; Hideyoshi Toyoda
    Agronomy MDPI AG 13 (7) 1954 - 1954 2023/07 [Refereed]
     
    This study developed an unattended electric weeder (UEW) to control floor weeds in an orchard greenhouse. The UEW was a motor-driven dolly equipped with a spark exposer. The spark exposer was constructed by applying an alternating voltage (10 kV) to a conductor net (expanded metal net). The charged conductor net (C-CN) discharged into the surrounding space. Wild oat and white clover were used as test weed species. Weed seedlings growing on the floor were grounded by the biological conductor and were subjected to a spark from the C-CN when they reached the discharge space. The spark-exposed seedlings were singed and shrunk instantaneously. In the present experiment, the UEW was remotely controlled to move on the soil-cover metal nets, which were laid on the floor to make a flat surface, in a stop-and-go manner, and to eject a spark to the weed seedlings that emerged from the floor. All of the mono- and dicotyledonous weed seedlings, which had been artificially sown on the floor, were completely eradicated using this method. Thus, this study provides an experimental basis for developing an unattended technique for controlling floor weeds in an orchard greenhouse.
  • Yoshinori Matsuda; Yoshihiro Takikawa; Kunihiko Shimizu; Shin-ichi Kusakari; Hideyoshi Toyoda
    Agronomy MDPI AG 13 (4) 1115 - 1115 2023/04 [Refereed]
     
    An electrostatic technique was developed to generate a simple physical method to eradicate weeds in crop fields. The proposed apparatus consisted of double-expanded metal nets connected to a pulse-charging type negative voltage generator and a grounded line. The two metal nets were arranged in parallel at an interval (6 mm) that caused no arc (spark) discharge between the negatively charged metal net (NC-MN) and the grounded metal net (G-MN). The paired nets were used as a soil cover to zap weed seedlings emerging from the ground. As plant seedlings are biological conductors, the seedling was subjected to an arc discharge from the upper metal net (NC-MN) when it emerged from the soil and passed through the lower net (G-MN). The discharge was strong enough to destroy the seedling with a single exposure. The arc treatment was highly effective for eradicating successively emerging mono- and dicotyledonous weed seedlings, regardless of the number of coexisting weeds or the area of the netted field. Thus, the present study provides a simple and reliable weed eradication method that could be integrated into a sustainable crop production system.
  • Koji Kakutani; Yoshinori Matsuda; Hideyoshi Toyoda
    Agronomy MDPI AG 13 (2) 310 - 310 2023/01 [Refereed]
     
    A simple electrostatic apparatus that generates an arc discharge was devised to control adult houseflies emerging from a soil bed in a greenhouse. Adult houseflies emerging from a soil bed in a greenhouse are a potential vector of pathogenic Escherichia coli O157, carried by animal manure used for soil fertilization. A simple electrostatic apparatus that generates an arc discharge was devised to control these houseflies. The apparatus consisted of two identical metal nets; one was linked to a negative-voltage generator to create a negatively charged metal net (NC-MN), and the other was linked to a grounded line to create a grounded metal net (G-MN). A square insulator frame was placed between the two nets, separating them by 6 mm, and a plastic grating with multiple cells was placed beneath the G-MN to provide a climbing path (54 mm in height) to the arcing sites of the apparatus for adult houseflies emerging on the soil surface. Houseflies that climbed up the wall of the grating and reached the arcing zone were subjected to arc-discharge exposure from the NC-MN and thrown down onto the soil by the impact of the arcing. The impact was destructive enough to kill the houseflies. The structure of this apparatus is very safe and simple, enabling ordinary greenhouse workers to fabricate or improve it according to their own requirements. This study developed a simple and safe tool that provides a physical method to manage houseflies.
  • Yoshinori Matsuda; Hideyoshi Toyoda
    Agronomy MDPI AG 13 (1) 125 - 125 2023/01 [Refereed]
     
    Two new electrostatic devices were developed to manage greenhouse insect pests. One was an electrostatic insect catcher (EIC) to trap small flying pests, and the other was an arc-discharge zapper (ADZ) to kill larger insects emerging from soil beds. The EIC consisted of negatively charged insulated conductor plates (NIPs) and grounded conductor plates (GCPs), which were alternately arrayed in parallel at defined intervals. The ADZ had the same framework as the EIC, except that the NIPs were replaced with negatively charged non-insulated iron plates (NNPs). The EIC formed a non-discharging electric field between the NIP and GCP to create an attractive force to capture insects. By contrast, the ADZ formed a discharge-generating electric field between the NNP and GCP that killed insects. The EIC was effectively applied to small pests, such as whiteflies, thrips, leaf miners, winged aphids, and shore flies, that can pass through the conventional insect-proof nets installed on greenhouse windows. The ADZ was effective for adult houseflies emerging from pupae in soil beds. Our electrostatic devices are useful for controlling insect pests of different sizes.
  • Shin-ichi Kusakari; Yoshinori Matsuda; Hideyoshi Toyoda
    Agronomy MDPI AG 13 (1) 23 - 23 2023/01 [Refereed]
     
    This article reviews the development of electrostatic apparatuses for controlling insect pests in greenhouses. The apparatuses control insects by repelling them, capturing them, and killing them by producing an arc discharge. The single-charged dipolar electric field screen (SD screen) repels insects due to insects’ inherent avoidance behavior toward entering the electric field produced. As this behavior is common to many insect pests, the SD screen effectively prevents many pests from entering a greenhouse. The double-charged dipolar electric field screen (DD screen) has a strong attractive force that captures insects entering its electric field. The DD screen is useful for capturing small insects that pass through a conventional insect net, and unique derivatives of this screen have been invented to trap various insect pests on-site in a greenhouse. An arc-discharge exposer was used as a soil cover to kill adult houseflies that emerged from underground pupae transferred along with cattle manure used for soil fertilization. The houseflies were subjected to arc discharge when they appeared at the soil surface. These apparatuses have the common characteristic of a simple structure, so ordinary workers can be encouraged to fabricate or modify them based on their own needs. This review provides an experimental basis for designing efficient physical measures for controlling insect pests in greenhouses.
  • Yoshihiro Takikawa; Yoshinori Matsuda; Koji Kakutani; Teruo Nonomura; Hideyoshi Toyoda
    Horticulturae MDPI AG 8 (9) 764 - 764 2022/08 [Refereed]
     
    An unattended pest control system was developed to eliminate whiteflies (Bemisia tabaci) that settled on greenhouse tomato plants. The system exploited the whitefly’s habit of flying up from a plant that was mechanically tapped and then heading toward yellow objects. Remote-controlled dollies with arms that tapped plants and yellow-colored double-charged dipolar electric field screens (YDD-EFSs) (oppositely electrified transparent insulator tubes filled with yellow-colored water) attracted and trapped the whiteflies. The whiteflies flew up when the plants were mechanically tapped with the dolly’s arms during reciprocating movements and were subsequently trapped by YDD-EFSs that were automatically translocated to the target plants. The system was applied to rows of whitefly-infested tomato plants. Almost all whiteflies transferred to plants were successfully recovered by two dollies moving on either side of the plants, approaching all plants individually (via programmed movement). In summary, we present an efficient unattended method for controlling whiteflies on tomato plants in greenhouses.
  • Koji Kakutani; Yoshinori Matsuda; Teruo Nonomura; Hideyoshi Toyoda
    Horticulturae MDPI AG 8 (6) 543 - 543 2022/06 [Refereed]
     
    Electrostatic devices generating an electric field (EF) are promising tools for greenhouse tomato cultivation. In these devices, an EF is generated in the space surrounding an insulated conductor (IC) that is charged by a voltage generator. Thus, a physical force is exerted on any insect that enters the EF, as a negatively charged IC (NC-IC) pushes a negative charge (free electrons) out of the insect body. The insect is polarized positively to be attracted to the NC-IC, and a grounded metal net (G-MN) repels the insect. This dual function of the apparatus (insect capture and repulsion) is the core of the electrostatic pest-exclusion strategy. In this study, we applied various innovative EF-based devices to evaluate their efficacy in greenhouse tomato cultivation. Our objective was to determine the optimal apparatus for simple, inexpensive construction by greenhouse workers. The results of this study will contribute to the development of sustainable pest-management protocols in greenhouse horticulture.
  • Yoshinori Matsuda; Teruo Nonomura; Hideyoshi Toyoda
    Insects MDPI AG 13 (3) 253 - 253 2022/03 [Refereed]
     
    In the present study, we analyzed negative electricity released from insects captured by an electric field (EF)-producing apparatus. Adult houseflies (Musca domestica) were used as the model insect. The EF producer consisted of a negatively charged polyvinyl chloride membrane-insulated iron plate (N-PIP) and a non-insulated grounded iron plate (GIP) paralleled with the N-PIP. An EF was formed in the space between the plates. A housefly placed on the GIP was physically attracted to the N-PIP, and electricity released from the fly was detected as a specific transient electric current at the time of attraction and during subsequent confinement of the fly to the N-PIP. The magnitude of the insect-derived electric current became larger as the voltage applied to the N-PIP increased. We determined the total amount of electric current and confinement time within the apparatus necessary to kill all captured flies. These results demonstrate the insecticidal function and insect-capturing ability of the EF-producing apparatus.
  • Yoshihiro Takikawa; Teruo Nonomura; Takahiro Sonoda; Yoshinori Matsuda
    Insects MDPI AG 12 (11) 960 - 960 2021/10 [Refereed]
     
    Our aim was to develop an electrostatic apparatus to lure and capture silverleaf whiteflies (Bemisia tabaci), vegetable leafminers (Liriomyza sativae), and western flower thrips (Frankliniella occidentalis) that invade tomato greenhouses. A double-charged dipolar electric field producer (DD-EFP) was constructed by filling water in two identical transparent soft polyvinyl chloride tubes arrayed in parallel with fixed separation, and then, inserting the probes of grounded negative and positive voltage generators into the water of the two tubes to generate negatively and positively charged waters, respectively. These charged waters electrified the outer surfaces of the opposite tubes via dielectric polarization. An electric field formed between the oppositely charged tubes. To lure these phototactic insects, the water was colored yellow using watercolor paste, then introduced into the transparent insulator tubes to construct the yellow-colored DD-EFP. This apparatus lured insects in a manner similar to commercially available yellow sticky traps. The yellow-colored DD-EFP was easily placed as a movable upright screen along the plants, such that invading pests were preferentially attracted to the trap before reaching the plants. Furthermore, pests settling on the plants were attracted to the apparatus, which used a plant-tapping method to drive them off the plants. Our study provided an experimental basis for developing an electrostatic device to attract and capture insects that enter greenhouses.
  • Yoshinori Matsuda; Teruo Nonomura; Hideyoshi Toyoda
    Insects MDPI AG 12 (7) 621 - 621 2021/07 [Refereed]
     
    This study analysed the mechanism of avoidance behaviour by adult Turkestan cockroaches (Shelfordella lateralis Walker) in response to a static electric field (S-EF) formed in the space between a negatively charged polyvinyl chloride-insulated iron plate (N-PIP) and a grounded metal net (G-MN). The negative surface charge supplied to the iron plate by a voltage generator caused the G-MN to polarise positively via electrostatic induction. In the S-EF, the negative charge of the N-PIP created a repulsive force that pushed free electrons in the field toward the ground via the G-MN. When insects released in the space surrounded by the S-EF inserted their antennae into the S-EF, they pulled them back reflexively and moved backward. The analysis indicated that an electric current flowed transiently toward the ground when an insect inserted its antennae into the S-EF. The insect became positively charged via this discharge and was attracted to the opposite pole (N-PIP). In response to this attractive force, the insect pulled its antennae back quickly. The positive electrification caused by the removal of free electrons from the antenna tip triggered the avoidance behaviour.
  • Koji Kakutani; Yoshihiro Takikawa; Yoshinori Matsuda
    Insects MDPI AG 12 (6) 522 - 522 2021/06 [Refereed]
     
    We developed an arc discharge exposer (ADE) that kills rice weevils nesting in dried rice. The ADE features multiple identical metal plates, half of these are linked to a voltage generator and the others are grounded. The plates were arrayed in parallel and an electric field formed between them. Any insect entering the field was arced from the negatively charged plate and killed. The ADE was placed on a vessel containing pest-infested rice grains; the insects were lured out of the grains by mechanically vibrating the vessel. When rice grains move, insects tend to climb upward, thus, the weevils were effectively removed. Our electrostatic apparatus is easy to construct and could be used to control pests in stored rice.
  • Koji Kakutani; Yoshinori Matsuda; Teruo Nonomura; Yoshihiro Takikawa; Takeshi Takami; Hideyoshi Toyoda
    International Journal of Environmental Research and Public Health MDPI AG 18 (9) 4934 - 4934 2021/05 [Refereed]
     
    The purpose of this study was to develop a simple electrostatic apparatus to precipitate virus particles spread via droplet transmission, which is especially significant in the context of the recent coronavirus disease 2019 (COVID-19) pandemic. The bacteriophage φ6 of Pseudomonas syringae was used as a model of the COVID-19 virus because of its similar structure and safety in experiments. The apparatus consisted of a spiked, perforated stainless plate (S-PSP) linked to a direct-current voltage generator to supply negative charge to the spike tips and a vessel with water (G-water) linked to a ground line. The S-PSP and G-water surface were paralleled at a definite interval. Negative charge supplied to the spike tips positively polarised the G-water by electrostatic induction to form an electric field between them in which ionic wind and negative ions were generated. Bacteriophage-containing water was atomised with a nebuliser and introduced into the electric field. The mist particles were ionised by the negative ions and attracted to the opposite pole (G-water). This apparatus demonstrated a prominent ability to capture phage-containing mist particles of the same sizes as respiratory droplets and aerosols regardless of the phage concentration of the mist particles. The trapped phages were successfully sterilised using ozone bubbling. Thus, the present study provides an effective system for eliminating droplet transmission of viral pathogens from public spaces.
  • Koji Kakutani; Yoshinori Matsuda; Teruo Nonomura; Yoshihiro Takikawa; Kazumi Osamura; Hideyoshi Toyoda
    Agriculture MDPI AG 11 (2) 176 1 - 176 16 2021/02 [Refereed]
     
    The purpose of the study was to construct an electrostatic insect-capturing apparatus that could be applied to a drone (quadcopter). For this purpose, a double-charged dipolar electric field screen (DD-screen) was constructed using oppositely charged insulator tubes that was then attached to a drone. For charging, the inner surface of the tubes was coated with a conductive paste and then linked to a negative or positive voltage generator. The opposite charges of the tubes formed an electric field between them and created an attractive force to capture insects that entered the field. The DD-screen constructed here was sufficiently light to enable its attachment to a drone. The screen was hung from the drone perpendicular to the direction of drone movement, so as to receive the longitudinal airflow produced by the movement of the drone. It was positioned 1.8 m below the drone body to avoid the influence of the downward slipstream generated by the rotating propellers. Eventually, the drone was able to conduct a stable flight, with sufficient endurance, and captured airborne insects carried by an airflow of 8 m/s during the flight. This study, therefore, provides an experimental basis for establishing a new method for conducting trap-based monitoring of airborne insects during remote-controlled flight through operation of a DD-screen attached to a drone.
  • Yoshinori Matsuda; Yoshihiro Takikawa; Koji Kakutani; Teruo Nonomura; Kiyotsugu Okada; Shin-ichi Kusakari; Hideyoshi Toyoda
    Agriculture 10 600 1 - 600 12 2020/12 [Refereed]
  • Yoshinori Matsuda; Kunihiko Shimizu; Takahiro Sonoda; Yoshihiro Takikawa
    Insects MDPI AG 11 (12) 861 1-15 - 861 1-15 2020/12 [Refereed]
     
    An electrostatic apparatus was developed to control weeds and houseflies emerging from ground soil in a greenhouse simultaneously. Identical iron plates were placed in parallel at a defined interval and fixed in an iron frame. Two sets of fixed iron plates were used, one for weed control and one for fly control. For weed control, all of the iron plates were negatively charged, and negative charges accumulated on the plates were released to weed shoots through arc discharge. Houseflies were introduced into the space between the negatively charged and grounded plates, then subjected to arc discharge from the charged plates. Both plant shoots and adult houseflies are electrically conductive; thus, they were killed by discharge-exposure in the electric field between the charged iron plate and the ground soil, and between the charged and grounded plates, respectively. In practical use, these two devices were assembled as a two-level apparatus for simultaneous control of both targets. Several apparatuses were linked together, which increased the total electricity charge on the plates and produced a stronger discharge force sufficient to kill all targets. Thus, this study provides an electrostatics-based pest-control method for pesticide-independent greenhouse farming.
  • Yoshinori Matsuda; Yoshihiro Takikawa; Koji Kakutani; Teruo Nonomura; Hideyoshi Toyoda
    Insects MDPI AG 11 (3) 187 1 - 187 10 2020/03 [Refereed]
     
    The present study was conducted to establish an electrostatic-based experimental system to enable new investigations of insect behavior. The instrument consists of an insulated conducting copper ring (ICR) linked to a direct current voltage generator to supply a negative charge to an ICR and a grounded aluminum pole (AP) passed vertically through the center of the horizontal ICR. An electric field was formed between the ICR and the AP. Rice weevil (Sitophilus oryzae) was selected as a model insect due to its habit of climbing erect poles. The electric field produced a force that could be imposed on the insect. In fact, the negative electricity (free electrons) was forced out of the insect to polarize its body positively. Eventually, the insect was attracted to the oppositely charged ICR. The force became weaker on the lower regions of the pole; the insects sensed the weaker force with their antennae, quickly stopped climbing, and retraced their steps. These behaviors led to a pole-ascending–descending action by the insect, which was highly reproducible and precisely corresponded to the changed expansion of the electric field. Other pole-climbing insects including the cigarette beetle (Lasioderma serricorne), which was shown to adopt the same behavior.
  • Yoshihiro Takikawa; Yoshinori Matsuda; Teruo Nonomura; Koji Kakutani; Shin-ichi Kusakari; Hideyoshi Toyoda
    Journal of Agricultural Science 12 (2) 50 - 60 1916-9752 2020/01 [Refereed]
  • Takikawa Y; Kakutani K; Matsuda Y; Nonomura T; Kusakari S; Toyoda H
    Journal of Agricultural Science 11 (18) 1 - 20 1916-9752 2019/11 [Refereed]
  • MATSUDA Yoshinori
    Instruments MDPI AG 2 (3) 13 - 22 2018/07 [Refereed]
     
    An electrostatic apparatus was constructed to capture tobacco sidestream smoke. This apparatus consisted of a perforated polypropylene plate with metal spikes and a grounded metal net arrayed in parallel at a defined interval. Spikes were negatively charged to positively polarize the net and an electric field was formed between the opposite charges of the spike tips and the grounded net. Discharge from the spike tips occurred, which depended on the pole distance and the voltage applied to the spikes. At lower voltages (<12.1 kV) that do not cause arc discharge from the tips, a corona discharge occurred with the generation of an ionic wind from the spiked plate to the net. This discharge increased in direct proportion to the applied voltage and relative humidity, while a larger corona discharge generated a stronger ionic wind. The ionic wind involved negative ions and the number of negative ions in the wind increased with increasing applied voltage. The optimal voltage (10 kV) generated sufficient negative ions to ionize smoke particles in the electric field, before the ionized smoke particles were successfully captured by the oppositely charged metal net. Thus, this study provides an experimental basis for the practical application of an electrostatic-based method to prevent the production of tobacco sidestream smoke that leads to passive smoking by non-smokers.
  • Kakutani K; Matsuda Y; Nonomura T; Takikawa Y; Okada K; Shibao M; Miyama K; Yokoo S; Kusakari S; Toyoda H
    International Journal of Scientific Research 7 (5) 47 - 50 2277-8179 2018/06 [Refereed]
  • Matsuda Y; Kakutani K; Nonomura T; Takikawa Y; Okada K; Shibao M; Miyama K; Yokoo S; Kusakari S; Toyoda H
    Journal of Food Technology and Preservation 2 (1) 15 - 20 2018/04 [Refereed]
  • Matsuda Y; Toyoda H
    Open Access Journal of Science 2 (5) 337 - 353 2018/01 [Refereed]
     
    Review
  • MATSUDA Yoshinori
    International Journal of Current Advanced Research 6 (8) 5517 - 5521 2319-6475 2017/08 [Refereed]
  • MATSUDA Yoshinori
    International Journal of Scientific Research in Environmental Sciences 5 (1) 17 - 23 2322-4983 2017/02 [Refereed]
     
    Our primary concern of air pollution problem was a transboundary movement of air pollutants to Japan from the pollutant generating country because of geographic and meteorological reasons. An electrostatic air purification screen (EAPS) was devised to capture particulate matter (PM) in smoke, which can cause health problems. The EAPS consisted of three layers of insulated conductor wires (ICWs) and two voltage generators that supplied negative charges to the two outer ICW layers and a positive charge to the middle ICW layer. The ICWs generated an attractive force that captured fine particles in the smoke passing through the EAPS. The attractive force was directly proportional to the applied voltage. At ≥4.5 kV, the EAPS exerted sufficient force to capture all fine particles carried in an airflow of 3 m/s. Results showed that the electrostatic barrier that formed inside the EAPS was effective at capturing PM, allowing homes and working environments to remain PM-free and healthy, despite continuous PM exposure.
  • Yoshihiro Takikawa; Yoshinori Matsuda; Teruo Nonomura; Koji Kakutani; Shin-Ichi Kusakari; Hideyoshi Toyoda
    INTERNATIONAL JOURNAL OF ENVIRONMENTAL RESEARCH AND PUBLIC HEALTH MDPI AG 14 (1) 1 - 5 1660-4601 2017/01 [Refereed]
     
    An electrostatic-barrier-forming window (EBW) was devised to capture airborne pollen, which can cause allergic pollinosis. The EBW consisted of three layers of insulated conductor wires (ICWs) and two voltage generators that supplied negative charges to the two outer ICW layers and a positive charge to the middle ICW layer. The ICWs generated an attractive force that captured pollen of the Japanese cedar, Cryptomeria japonica, from air blown through the EBW. The attractive force was directly proportional to the applied voltage. At >= 3.5 kV, the EBW exerted sufficient force to capture all pollen carried at an air flow of 3 m/s, and pollen-free air passed through the EBW. The findings demonstrated that the electrostatic barrier that formed inside the EBW was very effective at capturing airborne pollen; thus, it could allow a home to remain pollen-free and healthy despite continuous pollen exposure.
  • MATSUDA Yoshinori
    Journal of Agricultural Science 8 (4) 13 - 25 1916-9752 2016/03 [Refereed]
     
    The electrostatic nursery shelter reported in this work was a transparent film-covered rectangular box with three electric field screens on each of the long sides of the box. This arrangement prevents flying pests and airborne fungal pathogens from entering the nursery space. Insulated conducting wires (ICWs) were used as electrodes to create electric fields. The ICWs were arrayed in parallel, and linked to direct-current voltage sources. The ICW layers were negatively or positively charged with equal voltages to form dipoles; i.e., ICW(–) and ICW(+). The electric field screen consisted of three layers of the ICWs; i.e., an ICW(–) layer on either side of an ICW(+) layer. Four species of major tomato pests were used in a blowing assay: whiteflies (Bemisia tabaci), western flower thrips (Frankliniella occidentalis), green peach aphids (Myzus persicae) and tomato leaf miner flies (Liriomyza sativae). The ICWs were located to capture test pests that were mechanically blown into the electric-field screen. The electrostatic force to capture the insects was directly proportional to the applied voltage, and at voltages of 1.2 kV or greater, the screen exerted sufficient force to capture all of the test pests. An assay in a pest-infested greenhouse revealed that the ICWs captured all the pests that approached the screen, and the plants within the shelter remained pest-free. In addition, we show that the electric-field-screened shelter remained spore-free in the presence of continuous exposure to the conidia of tomato powdery mildew (Oidium neolycopersici).
  • MATSUDA Yoshinori
    Int. J. Adv. Agric. Res. 3 55 - 63 2053-1265 2015/10 [Refereed]
     
    Reliable information of pests dwelling in a warehouse is a prerequisite to construct an effective and realistic pest control strategy. Our new electrostatic pest-monitoring apparatus is presented for this purpose. The apparatus consists of an insect-discharge detector and electrostatic insect trap. The detector is an iron plate attached to a direct current (DC) positive voltage generator and the insect trap consists of a pair of opposite poles. Pests on the detector plate are stripped of free electrons and charged positively. This is recorded as a transient discharge signal from the insect, and the positively charged pests are trapped at the negative pole. Discharge signals from different checkpoints in a warehouse were monitored automatically and continuously to analyze the temporal and spatial movements of the pests. This monitoring system enabled us to apply most effective chemical and physical methods to the control of cigarette beetles dwelling in our warehouse.
  • Yoshihiro Takikawa; Yoshinori Matsuda; Koji Kakutani; Teruo Nonomura; Shin-Ichi Kusakari; Kiyotsugu Okada; Junji Kimbara; Kazumi Osamura; Hideyoshi Toyoda
    Insects MDPI AG 6 (2) 442 - 454 2075-4450 2015/05 [Refereed]
     
    Our greenhouse tomatoes have suffered from attacks by viruliferous whiteflies Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) over the last 10 years. The fundamental countermeasure was the application of an electric field screen to the greenhouse windows to prevent their entry. However, while the protection was effective, it was incomplete, because of the lack of a guard at the greenhouse entrance area in fact, the pests entered from the entrance door when workers entered and exited. To address this, we developed a portable electrostatic insect sweeper as a supplementary technique to the screen. In this sweeper, eight insulated conductor wires (ICWs) were arranged at constant intervals along a polyvinylchloride (PVC) pipe and covered with a cylindrical stainless net. The ICWs and metal net were linked to a DC voltage generator (operated by 3-V alkaline batteries) inside the grip and oppositely electrified to generate an electric field between them. Whiteflies on the plants were attracted to the sweeper that was gently slid along the leaves. This apparatus was easy to operate on-site in a greenhouse and enabled capture of the whiteflies detected during the routine care of the tomato plants. Using this apparatus, we caught all whiteflies that invaded the non-guarded entrance door and minimized the appearance and spread of the viral disease in tomato plants in the greenhouse.
  • Y. Matsuda; T. Nonomura; K. Kakutani; J. Kimbara; K. Osamura; S. Kusakari; H. Toyoda
    ELECTROSTATICS 2015 IOP PUBLISHING LTD 646 012003-1 - 012003-4 1742-6588 2015 [Refereed]
     
    An electric field screen is a physical device used to exclude pest insects from greenhouses and warehouses to protect crop production and storage. The screen consists of iron insulated conductor wires (ICWs) arrayed in parallel and linked to each other, an electrostatic DC voltage generator used to supply a negative charge to the ICWs, and an earthed stainless net placed on one side of the ICW layer. The ICW was negatively charged to polarize the earthed net to create a positive charge on the ICW side surface, and an electric field formed between the opposite charges of the ICW and earthed net. The current study focused on the ability of the screen to repel insects reaching the screen net. This repulsion was a result of the insect's behaviour, i.e., the insects were deterred from entering the electric field of the screen. In fact, when the screen was negatively charged with the appropriate voltages, the insects placed their antennae inside the screen and then flew away without entering. Obviously, the insects recognized the electric field using their antennae and thereby avoided entering. Using a wide range of insects and spiders belonging to different taxonomic groups, we confirmed that the avoidance response to the electric field was common in these animals.
  • Y. Matsuda; K. Kakutani; T. Nonomura; J. Kimbara; K. Osamura; S. Kusakar; H. Toyoda
    ELECTROSTATICS 2015 IOP PUBLISHING LTD 646 012002-1 - 012002-4 1742-6588 2015 [Refereed]
     
    An electric field screen can be used to keep mosquitoes out of houses with open windows. In this study, doubly charged dipolar electric field screens (DD-screens) were used to capture mosquitoes entering through a window. The screen had two components: three layers of insulated conductor iron wires (ICWs) in parallel arrays and two electrostatic direct current (DC) voltage generators that supplied negative or positive voltages to the ICWs. Within each layer, the ICWs were parallel at 5-mm intervals, and connected to each other and to a negative or positive voltage generator. The negatively and positively charged ICWs are represented as ICW(-) and ICW(+), respectively. The screen consisted of one ICW(+) layer with an ICW(-) layer on either side. The Asian tiger mosquito (Aedes albopictus) and house mosquito (Culex pipiens) were used as models of vectors carrying viral pathogens. Adult mosquitoes were blown into the space between the ICWs by sending compressed air through the tip of an insect aspirator to determine the voltage range that captured all of the test insects. Wind speed was measured at the surface of the ICW using a sensitive anemometer. The result showed that at >= 1.2 kV, the force was strong enough that the ICWs captured all of the mosquitoes, despite a wind speed of 7 m/s. Therefore, the DD-screen could serve as a physical barrier to prevent noxious mosquitoes from entering houses with good air penetration.
  • Yoshihiro Takikawa; Yoshinori Matsuda; Teruo Nonomura; Koji Kakutani; Junji Kimbara; Kazumi Osamura; Shin-ichi Kusakari; Hideyoshi Toyoda
    AEROBIOLOGIA SPRINGER 30 (4) 435 - 444 0393-5965 2014/12 [Refereed]
     
    Old books are highly susceptible to mould infection, and an effective method for avoiding moulding is needed to safely preserve valuable books in library stack rooms. Guarding a bookshelf with an electric field screen is a physical method that prevents airborne spores from entering the space used for book preservation. In this study, insulated conductor wires (ICWs) were used as electrodes to form electric fields. The ICWs were arrayed in parallel and linked to each other and to a direct current voltage generator. The electric field screen consisted of two layers of ICWs, which were negatively and positively charged with equal voltages to make dipoles, ICW(-) and ICW(+). Both ICWs generated an attractive force that captured airborne spores of Penicillium digitatum that were blown inside the screen. The attractive force was directly proportional to the applied voltage. At a parts per thousand 0.9 kV, the screen exerted sufficient force to capture all airflow-carried spores, but a few spores that were once captured were repulsed out of the electric field when subsequent spores were attracted to positions proximal to them. This phenomenon was explained by creeping discharge between spores located close to each other on the ICW surface. This spore-repulsion problem was resolved by adding an additional ICW layer to the electric field screen, namely an electric field screen with an ICW(-) layer on both sides of an ICW(+) layer. The present study demonstrated that the three-layered electric field screen remained mould-free inside a screen-guarded bookshelf, irrespective of continuous spore exposure.
  • MATSUDA Yoshinori
    Journal of Agricultural Science Canadian Center of Science and Education 6 (12) 1 - 13 1916-9752 2014/11 [Refereed]
     
    Two types of an electric field screen were used to exclude whiteflies from a greenhouse. Singly charged dipolar electric field screen had insulated conductor iron wires arrayed in parallel (ICW-layer), two earthed metal nets on both sides of the ICW-layer, and a direct current voltage generator. Screens were attached to the lateral windows of the greenhouse to repel whiteflies that approached the nets. To electrostatically guard the greenhouse entrance, doubly charged dipolar electric field screens (DD-screens) were used to capture whiteflies entering through the door. The ICWs, oppositely charged with equal voltages, were arrayed one after the other, and an insulator board or net was placed on one side of the ICW-layer. ICWs captured whiteflies entering an electric field of DD-screens. A small pre-entrance room was constructed at the entrance area, and three DD-screens (with yellow and gray board or gray net) were installed in the pre-entrance room taking into consideration the airflow inside the room, as most whiteflies were brought in by the air when the door was opened. Two DD-screens with the board were useful for directing the wind toward the wall into which the netted DD-screen was integrated. Insects were eliminated by this screen, and the pest-free air was circulated inside the greenhouse. The yellow-boarded DD-screen was highly attractive because of the photo-selective movement of the whiteflies and was effective for trapping the whiteflies when there was no wind. Our electrostatic method is effective at keeping the greenhouse pest-free throughout the entire period of tomato cultivation
  • Teruo Nonomura; Yoshinori Matsuda; Koji Kakutani; Junji Kimbara; Kazumi Osamura; Shin-ichi Kusakari; Hideyoshi Toyoda
    JOURNAL OF ELECTROSTATICS ELSEVIER SCIENCE BV 72 (1) 1 - 5 0304-3886 2014/02 [Refereed]
     
    A simple electrostatic apparatus was devised to measure dischargeable electricity and bioelectric potentials produced by flies. The apparatus involved two insulated electrodes, ICW(-) and ICW(+), oppositely charged with equal voltages supplied by two voltage-generators. In the electric field, the flies became net positive by instantaneously discharging their electricity and were attracted to negative surface charges on ICW(-). The tail-lifting movement by the attracted insect was an action creating electric potentials that could cause discharge of ICW(-). The discharge transiently appeared in response to individual movements and was larger when the tail was lifted at higher angles. (C) 2013 Elsevier B.V. All rights reserved.
  • T. Nonomura; Y. Matsuda; Y. Takikawa; K. Kakutani; H. Toyoda
    Ann. Rept. Kansai Pl. Prot 56 17 - 20 2014 [Refereed]
  • Yoshinori Matsuda; Koji Kakutani; Teruo Nonomura; Junji Kimbara; Shin-ichi Kusakari; Kazumi Osamura; Hideyoshi Toyoda
    JOURNAL OF APPLIED PHYSICS AMER INST PHYSICS 112 (11) 0021-8979 2012/12 [Refereed]
     
    An electric field screen was constructed to examine insect attraction mechanisms in multiple electric fields generated inside the screen. The screen consisted of two parallel insulated conductor wires (ICWs) charged with equal but opposite voltages and two separate grounded nets connected to each other and placed on each side of the ICW layer. Insects released inside the fields were charged either positively or negatively as a result of electricity flow from or to the insect, respectively. The force generated between the charged insects and opposite ICW charges was sufficient to capture all insects. (C) 2012 American Institute of Physics. [http://dx.doi.org/10.1063/1.4767635]
  • Teruo Nonomura; Yoshinori Matsuda; Koji Kakutani; Junji Kimbara; Kazumi Osamura; Shin-ichi Kusakari; Hideyoshi Toyoda
    EUROPEAN JOURNAL OF PLANT PATHOLOGY SPRINGER 134 (4) 661 - 670 0929-1873 2012/12 [Refereed]
     
    Dual functions (insect repelling and capturing) of a single-charged dipolar electric field screen were evaluated to successfully exclude whiteflies from a window-open greenhouse. The screen consisted of three parts: 1) insulated conductor wires (ICWs) arrayed in parallel at 5 mm intervals, 2) two earthed stainless nets placed within 3 mm of both sides of the ICW layer, and 3) a voltage generator for the negatively charged ICWs. The screen formed two electric fields between the ICW-layer and the ICW-side surface of the earthed net and between the ICWs. At negative charging of 1.5-2.5 kV, all whiteflies reaching the outer surface of the screen net avoided entering the electric field and flew away from the screen. This avoidance was disturbed by 3 m s(-1) wind, as the insects were compulsorily blown inside. However, almost all whiteflies (99.4 %) were captured with the ICW. These results indicate that the insect-capturing function is effective to complement a failure to repel. A greenhouse assay was conducted in the screen-attached and non-screened parts in which a greenhouse was divided with a partition. During the 3-month operation, the screen was durable and functional for excluding pests, and better air ventilation changed the climate conditions in the greenhouse. Thus, the present study demonstrated that our electric field screen can provide an airy condition for tomatoes in a window-open greenhouse and successfully exclude whiteflies using dual screen functions.
  • Teruo Nonomura; Yoshinori Matsuda; Koji Kakutani; Junji Kimbara; Kazumi Osamura; Shin-ichi Kusakari; Hideyoshi Toyoda
    EUROPEAN JOURNAL OF PLANT PATHOLOGY SPRINGER 134 (4) 661 - 670 0929-1873 2012/12 [Refereed]
     
    Dual functions (insect repelling and capturing) of a single-charged dipolar electric field screen were evaluated to successfully exclude whiteflies from a window-open greenhouse. The screen consisted of three parts: 1) insulated conductor wires (ICWs) arrayed in parallel at 5 mm intervals, 2) two earthed stainless nets placed within 3 mm of both sides of the ICW layer, and 3) a voltage generator for the negatively charged ICWs. The screen formed two electric fields between the ICW-layer and the ICW-side surface of the earthed net and between the ICWs. At negative charging of 1.5-2.5 kV, all whiteflies reaching the outer surface of the screen net avoided entering the electric field and flew away from the screen. This avoidance was disturbed by 3 m s(-1) wind, as the insects were compulsorily blown inside. However, almost all whiteflies (99.4 %) were captured with the ICW. These results indicate that the insect-capturing function is effective to complement a failure to repel. A greenhouse assay was conducted in the screen-attached and non-screened parts in which a greenhouse was divided with a partition. During the 3-month operation, the screen was durable and functional for excluding pests, and better air ventilation changed the climate conditions in the greenhouse. Thus, the present study demonstrated that our electric field screen can provide an airy condition for tomatoes in a window-open greenhouse and successfully exclude whiteflies using dual screen functions.
  • Koji Kakutani; Yoshinori Matsuda; Kayo Haneda; Dai Sekoguchi; Teruo Nonomura; Junji Kimbara; Kazumi Osamura; Shin-ichi Kusakari; Hideyoshi Toyoda
    JOURNAL OF ELECTROSTATICS ELSEVIER SCIENCE BV 70 (2) 207 - 211 0304-3886 2012/04 [Refereed]
     
    An insulated conductor wire (ICW) paralleled with an earthed net was used to observe movements by vinegar flies in relation to their electricity release. ICW was negatively charged to create a positive charge on the net. At particular voltages, flies were attracted to ICW. This attraction was triggered by the deprivation of the insect negative charge with the net. Eventually the insects became net positive and were drawn to the ICW negative charge. The attracted insects generated bioelectricity through skeletal muscular movements. However, the electricity produced was depleted by the net without neutralizing their positive charge in the insect body. (c) 2012 Elsevier B.V. All rights reserved.
  • MATSUDA Yoshinori
    Journal of Agricultural Science Canadian Center of Science and Education 5 (5) 51 - 60 1916-9752 2012/04 [Refereed]
  • Koji Kakutani; Yoshinori Matsuda; Kayo Haneda; Dai Sekoguchi; Teruo Nonomura; Junji Kimbara; Kazumi Osamura; Shin-ichi Kusakari; Hideyoshi Toyoda
    JOURNAL OF ELECTROSTATICS ELSEVIER SCIENCE BV 70 (2) 207 - 211 0304-3886 2012/04 [Refereed]
     
    An insulated conductor wire (ICW) paralleled with an earthed net was used to observe movements by vinegar flies in relation to their electricity release. ICW was negatively charged to create a positive charge on the net. At particular voltages, flies were attracted to ICW. This attraction was triggered by the deprivation of the insect negative charge with the net. Eventually the insects became net positive and were drawn to the ICW negative charge. The attracted insects generated bioelectricity through skeletal muscular movements. However, the electricity produced was depleted by the net without neutralizing their positive charge in the insect body. (c) 2012 Elsevier B.V. All rights reserved.
  • K. Kakutani; Y. Matsuda; T. Nonomura; H. Toyoda; J. Kimbara; S. Kusakari
    INTERNATIONAL SYMPOSIUM ON ADVANCED TECHNOLOGIES AND MANAGEMENT TOWARDS SUSTAINABLE GREENHOUSE ECOSYSTEMS: GREENSYS2011 INT SOC HORTICULTURAL SCIENCE 952 559 - 566 0567-7572 2012 [Refereed]
     
    In an attempt to control insect pests affecting greenhouse tomatoes, we evaluated an electric field screen to create an airy greenhouse condition that successfully excluded insect vectors (whiteflies, green peach aphids, western flower thrips, shore flies) of pathogens. The screen consisted of three parts: 1) insulated conductor wires (ICWs) arrayed in parallel at 5-mm intervals, 2) two stainless-steel nets that were grounded and placed on both sides of the ICWs, and 3) a DC voltage generator to negatively charge the ICWs. An electric field formed between the negative surface charge of the ICWs and the positive charge on the ICW-side surface of the grounded net. The ICWs captured insects that entered the field. Insects that contacted the outer surface of the screen net avoided the electric field and flew away from the screen. During continuous 3-month greenhouse operation, the screen was durable and functional in exerting stable pest exclusion and good air penetration for ventilation under changing greenhouse climate conditions. Thus, our electric field screen provided an airy condition for tomatoes in an open-window greenhouse that successfully excluded flying insect pests.
  • K. Kakutani; Y. Matsuda; K. Haneda; T. Nonomura; J. Kimbara; S. Kusakari; K. Osamura; H. Toyoda
    ANNALS OF APPLIED BIOLOGY WILEY-BLACKWELL 160 (3) 250 - 259 0003-4746 2012 [Refereed]
     
    An electric field screen (EF-screen) is a physical device for excluding pest insects from greenhouses and warehouses to protect crops during their production and storage periods. In this study, a simple version of the EF-screen, an insulated conductor iron wire (ICW) paralleled to an earthed net, was constructed to effectively observe the attraction of test insects in relation to their electricity release. The ICW was negatively charged to dielectrically polarise the insulator sleeve of the ICW: negatively on the outer surface and positively on the inner conductor wire surface of the sleeve. The negative surface charge of the ICW caused an electrostatic induction in the earthed net and a resultant positive charge at the ICW-side surface of the net. An electric field formed between the ICW (negative pole) and earthed net (positive pole). Insects were attracted to the ICW when they were placed onto the earthed net. A vital step for the attraction was the creation of a transient bioelectric discharge from an insect. During this discharge, an electric charge of the insect was transferred to the earthed net. Eventually, the insect became net positive and was then attracted to the ICW. The magnitude of the current increased in direct proportion to the increase in voltage applied to the ICW, and the attraction force was directly proportional to the increase in the electric current. Larger voltages were necessary to attract much larger insects because larger insects were stronger and therefore more able to escape from the ICW attraction. Similar results were obtained for a wide range of pest insects belonging to different taxonomic groups (8 orders and 15 families). This study demonstrated that transient bioelectric discharge is common in insects and can be utilised to create an electrostatic force capable of moving insects in a generated electric field.
  • K. Kakutani; Y. Matsuda; K. Haneda; T. Nonomura; J. Kimbara; S. Kusakari; K. Osamura; H. Toyoda
    ANNALS OF APPLIED BIOLOGY WILEY-BLACKWELL 160 (3) 250 - 259 0003-4746 2012 [Refereed]
     
    An electric field screen (EF-screen) is a physical device for excluding pest insects from greenhouses and warehouses to protect crops during their production and storage periods. In this study, a simple version of the EF-screen, an insulated conductor iron wire (ICW) paralleled to an earthed net, was constructed to effectively observe the attraction of test insects in relation to their electricity release. The ICW was negatively charged to dielectrically polarise the insulator sleeve of the ICW: negatively on the outer surface and positively on the inner conductor wire surface of the sleeve. The negative surface charge of the ICW caused an electrostatic induction in the earthed net and a resultant positive charge at the ICW-side surface of the net. An electric field formed between the ICW (negative pole) and earthed net (positive pole). Insects were attracted to the ICW when they were placed onto the earthed net. A vital step for the attraction was the creation of a transient bioelectric discharge from an insect. During this discharge, an electric charge of the insect was transferred to the earthed net. Eventually, the insect became net positive and was then attracted to the ICW. The magnitude of the current increased in direct proportion to the increase in voltage applied to the ICW, and the attraction force was directly proportional to the increase in the electric current. Larger voltages were necessary to attract much larger insects because larger insects were stronger and therefore more able to escape from the ICW attraction. Similar results were obtained for a wide range of pest insects belonging to different taxonomic groups (8 orders and 15 families). This study demonstrated that transient bioelectric discharge is common in insects and can be utilised to create an electrostatic force capable of moving insects in a generated electric field.
  • Yoshinori Matsuda; Teruo Nonomura; Koji Kakutani; Yoshihiro Takikawa; Junji Kimbara; Yoshihiro Kasaishi; Kazumi Osamura; Shin-ichi Kusakari; Hideyoshi Toyoda
    CROP PROTECTION ELSEVIER SCI LTD 30 (2) 155 - 162 0261-2194 2011/02 [Refereed]
     
    A bifunctional electric field screen was proposed to physically exclude insect pests from warehouses. The screen consists of insulated iron wires (ICW) arranged in parallel and two earthed conductor nets placed on both sides of the ICW. A negative charge (0.1-8.0 kV) was applied to the insulated wires with a voltage generator to polarize an insulator sleeve used to cover the wire, negatively on the outer surface and positively on the inner conductor wire surface of the sleeve. The negative surface charge of the ICW caused an electrostatic induction in the earthed nets and an opposite charge on the net surfaces facing the ICW. An electric field formed in a space between the ICW and the earthed net, and the field strength increased in direct proportion to increasing voltages applied to the ICW. Adults of the test insects (cigarette beetle (Lasioderma serricorne) and vinegar fly (Drosophila melanogaster)) reaching the outer surface of the earthed net were deterred from entering the inside of the charged screen, whereas all insects immediately passed through the screen when the ICW was not charged. This avoidance was directly proportional to the increase in the voltage. In addition, the capability of the screen to capture insects that enter inside the screen was proven by introducing insects into the space between the ICW and the earthed net. Strong capture was observed when the ICW was negatively charged with more than 4.1 kV, under which conditions a short-term electric current (peaking at 0.3-0.6 mu A, for 3 min) occurred transiently. This electric current was due to the release of electricity from the insects, giving a net overall positive charge to the insects, which therefore were attracted more strongly to the negatively charged ICW. A test using an attractant-set chamber showed that the insects were completely prevented from passing through the charged screen, in contrast to a rapid transfer of all insects when the screen was not charged. Thus, the present results show that the described screen is a promising physical tool for controlling insect pests in warehouses. (C) 2010 Elsevier Ltd. All rights reserved.
  • 温室内で発生したトマト根腐萎凋病(Fusarium crown and root rot disease)の感染拡大に関わるミギワバエScatella stagnalis(Diptera: Ephydridae)の影響.
    松田克礼; 角谷晃司; 野々村照雄; 瀧川義浩
    近畿大学農学部紀要 近畿大学農学部 42 237 - 242 0453-8889 2009/03
  • T. Nonomura; Y. Matsuda; K. Kakutani; Y. Takikawa; H. Toyoda
    CANADIAN JOURNAL OF PLANT PATHOLOGY-REVUE CANADIENNE DE PHYTOPATHOLOGIE CANADIAN PHYTOPATHOLOGICAL SOC 30 (4) 517 - 524 0706-0661 2008/10 [Refereed]
     
    We devised a cylindrical electrostatic discharge generator to physically eradicate tomato powdery mildew colonizing tomato leaves. The generator consists of a copper needle with a pointed tip, an insulating acrylic cylinder, and an electrostatic voltage generator. The needle is insulated with a vinyl sleeve, except for the pointed tip, and is coaxially fixed in the cylinder and connected to the voltage generator. The needle is negatively charged, and the treated plant is earthed. In initial tests, a corona, characterized by a blue glow, formed at the needle tip as the probe was brought closer to the leaf surface. The distance at which this Occurred increased from 16 to 50 mm as the voltage was increased from 5 to 30 kV. If the probe was brought too close to the leaf surface, an arc discharge Occurred that caused injury to the leaf. Powdery mildew colonies were destroyed by 2-second exposures at probe distances intermediate to where corona discharge was initiated and where arcing Occurred. A probe distance of 25 mm and 30 kV for a 2-second burst was selected to further test the efficacy of the probe for controlling powdery mildew in a greenhouse environment. Tomato plants were grown hydroponically in two open-window greenhouses under a first-truss cropping system. Colonies appeared on tomato leaves 10 to 14 days after transplanting. During the following 2 weeks, these colonies produced abundant progeny conidia that secondarily infected neighboring plants. Corona discharge treatment in one greenhouse, at the stage when colonies first became visible, completely Suppressed the spread of the disease compared with a non-treated greenhouse in which disease spread rapidly. The present discharge generator is portable and easy to operate on-site as a part of routine care of hydroponically Cultured tomatoes in greenhouses and provides a non-chemical method to control powdery mildew disease.
  • Norio Tanaka; Yoshinori Matsuda; Eiko Kato; Kiyoto Kokabe; Takeshi Furukawa; Teruo Nonomura; Ken-ichiro Honda; Shin-ichi Kusakari; Takeo Imura; Junji Kimbara; Hideyoshi Toyoda
    CROP PROTECTION ELSEVIER SCI LTD 27 (2) 215 - 221 0261-2194 2008/02 [Refereed]
     
    A new electrostatic insect-proof screen (electric dipolar screen) was developed using insulated conductor wire. Copper conductor wire is encased in a flexible transparent vinyl insulator sleeve and charged with an electrostatic voltage generator. Paired insulated conductor wires were placed in parallel with 5 mm of separation and oppositely charged with 15 kV DC using separate electrostatic voltage generators. A negatively charged conductor wire polarizes the insulator negatively at the outer surface and positively on the conductor side, and a positively charged conductor wire polarizes vice versa. A pair of insulated conductor wires with opposite surface charges was used as an electric dipole. An electric dipole creates an electrostatic force from positive to negative poles. This force was harnessed to trap whiteflies entering greenhouses. Dipolar wires were attached in parallel to a frame to construct an electric dipolar screen that could be fitted to greenhouse windows. The screen prevented adult whiteflies from passing through spaces (up to 30 mm) between the wires of the screen, and tomato plants inside remained free of whiteflies throughout the entire 3-week experiment, in contrast to heavy infestation of all plants in the uncharged area. Thus, the electric dipolar screen is a promising tool to physically exclude flying whiteflies from greenhouses. (C) 2007 Elsevier Ltd. All rights reserved.
  • K. Shimizu; Y. Matsuda; T. Nonomura; H. Ikeda; N. Tamura; S. Kusakari; J. Kimbara; H. Toyoda
    PLANT PATHOLOGY BLACKWELL PUBLISHING 56 (6) 987 - 997 0032-0862 2007/12 [Refereed]
     
    An ozone-generative electrostatic spore precipitator was developed to protect nursery-stage seedlings of tomato from both airborne conidia of powdery mildew (Oidium neolycopersici) and root-infecting pathogen propagules of bacterial wilt (Ralstonia solanacearum) and fusarium crown and root rot (Fusarium oxysporum f.sp. radicis-lycopersici). The device was a cylindrical electrostatic spore precipitator (S2 cylinder) in which a positively charged straight conductor wire insulated with a transparent acrylic cylinder originated from a spore-precipitation cylinder (S1 cylinder) designed to physically control airborne conidia of tomato powdery mildew in greenhouses. The S2 cylinder consisted of two sites for conidial attraction and ozone production. The site for ozone production was located at the end of the cylinder, where an earthed copper conductor ring (as a cathode) was attached to the edge of the cylinder, responding to the anodal tip of a positively charged central conductor wire. Distinct types of discharge (corona, corona-streamer, streamer and arc discharge) occurred between the two electrodes and were dependant on the voltages applied to the wire and the distances between the electrodes. The highest ozone production was observed through streamer discharge. The remaining portion of the S2 cylinder, which was dielectrically polarized by a positively charged wire, created a non-uniform electric field outside the cylinder to attract conidia that came into the generated field. Hydroponic culture troughs to raise tomato seedlings in a nursery greenhouse were paralleled with S2 cylinders. The aim was to control rhizosphere pathogens R. solanacearum and F. oxysporum f.sp. radicis-lycopersici and to prevent them entering the hydroponic system during cultivation, while at the same time trapping O. neolycopersici conidia in the spaces between the cylinders. The results indicated that susceptible tomato plants in culture troughs attached to the S2 cylinders remained uninfected by both rhizosphere and aerial pathogens throughout the experimental period (2 and 3 weeks, respectively). This suggests that the present system will enable the dual control of both these pathogens in hydroponic systems in greenhouses.
  • Yoshinori Matsuda; Hiroki Ikeda; Nobuyuki Moriura; Norio Tanaka; Kunihiko Shimizu; Wataru Oichi; Teruo Nonomura; Koji Kakutani; Shin-ichi Kusakari; Katsuhide Higashi; Hideyoshi Toyoda
    PHYTOPATHOLOGY AMER PHYTOPATHOLOGICAL SOC 96 (9) 967 - 974 0031-949X 2006/09 [Refereed]
     
    In an attempt to physically protect greenhouse tomato plants from the powdery mildew fungus Oidium neolycopersici, we developed a new electrostatic spore precipitator in which a copper wire conductor is linked to an electrostatic generator and covered with a transparent acrylic cylinder (insulator). The conductor was negatively charged by the generator, and the electrostatic field created by the conductor was used to dielectrically polarize the insulator cylinder. The dielectrically polarized cylinder also produced an electrostatic force without a spark discharge. This force was directly proportional to the potential applied to the conductor and was used to attract conidia of the pathogen. The efficacy of this spore precipitator in protecting hydroponically cultured tomato plants from powdery mildew was evaluated in the greenhouse. The hydroponic culture troughs were covered with a cubic frame installed with the spore precipitator, and the disease progress on precipitator-guarded and unguarded seedlings was traced after the conidia were disseminated mechanically from inoculum on tomato plants. Seedlings in the guarded troughs remained uninfected during the entire experiment, in spite of rapid spread of the disease to all leaves of the unguarded seedlings.
  • W Oichi; Y Matsuda; T Nonomura; H Toyoda; L Xu; S Kusakari
    PLANT DISEASE AMER PHYTOPATHOLOGICAL SOC 90 (7) 915 - 919 0191-2917 2006/07 [Refereed]
     
    The formation of conidial pseudochains by the tomato powdery mildew Oidium neolycopersici on tomato leaves was monitored using a high-fidelity digital microscope. Individual living conidiophores that formed mature conidial cells at their apex were selected for observation. The conidial cells were produced during repeated division and elongation by the generative cells of the conidiophores. Under weak wind conditions (0.1 m/s), these conidial cells did not separate from each other to produce a chain of conidial cells (pseudochain). The pseudochains dropped from the conidiophores once four conidial cells were connected. The conidiophores resumed conidium production. followed by another cycle of pseudochain formation. The formation of pseudochains by tomato powdery mildew was not influenced by the ambient relative humidity. On the other hand, the conidial cells produced were easily wind dispersed without forming pseudochains when conidiophores were exposed to stronger winds (1.0 m/s). The present study successfully demonstrated that the pathogen required wind to disperse progeny conidia from the conidiophores and produced conidial pseudochains when the wind was below a critical level, independent of high relative humidity as reported previously.
  • N. Moriura; Y. Matsuda; W. Oichi; S. Nakashima; T. Hirai; T. Nonomura; K. Kakutani; S. Kusakari; K. Higashi; H. Toyoda
    PLANT PATHOLOGY WILEY-BLACKWELL 55 (3) 367 - 374 0032-0862 2006/06 [Refereed]
     
    Conidia from living conidiophores of barley powdery mildew (Blumeria graminis f.sp. hordei) on host leaves were collected consecutively using an electrostatic spore collector. The collector consisted of an electrical conductor plate linked to an electrostatic voltage generator and insulator plates placed abreast on a timed conveyer. The conductor plate was negatively charged by the potential supplied from the voltage generator. The negatively charged conductor plate caused dielectric polarization of the insulator plate, and the surface charge on the insulator plate attracted mature conidia abstricted from conidiophores on colonies growing on leaves placed 2 cm from the insulator plate. The surface charge on the insulator plate was proportional to the voltage applied to the conductor plate. Under optimized conditions, abstricted conidia were attracted to the electrostatically activated insulator plates without any detriment to their survival. During a colony's life span of c. 460 h, conidia were released throughout the day and c. 12 x 10(4) conidia were collected during the lifetime of the colony. This is the first report on the direct quantification of progeny conidia produced by powdery mildew infecting host leaves.
  • N Moriura; Y Matsuda; W Oichi; S Nakashima; T Hirai; T Sameshima; T Nonomura; K Kakutani; S Kusakari; K Higashi; H Toyoda
    MYCOLOGICAL RESEARCH CAMBRIDGE UNIV PRESS 110 (1) 18 - 27 0953-7562 2006/01 [Refereed]
     
    Conidial formation and secession by living conidiophores of Blumeria graminis f. sp. hordei on barley leaves were consecutively monitored using a high-fidelity digital microscopic technique combined with electrostatic micromanipulation to trap the released conidia. Conidial chains formed on conidiophores through a series of septum-mediated division and growth of generative cells. Apical conidial cells on the conidiophores were abstricted after the conidial chains developed ten conidial cells. The conidia were electrically conductive, and a positive charge was induced in the cells by a negatively polarized insulator probe (ebonite). The electrostatic force between the conidia and the insulator was used to attract the abstricted conidia. from the conidiophores on leaves. This conidium movement from the targeted conidiophore to the rod was directly viewed under the digital microscope, and the length of the interval between conidial septation and secession, the total number of the conidia produced by a single conidiophore, and the modes of conidiogenesis were clarified. During the stage of conidial secession, the generative cells pushed new conidial cells upwards by repeated division and growth. The successive release of two apical conidia was synchronized with the successive septation and growth of a generative cell. The release ceased after 4-5 conidia were released without division and growth of the generative cell. Thus, the life of an individual conidiophore (from the erection of the conidiophore to the release of the final conidium) was shown to be 107 h and to produce an average of 33 conidia. To our knowledge, this is the first report on the direct estimation of life-long conidial production by a powdery mildew on host leaves. (c) 2005 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.
  • T Shinogi; Y Hamanishi; Y Otsu; YQQ Wang; T Nonomura; Y Matsuda; H Toyoda; Y Narusaka; Y Tosa; S Mayama
    PLANT SCIENCE ELSEVIER IRELAND LTD 168 (6) 1477 - 1485 0168-9452 2005/06 [Refereed]
     
    The process of host/non-host determination was dissected in interactions of Epilachna vigintioctopunctata, a specialist herbivore of solanaceous plants, with various plant species. On host plants (tomato and egg plant) the ladybird beetle started feeding within 5 min. On red pepper, another solanaceous plant, it also started feeding within 5 min, but did not continue the feeding as vigorously as on tomato or eggplant. This result suggests that the ladybird beetle recognizes red pepper as a host plant but does not overcome its constitutive resistance. On Chinese cabbage, the ladybird beetle did not start feeding as quickly as on the host plants, but once started, it continued feeding as vigorously as on the host plants. This result suggests that the ladybird beetle does not recognize Chinese cabbage as a host plant but overcomes its constitutive resistance. Subsequently, the effect of induced resistance in a host (tomato) and non-hosts (Chinese cabbage and Arabidopsis) was evaluated. The treatment with methyl jasmonate (MeJA) showed no effects in tomato but decreased the damaged area in Chinese cabbage and Arabidopsis. A feeding test with Arabialopsis mutants supported the idea that induced resistance via the jasmonic acid (JA) pathway is effective against the ladybird beetle on the cruciferous plants. We suggest that a specialist herbivore has to overcome not only constitutive resistance but also induced resistance to utilize the non-host plant as a host, and that induced resistance is one of the factors that determine host specificity of the specialist. (c) 2005 Elsevier Ireland Ltd. All rights reserved.
  • Direct RT-PCR amplification of mature mRNA in cytoplasm micropipetted from barley coleoptile epidermal cell ? A model system for analyzing gene expression in host cells attacked by powdery.
    豊田 秀吉; 松田 克礼; 角谷 晃司
    Mem. Fac. Agr. Kinki Univ 36 9 - 18 2005/03 
    本実験では、顕微鏡下で標的とした単一細胞からマイクロピペットを用いて細胞内容物を吸引し、そこに存在するmRNAを鋳型とした単一細胞RT-PCR法を適用することとした。病原菌の感染によって発現が誘導される遺伝子(CHI2、GLUなど)を検索した。誘導型遺伝子の発現を厳密に評価するため、RT-PCRを行う際に、先の構成的発現遺伝子を増幅するプライマーを混合し、指標遺伝子として同時に増幅することにより、誘導型遺伝子の発現検出を行った。その結果、病原菌の感染を受けていない細胞およびその感染を受けている両者の細胞において指標とした構成的発現遺伝子は検出された。誘導型遺伝子として使用したCHI2遺伝子およびGLU遺伝子は病原菌の感染を受けていない細胞においてその発現が検出され、うどんこ病菌の感染過程において抑制される傾向にあった。
  • M Kojima; T Yoshikawa; M Ueda; T Nonomura; Y Matsuda; H Toyoda; K Miyatake; M Arai; T Fukamizo
    JOURNAL OF BIOCHEMISTRY OXFORD UNIV PRESS 137 (2) 235 - 242 0021-924X 2005/02 [Refereed]
     
    Family 19 chitinase from Aeromonas sp. No.10S-24 (72.6 kDa) is composed of two chitin-binding domains (ChBDs), two proline- and threonine-rich (PT-rich) linkers, and a catalytic domain. The purified enzyme was labile in a standard buffer condition and spontaneously degraded into a 46-kDa fragment upon storage at 4 degrees C. The N-terminal sequence of the 46-kDa fragment was found to correspond to the sequence of the G terminal region of the second PT-rich linker, indicating that the 46-kDa fragment is produced by truncation of the two ChBDs and the two PT-rich linkers from the mature protein, and consists only of the catalytic domain. The hydrolytic activities toward insoluble and soluble substrates were significantly reduced by the truncation of two ChBDs. In addition, antifungal activity determined from the digestion rate of haustoria of powdery mildew was reduced by the ChBD truncation. Although the profile of the time-course of N-acetylglucosamine hexasaccharide [(GlcNAc)(6)] degradation catalyzed by the ChBD-truncated enzyme was similar to that of the mature enzyme protein, the specific activity of the ChBD-truncated enzyme determined from the rate of hexasaccharide degradation was lower than that of the mature enzyme. The two CBDs appear to be responsible for facilitating the hydrolytic reaction. The sugar residue affinities (binding free energy changes) at the individual subsites, (-2) (-1) (+1) (+2) (+3) (+4), were estimated by modeling the hexasaccharide hydrolysis by the mature and ChBD-truncated enzymes. The truncation of ChBDs was found to strongly affect the affinity at the (4) site. This situation seems to result in the lower enzymatic activity of the ChBD-truncated enzyme toward the chitinous substrates.
  • Yoshinori Matsuda; Yoshinori Mori; Yohei Sakano; Masayoshi Nishida; Koji Tarumoto; Teruo Nonomura; Hiroaki Nishimura; Shin-Ichi Kusakari; Hideyoshi Toyoda
    Breeding Science 55 (3) 355 - 360 1344-7610 2005
  • Wataru Oichi; Yoshinori Matsuda; Takeshi Sameshima; Teruo Nonomura; Koji Kakutani; Hiroaki Nishimura; Shin-Ichi Kusakari; Hideyoshi Toyoda
    Journal of General Plant Pathology 70 (6) 329 - 332 1345-2630 2004/12 
    Conidiogenesis by Oidium neolycopersici KTP-01 on tomato leaves was vitally monitored with a high-fidelity digital microscope. Conidiophores were initially formed 3 days after inoculation and then elongated to a maximum length within at least 12 h. The apical part was split into two cells after two successive septations, accompanied by apical expansion. These cells subsequently developed into primary and secondary conidia. An additional septation at the stem portion of the conidiophores produced a generative and a foot cell. Subsequent conidiation occurred during repeated cycles of splitting of the generative cell, maturation of the apical cell into a conidium, and abstriction of the conidium. To our knowledge, this report is the first on the developmental process of conidiogenesis by powdery mildew on host leaves as revealed with the digital microscope.
  • K Fujita; Y Matsuda; M Wada; Y Hirai; K Mori; N Moriura; T Nonomura; K Kakutani; H Toyoda
    PLANT CELL REPORTS SPRINGER 23 (7) 504 - 511 0721-7714 2004/12 
    Two-step PCR (RT-PCR and nested PCR) was used to detect gene expression in powdery mildew pathogen-infected cells of detached inner epidermis of barley coleoptiles. Cellular contents of infected cells were microscopically suctioned with a micropipette and subjected to PCR. Triosephosphate isomerase and glyceraldehyde-3-phosphate dehydrogenase genes involved in the glycolytic pathway and a stimulus-induced endochitinase gene were targeted, and their expression was determined by detecting cDNAs derived from spliced transcripts. The two gycolysis-related genes were constantly expressed in the tissue irrespective of pathogen inoculation. In contrast, chitinase gene expression was induced in non-infected inner epidermis after detachment. After inoculation, this expression was selectively suppressed in pathogen-invaded cells, in spite of continuous expression in non-invaded cells of the same epidermis. Thus, the present method enabled us to directly analyze transcripts in individual cells at the infection site and assess the capability of the pathogen to regulate host gene expression.
  • M Wada; Y Matsuda; K Fujita; A Nanjo; M Nishimura; T Nonomura; K Kakutani; H Toyoda
    PLANT CELL TISSUE AND ORGAN CULTURE KLUWER ACADEMIC PUBL 79 (1) 109 - 114 0167-6857 2004/10 
    RT-PCR was used to detect gene expression in situ in single selected cells of tomato callus aggregates. The cytoplasm from one cell was removed with a micropipette viewed under a light microscope and used directly for RT-PCR, followed by nested PCR. This method to remove cytosolic contents prevented the introduction of genomic DNA into the RT-PCR, and only intron-spliced products were amplified when intron-containing genes were used as PCR targets. In addition, transcription of the intron-free gene was possibly detected by simultaneously tracing the intron-containing and intron-free genes using mixed primers for the targeted genes. The present study indicated that some stimuli-activated genes, such as CHI3 and TLC1-LTR, were constitutively transcribed in tomato callus cells.
  • Y Otsu; Y Matsuda; H Mori; H Ueki; T Nakajima; K Fujiwara; M Matsumoto; N Azuma; K Kakutani; T Nonomura; Y Sakuratani; T Shinogi; Y Tosa; S Mayama; H Toyoda
    BIOCONTROL SCIENCE AND TECHNOLOGY CARFAX PUBLISHING 14 (5) 427 - 439 0958-3157 2004/08 
    An entomopathogenic bacterium was isolated from tomato leaves and used as a microbial agent to control larvae of phytophagous ladybird beetles Epilachna vigintioctopunctata. The isolate was identified as Pseudomonas fluorescens KPM-018P on the basis of its bacteriological characteristics. KPM-018P produced extracellular chitinase to form a transparent zone around their colonies by hydrolyzing chitin in a minimal medium. Pale-yellow colonies turned red after a change of incubation temperature. These characteristics were availed as markers for tracking KPM-018P. The bacteria produced biosurfactants that enabled the bacteria to stably colonize the hydrophobic leaf surface; they were recovered without any considerable decrease even after a suspension of KPM-018P was sprayed onto leaves. KPM-018P, transformed with the gfp gene and observed with fluorescence microscopy, stably dwelled in the junctions of epidermal cells of bacteria-sprayed leaves. Ingestion of KPM-018P-sprayed leaves by the larvae caused prompt death of these insects to eventually suppress their pupation. This method is thus effective for decreasing the population of larvae and adult insect pests in the subsequent generation. The study provides an experimental basis for the biocontrol of herbivorous insect pests using a leaf-inhabiting, entomopathogenic strain of P. fluorescens.
  • Takeshi Sameshima; Koichi Kashimoto; Keiko Kida; Yoshinori Matsuda; Teruo Nonomura; Koji Kakutani; Kengo Nakata; Shin-Ichi Kusakari; Hideyoshi Toyoda
    Journal of General Plant Pathology 70 (1) 7 - 10 1345-2630 2004/02 
    Leaves of tomato and barley were inoculated with conidia of Blumeria graminis f. sp. hordei race 1 (R1) or Oidium neolycopersici (KTP-01) to observe cytological responses in search of resistance to powdery mildew. Both conidia formed appressoria at similar rates on tomato or barley leaves, indicating that no resistance was expressed during the prepenetration stage of these fungi. On R1-inoculated tomato leaves, appressoria penetrated the papillae, but subsequent haustorium formation was inhibited by hypersensitive necrosis in the invaded epidermal cells. On the other hand, KTP-01 (pathogenic to tomato leaves) successfully developed functional haustoria in epidermal cells to elongate secondary hyphae, although the hyphal elongation from some conidia was later suppressed by delayed hypersensitive necrosis in some haustorium-harboring epidermal cells. Thus, the present study indicated that the resistance of tomato to powdery mildew fungi was associated with a hypersensitive response in invaded epidermal cells but not the prevention of fungal penetration through host papilla.
  • Takeshi Sameshima; Koichi Kashimoto; Keiko Kida; Yoshinori Matsuda; Teruo Nonomura; Koji Kakutani; Kengo Nakata; Shin-Ichi Kusakari; Hideyoshi Toyoda
    Journal of General Plant Pathology 70 (1) 7 - 10 1345-2630 2004/02 
    Leaves of tomato and barley were inoculated with conidia of Blumeria graminis f. sp. hordei race 1 (R1) or Oidium neolycopersici (KTP-01) to observe cytological responses in search of resistance to powdery mildew. Both conidia formed appressoria at similar rates on tomato or barley leaves, indicating that no resistance was expressed during the prepenetration stage of these fungi. On R1-inoculated tomato leaves, appressoria penetrated the papillae, but subsequent haustorium formation was inhibited by hypersensitive necrosis in the invaded epidermal cells. On the other hand, KTP-01 (pathogenic to tomato leaves) successfully developed functional haustoria in epidermal cells to elongate secondary hyphae, although the hyphal elongation from some conidia was later suppressed by delayed hypersensitive necrosis in some haustorium-harboring epidermal cells. Thus, the present study indicated that the resistance of tomato to powdery mildew fungi was associated with a hypersensitive response in invaded epidermal cells but not the prevention of fungal penetration through host papilla.
  • トマトうどんこ病ー動的解析法とその有効防除の確立をめざして 植物病の探究 In Pursuit of the Essence of Plant Pathogenesis
    角谷 晃司; 豊田 秀吉; 松田 克礼; 野々村 照雄
    「植物病の探究」出版会 2004
  • Y Otsu; Y Matsuda; H Shimizu; H Ueki; H Mori; K Fujiwara; T Nakajima; A Miwa; T Nonomura; Y Sakuratani; Y Tosa; S Mayama; H Toyoda
    JOURNAL OF APPLIED ENTOMOLOGY BLACKWELL VERLAG GMBH 127 (8) 441 - 446 1439-0418 2003/09 
    The chitinase secreting strain KPM-012A of Alcaligenes paradoxus was isolated from tomato leaves and vitally entrapped in sodium alginate gel beads to provide a new method for biocontrol of phytophagous ladybird beetles Epilachna vigintioctopunctata. First, the peritrophic membrane was dissected from the adult ladybird beetles that ingested the suspension of KPM-012A after starvation to observe degradation of the midgut surface by the bacteria under electron microscopy. The peritrophic membrane around the bacteria was degraded, suggesting the release of chitinase from the ingested bacteria. Large amounts of chitinase were successfully released from KPM-012A-entrapped calcium alginate beads. This chitinase release from the microbial beads was sustained for I week and was sufficient to digest the peritrophic membrane. Daily supply of tomato leaves treated with the microbial beads caused considerable suppression of leaf feeding and oviposition by the adult ladybird beetles, indicating that this method is effective for decreasing population of insect pests in the subsequent generation. Thus, the present study provided an experimental basis for the biocontrol measures of herbivorous insect pests by the chitinolytic bacteria entrapped in alginate beads.
  • Y. Otsu; Y. Matsuda; H. Shimizu; H. Ueki; H. Mori; K. Fujiwara; T. Nakajima; A. Miwa; T. Nonomura; Y. Sakuratani; Y. Tosa; S. Mayama; H. Toyoda
    Journal of Applied Entomology 127 (8) 441 - 446 0931-2048 2003/09 [Refereed]
     
    The chitinase secreting strain KPM-012A of Alcaligenes paradoxus was isolated from tomato leaves and vitally entrapped in sodium alginate gel beads to provide a new method for biocontrol of phytophagous ladybird beetles Epilachna vigintioctopunctata. First, the peritrophic membrane was dissected from the adult ladybird beetles that ingested the suspension of KPM-012A after starvation to observe degradation of the midgut surface by the bacteria under electron microscopy. The peritrophic membrane around the bacteria was degraded, suggesting the release of chitinase from the ingested bacteria. Large amounts of chitinase were successfully released from KPM-012A-entrapped calcium alginate beads. This chitinase release from the microbial beads was sustained for 1 week and was sufficient to digest the peritrophic membrane. Daily supply of tomato leaves treated with the microbial beads caused considerable suppression of leaf feeding and oviposition by the adult ladybird beetles, indicating that this method is effective for decreasing population of insect pests in the subsequent generation. Thus, the present study provided an experimental basis for the biocontrol measures of herbivorous insect pests by the chitinolytic bacteria entrapped in alginate beads.
  • Y Otsu; H Mori; K Komuta; H Shimizu; S Nogawa; Y Matsuda; T Nonomura; Y Sakuratani; Y Tosa; S Mayama; H Toyoda
    JOURNAL OF ECONOMIC ENTOMOLOGY ENTOMOL SOC AMER 96 (3) 555 - 563 0022-0493 2003/06 
    The chitinase gene-transformed strain KPM-007E/chi Of Enterobacter cloacae was vitally entrapped in sodium alginate gel beads with its specific virulent bacteriophage EcP-01 to provide a new method for microbially digesting chitinous peritrophic membranes of phytophagous ladybird beetles Epilachna vigintioctopunctata. First, chitinase SH1 from a gram-positive bacterium Kurthia zopfii was overproduced by Escherichia coli cells and purified by affinity column chromatography. The purified enzyme effectively digested peritrophic membranes dissected from the ladybird beetles to expose epithelial tissues beneath the peritrophic membrane, and the beetles that had ingested chitinase after submergence in chitinase solution had considerably reduced their feeding on tomato leaves. KPM-007E/chi, entrapped in the alginate beads, released the chitinase. More chitinase was released when KPM-007E/chi was present with their specific virulent bacteriophage EcP-01 in the beads because of lysis of bacterial cells infected with the bacteriophages. This chitinase release from the microbial beads (containing KPM-007E/chi and EcP-01) was sufficient to digest the peritrophic membrane as well as to suppress feeding of bead-sprayed tomato leaves by the ladybird beetles. A daily supply of tomato leaves treated with the microbial beads considerably suppressed leaf feeding and oviposition by the ladybird beetles, suggesting a possible application of chitinase-secreting bacteria for suppressing herbivorous insect pests.
  • Morphological and Molecular Characterization for a Japanese Isolate of Tomato Powdery Mildew Oidium neolycopersici and Its Host Range.
    角谷 晃司; 松田 克礼; 野々村 照雄; 豊田 秀吉; 近畿大学大学院農学研究科; 近畿大学大学院農学研究科; 近畿大学大学院農学研究科; 近畿大学大学院農学研究科; 大阪府立農林技術センター; 近畿大学大学院農学研究科; 近畿大学大学院農学研究科
    J. Gen. Plant Pathol. 69 179 - 185 2003
  • TAKIKAWA Yoshihiro; ISHII Yuuki; FUJIWARA Keiichi; MATSUDA Yoshinori; NONOMURA Teruo; KAKUTANI Koji; TOSA Yukio; MAYAMA Shigeyuki; TOYODA Hideyoshi
    J. Gen. Plant Pathol 69 (2) 131 - 137 1345-2630 2003
  • KASHIMOTO Koichi; SAMESHIMA Takeshi; MATSUDA Yoshinori; NONOMURA Teruo; OICHI Wataru; KAKUTANI Koji; NAKATA Kengo; KUSAKARI Shin-ichi; TOYODA Hideyoshi
    J. Gen. Plant Pathol 69 (6) 406 - 408 1345-2630 2003
  • Y Iida; Y Matsuda; R Saito; M Nakasato; T Nonomura; K Kakutani; Y Tosa; S Mayama; H Toyoda
    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY TAYLOR & FRANCIS LTD 67 (1) 198 - 202 0916-8451 2003/01 
    Overproduced proteins from Escherichia coli BL21(DE3) were efficiently released with virulent bacteriophages. Leviviridae-like bacteriophages were isolated from soil and used to lyse BL21(DE3) cells transformed with beta-glucosidase, chitinase, or chitosanase genes. This method caused lysis of bacterial cells similar to that by conventional sonication and enabled us to effectively recover and purify the enzymes.
  • Takeshi Fukumoto; Teruo Nonomura; Yoshinori Matsuda; Shin-ichirou Komaki; Nobuyuki Moriurai; Hideyoshi Toyoda; Koji Kakutani; Akiyoshi Sawabe
    Plant Biotechnology 20 (3) 257 - 261 1347-6114 2003 
    When leaf segments of a tomato cultivar 'Ponderosa' were inoculated with Agrobacterium rhizogenes MAFF07-20001 carrying the binary vectors pRi and pBI121/sGFP, adventitious roots were developed from calli formed at the edges of the segments. Primordial roots were obtained with green fluorescence under blue light and elongated vigorously on hormone-free medium without loss of the green fluorescence. They were easily distinguishable from the non-fluorescing roots on the same segments. Successful integration of the sGFP and rol C genes into the chromosome of tomato roots was confirmed by polymerase chain reaction and Southern hybridization. The present method enables us to evaluate the hairy root formation without subculture, isolation and DNA analysis. All commercial cultivars available in Japan (24 cultivars) and 14 breeding lines of tomato were tested by this method. All but two breeding lines produced the hairy roots. Thus, the present method is useful for hairy root production in tomato. © 2003, Japanese Society for Plant Cell and Molecular Biology. All rights reserved.
  • MATSUDA Yoshinori; ENMEIJI Toshiya; TOYODA Hideyoshi
    Memoirs of the Faculty of Agriculture of Kinki University Kinki University 36 131 - 136 0453-8889 2003 
    The occurrence of powdery mildew on natural vegetation in the Nara Campus of Kinki University was surveyed from April to September 2002. Totally 14 plants were infected with powdery mildew pathogens, and conidia produced on conidiophores were observed by microscope. Furthermore, cleistothecia formed on the mycelial colony of Quercus glauca were isolated by means of a micromanipulator, and asci were released from cleistothecium by crushing. The numbers of the asci in cleistothecium and the ascospores in ascus were confirmed by means of microscopic observation, and the pathogen was identified as a member of the Uncinula genus. These results would provide a model to aid in agricultural education.
  • Akiyoshi Sawabe; Yoshinori Matsuda; Hideyoshi Toyoda; Kazuhiko Maeda; Teruo Nonomura; Naoto Shimizu; Naoko Fujita; Seiji Ouchi
    Journal of Oleo Science 52 (3) 171 - 174 1347-3352 2003 
    Volatile oils were obtained from melon hairy roots produced by the infection with Agrobacterium rhizogenes, and their constituents were analyzed by GC and GC-MS spectroscopy. Characteristic major components were 1-nonanal, (Z)-6-nonenol, and (E, Z)-2, 6-nonadienal. The melon hairy roots are thus shown to produce volatile components with the fragrance of ripe melon fruit. © 2003, Japan Oil Chemists' Society. All rights reserved.
  • Aroma constituent production by the use of microorgnism gene and their development
    澤邊 昭義; 豊田 秀吉; 松田 克礼
    Foods Food Ingredients J. Jpn. 207 23 - 30 2002/12 
    微生物遺伝子導入によって作出したメロン毛状根の有効成分の培養条件検討および有効成分のGLC分析を行った。その結果、メロン毛状根は、メロン果実の香気成分を多量に生産することが明らかとなり、工業的生産に適合することが認められた。微生物遺伝子導入によって作出した毛状根の香気成分生産法は、ほとんど揮発性油(精油)をもっていない有用植物の香料を食品香料・香粧品香料として供給することに役立つことを紹介した。
  • K Kawamura; T Sugimoto; Y Matsuda; H Toyoda
    APPLIED ENTOMOLOGY AND ZOOLOGY JAPAN SOC APPL ENTOMOL ZOOL 37 (4) 645 - 648 0003-6862 2002/11 
    The polymorphic patterns of genomic DNA amplified by RAPD-PCR were detected in laboratory-cultured populations of sweet potato weevils, Cylas formicarius collected from the Southwest islands (Nansei-Shoto) of Japan. When three sets of primers, T13/T13, T05/T05 and T01/T07 were used for PCR, the polymorphic patterns of the amplified DNA were classified into nine types. Of these types, A(1)B(1)C(3) was common in all populations tested. The present study determined the useful primer sets that will enable the successful classification of sweet potato weevils based upon the polymorphic patterns of genomic DNA amplified by RAPD-PCR.
  • Y Takikawa; H Mori; Y Otsu; Y Matsuda; T Nonomura; K Kakutani; Y Tosa; S Mayama; H Toyoda
    JOURNAL OF APPLIED MICROBIOLOGY BLACKWELL PUBLISHING LTD 93 (6) 1042 - 1050 1364-5072 2002 
    Aims: To establish a rapid and efficient method for detecting Enterobacter cloacae based on chitinase gene transformation and lytic infection by virulent bacteriophages. Methods and Results: A phylloplane strain of E. cloacae was isolated from tomato leaves and transformed with a chitinase gene. Transformed bacteria were collected from single colonies and infected with newly isolated, virulent bacteriophages in the presence of the chitinase substrate 4-methylumbelliferon (4MU)-(GlcNac)(3). To assay chitinase activity in the lysates, the product 4MU was measured spectrofluorophotometrically or visibly detected under u.v. irradiation. Chitinase gene-transformed bacteria obtained from single colonies could be specifically identified in 30 min by the emission of 4MU fluorescence following lysis caused by phage infection. Conclusions: The chitinase gene was used as a reporter gene to construct a new system for easy and rapid monitoring of transgenic strains of E. cloacae released in the environment, in combination with specific recognition by virulent bacteriophages. Significance and Impact of the Study: The assay is simple, rapid, inexpensive, easy to perform and applicable to other strains. The system can be used for the routine monitoring of bacteria, which is important because of the increased use of transgenic strains of E. cloacae as an antagonistic biological control agent for plant diseases.
  • MATSUDA Yoshinori; IIDA Yuichiro; SHINOGI Takeshi; KAKUTANI Koji; NONOMURA Teruo; TOYODA Hideyoshi
    Journal of General Plant Pathology The Phytopathological Society of Japan 67 (4) 318 - 324 1345-2630 2001/12 
    A chitosan-degrading bacterium, isolated from field soil that had been amended with chitin, was identified as Sphingobacterium multivorum KST-009 on the basis of its bacteriological characteristics. The extracellular chitosanase (SM1) secreted by KST-009 was a 34-kDa protein and could be purified through ammonium sulfate precipitation, gel permeation column chromatography and SDS polyacrylamide gel electrophoresis. A chitosanase gene (csnSM1) was isolated from genomic DNA of the bacteria, and the entire nucleotide sequence of the gene and the partial N-terminal amino acid sequence of the purified SM1 were determined. The csnSM1 gene was found to encode 383 amino acids, 72 N-terminal amino acid residues were processed to produce the mature enzyme during the secretion process. Germinated microconidia of four formae speciales (lycopersici, radicis-lycopersici, melonis, and fragariae) of Fusarium oxysporum were treated with SM1. Chitosanase treatment caused morphological changes, such as swelling of hyphal cells or indistinctness of hyphal cell tips and cessation or reduction of mycelial elongation.
  • MATSUDA Yoshinori; KASHIMOTO Koichi; TAKIKAWA Yoshihiro; AIKAMl Rika; NONOMURA Teruo; TOYODA Hideyoshi
    Journal of General Plant Pathology The Phytopathological Society of Japan 67 (4) 294 - 298 1345-2630 2001/12 
    During a year-round survey on the occurrence of powdery mildew on greenhouse-cultivated tomato plants, the disease was most severe in June and July. All tomato plants (45 commercial cultivars and 11 breeding lines) tested were infected with the pathogen but had different degrees of susceptibility. The pathogen was epiphytic and produced white, round pustules mainly on leaves of tomato plants. The pathogen produced conidia singly on conidiophores and forked appressoria on inoculated tomato leaves and seemed to be an Oidium sp. of Erysiphe polygoni type.
  • 豊田 秀吉; 野々村 照雄; 松田 克礼; 津田元章; 浦中和人
    Journal of General Plant Pathology The Phytopathological Societ of Japan 67 (3) 224 - 227 2001/08 
    水耕栽培を利用して栽培トマト品種間における青枯病菌の感受性差異について検討した。 その結果、 栽培品種間において病徴発現するまでの期間および初期病徴から完全枯死に至るまでの期間に有異な差が認められた。
  • Teruo Nonomura; Yoshinori Matsuda; Miki Bingo; Maiko Onishi; Kazuhiko Matsuda; Satoshi Harada; Hideyoshi Toyoda
    Crop Protection ELSEVIER 20 (10) 935 - 939 0261-2194 2001 
    The chemical control agent 3-(3-indolyl)butanoic acid, previously reported as a control agent for the bacterial wilt pathogen Ralstonia solanacearum, was shown to suppress the growth of green algae during hydroponic culture of tomato. The algicidal activity of the compound was effective at 10 μg/ml, completely preventing generation of green algae under non-shaded greenhouse conditions. The algicidal effect was mainly due to suppression of the growth of motile unicellular algal cells tentatively identified as Chlamydomonas spp., which are commonly occurring in the hydroponic solution and vigorously multiply to form an algal mat on the sponge supports. The compound has potential as a non-phytotoxic algicide for hydroponically cultured crop plants. © 2001 Elsevier Science Ltd. All rights reserved.
  • NONOMURA Teruo; MATSUDA Yoshinori; SHIRATORI Satomi; MATSUDA Kazuhiko; HARADA Satoshi; TOYODA Hideyoshi
    Environmental Control in Biology Japanese Society of Agricultural, Biological and Environmental Engineers and Scientists 39 (2) 127 - 134 0582-4087 2001 
    Some Indole derivatives were examined for their algicidal activity to Chlamydomonas reinhardtii and the isolate of Chlamydomonas sp. which was isolated from the green algae-generating hydroponic solution in the hydroponic trough of the greenhouse. Of nine compounds tested, 3-indolepropionic acid, 3-indoleacrylic acid and 3- (3-indolyl) butanoic acid could suppress multiplication of C. reinhardtii cells. Although the first one exhibited the strong phytotoxicity, the latter two compounds were effective at lower concentrations which did not cause any toxic effect on the growth of tomato seedlings. In addition, 3-indoleacrylic acid and 3- (3-indolyl) butanoic acid inhibited the multiplication of Chlamydomonas green algae which were actively growing in our hydroponic facilities, and suppressed the algal emergence on the synthetic sponge cubes soaked in the hydroponic solution which had been inoculated with green algae. The present study suggests that these compounds have a potential to be used as an algicide for the hydroponics of crop plants.
  • Teruo Nonomura; Yukiko Ikegami; Yoshic Morikawa; Yoshinori Matsuda; Hideyoshi Toyoda
    Plant Biotechnology Japanese Society for Plant Cell and Molecular Biology 18 (3) 233 - 236 1347-6114 2001 [Refereed]
     
    The apical buds of lateral branches asexually multiplied by cutting were treated with some chemical mutagens, and the growth and differentiation or morphological changes of the mutagen- treated buds were traced in developed flowers. As a result, the variations in size, shape, color and number of petals were detected most frequently in the flowers that were developed from apical buds treated with N-methyl-N'-nitro-N-nitrosoguanidine at 100 μg ml-1. The variant petals were cultured on MS medium supplemented with NAA and BAP for in vitro isolation and multiplication of morphologically altered rose plants. Embryogenic calli were obtained via adventitious roots induced from the petals and successfully differentiated to intact plants. Consequently, the regenerated plants produced the varied flower different from that originally used for tissue culture. Thus, the present study suggested that our approach would provide an effective method for easily and rapidly inducing variations in flowers of rose and for in vitro multiplication of their regenerants.
  • Y Matsuda; H Toyoda; A Sawabe; K Maeda; N Shimizu; N Fujita; T Fujita; T Nonomura; S Ouchi
    JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY AMER CHEMICAL SOC 48 (4) 1417 - 1420 0021-8561 2000/04 [Refereed]
     
    Musk melon is the favorite fruit with a high market value in Japan, and the fragrance is one of the major factors determining the fruit quality of melon. In this study, mutant melon hairy roots which had been induced by means of the T-DNA insertion mutagenesis were found to produce volatile compounds with the fruity fragrance of mature melon. The volatile compounds were extracted and identified by GLC-mass spectrometry. Some essential oils such as (Z)-3-hexenol, (E)-2-hexenal, 1-nonanol, and (Z)-6-nonenol were stably synthesized by these hairy roots despite the increased number of subcultures. The productivity of these compounds by the best hairy root line was shown to be considerably higher than naturally ripened melon fruits.
  • MATSUDA Yoshinori; TOYODA Hideyoshi; UEDA Ayumi; TAMAKI Sae; HOSOI Yoshiyuki; OUCHI Seiji
    Environmental Control in Biology Japanese Society of Agricultural, Biological and Environmental Engineers and Scientists 35 (2) 131 - 134 0582-4087 1997 
    The potential traits of photosynthetic and photoautotrophic growth of hairy root derived from co-cultures of Cucumis melo L. with Agrobacterium rhizogenes were investigated. Thirty-two different lines of hairy roots induced from leaf explants of the plant were used to determine the conditions which favor the induction of photoautotrophy. Although all of the hairy root lines remained white under dark condition, some of them turned yellow green to pale green when incubated in MS medium containing low concentration of sucrose and supplemented with a buffer releasing CO2. One of the initially heterotrophic hairy root cultures, KMH-18, turned green rapidly, produced chlorophyl, and grew well depending on CO2 fixation under these conditions.
  • Hideyoshi Toyoda; Nobuhiro Kita; Koji Kakutani; Yoshinori Matsuda; Mitsue Dogo; Yasunari Kato; Tsuyoshi Nomura; Miki Bingo; Hiroyuki Tampo; Kazuyuki Chatani; Kunihiko Shimizu; Seiji Ouchi
    Plant Biotechnology Japanese Society for Plant Cell and Molecular Biology 14 (2) 105 - 110 1347-6114 1997 [Refereed]
     
    The bacterial wilt resistant line LNSR-7 of tomato was isolated from self-pollinated progenies of leaf-callus derived regenerants by directly inoculating a bacterial wilt pathogen Pseudomonas solanaceanum into injured roots of tested plants. The subsequent self-pollinated progenies of the line were examined for their fruit quality and resistance expression under natural cultivation conditions in a pathogen-infested tomato field. During three generations of progenies, the tomato plants showing both the bacterial wilt resistance and the high fruit qualities comparable to the parental cultivar were selected in order to fix commercial characteristics of the line. The stable inheritance of the resistance in the subsequent self-pollinated progenies was further examined by directly inoculating the pathogen into the roots of test plants. Inoculated plants were planted in soil heavily infested with the pathogen to ensure exposure to the pathogen. Under these artificial inoculation conditions, the selected line was shown to be highly resistant to the disease. The resistance mechanism in the line was analyzed by examining multiplication and translocation of the pathogen in planta. The precise monitoring of infection behavior of the pathogen was successfully achieved using the genetically marked P. solanacearum. Consequently the present line LNSR-7 strictly limited secondary multiplication and translocation of the pathogen and suppressed the wilt induction by the pathogen,.
  • Hideyoshi Toyoda; Yoshinori Matsuda; Mitsue Dogo; Hiroyuki Tanpo; Keiko Sekimoto; Seiji Ouchi
    Plant Biotechnology Japanese Society for Plant Cell and Molecular Biology 14 (2) 111 - 116 1347-6114 1997 [Refereed]
     
    To examine the mechanism for in planta detoxification of phytotoxicity of indole-3-propionic acid, the gene expression for glucosylation of this compound was detected in treated tomato seedlings by Northern and in situ hybridization with the specific probe. The probe was obtained by polymerase chain reaction of tomato chromosomal DNA using the primers designed on the basis of amino acid sequences which were highly conserved in several enzymes catalyzing glucosylation. The positive hybridization was intensively detected in stems of the treated tomato seedlings. The increase of hybridized transcript was well coincident with the accumulation of glucosyl indole-3-propionic acid analyzed by thin layer chromatography. The result suggests that the PCR clone obtained from tomato chromosome encodes partial sequences of a gene for glucosyl conjugation of the compound.
  • CHATANI Kazuyuki; TOYODA Hideyoshi; MORIKAWA Yoshie; OGATA Yoko; KOREEDA Kazuharu; YOSHIDA Kenji; HAGI Tomohiro; MATSUDA Yoshinori; OUCHI Seiji
    Plant Biotechnology Japanese Society for Plant Cell and Molecular Biology 13 (2) 185 - 188 0289-5773 1996
  • TOYODA Hideyoshi; MATSUDA Yoshinori; CHATANI Kazuyuki; OUCHI Seiji
    Plant Biotechnology Japanese Society for Plant Cell and Molecular Biology 13 (2) 211 - 214 0289-5773 1996
  • CHATANI Kazuyuki; TOYODA Hideyoshi; MATSUDA Yoshinori; SHIMIZU Kunihiko; OUCHI Seiji
    Plant Biotechnology Japanese Society for Plant Cell and Molecular Biology 13 (1) 87 - 89 0289-5773 1996
  • CHATANI Kazuyuki; TOYODA Hideyoshi; MATSUDA Yoshinori; OUCHI Seiji
    Plant Biotechnology Japanese Society for Plant Cell and Molecular Biology 11 (1) 71 - 73 0289-5773 1994
  • MATSUDA Yoshinori; TOYODA Hideyoshi; NOGI Yasushi; TAMAI Takayuki; OUCHI Seiji
    Plant Biotechnology Japanese Society for Plant Cell and Molecular Biology 9 (3) 154 - 163 0289-5773 1992 
    The foreign genes, β-glucuronidase gene and TMV coat protein gene both ligated with the CaMV 35S promoter and nopalinsynthase terminator, were introduced into the inner epidermal cells of this tissue, using a barley coleoptile/microinjection system. The expression of the introduced genes was successfully examined by cytochemical identification of the transcription and translation products of the genes. Transcripts of the introduced genes were in situ hybridized to photobiotin conjugated antisense RNAs which were secondarily introduced by a pricking microinjection, and the translation products were detected by a complex formation with injected enzyme-conjugated antibodies. With a dual microinjection method, sense RNAs of these genes could be introduced and their translation products were detected at the same frequency (more than 80% of successfully injected cells) as those in DNA-injected cells. Thus, the present system, a combination of microinjection and in situ cytochemical techniques, allows introduction of foreign genes into barley coleoptile epidermal cells and precise detection of foreign gene expression in a higher plant cell system.
  • Hideyoshi Toyoda; Takako Yamaga; Yoshinori Matsuda; Seiji Ouchi
    Plant Cell Reports Springer-Verlag 9 (6) 299 - 302 0721-7714 1990/10 [Refereed]
     
    A β-glucuronidase gene was introduced directly into barley (Hordeum vulgare L. cv. Kobinkatagi) coleoptile cells by microinjection and transient expression of the gene was examined. Inner epidermis tissue of coleoptiles was excised and injected with plasmid DNA, pBI221, carrying cauliflower mosaic virus 35S promoter, β-glucuronidase gene, and a nopaline synthase polyadenylation region. Histochemical assay for β-glucuronidase production showed positive enzyme activity only in coleoptile cells injected with plasmid DNA. Expression of the β-glucuronidase gene was examined chronologically using honogenates of injected coleoptile tissues. Glucuronidase activity first appeared after 6 hr, reached the maximum level 24 hr after injection, and decreased afterwards. These results suggest that microinjection of coleoptile tissues may be a useful approach for the genetic engineering of Gramineae plants in which protoplast regeneration is difficult. © 1990 Springer-Verlag.
  • Hideyoshi Toyoda; Kazuyuki Chatani; Yoshinori Matsuda; Seiji Ouchi
    Plant Cell Reports Springer-Verlag 8 (8) 433 - 436 0721-7714 1989/12 [Refereed]
     
    Tobacco mosaic virus-resistant tobacco was selected in vitro using callus tissues induced from axillary buds of systemically infected tobacco plants. Callus lines in which the virus was continuously multiplying were first isolated and redifferentiated into shoots. By the procedure, non-diseased, healthy shoots were successfully isolated from diseased shoots, which showed typical mosaic symptoms of the virus, and regenerated into intact plants. These regenerated plants showed resistance to virus inoculation, and selfed progeny of virus-resistant regenerants segregated the resistance and susceptibility according to the Mendelian system. © 1989 Springer-Verlag.
  • Hideyoshi Toyoda; Kunihiko Shimizu; Kazuyuki Chatani; Nobuhiro Kita; Yoshinori Matsuda; Seiji Ouchi
    Plant Cell Reports Springer-Verlag 8 (6) 317 - 320 0721-7714 1989/06 [Refereed]
     
    Bacterial wilt-resistant plants were obtained using a tomato tissue culture system. A virulent strain of Pseudomonas solanacearum secreted some toxic substances into the culture medium. Leaf explant-derived callus tissues which were resistant to these toxic substances in the culture filtrate were selected in vitro and regenerated into plants. These plants expressed bacterial wilt resistance at the early infection stage to suppress or delay the growth of the inoculated bacteria. On the other hand, complete resistance was obtained in self-pollinated progeny of regenerants derived from non-selected callus tissues. These plants showed a high resistance when inoculated with this strain, and were also resistant when planted in a field infested with a different strain of the pathogen. © 1989 Springer-Verlag.
  • MATSUDA Yoshinori; TOYODA Hideyoshi; OUCHI Seiji
    Plant Biotechnology Japanese Society for Plant Cell and Molecular Biology 6 (1) 33 - 34 0289-5773 1989
  • TOYODA Hideyoshi; OKI Tomoya; MATSUDA Yoshinori; KATSURAGI Kiyonori; NISHIGUCHI Tsutomu; OUCHI Seiji
    Plant Biotechnology Japanese Society for Plant Cell and Molecular Biology 6 (2) 95 - 97 0289-5773 1989 
    トマト葉由来の friable カルスを使用し, カルス集塊を構成する細胞の形質転換を試みた. 導入する外来遺伝子としては, 植物形質転換ベクター, pBI 121を使用し, マイクロインジェクション法で特定標的細胞の細胞核に注入した. 形質転換細胞はカナマイシン添加培地で選抜するか, 5-bromo-4-chloro-3 indolyl glucuronide を基質として, 注入細胞におけるβ-glucuronidase 活性を細胞化学的に検出した, その結果, 約10%の注入細胞がこれらの活性を示し, 本法でカルス集塊の特定細胞が形質転換されることが明らかとなった.
  • Y. Matsuda; H. Toyoda; T. Nishiguchi; S. Ouchi
    Journal of Phytopathology 125 (1) 89 - 96 1439-0434 1989 [Refereed]
     
    An electroporation procedure for the introduction of fluorescein isothiocyanate‐conjugated albumin into intact macroconidia of Fusarium oxysporum f. sp. lycopersici was performed in this paper. FITC‐albumin was used to establish an efficient electroporation procedure because its presence in spores could be easily detected by fluorescence microscopy. The uptake of fluorescein isothiocyanate‐conjugated albumin into spores was successfully mediated by high voltage electrical pulses at field strength of 10 KV/cm. In electroporation of 106 macroconidia per ml with the present system, 1.2 × 105 spores incorporated the protein, and then 3.1 × 104 normally germinated, elongated vegetative hyphae, and produced spores. Copyright © 1989, Wiley Blackwell. All rights reserved
  • Hideyoshi Toyoda; Yoshinori Matsuda; Ryutaro Utsumi; Seiji Ouchi
    Plant Cell Reports Springer-Verlag 7 (5) 293 - 296 0721-7714 1988/08 [Refereed]
     
    An efficient method, called the culture plate method, was devised for microinjection of foreign materials into nuclei of tomato callus cells. The culture plate method, used in this study, is advantageous because cells suitable for microinjection can be selected microscopically and the injected cells subsequently cultured in the same plate. With this microinjection system, some foreign materials were injected into nuclei of callus cells without causing detrimental effects. Kanamycin-resistant callus clones were obtained 1 month after injection from single cells whose nuclei were microinjected with a NPT II DNA fragment of the pE2KX plasmid. © 1988 Springer-Verlag.
  • TOYODA Hideyoshi; MATSUDA Yoshinori; SHIMIZU Kunihiko; OGATA Hiroshi; HASHIMOTO Hisako; OUCHI Seiji
    Plant Biotechnology Japanese Society for Plant Cell and Molecular Biology 5 (2) 66 - 71 0289-5773 1988 
    Fusaric acid-resistant regenerants were selected in vitro by using the tissue culture system of tomato. The callus tissues were induced from the leaf explants of tomato and subcultured in a MS (0.01mg/l IAA and 0.1mg/l BAP) medium. After incubation for two weeks, a lot of green spots were produced in the callus tissues. The tissues were cut into small callus clumps so as to contain one green spot and transferred to a MS medium (0.1mg/l IAA and 1.0mg/l BAP) containing 50μg/ml fusaric acid. Fusaric acid-resistant regenerants were effectively developed from enlarged green spots and rooted in a hormone-free MS medium. For examining the resistance in regenerants at one month after transplantation to soil, their upper branches were cut and dipped in the solution containing 300μg/ml fusaric acid. In the branches of regenerants grown in the absence of fusaric acid, the resistance was fully maintained and expressed.
  • TOYODA Hideyoshi; MATSUDA Yoshinori; HIRAI Tokuzo
    Plant Biotechnology Japanese Society for Plant Cell and Molecular Biology 3 (1) 22 - 27 0289-5773 1986 
    Tobacco mosaic virus (TMV) was microinjected into cell-aggregates (2 to 4 cells-aggregates) prepared from friable tomato callus and multiplication and translocation of TMV in multi-cellular system were examined in the present study. Only a cell positioned at the outside of the aggregates was injected with TMV and other constituent cells of the same aggregates were not injected. Multiplication of TMV in the aggregates was detected by staining with fluorescent anti-TMV antibody. The rates of aggregates, where all constituent cells were stained with fluorescent antibody, were highest when an inoculum solution (concentrations of TMV, 100μg/ml) was injected into cytoplasm for 10sec. Multiplication of TMV simultaneously occurred in both TMV-injected and non-injected cells of the aggregates within 48hr after the injection. These results suggest that TMV might be translocated to non-injected cells soon after the injection and simultaneously multiplied in both injected and non-injected cells.
  • 豐田 秀吉; 松田 克礼; 平井 篤造
    植物組織培養 Japanese Society for Plant Cell and Molecular Biology 2 111 - 111 0289-5773 1985
  • 豊田 秀吉; 茶谷 和行; 松田 克礼; 平井 篤造
    植物組織培養 Japanese Society for Plant Cell and Molecular Biology 2 169 - 169 0289-5773 1985
  • TOYODA Hideyoshi; OGATA Hiroshi; MATSUDA Yoshinori; CHATANI Kazuyuki; HIRAI Tokuzo
    Plant Biotechnology Japanese Society for Plant Cell and Molecular Biology 2 (2) 70 - 73 0289-5773 1985 
    Plant regeneration from tomato leaf-explants was investigated in the study. Shoots were induced from explant-derived callus tissues in the medium containing plant hormones, IAA and BAP. Highly efficient and synchronous formation of shoots was observed when shoot-forming tissues were excised and transferred to the medium with 0.1mg/l IAA and 1mg/l BAP. Roots were developed in culturing shoots in hormone-free medium.
  • H. Toyoda; Y. Oishi; Y. Matsuda; K. Chatani; T. Hirai
    Journal of Phytopathology 114 (2) 126 - 133 1439-0434 1985 [Refereed]
     
    In somaclonal tissues obtained from systemically TMV‐infected tobacco plants, a relation between changes of TMV amounts and the callus growth was examined. The culture medium was suitable for maintaining a constant concentration of TMV as well as active callus growth. By using the shake‐culture method, somaclonal tissues were separated into two classes on the basis of callus sizes. In large callus tissues, TMV amounts were constant during subculturing but the tissues did not either grow or release the newly divided cells after the last subculture. On the other hand, smaller callus tissues grew markedly and the TMV amounts were conspicuously lowered. After shake‐subculture of smaller tissues, they were successfully regenerated to plantlets. None of the plantlets expressed any mosaic symptoms, while plantlets from the original somaclones showed severe mosaic symptoms of TMV in leaflets. Thus, the present report describes the successful production of virus‐free plantlets from infected somaclonal callus cultures. Copyright © 1985, Wiley Blackwell. All rights reserved
  • TOYODA Hideyoshi; MATSUDA Yoshinori; MOCHIZUKI Toshihiko; OISHI Yasuharu; HIRAI Tokuzo
    Plant Biotechnology Japanese Society for Plant Cell and Molecular Biology 1 (2) 53 - 56 0289-5773 1984 
    The system devised in the present study was equipped with plural culture vessels and some devices for changing chemical and physical culture conditions. The operation for the system was very easy and the sterility was kept over long periods. Conditions suitable for tomato cell culture were rapidly and easily established by using the system.

Books etc

  • An illustrated Manual of Electric Field Screens - Their Structures and Functions
    MATSUDA Yoshinori (Joint work)RAEFSS Publishing Department 2019/02
  • 図解静電場スクリーン その構造と機能
    松田克礼 (Joint work)2019/01 110
  • Electric field screen; principles and applications
    MATSUDA Yoshinori (Joint work)Nobunkyo Production Ltd 2015/03
  • 静電場スクリーンによる農作物防除システム
    松田 克礼 (Joint work)農山漁村文化協会 2014/03
  • 植物防疫, 静電場スクリーンの開発と実用化
    豊田 秀吉; 野々村 照雄; 松田 克礼; 金原淳司; 草刈眞一 (Joint work)社団法人 日本植物防疫協会 2008/10
  • 「農林水産技術 研究ジャーナル」 , 静電気で病害虫を絶つ ―静電場スクリーンの開発と実用化―
    豊田 秀吉; 野々村 照雄; 松田 克礼; 金原淳司; 草刈眞一 (Joint work)社団法人 農林水産技術情報協会 2008
  • クリーンテクノロジー, 誘電分極体スクリーンによる病害虫遮蔽システム 屋内への花粉侵入と栽培施設への病害虫侵入を遮蔽する新しい環境保全システム
    豊田 秀吉; 野々村 照雄; 松田 克礼; 本多健一郎; 草刈眞一; 井村岳男; 金原淳司 (Joint work)日本工業出版 2007/05
  • ニューカントリー, 静電場で病害虫のハウス侵入をシャットアウト
    豊田 秀吉; 野々村 照雄; 松田 克礼; 本多健一郎; 草刈眞一; 井村岳男; 金原淳司 (Joint work)北海道協同組合通信社 2007/03
  • 農業電化, 静電場を利用した病害虫の捕捉と施設栽培への応用
    豊田 秀吉; 野々村 照雄; 松田 克礼; 本多健一郎; 草刈眞一; 井村岳男; 金原淳司 (Joint work)農業電化協会 2007
  • ビニールと農園芸, 静電場スクリーンの開発と栽培施設への応用
    豊田 秀吉; 野々村 照雄; 松田 克礼; 本多健一郎; 草刈眞一; 井村岳男; 金原淳司 (Joint work)MKVプラテック株式会社 2007
  • 農業および園芸, 静電場スクリーンを利用した病害虫捕捉技術の開発と施設栽培への応用
    豊田 秀吉; 野々村 照雄; 松田 克礼; 本多健一郎; 草刈眞一; 井村岳男; 金原淳司 (Joint work)2007
  • 施設と園芸, 静電場を利用した画期的病害虫侵入防止技術の開発
    豊田 秀吉; 野々村 照雄; 松田 克礼; 本多健一郎; 大阪府立食とみどり; 総合技術センター; 奈良県農業総合センター; 金原淳司 (Joint work)日本施設園芸協会 2007
  • 植物防疫, トマトうどんこ病菌はどのように偽鎖生分生子柄をつくるのか
    豊田 秀吉; 松田 克礼 (Joint work)日本植物防疫協会 2007
  • 植物病の探究, トマトうどんこ病―動的解析法とその有効防除の確立をめざして
    豊田 秀吉; 松田 克礼; 野々村 照雄 (Joint work)「植物病の探究」出版会 2004
  • 新農業環境工学 21世紀のパースペクティブ, 植物を利用した有用物質の生産 ~1.工業原料の生産と利用~
    豊田 秀吉; 松田 克礼; 野々村 照雄 (Joint work)養賢堂 2004
  • Direct RT-PCR amplification of mature mRNAs in single trichome cells of plant leaves
    Recent Res. Devel. Cell Biol. 2003
  • Cloning of a chitinase gene from a chitin-degrading bacterium and its application to control of plant disease
    CHITIN ENZYMOLOZY 1993
  • Application of microinjection technique for the analysis of gene expression during host-parasite infraction
    Advances in Molecular Genetics of Plant-Microbe Interactions 1993

Conference Activities & Talks

  • Control of insect pests by electric field screen  [Invited]
    MATSUDA Yoshinori
    MY003915  2015/09
  • Electric field screen - Principles and applications  [Invited]
    MATSUDA Yoshinori
    MY003915  2015/05
  • Avoidance of an electric field by insects: Fundamental biological phenomenon for an electrostatic pest-exclusion strategy  [Invited]
    MATSUDA Yoshinori
    Electrostatics 2015  2015/04
  • Safe housing ensured by an electric field screen that excludes insect-net permeating haematophagous mosquitoes carrying human pathogens  [Invited]
    MATSUDA Yoshinori
    Electrostatics 2015  2015/04
  • Micropipette extraction-based RT-PCR amplification of mature mRNAs in single trichome cells of tomato leaves  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    Biotechnology and other omics in vegetable science  2012/05  トルコ  Biotechnology and other omics in vegetable science
  • Aroma compounds are produced in hairy root cultures of musk melon  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    Biotechnology and other omics in vegetable science  2012/05  トルコ  Biotechnology and other omics in vegetable science
  • Micropipette extraction-based RT-PCR amplification of mRNAs in nuclei or cytosol of single cells of tomato calli  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    Biotechnology and other omics in vegetable science  2012/05  トルコ  Biotechnology and other omics in vegetable science
  • A field investigation of the bryophyte distribution in the mountainous campus of Kinki University: Collection and cytological observation of liverworts as a potential source of nutraceuticals  [Not invited]
    瀧川 義浩; 角谷 晃司; 松田 克礼; 野々村 照雄; 豊田 秀吉
    MOSS 2011, The 14th annual international conference  2011/09  ドイツ  MOSS 2011, The 14th annual international conference
  • The flat liverwort thallus surface is the site of interactions with falling airborne fungal spores: Specification of conidial germination by powdery mildew on a thalloid surface  [Not invited]
    瀧川 義浩; 松田 克礼; 角谷 晃司; 野々村 照雄; 豊田 秀吉
    MOSS 2011, The 14th annual international conference  2011/09  ドイツ  MOSS 2011, The 14th annual international conference
  • Leaf surface of the moss is the site for trapping airborne fungal spores as nutritional targets  [Not invited]
    瀧川 義浩; 角谷 晃司; 松田 克礼; 野々村 照雄; 豊田 秀吉
    MOSS 2011, The 14th annual international conference  2011/09  ドイツ  MOSS 2011, The 14th annual international conference
  • AN ELECTRIC FIELD SCREEN CAN CREATE PEST-FREE SPACE WITH BETTER AIR PENETRATION IN OPEN-WINDOW GREENHOUSES  [Not invited]
    MATSUDA Yoshinori
    International Symposium on Advanced Technologies and Management Towards Sustainable Greenhouse Ecosystems: Greensys2011  2011/06
  • A newly devised electric field screen for avoidance and capture of greenhouse insect pests  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄; 金原淳司; 草刈眞一
    GreenSys2011  2011/06  ギリシア  GreenSys2011
  • An electrostatic insect exclusion technique enables germfree cultivation of tomato plants in open greenhouses  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄; 金原淳司; 草刈眞一
    GreenSys2011  2011/06  ギリシア  GreenSys2011
  • Suppression of leaf-surface germination of Oidium neolycopersici conidia by trichome exudates of Lycopersicon pennellii  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    9th International Mycological Congress: The Biology of Fungi  2010/08  スコットランド  9th International Mycological Congress: The Biology of Fungi
  • Instantaneous eradication of powdery mildew Oidium neolycopersici on tomato leaves by exposure to corona discharge  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    9th International Mycological Congress: The Biology of Fungi  2010/08  スコットランド  9th International Mycological Congress: The Biology of Fungi
  • Electrostatic attraction of highly germinative pseudochain conidia on conidiophores of tomato powdery mildew Oidium neolycopersici  [Not invited]
    豊田 秀吉; 野々村 照雄; 松田 克礼
    9th International Mycological Congress: The Biology of Fungi  2010/08  スコットランド  9th International Mycological Congress: The Biology of Fungi
  • Pseudochain formation and lifelong production of highly germinative conidia on conidiophores by highly virulent isolates of Oidium neolycopersici  [Not invited]
    豊田 秀吉; 野々村 照雄; 松田 克礼
    A Special Interest Group Meeting of the 9th International Mycological Congress  2010/08  スコットランド  A Special Interest Group Meeting of the 9th International Mycological Congress
  • A New Spore Precipitator with Polarized Dielectric Insulators for Physical Control of Tomato Powdery Mildew  [Not invited]
    MATSUDA Yoshinori
    International Symposium on Tomato Diseases  2010/07  イタリア  3rd International Symposium on Tomato Diseases
  • Protection of nursery stage tomato seedlings from rhizosphere pathogens Ralstonia solanacearum and Fusarium oxysporum f. sp. radicis-lycopersici  [Not invited]
    豊田 秀吉; 野々村 照雄; 松田 克礼
    3rd International Symposium on Tomato Diseases  2010/07  イタリア  3rd International Symposium on Tomato Diseases
  • 捕捉・忌避型静電場スクリーンを利用した施設栽培トマトの病虫害防除  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄; 金原淳司; 草刈眞一
    平成22年度日本植物病理学会大会  2010/04  京都  平成22年度日本植物病理学会大会
  • トマトうどんこ病菌の分生子柄に形成された偽鎖生分生子は高い発芽力を有し同調的に分化する  [Not invited]
    豊田 秀吉; 野々村 照雄; 松田 克礼; 瀧川義浩; 角谷晃司
    平成22年度日本植物病理学会大会  2010/04  京都  平成22年度日本植物病理学会大会
  • ショウジョウバエを用いた各種ポリフェノール化合物の抗加齢効果の検討  [Not invited]
    角谷 晃司; 瀧川 義浩; 野々村 照雄; 松田 克礼; 豊田 秀吉; 早川 堯夫; 近畿大学大学院農学研究科
    第59回日本薬学会近畿支部例会  2009/10  大阪  第59回日本薬学会近畿支部例会
  • トマトに含まれるポリフェノール類の動物培養細胞に及ぼす影響  [Not invited]
    角谷 晃司; 瀧川 義浩; 早川 堯夫; 野々村 照雄; 松田 克礼; 豊田 秀吉; 近畿大学農学部; 近畿大学農学部; 近畿大学農学部
    第59回日本薬学会近畿支部例会  2009/10  大阪  第59回日本薬学会近畿支部例会
  • A DIELECTRIC DIPOLAR SCREEN WITH OPPOSITELY POLARIZED INSULATORS FOR PROTECTING TOMATO SEEDLINGS ON A NURSERY HYDROPONIC CULTURE BED FROM WHITEFLIES  [Invited]
    MATSUDA Yoshinori
    International Symposium on Seed, Transplant and Stand Establishment of Horticultural Crops  2009/09
  • A new spore precipitator with polarized dielectric insulators for physical control of tomato powdery mildew.  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    5th International Symposium on Seed, Transplant and Stand Establishment of Horticultural Crops  2009/09  スペイン  5th International Symposium on Seed, Transplant and Stand Establishment of Horticultural Crops
  • A dielectric dipolar screen with oppositely polarized insulators for tomato seedlings on a nursery hydroponic culture bed from whiteflies.  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    5th International Symposium on Seed, Transplant and Stand Establishment of Horticultural Crops  2009/09  スペイン  5th International Symposium on Seed, Transplant and Stand Establishment of Horticultural Crops
  • Protection of nursery stage tomato seedlings from rhizosphere pathogens Ralstonia solanacearum and Fusarium oxysporum f. sp. radicis-lycopersici.  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    5th International Symposium on Seed, Transplant and Stand Establishment of Horticultural Crops  2009/09  スペイン  5th International Symposium on Seed, Transplant and Stand Establishment of Horticultural Crops
  • 静電場スクリーンを用いた施設栽培トマトの病害虫防除技術の開発.  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    日本生物環境工学会2009年福岡大会  2009/09  福岡  日本生物環境工学会2009年福岡大会
  • 静電場スクリーンを用いた温室トマトうどんこ病の物理的防除.  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    日本生物環境工学会2009年福岡大会  2009/09  福岡  日本生物環境工学会2009年福岡大会
  • オゾン生成型静電場スクリーンを用いた施設栽培トマトの地上および根圏病害の物理的防除.  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    日本生物環境工学会2009年福岡大会  2009/09  福岡  日本生物環境工学会2009年福岡大会
  • An electric dipolar screen with oppositely polarized insulators for excluding whiteflies from greenhouses.  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    GreenSys 2009  2009/06  カナダ  GreenSys 2009
  • Dual protection of hydroponic tomatoes from rhizosphere pathogens Ralstonia solanacearum and Fusarium oxysporum f. sp. radicis-lycopersici and airborne conidia of Oidium neolycopersici with an ozone-generative electrostatic spore precipitator.  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    GreenSys 2009  2009/06  カナダ  GreenSys 2009
  • A new spore precipitator with polarized dielectric insulators for physical control of tomato powdery mildew.  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    GreenSys 2009  2009/06  カナダ  GreenSys 2009
  • An ozone generative dipolar screen protects hydroponic tomatoes from rhizosphere and aerial pathogens  [Not invited]
    豊田 秀吉; 野々村 照雄; 松田 克礼
    9th International Congress of Plant Pathology  2008/08  イタリア(トリノ)  9th International Congress of Plant Pathology
  • Physical control of tomato powdery mildew Oidium neolycopersici with an electrostatic dipolar screen  [Not invited]
    豊田 秀吉; 野々村 照雄; 松田 克礼
    9th International Congress of Plant Pathology  2008/08  イタリア(トリノ)  9th International Congress of Plant Pathology
  • Bacteria colonizing melon fruit surface act as biocontrol agents to postharvest disease pathogens  [Not invited]
    豊田 秀吉; 松田 克礼; Xu, L
    9th International Congress of Plant Pathology  2008/08  イタリア(トリノ)  9th International Congress of Plant Pathology
  • Oidium neolycopersici: Intra-specific variability inferred from AFLP analysis and relationship with closely related powdery mildew fungi infecting various plant species  [Not invited]
    豊田 秀吉; 松田 克礼; Jankovics, T; Kovacs, G.M; Kiss, L; Bai, Y; Niks, R.E; Kovacs, G.M; Bardin, M; Nicot, P.C
    9th International Congress of Plant Pathology  2008/08  イタリア(トリノ)  9th International Congress of Plant Pathology
  • Comprehensive strategies for controlling tomato powdery mildew Oidium neolycopersici (1) Breeding of powdery mildew resistant cultivars of tomato by interspecific hybridization between common and wild Lycopersicon species  [Not invited]
    豊田 秀吉; 野々村 照雄; 松田 克礼
    平成20年度日本植物病理学会大会  2008/04  島根(くにびきメッセ)  平成20年度日本植物病理学会大会
  • Comprehensive strategies for controlling tomato powdery mildew Oidium neolycopersici (2) Biocontrol of the tomato powdery mildew with conidia of barley powdery mildew  [Not invited]
    豊田 秀吉; 野々村 照雄; 松田 克礼
    平成20年度日本植物病理学会大会  2008/04  島根(くにびきメッセ)  平成20年度日本植物病理学会大会
  • Coprehensive strategies for controlling tomato powdery mildew Oidium neolycopersici (3) Electrostatic eradication of Oidium neolycopersici colonizing tomato leaves by a plasma stream exposure  [Not invited]
    豊田 秀吉; 野々村 照雄; 松田 克礼
    平成20年度日本植物病理学会大会  2008/04  島根(くにびきメッセ)  平成20年度日本植物病理学会大会
  • Comprehensive strategies for controlling tomato powdery mildew Oidium neolycopersici (4) Physical control of the tomato powdery mildew with an electrostatic dipolar screen  [Not invited]
    豊田 秀吉; 野々村 照雄; 松田 克礼; 金原淳司; 草刈眞一
    平成20年度日本植物病理学会大会  2008/04  島根(くにびきメッセ)  平成20年度日本植物病理学会大会
  • Mechanisms for aerial water collection by foliar trichomes of the wild tomato Lycopersicon pennellii can be functional for suppressing conidial germination by Oidium neolycopersici  [Not invited]
    豊田 秀吉; 野々村 照雄; 松田 克礼
    平成20年度日本植物病理学会大会  2008/04  島根(くにびきメッセ)  平成20年度日本植物病理学会大会
  • RAPD分析による大和トウキと北海トウキの識別  [Not invited]
    角谷 晃司; 野々村 照雄; 松田 克礼; 豊田 秀吉; 近畿大学農学部; 福田商店; 福田商店
    日本生薬学会第53回年会  2007/09  埼玉  日本生薬学会第53回年会
  • Resveratrol合成関連酵素を利用したResvertarolおよびその前駆体4-coumaroyl CoAの生合成  [Not invited]
    角谷 晃司; 瀧川 義浩; 松田 克礼; 豊田 秀吉; 野々村 照雄; 近畿大学大学院農学研究科
    日本生薬学会第54回年会  2007/09  名古屋  日本生薬学会第54回年会
  • わが国に自生するツユクサの遺伝子多型解析-オオボウシバナの識別  [Not invited]
    角谷 晃司; 野々村 照雄; 松田 克礼; 豊田 秀吉; 近畿大学農学部
    第24回日本植物細胞分子生物学会(つくば)大会  2007/07  筑波  第24回日本植物細胞分子生物学会(つくば)大会
  • Discrimination of two powdery mildew fungi infecting leaves using its sequences amplified from their individual propagules by PCR  [Not invited]
    豊田 秀吉; 野々村 照雄; 松田 克礼
    XIII International congress on Molecular Plant-Microbe Interactions  2007/07  イタリア(ソレント)  XIII International congress on Molecular Plant-Microbe Interactions
  • Symptomatic evidence for differential root invasion by Fusarium crown and root rot pathogens between common tomato Lycopersicon esculentum and its varieties  [Not invited]
    豊田 秀吉; 野々村 照雄; 松田 克礼
    XIII International congress on Molecular Plant-Microbe Interactions  2007/07  イタリア(ソレント)  XIII International congress on Molecular Plant-Microbe Interactions
  • Direct RT-PCR amplification of mature mrnas in cytoplasm micropipetted from barley coleoptile epidermal cell ?A model system for analyzing gene expression in host cells attacked by powdery mildew  [Not invited]
    豊田 秀吉; 野々村 照雄; 松田 克礼
    XIII International congress on Molecular Plant-Microbe Interactions  2007/06  イタリア(ソレント)  XIII International congress on Molecular Plant-Microbe Interactions
  • 腫瘍細胞表面で発現するCD98hc抗原領域の探索  [Not invited]
    角谷 晃司; 野々村 照雄; 松田 克礼; 豊田 秀吉; 瀧川 義浩; 益子 高; 近畿大学大学院農学研究科
    2007年度日本農芸化学会大会  2007/03  東京  2007年度日本農芸化学会大会
  • 酵母細胞膜表面で固定化したResvertrol合成関連酵素による、 Resvertrolおよびその前駆体4-coumaroyl CoAの生合成  [Not invited]
    角谷 晃司; 野々村 照雄; 松田 克礼; 豊田 秀吉; 近畿大学大学院農学研究科; 近畿大学農学部
    2007年度日本農芸化学会大会  2007/03  東京  2007年度日本農芸化学会大会
  • アルギニントランスポーターを利用した出芽酵母細胞膜上における固定化酵素の作出  [Not invited]
    角谷 晃司; 野々村 照雄; 松田 克礼; 豊田 秀吉; 近畿大学大学院農学研究科
    第126回日本薬学会年会  2007/03  仙台  第126回日本薬学会年会
  • Electrostatic spore precipitation of tomato powdery mildew conidia with an electrostatic dipolar screen  [Not invited]
    豊田 秀吉; 野々村 照雄; 松田 克礼; 金原淳司; 草刈眞一
    平成19年度日本植物病理学会大会  2007/03  宇都宮大学  平成19年度日本植物病理学会大会
  • Exclusion of tobacco whiteflies by an electrostatic dipolar screen from greenhouse  [Not invited]
    豊田 秀吉; 野々村 照雄; 松田 克礼; 金原淳司; 井村岳男; 野菜茶業研究所; 草刈眞一
    平成19年度日本植物病理学会大会  2007/03  宇都宮大学  平成19年度日本植物病理学会大会
  • Suppression of colonies of tomato powdery mildew with an electrostatic ozone discharge  [Not invited]
    豊田 秀吉; 野々村 照雄; 松田 克礼; 草刈眞一; 金原淳司
    平成19年度日本植物病理学会大会  2007/03  宇都宮大学  平成19年度日本植物病理学会大会
  • An ozone generative electrostatic spore precipitator for dual control of rhizosphere and phyllosphere pathogens of hydroponic tomatoes  [Not invited]
    豊田 秀吉; 野々村 照雄; 松田 克礼; 草刈眞一; 金原淳司
    平成19年度日本植物病理学会大会  2007/03  宇都宮大学  平成19年度日本植物病理学会大会
  • Consecutive monitoring of lifelong production of conidia by individual conidiophores of Blumeria graminis f. sp. hordei on barley leaves by digital microscopic techniques with electrostatic micromanipulation.  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    8th International Mycological Congress  2006/08  オーストラリア ケアンズ  8th International Mycological Congress
     
    筆者らは、高解像能型デジタルマイクロスコープを用いてうどんこ病菌分生子の形成過程を観察し、その形成機構を解析してきた。そこで本実験では、デジタルスコープを用いてうどんこ病菌分生子の放出過程を連続観察することとした。また、不均一電場に置かれたうどんこ病菌分生子が電気力線に従って移動することに着目し、分生子柄から放出される分生子を連続的に誘電分極絶縁体へ捕獲することを試みた。まず、エボナイト製のマイクロプローブを作製し、静電気発生装置と連結させ、デジタルスコープのマニピュレーターに取り付けた。次に、オオムギうどんこ病菌Blumeria graminis f. sp. hordeiの菌叢を観察し、形成直後の分生子柄上部に誘電分極したプローブを近づけ、成熟して放出される分生子を捕獲した。独立した分生子柄から順次放出される分生子を捕獲し、その放出数を測定したところ、ひとつの分生子柄からは、2.5時間間隔で33個の分生子が放出されることが明らかとな
  • Symptomatic evidence for differential root invasion by Fusarium crown and root rot pathogens between common tomato Lycopersicon esculentum and its varieties.  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    8th International Mycological Congress  2006/08  オーストラリア ケアンズ  8th International Mycological Congress
     
    本実験では、トマト(Lycopersicon esculentum var. pyriforme cv. Yellow Pear)の幼苗を用い、日本各地で単離されたトマト根腐萎凋病菌(13株)の発病差異を解析した。まず、主根部に100μlの胞子懸濁液(1.0×107胞子/ml)を滴下接種し、接種個体の根部病徴を詳細に観察したところ、強度病原性菌株では、側根の先端部が速やかに褐変化するとともに、導管部への褐変化が広がり、最終的に地際部へ伸展することが明らかとなった。一方、弱度病原性菌株では、側根形成部位周辺で表皮細胞の壊死が観察されたが、その病徴伸展は遅いものであった。また、中度病原性菌株は、側根の出現時に形成された皮層の裂け目から侵入し、導管褐変を引き起こす現象が観察された。
  • An apparatus for collecting total conidia of Blumeria graminis f. sp. hordei from leaf colonies using electrostatic attraction.  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    8th International Mycological Congress  2006/08  オーストラリア ケアンズ  8th International Mycological Congress
     
    筆者らは、不均一電場に置かれたうどんこ病菌分生子が電気力線に従って移動することに着目し、誘電分極絶縁体を用いた分生子捕獲技術を開発した。そこで本実験では、実際の栽培環境に対応した分生子モニタリングシステムの開発を目的とし、ひとつの菌叢からその生涯を通して放出される総胞子数の測定技術を確立することとした。分生子の捕獲には、誘電分極体としてエボナイトプレート(3cm×18cm)を使用し、オオムギ葉上に形成された菌叢(Blumeria graminis f. sp. hordei)から成熟して放出される分生子をプレート上に捕獲した。誘電分極プレートの移動を自動化することにより、10分間隔で3週間(3,024枚)、連続的に分生子を捕獲し、分生子数を測定した。その結果、ひとつの菌叢から1.2×105個の胞子が放出され、その放出は約460時間(20日間)継続されることが明らかとなった。今後、分生子モニタリングシステムを栽培施設に設置し、分生子生産の概日リズムや異なる環境条件に
  • Identification of leaf epicuticular amphiphiles produced by the wild tomato species Solanum pennellii  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄; 東勝秀; 西村浩明
    平成18年度日本植物病理学会大会  2006/06  北海道 札幌  平成18年度日本植物病理学会大会
     
    筆者らの研究室では、野生種トマトSolanum pennellii TK4560の葉上に分泌される両親媒性物質が空気中の水分をトラップし、形成された水滴が葉上に広がること、およびその物質が葉面生息細菌の栄養源になることを明らかにしてきた。そこで本実験では、S. pennelliiの葉上分泌物質の化学的構造を決定することとした。まず、S. pennellii TK4560の種子を播種し、3ヶ月間、ガラス温室内で栽培した後、葉面を滅菌水で洗浄し、フィルターろ過した洗液を凍結乾燥した。得られた乾燥粉末(70mg)を20mlのジエチルエーテルと0.5%炭酸ナトリウム混液に溶解し、有機溶媒相を分取した。この画分(10mg)について超臨界MeOH分解を行い、さらにNMR、IR、GCおよびGC/MS分析を行った。その結果、S. pennellii葉上に分泌される両親媒性物質は、少なくとも9種類のアシル基を有するトリアシルグルコース混合物であることが明らかとなった。
  • Antifungal activity of glandular trichome exudates of the wild tomato Solanum pennelli against Oidium neolycopersici conidia  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄; 西村浩明
    平成18年度日本植物病理学会大会  2006/06  北海道 札幌  平成18年度日本植物病理学会大会
     
    筆者らの研究室では、トマトうどんこ病に対する抵抗性因子を検索するため、8種120系統の野生種トマトをスクリーニングしたところ、それら野生種トマトの中でSolanum pennelliが他のトマトに比べて、葉上に形成されたうどんこ病菌菌叢が少ないことを見出した。そこで本実験では、葉上トリコームによるトマトうどんこ病菌分生子の捕獲と分泌物による分生子の抗菌活性を検討することとした。まず、S. pennellii葉上にトマトうどんこ病菌分生子を接種したところ、粘性のあるIV型トリコーム先端部位で分生子が捕獲されるとともに、分泌液中に埋没し、発芽の抑制が観察された。次に、顕微鏡下でマイクロピペットを用い、IV型トリコーム分泌液を直接回収し、メタノールに溶解した後、種々の濃度に希釈し、分生子の発芽に及ぼす影響を調べた。その結果、10μg/ml以上の濃度で分生子の発芽が完全に抑制され、分生子内構造の崩壊が観察された。以上の結果から、S. pennellii TK4560のIV型トリ
  • Expression of powdery mildew resistance in crosses between the powdery mildew-resistant wild tomato species Lycopersicon hirsutum and common tomato L. esculentum  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄; 西村浩明; 草刈眞一
    平成18年度日本植物病理学会大会  2006/06  北海道 札幌  平成18年度日本植物病理学会大会
     
    筆者らは、トマトうどんこ病菌Oidium neolycopersici Kissに対する抵抗性因子を検索するため、野生種トマト8種120系統から抵抗性系統の選抜を実施し、本菌に対して抵抗性を示す8系統のLycopersicon hirsutum を選抜した。また、これら8系統に分生子を人為的に接種し、2週間後に標徴の有無を観察したところ、4系統は強度抵抗性を示し、他の4系統は壊死を伴う中度抵抗性を示した。完全な抵抗性を示す4系統に本菌分生子を接種し、24時間後に表皮細胞を観察したところ、過敏感反応によって、うどんこ病菌の吸器形成は完全に抑制されていた。そこで本実験では、強度抵抗性個体と栽培種(L. esculentum cv. Moneymaker)の種間交雑を行い、うどんこ病に対する抵抗性を検討することとした。得られた交雑系統に本菌分生子を接種し、24時間後に表皮細胞を観察したところ、うどんこ病菌の吸器形成は完全に抑制されず、その抵抗性レベルは栽培種との中間に位置した。現在、種間交雑系統から得た
  • Counting ot total conidia produced by barley powdery mildew colonizing host leaves  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄; 東勝秀; 草刈眞一
    平成18年度日本植物病理学会大会  2006/06  北海道 札幌  平成18年度日本植物病理学会大会
     
    筆者らは、不均一電場に置かれたうどんこ病菌分生子が電気力線に従って移動することに着目し、誘電分極絶縁体を用いた分生子捕獲技術を開発した。そこで本実験では、実際の栽培環境に対応した分生子モニタリングシステムの開発を目的とし、ひとつの菌叢からその生涯を通して放出される総胞子数の測定技術を確立することとした。分生子の捕獲には、誘電分極体としてエボナイトプレート(3cm×18cm)を使用し、オオムギ葉上に形成された菌叢(Blumeria graminis f. sp. hordei)から成熟して放出される分生子をプレート上に捕獲した。誘電分極プレートの移動を自動化することにより、10分間隔で3週間(3,024枚)、連続的に分生子を捕獲し、分生子数を測定した。その結果、ひとつの菌叢から1.2×105個の胞子が放出され、その放出は約460時間(20日間)継続されることが明らかとなった。今後、分生子モニタリングシステムを栽培施設に設置し、分生子生産の概日リズムや異なる環境条件に
  • Consecutive monitoring of lifelong conidial production by barley powdery mildew  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄; 東勝秀; 草刈眞一
    平成18年度日本植物病理学会大会  2006/06  北海道 札幌  平成18年度日本植物病理学会大会
     
    筆者らは、高解像能型デジタルマイクロスコープを用いてうどんこ病菌分生子の形成過程を観察し、その形成機構を解析してきた。そこで本実験では、デジタルスコープを用いてうどんこ病菌分生子の放出過程を連続観察することとした。また、不均一電場に置かれたうどんこ病菌分生子が電気力線に従って移動することに着目し、分生子柄から放出される分生子を連続的に誘電分極絶縁体へ捕獲することを試みた。まず、エボナイト製のマイクロプローブを作製し、静電気発生装置と連結させ、デジタルスコープのマニピュレーターに取り付けた。次に、オオムギうどんこ病菌Blumeria graminis f. sp. hordeiの菌叢を観察し、形成直後の分生子柄上部に誘電分極したプローブを近づけ、成熟して放出される分生子を捕獲した。独立した分生子柄から順次放出される分生子を捕獲し、その放出数を測定したところ、ひとつの分生子柄からは、2.5時間間隔で33個の分生子が放出されることが明らかとな
  • Comparative study for conidium formation by tomato and barley powdery mildews  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄; 許玲; 草刈眞一
    平成18年度日本植物病理学会大会  2006/06  北海道 札幌  平成18年度日本植物病理学会大会
     
    トマトうどんこ病菌Oidium neolycopersici Kissは,単生型うどんこ病菌であるが、頻繁に偽鎖生状態を形成することから鎖生型うどんこ病菌との識別を困難にしている。そこで本実験では、近畿大学農学部で分離されたトマトうどんこ病菌O. neolycopersici KTP-01(Kashimoto et al)の分生子形成過程およびオオムギうどんこ病菌Blumeria graminis f. sp. hordeiの分生子形成過程を連続的に観察することとした。その結果、トマトうどんこ病菌の分生子は、分生子柄上に単一で形成され、その後分生子柄から離脱することなく、新たな分生子形成がくりかえされ、偽鎖生状態となった。また、偽鎖生を形成する分生子が4個になるとその重みによって傾斜し、分生子柄から離脱して菌叢上に落下した。一方、オオムギうどんこ病菌の分生子は、分生子柄上に鎖状で形成され、鎖を形成する分生子が10個になると、成熟した最先端の分生子のみ、分生子柄から離脱・放出されることが明らかとなった。
  • Instantaneous breakdown of leaf-surface colonizing powdery mildew by an electrostatic streamer discharge  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄; 東勝秀; 草刈眞一
    平成18年度日本植物病理学会大会  2006/06  北海道 札幌  平成18年度日本植物病理学会大会
     
    針状プローブにプラス電荷を与えると、アースされている導体との間で放電が発生し、それらは電位差によってコロナ放電、ストリーマ放電、アーク放電の3つのパターンに分類される。また、植物が導体特性を示すことから、土壌に定植された植物はアースされた状態となり、類似の放電現象を起こすことが可能となる。そこで本実験では、植物葉上のうどんこ病菌菌叢にプローブを接近させ、植物体に被害を及ぼすことなく病原菌を物理的に駆除するシステムを確立することとした。まず、絶縁体のアクリルシリンダー内にステンレス製のプローブを固定し、Streamer dischargerを作製した。次に、それを用いてトマトうどんこ病菌Oidium neolycopersici菌叢とプローブ先端部との間に発生する放電現象およびうどんこ病菌の状態を観察した。その結果、ストリーマ放電が発生している場合には分生子柄および菌糸の瞬間的崩壊が確認され、植物体への影響は全く認められなかった。また、駆除に要
  • Development of a new spore precipitator with polarized dielectric insulators for physical control of tomato powdery mildew  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄; 東勝秀; 草刈眞一
    平成18年度日本植物病理学会大会  2006/06  北海道 札幌  平成18年度日本植物病理学会大会
     
    施設栽培におけるうどんこ病の発生は、外部から施設内への分生子の侵入に起因している。そこで本実験では、うどんこ病菌分生子が不均一電場を移動することに着目し、誘電分極絶縁体を用いた新しい物理的遮蔽システムを確立することとした。まず、透明なアクリル製シリンダー(直径1cm)に銅線を通過させ、それらを平行に配列することにより誘電分極遮蔽スクリーンを作製した。そのスクリーンを立方体フレームに装着し、ガラスハウス内に設置して静電気発生装置と連結した後、その内部でトマト苗を育成した。また、同時に遮蔽装置の外部には、うどんこ病の感染を受けたトマト苗を放置し、大型ファンによりハウス内の空気を循環させた。3ヶ月間栽培を継続したところ、遮蔽装置外部のトマトは全て激しくうどんこ病菌の感染を受けていた。しかしながら、遮蔽装置内部のトマトには、栽培期間を通してうどんこ病の発生は全く認められず、健全な状態を維持していた。今
  • Electrostatic attraction of powdery mildew conidia with polarized dielectric insulators  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄; 東勝秀; 草刈眞一
    平成18年度日本植物病理学会大会  2006/06  北海道 札幌  平成18年度日本植物病理学会大会
     
    不均一電場に置かれたうどんこ病菌分生子は、静電誘導されて分極状態となり、不均一電場を電気力線に従って移動する。そこで本実験では、誘電分極絶縁体を用いて不均一電場を形成し、分生子柄上で成熟した分生子および空中を浮遊落下する分生子の捕獲を試みた。まず、絶縁体であるエボナイト棒(直径2mm)の先端を20μmに加工し、誘電分極体マイクロプローブを作製した。次に、作製したプローブに電気コードを接続し、静電気発生装置と連結させ、デジタルスコープに設置したマニピュレーターに取り付けた。顕微鏡下においてトマトうどんこ病菌(Oidium neolycopersici KTP-01)の分生子柄を観察し、成熟した分生子にプローブを近づけた後、静電気発生装置のスイッチを入れ、プローブ先端を誘電分極させた。その結果、成熟した分生子は不均一電場を移動し、誘電分極体プローブに捕獲された。また、分生子を自然落下させた場合にも、不均一電場を移動し、誘電分極体プローブ
  • 高解像能デジタルマイクロスコープによるトマトうどんこ病菌分生胞子発芽の連続観察  [Not invited]
    角谷 晃司; 野々村 照雄; 松田 克礼; 豊田 秀吉; 近畿大学農学部; 近畿大学大学院農学研究科; 近畿大学農学部; 大阪府立農林技術センター
    平成17 年度日本植物病理学会大会  2006/03  静岡  平成17 年度日本植物病理学会大会
  • オムギうどんこ病菌による子葉鞘細胞において傷害刺激で誘導されるキチナーゼ遺伝子の能動的抑制  [Not invited]
    角谷 晃司; 野々村 照雄; 松田 克礼; 豊田 秀吉; 近畿大学農学部; 近畿大学農学部; 近畿大学農学部
    平成17 年度日本植物病理学会大会  2006/03  静岡  平成17 年度日本植物病理学会大会
  • エンド型およびエキソ型ポリガラクツロナーゼ遺伝子の塩基置換を利用したトマト根腐萎凋病菌菌株間での識別マーカーの開発  [Not invited]
    角谷 晃司; 野々村 照雄; 松田 克礼; 豊田 秀吉; 近畿大学農学部; 近畿大学農学部; 近畿大学農学部
    平成17 年度日本植物病理学会大会  2006/03  静岡  平成17 年度日本植物病理学会大会
  • Development of DNA markers for discriminating isolates of Fusarium oxysporum f. sp. radicis-lycopersici on the basis of single nucleotide polymorphism of polygalacuturonase genes.  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄; 角谷晃司
    日本植物病理学会関西部会  2005/09  名古屋  日本植物病理学会関西部会
     
    本実験では、トマト根腐萎凋病菌がもつエンド型およびエキソ型のポリガラクツロナーゼ(PG)遺伝子の塩基置換に着目し、単離菌株を明確に識別するDNAマーカーの開発を試みた。まず、既報の外国産分離菌株から4種類のPG遺伝子を検索し、それらの塩基配列を基にそれぞれPG遺伝子を特異的に増幅するプライマーを構築した。次に、幼苗トマト接種法で強度、中度および弱度病原性を示した根腐萎凋病菌5菌株および非病原性F. oxysporumからPG遺伝子をPCRで増幅し、全塩基配列を決定した後、アライメントを作製した。その結果、4種類のPG遺伝子にそれぞれ複数箇所の塩基置換が確認されたことから、それらの塩基置換を利用した単離菌株特異的プライマーを構築した。次に、作製したプライマーを組み合わせてPCRを行ったところ、供試した単離菌株を明確に識別できた。
  • Effect of relative humidity on pseudochain formation by Oidium neolycopersici on tomato leaves.  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄; 西村浩明; 草刈眞一
    日本植物病理学会関西部会  2005/09  名古屋  日本植物病理学会関西部会
     
    本実験では、近畿大学農学部で分離されたO. neolycopersici KTP-01を異なる湿度条件下(相対湿度20~99%)で培養し、偽鎖生形成に及ぼす影響を調査した。その結果、設定したすべての湿度条件下において2~4個の分生子で偽鎖生が形成され、定義されている高湿度条件下に限定されないことが明らかとなった。
  • Possible amplification of mature mRNAs in single cells of tomato callus aggregates by direct RT-PCR of cytosolic contents suctioned out with a micropipette  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    XVII International Botanical Congress  2005/08  オーストリア  XVII International Botanical Congress
     
    本実験では、単一細胞から核以外の細胞質をマイクロピペットで吸引する単一細胞でRT-PCR法を確立することとした。トマトで構成的に発現する遺伝子を検索し、glyceraldehyde- 3-phosphate dehydrogenaseを指標遺伝子として選抜した。トマトカルス細胞において核を細胞内に残存させ、それ以外の内容物をガラス針で吸引後、反応液へと排出してRT-PCRを行った。その結果、mRNA由来の産物のみ増幅され、本法を用いれば転写された遺伝子のみを特異的に検出できることが明らかとなった。
  • Direct RT-PCR amplification of mature mRNAs in cytoplasm micropipetted from barley coleoptile epidermal cell ?A model system for analyzing gene expression in host cells attacked by powdery mildew  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    XVII International Botanical Congress  2005/08  オーストリア  XVII International Botanical Congress
     
    本実験では、顕微鏡下で標的とした単一細胞からマイクロピペットを用いて細胞内容物を吸引し、そこに存在するmRNAを鋳型とした単一細胞RT-PCR法を適用することとした。病原菌の感染によって発現が誘導される遺伝子(CHI2、GLUなど)を検索した。誘導型遺伝子の発現を厳密に評価するため、RT-PCRを行う際に、先の構成的発現遺伝子を増幅するプライマーを混合し、指標遺伝子として同時に増幅することにより、誘導型遺伝子の発現検出を行った。その結果、病原菌の感染を受けていない細胞およびその感染を受けている両者の細胞において指標とした構成的発現遺伝子は検出された。誘導型遺伝子として使用したCHI2遺伝子およびGLU遺伝子は病原菌の感染を受けていない細胞においてその発現が検出され、うどんこ病菌の感染過程において抑制される傾向にあった。
  • Gene expression detection in single trichome cells of plant leaves by direct RT-PCR amplification of mature mRNAs in response to external stimuli  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    XVII International Botanical Congress  2005/08  オーストリア  XVII International Botanical Congress
     
    本実験では、単一細胞RT-PCR法をトリコーム細胞に適用し、揮発性物質により誘導される遺伝子の発現検出を行うこととした。シグナル伝達に関与する遺伝子(Pto、MEK1など)を選抜した。遺伝子を活性化させる物質としてサリチル酸(SA)とジャスモン酸(JA)を適用し、標的としたトリコームに暴露処理した後、速やかに細胞内容物を吸引してRT-PCRを行った。その結果、PtoはJA処理によって特異的に誘導され、MEK1はSAおよびJAによって誘導されることが明らかとなった。
  • Production of wholesome plants by introduction of genes for tumor-specific protein (CD98),  [Not invited]
    角谷 晃司; 野々村 照雄; 松田 克礼; 豊田 秀吉; 近畿大学大学院農学研究科
    XVII Interntional Botanical Congress  2005/06  Austria  XVII Interntional Botanical Congress
  • Possible suppression of chitinase gene expression induced in detached inner epidermis of barley coleoptile by powdery mildew pathogen  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    The Second Asian Conference on Plant Pathology 2005  2005/06  シンガポール  The Second Asian Conference on Plant Pathology 2005
     
    オオムギうどんこ病菌の感染を受けた表皮細胞は、その感染過程の進行に対応し、異なる反応を示すと考えられる。しかしながら、これらの宿主細胞の反応が感染を受けている細胞に限定されること、また、その反応が数分単位で進行することから、病原菌の感染過程で誘導される遺伝子の発現を解析するためには、顕微鏡下で感染過程を連続的に観察し、感染を受けている特定細胞において遺伝子発現を検出する細胞レベルの解析系が必須となる。本実験では、顕微鏡下で標的とした単一細胞からマイクロピペットを用いて細胞内容物を吸引し、そこに存在するmRNAを鋳型とした単一細胞RT-PCR法を適用することとした。
  • Symptomatic analysis of Fusarium crown and root rot in pear tomato Lycopersicon esculentum var. pyriforme inoculated with different isolates of Fusarium oxysporum f. sp. radicis-lycopersici  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    The Second Asian Conference on Plant Pathology 2005  2005/06  シンガポール  The Second Asian Conference on Plant Pathology 2005
     
    本実験では、トマト根腐萎凋病菌の病原性を迅速に検定する方法を考案し、幼苗スラント法と命名した。本法によって、接種個体の根部病徴を14日間経時的に観察し、個体ごとの発病指数を算出して各品種における罹病性程度を解析した。まず、三重県分離株を用いて市販トマト20品種の罹病性差異を比較検討したところ、特に感受性程度の高い品種が存在し、95%以上の個体で根部の褐変・壊死が観察された。そこで、これら高度感受性品種を指標として日本各地で単離されたトマト根腐萎凋病菌の病原性検定を行ったところ、各分離菌株間で発病に差異がみられ、栃木県分離株では95%以上の枯死率が確認された。
  • Symptomatic analysis of Fusarium crown and root rot i pear tomato Lycopersicon esculentum var. pyriforme inoculated with different isolates of Fusarium oxysporum f. sp. radicis-lycopersici.  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄; 岡田清嗣; 草刈
    日本植物病理学会大会  2005/03  静岡  日本植物病理学会大会
     
    本実験では、トマト(Lycopersicon esculentum var. pyriforme cv. Yellow Pear)の幼苗を用い、日本各地で単離されたトマト根腐萎凋病菌(13株)の発病差異を解析した。まず、主根部に100μlの胞子懸濁液(1.0×107胞子/ml)を滴下接種し、接種個体の根部病徴を詳細に観察したところ、強度病原性菌株では、側根の先端部が速やかに褐変化するとともに、導管部への褐変化が広がり、最終的に地際部へ伸展することが明らかとなった。一方、弱度病原性菌株では、側根形成部位周辺で表皮細胞の壊死が観察されたが、その病徴伸展は遅いものであった。また、中度病原性菌株は、側根の出現時に形成された皮層の裂け目から侵入し、導管褐変を引き起こす現象が観察された。
  • Development of DNA markers for discriminating isolates of Fusarium oxysporum f. sp. radicis-lycopersici on the basis of nucleotide substitution of endo- and exo-polygalacuturonase genes.  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄; 角谷晃司
    日本植物病理学会大会  2005/03  静岡  日本植物病理学会大会
     
    本実験では、トマト根腐萎凋病菌(Fusarium oxysporum f. sp. radicis-lycopersici)がもつエンド型およびエキソ型のポリガラクツロナーゼ(PG)遺伝子の塩基置換に着目し、単離菌株を識別するDNAマーカーの開発を試みた。まず、既報の外国産分離菌株から4種類のPG遺伝子(pg1、pg2、exoPG1およびpgx1)を検索し、それらの塩基配列を基にそれぞれPG遺伝子を特異的に増幅するプライマーを構築した。次に、幼苗トマト接種法で強度、中度および弱度病原性を示したトマト根腐萎凋病菌5菌株および非病原性F. oxysporumからPG遺伝子をPCRで増幅し、それらの塩基配列を決定した後、アライメントを作製した。その結果、4種類のPG遺伝子にそれぞれ複数箇所の塩基置換が確認されたことから、それらの塩基置換を利用した単離菌株特異的プライマーを構築した。作製したプライマーを組み合わせて使用することにより、本実験で供試した単離菌株を明確に識別できることが明らかとなった。
  • Consecutive observation of germination by tomato powdery mildew Oidium neolycopersici with a high-fidelity digital microscope.  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄; 角谷晃司; 西村浩明; 草刈眞一
    日本植物病理学会大会  2005/03  静岡  日本植物病理学会大会
     
    筆者らは、高解像能デジタルマイクロスコープ(HFDM)を用いて、トマト葉に接種したうどんこ病菌の分生胞子形成過程を連続的に観察してきた。本実験では、HFDMを用いて疎水もしくは親水面上に接種した分生胞子を観察し、その形態変化を追跡した。まず、疎水面としてプラスチック、親水面としてカバーグラスおよびセルロース膜を用い、その上に分生胞子を接種して発芽過程に要する時間とその形態変化を観察した。その結果、いずれの接種面においても3時間後から発芽の開始が観察され、接種面の相違による差異は認められなかった。
  • Screening of wild Lycopersicon species for resistance to Japanese isolate of tomato podery mildew Oidium neolycopersici.  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄; 角谷晃司; 西村浩明; 草刈眞一
    平成17年度日本植物病理学会大会  2005/03  静岡  平成17年度日本植物病理学会大会
     
    トマトうどんこ病菌Oidium neolycopersici KTP-01に対する抵抗性遺伝子を検索するため、野生型トマト8種96系統から抵抗性系統の選抜を試みた。まず、野生種トマトおよび高度感受性品種Moneymakerの苗を温室内に定植し、本菌に罹病したトマト個体から分生胞子を拡散・接種した(1次選抜)。その結果、本菌に対して抵抗性を示す8系統のL. hirusutumが確認された。
  • Suppression of chitinase gene expression iduced in detached inner epidermis by invading conidia of barley powdery mildew.  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄; 角谷晃司
    平成17年度日本植物病理学会大会  2005/03  静岡  平成17年度日本植物病理学会大会
     
    本実験では、うどんこ病菌の侵入を受けている細胞にRT-PCR/Nested-PCRを適用し、キチナーゼ遺伝子の発現推移を検討した。また、遺伝子の発現検出をより厳密に行うため構成的発現遺伝子を指標としたmultiplex PCRを適用した。その結果、同一子葉鞘組織の感染および非感染細胞の両者において指標遺伝子の発現は構成的に検出されたが、キチナーゼ遺伝子の発現は、非感染細胞からは検出されたものの、病原菌侵入細胞からは検出されなかった。
  • Biological control of phytophagous ladybird beetles Epilachna vigintioctopunctata (Coleoptera: Coccinellidae) by chitinolytic phylloplane bacteria Alcaligenes paradoxus entrapped in alginate beads  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    12th Symposium Insect-Plant Relationships  2004/09  ベルリン  12th Symposium Insect-Plant Relationships
  • Suppression of leaf feeding and oviposition of phytophagous ladybird beetles (Coleoptera: Coccinellidae) by chitinase gene-transformed phylloplane bacteria and their specific bacteriophages entrapped in alginate gel beads  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    12th Symposium Insect-Plant Relationship  2004/09  ベルリン  12th Symposium Insect-Plant Relationship
  • ミヤコグサ(Lotus japonicus)におけるトランスポーター遺伝子の発現解析  [Not invited]
    角谷 晃司; 瀧川 義浩; 野々村 照雄; 松田 克礼; 豊田 秀吉; 近畿大学大学院農学研究科; 近畿大学農学部
    第45回日本植物生理学会年会  2004/08  第45回日本植物生理学会年会
  • ミヤコグサにおけるトランスポーター遺伝子の発現解析  [Not invited]
    角谷 晃司; 瀧川 義浩; 野々村 照雄; 松田 克礼; 豊田 秀吉; 益子 高; 近畿大学大学院農学研究科; 近畿大学農学部; 近畿大学農学部; かずさDNA研; かずさDNA研
    日本農芸化学学会平成15年度大会  2004/08  日本農芸化学学会平成15年度大会
  • トマト栽培品種における迅速かつ効率的な再分化条件の確立  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    平成16年度日本植物細胞分子生物学会大会  2004/08  秋田  平成16年度日本植物細胞分子生物学会大会
  • ミヤコグサにおける硝酸およびアミノ酸トランスポーター遺伝子の発現解析  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    平成16年度日本植物細胞分子生物学会大会  2004/08  秋田  平成16年度日本植物細胞分子生物学会大会
  • Stable phylloplane colonization by entomopathogenic bacterium Pseudomonas fluorescens KPM-018P and biological control of phytophagous ladybird beetles Epilachna vigintioctopunctata  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    12th Symposium Insect-Plant Relationships  2004/08  ベルリン  12th Symposium Insect-Plant Relationships
  • GFP標識トマト根腐れ萎凋病菌を用いた糸状菌捕食性水生生物の効果的な選抜  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    平成16年度日本植物病理学会大会  2004/03  福岡  平成16年度日本植物病理学会大会
  • 異なる培養条件でのトマト根腐れ萎凋病菌のペクチン分解性検定  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    平成16年度日本植物病理学会大会  2004/03  福岡  平成16年度日本植物病理学会大会
  • フザリウム病菌の4つのポリガラクチュロナーゼ遺伝子を利用した分子系統解析  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    平成16年度日本植物病理学会大会  2004/03  福岡  平成16年度日本植物病理学会大会
  • 根腐れ萎凋病菌接種トマト根における根部病徴進展の解析  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    平成16年度日本植物病理学会大会  2004/03  福岡  平成16年度日本植物病理学会大会
  • 5-フルオロインドール(5-FI)によるフザリウム病菌の抑制と5-FI耐性植物の作出(V) シロイヌナズナおよびトマトへのアントラニル酸突然変異遺伝子の導入と発現解析  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    平成16年度日本植物病理学会大会  2004/03  福岡  平成16年度日本植物病理学会大会
  • デジタルマイクロスコープを利用したトマト葉上におけるOidium neolycopersiciによる分生胞子形成の連続観察  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    平成16年度日本植物病理学会大会  2004/03  福岡  平成16年度日本植物病理学会大会
  • デジタルマイクロスコープを利用した単一胞子PCRによるうどんこ病菌ITS領域の解析  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    平成16年度日本植物病理学会大会  2004/03  福岡  平成16年度日本植物病理学会大会
  • デジタルマイクロスコープを用いたうどんこ病菌の感染挙動と単一胞子RT-PCR/Nested PCRによる遺伝子発現の動的解析  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    平成16年度日本植物病理学会大会  2004/03  福岡  平成16年度日本植物病理学会大会
  • トマトうどんこ病菌Oidium neolycopersiciの薬剤感受性検定  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    平成16年度日本植物病理学会大会  2004/03  福岡  平成16年度日本植物病理学会大会
  • トマトうどんこ病菌Oidium neolycopersiciKTP-01に対する抵抗性系統の検索  [Not invited]
    豊田 秀吉; 松田 克礼; 野々村 照雄
    平成16年度日本植物病理学会大会  2004/03  福岡  平成16年度日本植物病理学会大会
  • トマトgypsy 様レトロトランスポゾンの逆転写酵素領域における変異の解析  [Not invited]
    角谷 晃司; 豊田 秀吉; 田中正起; 笹尾真理; 戸田亜希子; 南條綾子; 玉井智子; 樫尾智子; 野々村 照雄; 松田 克礼
    日本植物病理学会関西支部会(奈良)  2003/10  日本植物病理学会関西支部会(奈良)
     
    トマトのgypsy 型レトロトランスポゾンの逆転写酵素領域の遺伝子解析を行ったところ、様々な塩基置換や欠失等の変異が認められた。また、最大節約法により系統樹を作製したところ、各塩基配列の進化系列を明らかにすることが出来た。
  • トマトにおけるレトロトランスポゾンの発現解析  [Not invited]
    角谷 晃司; 豊田 秀吉; 松田 克礼; 野々村 照雄; 南條綾子; 笹尾真理
    日本農芸化学関西支部大会(奈良)  2002/10  日本農芸化学関西支部大会(奈良)
     
    トマトのレトロトランスポゾンの発現をRT?PCR法により検出した。Ty1/copia型およびTy3/gypsy型レトロトランスポゾンの逆転写酵素領域は、葉ならびにカルス細胞で発現していた。
  • トマトにおけるTy1/copia型およびTy3/gypsy型レトロトランスポゾンのクローニング  [Not invited]
    角谷 晃司; 豊田 秀吉; 松田 克礼; 野々村 照雄; 南條綾子; 笹尾真理; 玉井智子
    日本育種学会第102回年会(帯広)  2002/08  日本育種学会第102回年会(帯広)
     
    トマトのレトロトランスポゾンをゲノムからクローニングし、その遺伝子解析を試みた。ゲノム上には多数のレトロトランスポゾンが存在しており、また、培養栽培において発現している新規のTy1/copia型レトロトランスポゾンをクローニングした。
  • トマトレトロトランスポゾンの発現解析  [Not invited]
    角谷 晃司; 豊田 秀吉; 松田 克礼; 野々村 照雄; 南條綾子; 中川真樹; 笹尾真理
    第20回日本植物細胞分子生物学会大会(奈良)  2002/07  第20回日本植物細胞分子生物学会大会(奈良)
     
    Ty1/copia型およびTy3/gypsy型レトロトランスポゾンの逆転写酵素領域で保存されているアミノ酸配列からプライマーを合成し、RT?PCR分析に用いた。トマト葉およびカルスにおいて、これらは発現しており、既知Ty1/copia型とは異なる断片がカルスより増幅した。
  • ガン特異的タンパクCD98遺伝子を導入した形質転換体の作出  [Not invited]
    角谷 晃司; 豊田 秀吉; 松田 克礼; 野々村 照雄; 森浦展行; 益子 高
    第20回日本植物細胞分子生物学会大会(奈良)  2002/07  第20回日本植物細胞分子生物学会大会(奈良)
     
    経口ガンワクチンの開発を目的とし、ガン細胞特異的タンパク質CD98遺伝子を植物へ導入した。その遺伝子発現をRT?PCRで検出し、さらにタンパク質の翻訳をウエスタンブロット分析で調べた。その結果、組換え植物はCD98タンパク質を生産しており経口ワクチンの可能性が示唆された。

MISC

Industrial Property Rights

  • 特願2024-29724:雑草抑制装置  2024年/02/29
    松田克礼, 角谷晃司, 野々村照雄, 瀧川義浩  近畿大学, トワロン株式会社
  • 特願2023-049449:雑草抑制装置  2023年/03/27
    松田克礼, 角谷晃司, 野々村照雄, 瀧川義浩  近畿大学、トワロン株式会社
  • 特許7146186:微小粒子状物質捕捉装置    2022/09/26
    松田克礼, 角谷晃司, 野々村照雄, 瀧川義浩
  • 特許7090276:害虫防除用静電場スクリーン    2022/06/16
    松田克礼, 角谷晃司, 野々村照雄, 瀧川義浩
  • 松田克礼, 角谷晃司, 野々村照雄, 瀧川義浩  近畿大学、大阪府立環境農林水産総合研究所、トワロン株式会社、株式会社園田製作所
  • 松田克礼, 角谷晃司, 野々村照雄, 瀧川義浩
  • 松田克礼, 角谷晃司, 野々村照雄, 瀧川義浩
  • 特開2017-063655:静電苗シェルター及びこれに使用される静電場スクリーン発生装置  2017/04/06
    松田克礼, 角谷晃司, 野々村照雄, 瀧川義浩
  • 特開2016-220629:携帯型害虫誘引捕捉装置及び携帯型静電場スクリーン発生装置  2016/12/28
    松田克礼, 角谷晃司, 野々村照雄, 瀧川義浩
  • 特開2016-209783:静電場スクリーン発生装置、食材保管装置及びコンテナ  2016/12/15
    松田克礼, 角谷晃司, 野々村照雄, 瀧川義浩
  • 松田克礼, 角谷晃司, 野々村照雄, 瀧川義浩
  • 特許第5252449号:静電場発生装置及びそれを用いた飛動生物除去方法    2013/04/26
    松田克礼, 角谷晃司, 野々村照雄, 瀧川義浩
  • 特許4771310号:飛動生物除去装置及び植物保護装置    2011/07/01
    松田克礼, 角谷晃司, 野々村照雄, 瀧川義浩
  • 特許US8,105,418B2:Fungi preventing method, flying organism removing apparatus and plant protecting apparatus by adsorption of conidia using dielectric polarization    2009/07/09
    MATSUDA Yoshinori
  • 特開2007-195405:植物病害の治療方法および治療装置  2007/08/09
    松田克礼, 角谷晃司, 野々村照雄, 瀧川義浩

Research Grants & Projects

  • Kinki Kensetsu Kyokai (Kinki Road Construction Association) grant:Kinki Kensetsu Kyokai (Kinki Road Construction Association) grant
    Date (from‐to) : 2023/04 -2024/03 
    Author : Yoshinori Matsuda
  • 日本学術振興会:科学研究費助成事業
    Date (from‐to) : 2019/04 -2022/03 
    Author : 松田克礼
  • 日本学術振興会:科学研究費助成事業
    Date (from‐to) : 2016/04 -2019/03 
    Author : 松田 克礼
  • 日本学術振興会:科学研究費助成事業
    Date (from‐to) : 2013/04 -2016/03 
    Author : 松田 克礼
  • 日本学術振興会:科学研究費助成事業
    Date (from‐to) : 2010/04 -2013/03 
    Author : 松田克礼
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research
    Date (from‐to) : 2000 -2003 
    Author : TOYODA Hideyoshi; NONOMURA Teruo; MATSUDA Yoshinori
     
    When leaf segments of a tomato cultivar 'Ponderosa' were inoculated with Agrobacterium rhizogenes MAFF07-20001 carrying the binary vectors pRi and pBI121/sGFP, adventitious roots were developed from calli formed at the edges of the segments. Primordial roots that showed green fluorescence under blue light and elongated vigorously on hormone-free medium without loss of the green fluorescence were obtained. They were easily distinguishable from the non-fluorescing roots on the same segments. Successful integration of the sGFP and rol C genes into the chromosome of tomato roots was confirmed by polymerase chain reaction and Sourhem hybridization. The present method enables us to evaluate the hairy root formation without subculture, isolation and DNA analysis. This method was tested on all of commercial cultivars available in Japan(42 cultivars) and 14 breeding lines of tomato. All but two breeding lines produced the hairy roots. Thus, the present method is useful for hairy root production in tomato. RT-PCR was used to detect gene expression in situ in single selected cells from tomato callus aggregates. The cytoplasm from one cell was removed with a micropipette viewed under a light microscope and used directly for RT-PCR, followed by nested PCR. This method of removing cytosolic contents prevented the introduction of genomic DNA into the RT-PCR, and only intron-spliced products were amplified when intron-containing genes were used as PCR targets. In addition, transcription of the intron-free gene was possibly detected by simultaneously tracing the intron-containing and intron-free genes using mixed primers for the targeted genes. The present study indicated that some stimuli-activated genes, such as CHI3 and TLC1-retrotransposon long terminal repeat, were constitutively transcribed in tomato callus cells. A single-cell RT-PCR was conducted to detect gene expression in situ in pinpointed trichome cells of tomato leaves. The cytoplasm was removed with the micropipette using a light microscope and directly used for RT-PCR, followed by nested PCR. Two intron-containing genes, glyceraldehydes 3-phosphate dehydrogenase gene and plasma membrane H^+-ATPas gene were constantly expressed in this tissues and therefore used as the indicator, because of easy detection of shorter-size PCR-products produced by splicing. In addition, the sucking of nucleus-free cellular contents was effective to prevent contamination of genomic DNA led to miss-amplification of corresponding genomic DNA sequences of the intron-less genes in the process of RT-PCR and subsequent nested PCR. Thus, the present technique could be applicable to single trichome cells of tomato leaves for directly detecting their gene expression in response to chemical and physical stimulation.
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research
    Date (from‐to) : 1989 -1990 
    Author : TOYODA Hideyoshi; YOSHIDA Motonobu
     
    In a first year, we established an efficient system for direct gene transfer to macrospores of Fusarium oxysporum f. sp. lycopersici (race I), which causes the wilting in tomato plant, by electroporation. This work has been published in Journal of Phytopathology (1989). This method enabled us to introduce foreign genes into plant pathogic fungus without any special treatments such as protoplast formation and its regeneration. In a last year, therefore, we attempted to construct the vector system functional in this fungus. For this purpose. We first isolated the genes encoding rRNA in fungal chromosome by synthesizing cDNAs from rRNAs extracted from mycelia of this fungus. These cDNAs were hybridized with genomic DNAs which had been prepared from chromosomal DNA, and the cDNA specific to 28S rRNA was isolated. The DNA sequence of this clone was determined by dideoxy method and cleaved into two parts of the fragment. In order to construct the vector, the marker gene (hygromycin B) was flanked between these two fractions. This plasmid vector allowed us to ensure the homologous recombination of DNA sequences between the vector and chromosome. With this vector, macroconidia were successfully transformed by electroporation. In addition, we developed the method for in situ hybridization of mRNA with the probe clarified in this study. The probe DNA was introduced into mycelia of this fungus by microinjection in order to ensure cytoplasmic hybridization in mycelial cytoplasm. These results were submitted to the Journal, Agricultural and Biological Chemistry.

Others

  • 2020/04 -2021/03  “オール近大”新型コロナウイルス感染症対策支援プロジェクト 
    コロナウイルス対策における誘電体空間遮蔽装置「静電ブラインド」の提案


Copyright © MEDIA FUSION Co.,Ltd. All rights reserved.