In this study, we evaluated the effectiveness of hyperparasitic fungi in controlling powdery mildew (PM). In a greenhouse, we spray-inoculated single colonies of the melon PM-causing fungus Podosphaera xanthii strain KMP-6N at three different fungal developmental stages (i.e., 5, 10, and 15 days old) with spores of the hyperparasitic fungus Ampelomyces sp. strain Xs-q. After spray inoculation, we collected and counted KMP-6N conidia produced as asexual progeny from PM colonies using an electrostatic rotational spore collector. Collector insulator films were replaced at 24 h intervals until KMP-6N ceased to release additional progeny conidia. Conidial releases from each of the single Xs-q-inoculated KMP-6N colonies gradually reduced, then stopped within ca. 4 and 8 days of the first treatment in 5- and 10-day-old KMP-6N colonies, and within ca. 20 days of the second spray treatment in 15-day-old KMP-6N colonies, respectively. The total numbers of asexual progeny conidia collected from single 5-, 10-, and 15-day-old colonies were ca. 156, 1167, and 44,866, respectively. After electrostatic spore collection, conidiophores in Xs-q-uninoculated KMP-6N colonies appeared normal, whereas almost all conidiophores in 5- and 10-day-old Xs-q-inoculated KMP-6N colonies were completely deformed or collapsed due to the infection of the hyperparasitic fungus. This is the first study to apply electrostatic and digital microscopic techniques to clarify the impact of fungal hyperparasitism on mycohost survival, and, in particular, to assess quantitatively and visually the suppression of conidial release from any PM colonies infected with Ampelomyces.

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.

Powdery mildew fungi produce progeny conidia on conidiophores, and promote the spread of powdery mildew diseases by dispersal of the conidia from conidiophores in the natural environment. To gain insights and devise strategies for preventing the spread of powdery mildew infection, it is important to clarify the ecological mechanism of conidial dispersal from conidiophores. In this study, all of the progeny conidia released from single colonies of strawberry powdery mildew fungus (Podosphaera aphanis (Wallroth) U. Braun and S. Takamatsu var. aphanis KSP-7N) on true leaves of living strawberry plants (Fragaria × ananassa Duchesne ex Rozier cv. Sagahonoka) were consecutively collected over the lifetime of the colony with an electrostatic rotational spore collector (insulator drum) under greenhouse conditions, and counted under a high-fidelity digital microscope. The insulator drum consisted of a round plastic container, copper film, thin and transparent collector film, electrostatic voltage generator, and timer mechanism. When negative charge was supplied from the voltage generator to the copper film, the collector film created an attractive force to trap conidia. The electrostatically activated collector film successfully attracted progeny conidia released from the colony. Experiment was carried out at just one colony on one leaf for each month (in February, May, July, October, November, and December in 2021), respectively. Each collector film was exchanged for a new collector film at 24 h intervals until KSP-7N ceased to release progeny conidia from single colonies. Collection experiments were carried out to estimate the total number of conidia released from a single KSP-7N colony over a 35–45-day period after inoculation. During the fungal lifetime, KSP-7N released an average of 6.7 × 104 conidia from each of the single colonies at approximately 816 h. In addition, conidial release from KSP-7N colonies was largely affected by the light intensity and day length throughout a year; the number of conidia released from single KSP-7N colonies in night-time was clearly smaller than that in daytime, and the time of conidial release from single KSP-7N colonies was shorter by approximately 2 to 4 h in autumn and winter than in spring and summer. The ecological characteristics related to conidial releases from KSP-7N colonies will be helpful information for us to successfully suppress the spread of strawberry powdery mildews onto host plants under greenhouse conditions.

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.

Type I trichomes of tomato leaves (Solanum lycopersicum Mill. cv. Moneymaker), as outgrowths of the plant epidermis, are suitable for monitoring infection processes of powdery mildew species using a high-fidelity digital microscope (DM) without fungal staining. On the trichomes, tomato powdery mildew (Pseudoidium neolycopersici L. Kiss) isolate KTP-03 produced a maximum of four vigorously elongated hyphae per conidium, which stopped growth approximately 12 days after inoculation. Single trichome cells, invaded by fungal hyphae at various fungal infection stages during the 12-day period after the inoculation of single conidia, were cut at the bases and directly collected with small precision scissors (i.e., microscissors) held by the manipulator under a DM. Subsequently, suc-polymerase chain reaction (PCR) (reverse transcription (RT)-PCR followed by nested (N)-PCR) was conducted to explore gene expression in the infected trichome. We selected intron-containing genes from tomatoes and powdery mildew fungi for the detection of constitutive gene transcripts, namely plasma membrane H+-ATPase (LHA2) and β-tubulin 2 (TUB2) genes. In suc-PCR, a single band from spliced mRNAs of both LHA2 and TUB2 genes were detected, suggesting that both genes were successfully transcribed in single KTP-03-infected trichomes. With combined primers for both LHA2 and TUB2 (multiplex RT-PCR/N-PCR), two bands were detected through the amplification of intron-spliced mRNAs of both genes. Therefore, our single-trichome cell PCR amplification method is effective for detecting the expression patterns of genes from both tomato and powdery mildew fungus. Combinations of digital microscopy, microscissors, and multiplex RT-PCR/N-PCR amplification techniques will be useful for simultaneously analysing the molecular interactions between plants and powdery mildew fungi at the level of single tomato leaf trichome cells. Also, this employed technique will be of benefit in other plant species and crops, possessing leaf trichome cells, to elucidate the molecular interactions between plants and pathogens.

Powdery mildew fungi infect plant leaves, reducing the yield of infected melon plants. Therefore, an eco-friendly method of controlling powdery mildew in melon plants needs to be developed. A previous study described how the morphological characteristics of the conidiophores of the melon powdery mildew fungus Podosphaera xanthii Pollacci (designated KMP-6N) grown under greenhouse (natural) conditions and red light-emitting diode (LED) irradiation differed from those grown under growth chamber conditions and blue LED irradiation. In the present study, conidiophores with unconstricted conidia under blue light were collected and inoculated onto host leaves through micromanipulation; the unconstricted conidia germinated and infected the leaves, producing vigorously elongated hyphae. The number of conidia collected, the initial times of conidial release from single colonies, and the number of conidia remaining in chains on conidiophores were examined with electrostatic techniques. Under red light, the number of collected conidia gradually increased with the light irradiation period. The initial conidial release occurred between 2 to 4 h; the number of conidia remaining on the conidiophores gradually decreased and, eventually, the conidiophore lengths became shorter. In contrast, under blue light, few conidia were collected at any given time; the number of conidia on the conidiophores gradually increased and, eventually, the conidiophore lengths became longer. Next, the effects of red and blue light on the spread of powdery mildew infection by placing a KMP-6N-infected melon seedling at the centre of a tray containing healthy melon seedlings were examined. Almost all healthy seedlings caused powdery mildew symptoms at ca. 21 days after red light irradiation, whereas only healthy seedlings near the infected seedlings showed symptoms after blue light irradiation. Thus, the spread of melon powdery mildew infection clearly differed between red and blue light irradiation. This is the first report describing the effects of red and blue light on the spread of P. xanthii infection from a single infected seedling to healthy host seedlings; their results provide insight into the ecological mechanisms of powdery mildew conidial scatter from conidiophores.

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.

A total of 26 Ampelomyces strains were isolated from mycelia of six different powdery mildew species that naturally infected their host plants in Japan. These were characterized based on morphological characteristics and sequences of ribosomal DNA internal transcribed spacer (rDNA-ITS) regions and actin gene (ACT) fragments. Collected strains represented six different genotypes and were accommodated in three different clades of the genus Ampelomyces. Morphology of the strains agreed with that of other Ampelomyces strains, but none of the examined characters were associated with any groups identified in the genetic analysis. Five powdery mildew species were inoculated with eight selected Ampelomyces strains to study their mycoparasitic activity. In the inoculation experiments, all Ampelomyces strains successfully infected all tested powdery mildew species, and showed no significant differences in their mycoparasitic activity as determined by the number of Ampelomyces pycnidia developed in powdery mildew colonies. The mycoparasitic interaction between the eight selected Ampelomyces strains and the tomato powdery mildew fungus (Pseudoidium neolycopersici strain KTP-03) was studied experimentally in the laboratory using digital microscopic technologies. It was documented that the spores of the mycoparasites germinated on tomato leaves and their hyphae penetrated the hyphae of Ps. neolycopersici. Ampelomyces hyphae continued their growth internally, which initiated the atrophy of the powdery mildew conidiophores 5 days post inoculation (dpi); caused atrophy 6 dpi; and complete collapse of the parasitized conidiphores 7 dpi. Ampelomyces strains produced new intracellular pycnidia in Ps. neolycopersici conidiophores ca. 8–10 dpi, when Ps. neolycopersici hyphae were successfully destroyed by the mycoparasitic strain. Mature pycnidia released spores ca. 10–14 dpi, which became the sources of subsequent infections of the intact powdery mildew hyphae. Mature pycnidia contained each ca. 200 to 1,500 spores depending on the mycohost species and Ampelomyces strain. This is the first detailed analysis of Ampelomyces strains isolated in Japan, and the first timing and quantification of mycoparasitism of Ps. neolycopersici on tomato by phylogenetically diverse Ampelomyces strains using digital microscopic technologies. The developed model system is useful for future biocontrol and ecological studies on Ampelomyces mycoparasites.

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.

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.

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.

In the present study, the relationship between body water loss and conductivity was examined in adult houseflies (Musca domestica). The events an insect experiences in an electric field are caused by the conductive nature of the insect body (i.e., movement of electricity within or its release from the insect). After houseflies were dehydrated, rehydrated, refrigerated, and frozen and thawed, they were placed in static and dynamic electric fields. Untreated houseflies were deprived of their free electrons to become positively charged and then attracted to the insulated negative pole in the static electric field and were exposed to corona and arc discharge from non-insulated negative pole in the dynamic electric field. There was no current in the bodies of dehydrated and frozen flies; hence, there was no attractive force or discharge exposure. In the remaining insects, the results were identical to those in the untreated control insects. These results indicated that the reduction of body water conductivity inhibited the release of electricity from the body in the static electric field and the discharge-mediated current flow through the body in the dynamic electric field. The insect was affected by the electric fields because of its conductivity mediated by body water.

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.

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.

The lengths of conidiophores in fungal colonies of the melon powdery mildew pathogen Podosphaera xanthii Pollacci KMP-6 N cultured under greenhouse (natural) conditions differed markedly from those cultured in a growth chamber. We hypothesized that light wavelength was responsible for the differences in conidiophore length. In this study, we examined the effects of light-emitting diode (LED) irradiation (purple, blue, green, orange, and red light) and white light on colony development and conidiophore formation in KMP-6 N using a stereomicroscope and a high-fidelity digital microscope. Colonies on leaves were flat under greenhouse conditions and under red LED light irradiation but were stacked under growth chamber conditions and under purple, blue, green, and orange LED light irradiation. In addition, KMP-6 N formed catenated conidia comprising six conidia per conidiophore under greenhouse conditions and red light but more than seven conidia per conidiophore under growth chamber conditions and purple, blue, green, and orange light. Furthermore, almost none of the conidia on top of the conidiophores grown under blue light were fully constricted. Therefore, these fungi could not scatter their conidia and spread infection. This is the first report of the effects of LED lights on conidiophore formation in the melon powdery mildew fungus P. xanthii. The results provide insight into the mechanisms underlying the responses of conidiophores to light of specific wavelengths and conidial scatter from conidiophores of melon powdery mildew fungi.

The present study aimed to explore the possibility of using the type I trichomes of tomato (Solanum lycopersicum) to monitor the infection processes of powdery mildews by microscopy. Individual trichome cells of two tomato genotypes were inoculated with pathogenic and non-pathogenic powdery mildew species, Pseudoidium neolycopersici, Erysiphe trifoliorum and Podosphaera xanthii. On the trichome cells of the tomato cultivar Moneymaker, hyphae of the pathogenic Pseudoidium neolycopersici (isolates KTP-03 and KTP-04) grew vigorously whereas hyphal growth of the non-pathogenic Erysiphe trifoliorum and Podosphaera xanthii ceased after appressorium formation, which was associated with papilla formation and hypersensitive cell death, respectively. Similar infection processes of the tested powdery mildews were seen in Moneymaker epidermal cells. Therefore, tomato trichomes are suitable for analysing, at individual cell level, the infection processes of different pathotypes of powdery mildews and for observing the cytological responses of plants by non-pathogenic powdery mildews. On the other hand, it was observed that both isolates KTP-03 and KTP-04 failed to produce conidiophores on the hyphae elongating on Moneymaker trichomes. Similarly, no conidiophores were produced on the hyphae elongating on trichomes of Solanum peruvianum LA2172, which is resistant to KTP-03 and susceptible to KTP-04. Interestingly, delayed cell death occurred in LA2172 epidermal cells, which were attacked by KTP-03 hyphae elongating from trichomes and conidiophores were formed on new hyphae growing from the leaf epidermal cells. Thus, leaf trichomes and epidermal cells of the wild tomato species LA2172 reacted differently to the avirulent isolate KTP-03.

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.

This study examined whether Agrobacterium-mediated T-DNA insertion was useful for obtaining mutant melon plants. For this purpose, we used Agrobacterium rhizogenes because of its frequent hairy root production when inoculated into plants and the easy detection of fruit odours produced by the hairy roots. Of the 6534 hairy root clones obtained, five clones were mutants emitting fruit odours, and the KMH-4196 clone produced the strongest melon fruit odours. These odours were due to the production of at least four compounds that were not detected in non-aromatic hairy root clones: (Z)-3-hexenol, (E)-2-hexenal, 1-nonanoland (Z)-6-nonenol. (Z)-6-nonenol was the most important and was produced stably during successive subculture for 3 years. Molecular analyses indicated that a single copy of T-DNA from the inoculated bacteria was integrated into a chromosome in the clone, suggesting that Agrobacterium-mediated T-DNA insertion is an effective mutagenesis method in higher plants.

A microscopy-based needle micromanipulation technique for direct injection of foreign material into target cells was refined as a novel method of aspiration of cellular contents from a single target cell using a micropipette. Subsequently, we performed direct PCR amplification of mRNAs transcribed in target tomato callus cells. Friable calli were induced from leaf explants, and single cells from active small cell aggregates were used as targets for aspiration. The aspirated cellular contents were subjected to RT-PCR and subsequent nested PCR amplification analyses. Five tomato genes (LHA2, GAPDH, CHI3, PI2 and TLC1) were selected from the cDNA database as PCR targets. The GAPDH and LHA2 genes were amplified from all aspirated samples and were used as indicators of successful PCR amplification. Aspiration of only the cytosol (excluding the nucleus), facilitated amplification of mature target gene mRNA that was not contaminated with PCR products derived from the genomic DNA sequences of the target genes. We identified novel stimulus activated genes, such as CHI3 and TLC1, which were constitutively transcribed in tomato callus cells.

Single-cell polymerase chain reaction (PCR) was conducted to detect in situ gene expression in targeted cells of tomato leaf trichomes. The cytoplasm was removed with a micropipette under a light microscope and subsequently used for reverse transcription PCR (RT-PCR), followed by nested PCR. Two intron-containing genes, a glyceraldehyde 3-phosphate dehydrogenase gene and a plasma membrane H+-ATPase gene, were constantly expressed in these cells and therefore used as indicators of successful PCR reactions. In addition, the use of nucleus-free cellular contents for the RT-PCR and subsequent nested PCR analyses was effective for preventing contamination with the products derived from misamplification of corresponding genomic DNA sequences. Using this method, we detected the expression of certain stimuli-activated genes, following the exposure of trichome cells to volatile chemicals. Therefore, the present technique can be used to directly detect gene expression in single trichome cells of tomato leaves in response to external stimulation.

Melons (Cucumis melo L.) grown hydroponically in a greenhouse were heavily infested with powdery mildew. We isolated powdery mildew pathogens from the melon leaves and identified the isolate as Podosphaera xanthii KMP-6N, based on morphological characteristics and sequences of ribosomal DNA internal transcribed spacer (rDNA-ITS) regions. Host ranges of KMP-6N were determined by estimating the infectivity or pathogenicity after inoculating the conidia onto multiple plant species. The fungi caused severe powdery mildew symptoms on Cucurbitaceae plants, producing scattered conidia on conidiophores. The goal of this study was to observe KMP-6N conidiogenesis on melon leaves. The pathogen formed completely catenated conidiophores approximately 24 h from conidiophore erection to release of mature conidia. Six conidia were produced on the conidiophores and only the conidia at the apex reached maturity. The cycles of conidial release were repeated on melon leaves 14 to 18 times, at approximately 6-h intervals. In the final stage, conidia were released without causing growth and septation of generative cells. Conidiophores produced an average of 36 conidia during a 90-h period. In our study, the modes of conidiogenesis, lifetime of conidiophores and productivity of conidia on a conidiophore were described for powdery mildew fungi.

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.

We examined the defensive responses of leaf surfaces, rhizoids, and protonemata of the moss Aphanoregma patens (Hedw.) Lindb. to inoculation with zoospores and encysted zoospores (cysts) of the oomycete Pythium aphanidermatum OPU849. Aphanoregma patens infected with Pythium exhibited extensive browning of the leaves and stems 4 days after inoculation. The zoospore infection sites were rhizoids and protonemata. Cysts on the rhizoids and protonemata germinated and formed elongated hyphae that invaded the cells of the moss, forming oospores and lobed sporangia. The subsequent production of new zoospores on infected plants indicates that A. patens is a suitable host for manipulating the life cycle of Pythium. However, cysts inoculated onto leaf surfaces did not germinate, and they began to show signs of disruption after only 3 hours. After 2 days, more than 80% of the cysts on leaf surfaces were completely disrupted, possibly due to the secretion of degrading enzymes. Cyst disruption was not observed on rhizoids nor on protonemata.

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.

Studies on extracellular proteins (ECPs) contribute to understanding of the multifunctional nature of apoplast. Unlike vascular plants (tracheophytes), little information about ECPs is available from nonvascular plants, such as mosses (bryophytes). In this study, moss plants (Physcomitrella patens) were grown in liquid culture and treated with chitosan, a water-soluble form of chitin that occurs in cell walls of fungi and insects and elicits pathogen defense in plants. ECPs released to the culture medium were compared between chitosan-treated and nontreated control cultures using quantitative mass spectrometry (Orbitrap) and 2-DELC-MS/MS. Over 400 secreted proteins were detected, of which 70% were homologous to ECPs reported in tracheophyte secretomes. Bioinformatics analyses using SignalP and SecretomeP predicted classical signal peptides for secretion (37%) or leaderless secretion (27%) for most ECPs of P. patens, but secretion of the remaining proteins (36%) could not be predicted using bioinformatics. Cultures treated with chitosan contained 72 proteins not found in untreated controls, whereas 27 proteins found in controls were not detected in chitosan-treated cultures. Pathogen defense-related proteins dominated in the secretome of P. patens, as reported in tracheophytes. These results advance knowledge on protein secretomes of plants by providing a comprehensive account of ECPs of a bryophyte.

In this study, we observed the germination behaviour of airborne conidia from powdery mildews that settle on thalloid surfaces. We inoculated thalli (flat, sheet-like leaf tissues) and gemmae (small, flat, sheet-like leaf tissues that propagate asexually via bud-like structures) of the common liverwort (Marchantia polymorpha) with conidia from tomato powdery mildew (Oidium neolycopersici; KTP-02) and red clover powdery mildew (Erysiphe trifoliorum; KRCP-4N) and examined their germination and subsequent appressorium formation under a high-fidelity digital microscope. Conidial bodies and germ tubes of the inoculated KRCP-4N conidia were destroyed on both the thalli and gemmae. The destruction of these fungal structures was observed only for KRCP-4N conidia inoculated onto M.polymorpha on both leaf surfaces. No differences in destruction of the KRCP-4N fungal structures between thalli and gemmae were observed. At 4h post-inoculation, destruction of the germ tube tip was observed when it reached the gemmae leaf surface. At 6h post-inoculation, the conidial bodies and germ tubes were destroyed. In contrast, KTP-02 conidia were not destroyed and formed normal, well-lobed appressoria on the surface of M.polymorpha gemmae.

In our routine surveys for the powdery mildew disease in greenhouse tomatoes, we detected a new pathogen that forms pseudochains consisting of 12 conidia. To identify the original plant that dispersed this pathogen, wild plants infected with powdery mildew were monitored. The pathogen on Japanese mallotus, Mallotus japonicus, produced a similar type of pseudochain, and conidia were infectious to tomatoes. Inversely, the conidia on the tomato leaves infected M. japonicus. Infectivity assays and internal transcribed spacer (ITS)-based phylogenetic analyses indicated that the two pathogens on the tomato and M. japonicus were identical. These results suggest that the conidia on M. japonicus can be transmitted to greenhouse tomatoes. This work documents the ecological transmission of conidia between wild plants and greenhouse tomatoes.

An insulated conductor wire (ICW) paralleled with an earthed net was used to observe movements by vinegar flies in relation totheir electricity release. ICW was negativelycharged 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.

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.

Anelectricfieldscreen(EF-screen)isaphysicaldeviceforexcludingpestinsects from greenhousesand warehousesto 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.

The emergence of germ tubes from the conidia of powdery mildew fungi is the first morphological event of the infection process, preceding appressoria formation, peg penetration and primary haustoria formation. Germination patterns of the conidia are specific in powdery mildew fungi and therefore considered useful for identification. In the present study, we examined conidial germination of the tomato powdery mildew Oidium neolycopersici KTP-01 in order to clarify whether germ tube emergence site in KTP-01 conidia is determined by the first contact of the conidia to leaves (as found for the conidia of barley powdery mildew), or alternatively is predetermined and is unrelated to contact stimulus. Highly germinative conidia of KTP-01 were collected from conidial pseudochains on conidiophores in colonies on tomato leaves using two methods involving an electrostatic spore attractor and a blower. In the electrostatic spore attraction method, the conidia were attracted to the electrified insulator probe of the spore collector-this being the first contact stimulus for the conidia. In addition, the blowing method was used as a model of natural infection; pseudochain conidia were transferred to detached leaves by air (1 m/s) from a blower. Thus, landing on the leaves was the first contact for the conidia. Furthermore, conidia were also blown onto an artificial membrane (Parafilm-coated glass slides forming a hydrophobic surface) or solidified agar plates in Petri dishes (hydrophilic surface). Eventually, almost all conidia on the probe and on tomato leaves or artificial hydrophobic and hydrophilic surfaces synchronously germinated within 6 h of incubation, indicating that the first contact of the conidia with any of the aforementioned substrata was an effective germination induction signal. Germ tube emergence sites were exclusively subterminal on the conidia. Moreover, the germ tubes emerged without any relation to the sites touched first on the conidia. Thus, the present study strongly indicates that conidia of O. neolycopersici produce germ tubes at a predetermined site.

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.

In the present study, using a high-fidelity digital microscope, we observed the sequence of appressorial development on the germ tubes of a powdery mildew fungus isolated from red clover leaves. Based on its morphological characteristics and rDNA internal transcribed spacer (ITS) sequences, the fungus was identified as Erysiphe trifoliorum, and one of its isolates, designated as KRCP-4N, was used in this work. The conidial germination of isolate KRCP-4N was studied on host (red clover) and non-host (barley) leaves, as well as on an artificial hydrophobic membrane (Parafilm). More than 90% of conidia germinated synchronously and developed dichotomous appressoria (symmetrical double-headed appressoria) on all substrata used. On host leaves, all appressorium-forming conidia developed hyphae (colony-forming hyphae) from conidial bodies without extending germ tubes from the tips of the appressoria. On non-host leaves and on Parafilm-covered glass slides, however, all conidia extended germ tubes from one side of dichotomous appressoria (two-step germination). In addition to the dichotomous appressoria, we detected a few conidia that produced hooked appressoria and extended germ tubes from the tip of the appressorium. Penetration attempts by KRCP-4N conidia on barley leaves were impeded by papillae formed at penetration sites beneath these two types of appressorium. From these results, we conclude that the "two-step germination" of E. trifoliorum KRCP-4N conidia is the result of an unsuccessful penetration attempt, causing diversity in appressorial shape. © 2011 The Mycological Society of Japan and Springer.

The appressorial shapes of the powdery mildews are an important clue to the taxonomy of the powdery mildew fungi, but the conidia of the tomato powdery mildew Oidium neolycopersici KTP-01 develop non-lobed, nipple-shaped, and moderately lobed or multilobed appressoria on the same leaves. To remove this ambiguity, we performed consecutive observations of sequential appressorial development of KTP-01 conidia with a high-fidelity digital microscope. Highly germinative conidia of KTP-01, collected from conidial pseudo-chains formed on the tomato leaves, were inoculated into host tomato and nonhost barley leaves or an artificial hydrophobic membrane (Parafilm). Events from germination initiation to appressorium formation were synchronous in all conidia on all materials used for inoculation, but post-appressorial behaviors varied among the materials. Appressoria on the membrane-stuck glass slide formed several projections at different portions of the appressoria to repeat unsuccessful penetration attempts. Similar unsuccessful penetration behavior by KTP-01 conidia was observed in the inoculations into leaves of barley plants, wild tomato species Lycopersicon peruvianum LA2172 (carrying the Ol-4 gene for powdery mildew resistance), and a susceptible host tomato (Lycopersicon esculentum) that had been inoculated with the barley powdery mildew (Blumeria graminis f. sp. hordei, race 1) conidia. On the barley leaves, all penetrations of KTP-01 were impeded by the papillae formed beneath the sites of the appressorial projections. On both the wild tomato and the race 1-inoculated cultivated tomato plants, KTP-01 conidia were prevented from forming functional haustoria by hypersensitive epidermal cell death; this hypersensitive reaction involved the Ol-4 gene in the wild tomato plants or the 'induced resistance' acquired by the nonpathogenic conidia previously inoculated into the cultivated tomato plants. All these KTP-01 conidia produced several projections on the appressoria during the repeated unsuccessful penetration attempts and eventually exhibited multilobed appressoria. On the host tomato leaves inoculated singly with KTP-01 conidia, fewer than 20 % of the conidia located appressoria on the central part of target epidermal cells and succeeded in forming functional haustoria at the first penetration attempt without forming an appressorial projection. These conidia exhibited non-lobed appressoria. The remaining conidia, however, whose appressoria were located on/near the border of the target epidermal cells, were more likely to fail to penetrate at the first penetration, and then to develop additional projections for subsequent penetrations. Most conidia succeeded in forming functional haustoria at the second to fourth penetration attempts, but a few conidia failed to produce haustoria at all attempted penetrations. Eventually, the conidia that succeeded at the second penetration possessed a single appressorial projection (exhibiting the nipple-shaped appressoria), whereas the remaining conidia exhibited moderately lobed appressoria with two to four appressorial projections and multilobed appressoria, with more projections. Thus, the present study revealed that the basic shape of appressoria of KTP-01 was the non-lobed type, and that polymorphic changes of the appressoria occurred as a result of successive production of projections during repeated unsuccessful penetration attempts. (C) 2010 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

The feedback-insensitive anthranilate synthase (AS) gene was used as a selection marker for transformants of Arabidopsis thaliana. The mutant gene (mAS1-2) was constructed by substituting nucleotide at the effector-binding site of the intrinsic AS gene via PCR-mediated site-directed mutagenesis and flanked with the myrosinase promoter pyk10 to drive its expression during initial root elongation. This inducible gene cassette was first introduced into Agrobacterium tumefaciens and then delivered into A. thaliana by floral-dip inoculation. 5-methyltryptophan (5-MT) inhibited AS and suppressed seedling growth of wild type plants as a result of tryptophan starvation. With the addition of sucrose (10 mg/ml), 5-MT inhibited cotyledon opening and caused anthocyanin to accumulate in juvenile seedlings. The present mutant reversed the tryptophan starvation caused by 5-MT and blocked subsequent sugar responses. The sugar responses were detected in non-transformed plants grown on a selection medium containing 10 mg/ml of sucrose and 10 mu g/ml of 5-MT after 3 days of incubation. Thus, true transformants could be selected after a short incubation, compared to the conventional kanamycin-selection method that did not eliminate all non-transformed plants.

A population of simultaneously germinating conidia is an ideal inoculum of the powdery mildew pathogen, Oidium neolycopersici. In conditions of no or low wind velocity, O. neolycopersici successively stacks mature conidia on conidiophores in a chain formation (pseudochain), without releasing the precedent mature conidia. These pseudochain conidia represent a perfect inoculum, in which all conidia used for inoculation germinate simultaneously. However, we found that conidia must be collected before they fall to the leaf surface, because the germination rate was lower among conidia deposited on the leaf surface. We used an electrostatic spore collector to collect the pseudochain conidia, and their high germination rate was not affected by this treatment. The spore collector consisted of an electrified insulator probe, which created an electrostatic field around its pointed tip, and attracted conidia within its electric field. The attractive force created by the probe tip was directly proportional to voltage, and was inversely proportional to the distance between the tip and a target colony on a leaf. Pseudochain conidia were successfully collected by bringing the electrified probe tip close to target colonies on leaves. In this way, conidia were collected from colonies at 3-d intervals. This effectively collected all conidia from conidiophores before they dropped to the leaf surface. A high germination rate was observed among conidia attracted to the probe tip (95.5 +/- 0.6 %). Conidia were easily suspended in water with added surfactant, and retained their germination ability. These conidia were infective and produced conidia in pseudochains on conidiophores after inoculation. The electrostatic spore collection method can be used to collect conidia as they form on conidiophores, thus obtaining an inoculum population in which all of the conidia germinate simultaneously. (c) 2008 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Secretion by glandular trichomes is thought to be a strategy to retain moisture water on leaves under drought conditions. In the wild tomato species Lycopersicon pennellii exudates accumulate at the apical terminal of the trichome, forming a bulbous head. During a 30 s mist treatment, the exudates condensed moisture water to form water drops on the trichome heads, and water drops were absorbed through trichomes by the plants. However, a longer mist treatment (40 s) caused the drops on all trichome heads to drop down to the leaf surface. Water drops containing the exudates were amphiphilic and spread over the leaf surface in a thin layer. This is an additional mechanism of the glandular trichomes for covering leaves with the exudates as a chemical boundary layer. Exudates of type IV glandular trichomes of L. pennellii showed antifungal activity against conidia of the tomato powdery mildew pathogen Oidium neolycopersici. The exudates covering the leaves completely suppressed germination of inoculated conidia, which prevented subsequent infection events such as formation of appressoria and haustoria. On non-misted leaves almost all conidia germinated and formed appressoria, resulting in successful infection. L. pennellii is susceptible to tomato powdery mildew, but the mechanisms for condensing mist water by type IV trichome-derived exudates can contribute to suppressing foliar infection by the powdery mildew pathogen. Crown Copyright (C) 2008 Published by Elsevier Ireland Ltd. All rights reserved.

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.

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 5mm of separation and oppositely charged with 15kV 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 30mm) 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.

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.

低コストオープンソースハードウェアによるコンピュータ基板を使って、施設植物生産をスマート化する小型高性能情報機器(ガジェット)を提案した。計測位置を固定したこれまでの静的・硬直的な環境計測ではなく、気になる箇所に携帯性の高い小型機器を局所的に設置することにより、機動的・柔軟な環境計測による生産性改善を行う新たな視点の手法である。事例研究として、局所分光環境ガジェットと、局所植物成育評価モデリングガジェットを設計・試作し、植物生産施設で試験して有用性を評価した。

本研究では、ゼニゴケをモデルとする遺伝子機能解析を促進するため、CRISPR RNAおよびCas9タンパク質の複合体のマイクロインジェクションによるゲノム編集法の開発を目指した。標的とする遺伝子として、気室形成制御因子NOPPERABO1 (NOP1) を選択した。NOP1が欠失すると、葉状体に気室が形成されず、葉状体表面の緑色が薄くなるとともに表面が滑らかになるので、変異表現型を容易に観察できる。NOP1遺伝子を標的とするCRISPR/Cas9複合体を導入したところ、nop1変異表現型を示す株を得た。この株の標的配列中に5塩基対の欠失が認められ、ゲノム編集を確認した。