Method for processing plants, method for producing plants infected with microorganisms, method for producing plant fermentation products, and plant processing apparatus.

Abstract

To provide a method for infecting a plant with useful microorganisms efficiently.SOLUTION: The present disclosure provides a method for treating a plant, comprising: an irradiation step of irradiating a plant with light in the wavelength range of 300 to 325 nm so that the irradiation amount is 4000 μmol / m2 or more and 50000 μmol / m2 or less, and light in the wavelength range of 290 nm or less is not applied, or the amount of irradiation is less than 20% of the amount of light in the wavelength range of 300 to 325 nm; and an infection step of infecting the plant irradiated with the light with a microorganism.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] This disclosure relates to methods and apparatus for processing plants. More specifically, this disclosure relates to methods for infecting plants with microorganisms, methods for producing plants infected with microorganisms, methods for producing plant fermentation products, and apparatus for infecting plants with microorganisms. [Background technology]

[0002] For example, techniques exist for introducing foreign genes by infecting plants with infectious agents or microorganisms containing those foreign genes, such as the Agrobacterium method and the plant virus vector method. However, conventionally known methods limit the infection of plants by microorganisms due to the plant's resistance to microbial infection. [Overview of the project] [Problems that the invention aims to solve]

[0003] There is a need for a simpler and / or more efficient method to infect plants with beneficial microorganisms. [Means for solving the problem]

[0004] This disclosure, from one perspective, describes how to expose plants to light in the wavelength range of 300-325 nm at an irradiation dose of 4000 μmol / m³. 2 More than 50000μmol / m 2 The present invention provides a method for treating plants (hereinafter also referred to as "the plant treatment method of this disclosure") which includes an irradiation step in which light in the wavelength range of 290 nm or less is not irradiated or the amount of such irradiation is less than 20% of the amount of light in the wavelength range of 300 to 325 nm is irradiated, and an infection step in which microorganisms are infected into the plants that have been irradiated with the light. This disclosure, from another perspective, describes how to expose plants to light in the wavelength range of 300-325 nm at an irradiation dose of 4000 μmol / m³. 2 More than 50000μmol / m 2The present invention provides a method for producing plants infected with microorganisms (hereinafter also referred to as "the method for producing microorganism-infected plants of this disclosure"), which includes an irradiation step in which light in the wavelength range of 290 nm or less is not irradiated or the amount of such irradiation is less than 20% of the amount of light in the wavelength range of 300 to 325 nm, and an infection step in which microorganisms are infected into the plants that have been irradiated with the light.

[0005] This disclosure, from yet another perspective, describes how to expose plants to light in the wavelength range of 300-325 nm at an irradiation dose of 4000 μmol / m³. 2 More than 50000μmol / m 2 The present invention provides a method for producing a plant fermentation product (hereinafter also referred to as "the method for producing a plant fermentation product of this disclosure"), comprising: an irradiation step in which light in the wavelength range of 290 nm or less is not irradiated or the amount of such irradiation is less than 20% of the amount of light in the wavelength range of 300 to 325 nm; an infection step in which microorganisms are infected into the plant irradiated with the light; and a step in which the infected plant is fermented.

[0006] This disclosure provides a plant immersion apparatus comprising: a plant holding section configured to hold a plant; an ultraviolet light irradiation section configured to irradiate at least a portion of the plant held in the plant holding section with ultraviolet light in the wavelength range of 300 to 325 nm, such that light in the wavelength range of 290 nm or less is not irradiated or the amount of such irradiation is less than 20% of the amount of light in the wavelength range of 300 to 325 nm; and a storage tank configured to contain a liquid containing microorganisms in a liquid storage area partitioned within its internal cavity, wherein the portion of the plant irradiated with ultraviolet light by the ultraviolet light irradiation section is located within the liquid storage area, or a liquid spraying section configured to spray a liquid containing microorganisms towards the portion of the plant irradiated with ultraviolet light by the ultraviolet light irradiation section. [Effects of the Invention]

[0007] The plant treatment method and / or plant treatment apparatus of this disclosure makes it possible to infect plants with useful microorganisms more easily and / or more efficiently. [Brief explanation of the drawing]

[0008] [Figure 1] This is a schematic diagram illustrating one embodiment of the apparatus of the present disclosure. [Figure 2] This is a schematic diagram illustrating another embodiment of the apparatus of the present disclosure. [Figure 3] The emission spectra of the three LEDs used in the example are shown. [Figure 4] This is a photograph of tobacco benthamiana before irradiation, showing that each plant has two leaves equipped with LED light irradiation and non-LED light irradiation areas as treatment areas. [Figure 5] These are photographs of tobacco plants (Benthamiana citrus) immediately after irradiation with 280nm-LED light (left) and 24 hours later (right). [Figure 6] This image shows the luminescence of tobacco leaves used in a 310nm-LED light irradiation test at an irradiation dose of 6750 μmol / m2. The area enclosed by the frame is the unirradiated area. Top row: An image of tobacco leaves superimposed with the luminescence image of firefly luciferase to identify the luminescent area. Bottom row: Luminescence image of firefly luciferase. [Figure 7] This image shows the luminescence of tobacco leaves used in a 340nm-LED light irradiation test at an irradiation dose of 6750 μmol / m2. The area enclosed by the frame is the unirradiated area. Top row: An image of tobacco leaves superimposed with the luminescence image of firefly luciferase to identify the luminescent area. Bottom row: Luminescence image of firefly luciferase. [Figure 8] The amount of light emitted in the un-irradiated and irradiated areas of each leaf of the tobacco plant used in the 310nm-LED light irradiation test is shown. [Figure 9] The amount of light emitted in the un-irradiated and irradiated areas of each leaf of the tobacco plant used in the 340nm-LED light irradiation test is shown. [Figure 10] The luminescence levels in the unirradiated and irradiated areas of each leaf of tobacco plants (Cigarette benthamiana) irradiated with 310nm-LED light at an irradiation dose of 675 μmol / m2 are shown. [Figure 11] The luminescence levels in the unirradiated and irradiated areas of each leaf of tobacco plants (Cigarette benthamiana) irradiated with 310nm-LED light at an irradiation dose of 13,500 μmol / m2 are shown. [Figure 12] The luminescence levels in the unirradiated and irradiated areas of each leaf of tobacco plants (Cigarette benthamiana) irradiated with 310nm-LED light at an irradiation dose of 33,750 μmol / m2 are shown for each test section. [Figure 13] The luminescence levels in the unirradiated and irradiated areas of each leaf of tobacco plants (Cigarette benthamiana) irradiated with 310nm-LED light at an irradiation dose of 54,000 μmol / m2 are shown. [Figure 14] This image shows the luminescence of tobacco leaves used in a 310nm-LED light irradiation test at an irradiation dose of 13,500 μmol / m2. The area enclosed by the frame is the unirradiated area. Top row: An image of tobacco leaves superimposed with the luminescence image of firefly luciferase to identify the luminescent area. Bottom row: Luminescence image of firefly luciferase. [Figure 15] The luminescence levels in the unirradiated and irradiated areas of each leaf of tobacco plants (Cigarette benthamiana) that were infiltrated with Agrobacterium using a needleless syringe immediately after irradiation with 310nm-LED light at a dose of 6750 μmol / m2 are shown. [Modes for carrying out the invention]

[0009] In this specification, the numerical range "a~b" (where a and b are specific numerical values) means the range that includes the values ​​"a" and "b" at both ends. In other words, "a~b" is synonymous with "a or greater and b or less".

[0010] This disclosure is based on new findings that, as shown in the examples described below, irradiating plants with light in the wavelength range of 300 to 325 nm (also referred to as "ultraviolet light irradiation treatment according to this disclosure") can transiently reduce the resistance of plants to microbial infection.

[0011] <Plant treatment methods and methods for producing microbially infected plants> The plant treatment method of the present disclosure is to irradiate a plant with light in the wavelength range of 300 to 325 nm, with an irradiation amount of 4000 μmol / m 2 or more and 50000 μmol / m 2 or less, and the light in the wavelength range of 290 nm or less is not irradiated or is irradiated so that its irradiation amount is less than 20% of the irradiation amount of the light in the wavelength range of 300 to 325 nm; an irradiation step an infection step of infecting the plant irradiated with the light with a microorganism and includes

[0012] In addition, the method for producing a microorganism-infected plant of the present disclosure is to irradiate a plant with light in the wavelength range of 300 to 325 nm, with an irradiation amount of 4000 μmol / m 2 or more and 50000 μmol / m 2 or less, and the light in the wavelength range of 290 nm or less is not irradiated or is irradiated so that its irradiation amount is less than 20% of the irradiation amount of the light in the wavelength range of 300 to 325 nm; an irradiation step an infection step of infecting the plant irradiated with the light with a microorganism and includes

[0013] Hereinafter, the "plant treatment method of the present disclosure", the "method for producing a microorganism-infected plant of the present disclosure", and the "method for producing a plant ferment of the present disclosure" may also be collectively referred to as the "method of the present disclosure".

[0014] (Plant) In this specification, plants are not particularly limited as long as they can be infected with the microorganisms or infectious agents used. Plants may be dicotyledonous or monocotyledonous. Examples of dicotyledonous plants include those of the Solanaceae family, especially the genera Tobacco and Solanum; the Brassicaceae family, especially the genera Brassica, Raphanus, and Arabidopsis; the Rosaceae family, especially the genera Fragaria; the Asteraceae family, especially the genera Lactuca; the Cucurbitaceae family, especially the genera Cucumis and Pumpkin; the Fabaceae family, especially the genera Glycine, Vigna, Nelumbo, Vicia, and Peanut; the Rutaceae family, especially the genera Citrus; the Vitaceae family, especially the genera Vitis; the Lamiaceae family, especially the genera Lamiaceae; and the Theaceae family, especially the genera Camellia. Examples of monocotyledonous plants include those of the Poaceae family, especially the genera Poa, Wheat, Barley, Rye, Maize, and Sugarcane. The ultraviolet light irradiation treatment described herein transiently reduces the microbial resistance of plants, or in other words, transiently improves the infection efficiency of plants. As a result, plants that are inefficiently infected by conventional microbial infection methods, particularly the Agrobacterium method, such as grasses and legumes, become more efficiently infected by microorganisms. Therefore, these plants can be used as plants in the method described herein.

[0015] The plants are preferably selected from Solanaceae, Brassicaceae, Asteraceae, Rosaceae, Fabaceae and Poaceae, more preferably selected from Solanaceae, Rosaceae, Fabaceae and Poaceae, more preferably selected from Solanaceae and Rosaceae, and most preferably Solanaceae, with tobacco plants being even more preferred. Specific examples of plants in the genus Nicotiana include tobacco (Nicotiana tabacum), benthamiana tobacco (N. benthamiana), flower tobacco (N. alata), tree tobacco (N. glauca), and long-flowered tobacco (N. longiflora), with Nicotiana tabacum being the preferred choice among them. Specific examples of plants other than tobacco plants that can be used in the method disclosed herein include eggplant (Solanum melongena), tomato (Solanum lycopersicum), potato (Solanum tuberosum), cauliflower (Brassica oleracea var. botrytis), cabbage (Brassica oleracea var. capitata), broccoli (Brassica oleracea var. italica), komatsuna (Brassica rapa var. perviridis), bok choy (Brassica rapa var. chinensis), mizuna (Brassica rapa var. nipposinica), turnip (Brassica rapa var. rapa), Chinese cabbage (Brassica rapa var. glabra), rapeseed (Brassica campestris), European rapeseed (Brassica napus), mustard (Brassica juncea), and radish (Raphanus sativa). Arabidopsis thaliana, cucumber (Cucumis sativus), pumpkin (Cucurbita moschata, Cucurbita mamima), zucchini (Cucurbita pepo), strawberry (Fragaria x ananassa Duchesne ex Rozier), lettuce (Lactuca sativa), soybean (Saccharum officinarum), adzuki bean (Vigna angularis), green bean (Phaseolus vulgaris), broad bean (Vicia faba), peanut (Arachis hypogaea), perilla (Perilla frutescens var. crispa), rice (Oryza sativa), wheat (Triticum aestivum), durum wheat (Triticum durum), barley (Hordeum vulgare), rye (Secale) Cereale, corn (Zea mays), sugarcane (Saccharum officinarum), grapes (Vitis spp.)Examples include the tea plant (Camellia sinensis).

[0016] In some embodiments, the plant is a plant that can be infected with Agrobacterium. In some other embodiments, the plant is a plant that can be infected with a plant virus. In this specification, a plant may be the entire plant body, or, unless otherwise evident from the context, a part of a plant, cultured plant cells, such as cultured plant cells in the form of callus, or a fragment of the whole plant or a part thereof. A part of a plant may be a root, stem, leaf, flower, bud, seed, or any combination thereof. The plants may be plants being cultivated or plants whose cultivation has been temporarily suspended. Here, cultivation may be soil cultivation, more specifically open field cultivation or greenhouse cultivation, or hydroponics, more specifically hydroponics, spray cultivation or solid substrate cultivation. Plants whose cultivation has been temporarily suspended may be resumed during or after the application of the method of this disclosure. The plants may also be plants that have been harvested.

[0017] (irradiation process) In the irradiation process, the plants are irradiated with light in the wavelength range of 300 to 325 nm. The lower limit of the light irradiation dose in the wavelength range of 300-325 nm is 4,000 μmol / m³. 2 That's all, more specifically 4,500 μmol / m³ 2 That's all, more specifically 5,000 μmol / m 2 That's all, more specifically 5,500 μmol / m² 2 That's all, more specifically 6,000 μmol / m 2 That's all, more specifically 6,500 μmol / m³ 2 That's all, more specifically 6,750 μmol / m² 2 This is possible. The irradiation dose of light in the wavelength range of 300-325 nm is 4,000 μmol / m³. 2 If the value is less than the target value, it may not be possible to reduce the microbial infection resistance of the irradiated plants.

[0018] The upper limit for the amount of light irradiation in the wavelength range of 300-325 nm is 50,000 μmol / m³. 2 More specifically, 47,500 μmol / m² 2 More specifically, 45,000 μmol / m² 2 More specifically, 42,500 μmol / m² 2 More specifically, 40,000 μmol / m² 2 More specifically, 37,500 μmol / m² 2 More specifically, 35,000 μmol / m² 2 More specifically, 33,750 μmol / m² 2 This is possible. The irradiation dose of light in the wavelength range of 300-325 nm is 50,000 μmol / m³. 2 If the radiation level exceeds a certain point, the plants being irradiated are more likely to be damaged.

[0019] The range of light irradiation dose within the wavelength range of 300-325 nm can be any combination of the above lower and upper limits. A specific example of the irradiation dose range is 4,000 μmol / m³. 2 More than 50,000 μmol / m 2 Below, 4,000μmol / m 2 More than 47,500μmol / m 2 Below, 4,500μmol / m 2 More than 47,500μmol / m 2 Below, 4,500μmol / m 2 More than 45,000μmol / m 2 Below, 5,000μmol / m 2 More than 45,000μmol / m 2 Below, 5,000μmol / m 2 More than 42,500 μmol / m 2 Below, 5,500μmol / m 2 More than 42,500 μmol / m 2 Below, 6,000μmol / m 2 More than 42,500 μmol / m 2 Below, 6,000μmol / m 2 More than 40,000 μmol / m 2 Below, 6,500μmol / m 2More than 40,000 μmol / m 2 Below, 6,500μmol / m 2 More than 37,500 μmol / m 2 Below, 6,750μmol / m 2 More than 37,500 μmol / m 2 Below, 6,750μmol / m 2 More than 35,000μmol / m 2 Below, 6,750μmol / m 2 More than 33,750μmol / m 2 The following are some examples, but are not limited to these.

[0020] On the other hand, since the absorption maximum wavelength for DNA and RNA is around 260 nm, there are concerns that light with wavelengths around 260 nm may have a significant adverse effect on plants. In addition, light in the 280-290 nm wavelength range may show an inhibitory effect on viral infection in tomato seedlings, for example. Therefore, while irradiating with light in the 300-325 nm wavelength range, the plants should not be irradiated with light in the 290 nm wavelength range or lower, or if irradiated, the amount of such light should be less than 20%, preferably less than 10%, more preferably less than 5%, and more preferably less than 1% of the amount of light in the 300-325 nm wavelength range.

[0021] In addition, light in the wavelength range of 330 nm or higher does not contribute to a decrease in the resistance of plants to microbial infection. Therefore, from the viewpoint of energy efficiency, the irradiation amount of light in the wavelength range of 330 nm or higher may be, for example, less than 50%, preferably less than 30%, more preferably less than 20%, more preferably less than 10%, more preferably less than 5%, and more preferably less than 1% of the irradiation amount of light in the wavelength range of 300 to 325 nm.

[0022] Light in the wavelength range of 300-325 nm has a molecular weight of, for example, 0.05-300 μmol / m³. 2 Irradiated at / s. 0.05 μmol / m³ 2 If the value is less than / s, it may not be possible to reduce the microbial infection resistance of the irradiated plants. 300 μmol / m 2If the radiation level exceeds / s, it may induce damage to the irradiated plants. Light in the wavelength range of 300-325 nm is preferably 0.1-300 μmol / m³. 2 / s, more preferably 0.2~200 μmol / m³ 2 / s, more preferably 0.5~100 μmol / m³ 2 / s, more preferably 0.5~50 μmol / m³ 2 / s, more preferably 1-20 μmol / m³ 2 / s, more preferably 1-15 μmol / m³ 2 / s, more preferably 2-15 μmol / m³ 2 / s, more preferably 2-10 μmol / m³ 2 It is irradiated with a photon flux density of / s.

[0023] The light source used in the method disclosed herein is not particularly limited as long as it can emit light in the wavelength range of 300 to 325 nm. For example, commonly used ultraviolet light sources such as UV lamps can be used. Examples of UV lamps include light-emitting diodes (LEDs), laser diodes (LDs), xenon lamps, fluorescent lamps, incandescent lamps, metal halide lamps, and high-pressure mercury lamps. Alternatively, ultraviolet light extracted from sunlight using an optical filter or the like may be used. If the light source used emits light in the 300-325nm wavelength range along with light in the 290nm wavelength range or less at a photon flux density of 20% or more of the light in the 300-325nm wavelength range, then by using a filter whose transmittance for light in the 300-325nm wavelength range is greater than its transmittance for light in the 290nm wavelength range or less, the photon flux density of light in the 290nm wavelength range or less irradiated onto the plant may be less than 20% of the photon flux density of light in the 300-325nm wavelength range. More specifically, it may be less than 15%, less than 10%, less than 5%, or less than 1%. Additionally, or alternatively, if the light source used emits light in the wavelength range of 330 nm or higher, along with light in the wavelength range of 300 to 325 nm, at a photon flux density of 50% or more of the light in the wavelength range of 300 to 325 nm, then by using a filter whose transmittance for light in the wavelength range of 300 to 325 nm is greater than its transmittance for light in the wavelength range of 330 nm or higher, the photon flux density of light in the wavelength range of 330 nm or higher irradiated onto the plant may be less than 50% of the photon flux density of light in the wavelength range of 300 to 325 nm. More specifically, it may be less than 40%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, or less than 5%.

[0024] From the viewpoint of energy efficiency, light in the wavelength range of 300 to 325 nm is irradiated as light having a main peak wavelength, for example, within 310 ± 7 nm, preferably within 310 ± 5 nm, and more preferably within 310 ± 3 nm. In the wavelength range of 300 to 325 nm, it is preferable that there is no second peak, or if there is, its intensity is 1 / 10 or less of the main peak. In this specification, "main peak wavelength" refers to the peak wavelength at which the intensity is maximum. Note that for light whose spectrum exhibits a single peak, such as LED light, "peak wavelength" is synonymous with "main peak wavelength". The full width at half maximum (FMAX) of the main peak within 303-317 nm, preferably 305-315 nm, and more preferably 303-313 nm, is preferably, for example, 1-15 nm, preferably 5-15 nm, and more preferably 5-10 nm. By having a FMAX of 15 nm or less of the main peak, it becomes possible to selectively irradiate with light in a wavelength range that is effective in reducing the plant's resistance to microbial infection while avoiding irradiation with light in a wavelength range that does not contribute to reducing the plant's resistance to microbial infection, and energy efficiency is also further improved. Light with a FMAX of less than 1 nm of the main peak can also be used in the method disclosed herein, but from the viewpoint of cost-effectiveness, it is currently preferable to use light with a FMAX of 1 nm or more of the main peak. Therefore, specific examples of light in the 300-325 nm wavelength range may include light having a wavelength spectrum with a peak wavelength of 310 ± 7 nm and a full width at half maximum (FMAX) of 1-15 nm, more specifically 5-15 nm, or more specifically 5-10 nm; light having a wavelength spectrum with a peak wavelength of 310 ± 5 nm and a FMAX of 5-15 nm, more specifically 5-15 nm, or more specifically 5-10 nm; and light having a wavelength spectrum with a peak wavelength of 310 ± 3 nm and a FMAX of 5-15 nm, more specifically 5-15 nm, or more specifically 5-10 nm.

[0025] As a light source in the 300-325 nm wavelength range, light-emitting diodes (LEDs) or laser diodes (LDs) having a single peak in their emission spectrum are particularly preferred. When using LEDs or LDs as light sources, it becomes easy to selectively irradiate plants with light in a wavelength range effective in reducing microbial infection resistance, while avoiding irradiation with light in wavelength ranges that not only do not contribute to reducing plant resistance but may also adversely affect plants. Furthermore, the use of LEDs or LDs is also preferable from the viewpoint of energy efficiency and economics due to their low heat generation, low power consumption, and long lifespan. In addition, it becomes easier to control / manage the irradiation dose and / or photon flux density. LEDs or LDs capable of emitting light in the wavelength range of 300-325 nm may, for example, be made of AlGaN-based materials or InAlGaN-based materials.

[0026] The amount of light irradiated onto plants in the 300-325 nm wavelength range can be controlled, for example, by controlling the on / off state of the light source (e.g., when the plants are stationary) or by controlling the time required for the plants to pass through the irradiated area (e.g., when the plants are transported by a transport device), resulting in a dose of 4,000-50,000 μmol / m³. 2 It can be set to this.

[0027] Light in the wavelength range of 300 to 325 nm may be irradiated to plants as continuous light, intermittent light, or a combination thereof. It is preferable that the light in the wavelength range of 300 to 325 nm be irradiated as intermittent light. By using intermittent light, the temperature rise of the plants and / or the light source can be avoided or reduced. Specific examples of intermittent light include a pulse width of 100 ms or less, more specifically 50 ms or less, more specifically 20 ms or less, more specifically 10 ms or less, more specifically 5 ms or less, and a duty cycle of 50% or less, more specifically 40% or less, more specifically 30% or less, more specifically 20% or less, more specifically 10% or less, more specifically 5% or less.

[0028] During irradiation with light in the wavelength range of 300-325 nm, the plants may be stationary or in motion. If the plants are parts of a plant body or crushed material of the whole or parts of a plant body, they may, for example, rotate, vibrate, or float, or be agitated. The plants may be irradiated with light during transport. The irradiation may come from one direction, or from two or three or more directions. Light in the 300-325nm wavelength range does not need to irradiate the entire plant held in the plant holder; it may be irradiated to one or more parts or regions of the plant, as long as the required amount of light is applied. By limiting the irradiation of light in the 300-325nm wavelength range to desired parts of the plant, such as flower buds, flowers and / or blossoms, or roots, the resistance to microbial infection in other parts will not be affected. Thus, it is possible to avoid an unnecessary increase in the risk of infection in the plant. By limiting the irradiation to desired parts, wilting can also be reduced.

[0029] Irradiation with light in the wavelength range of 300-325 nm is preferably carried out in the dark. In this specification, "dark" means a level at which the photosynthetically effective wavelength range, i.e., the photon flux density in the wavelength range of 400-700 nm (hereinafter referred to as "photosynthetically effective photon flux density"), does not cause photosynthesis in the cells of the plant, more specifically, a photosynthetically effective photon flux density ≤ 10 μmol / m³ 2 This refers to a location where / s is used. By placing plants in the dark while irradiated with light in the 300-325 nm wavelength range, that is, by preventing photosynthesis in the plant's cells, a reduction in the plant's resistance to microbial infection can be efficiently induced.

[0030] (infection process) In the infection process, plants are irradiated with light in the wavelength range of 300-325 nm, and then infected with microorganisms. The microorganisms used in the methods of this disclosure are not particularly limited, as long as they are plant cells that have been irradiated with light in the wavelength range of 300–325 nm and have reduced resistance to microbial infection than before irradiation. Naturally, the microorganisms may also be microorganisms that can infect cells that have normal resistance to microbial infection before being irradiated with light in the wavelength range of 300–325 nm. In this specification, “microorganisms” include bacteria, fungi such as yeasts and filamentous fungi, viruses and viral vectors (these may also be called “infectious agents”). In this specification, “microorganisms” and “infectious agents” are used interchangeably with respect to infection of plants.

[0031] In some embodiments, the microorganism is an endophyte or endosymbiotic fungus. The endophyte is not particularly limited as long as it is capable of endosymbiosis with the plant used, and can be either a bacterial endophyte or a fungal endophyte. Bacterial endophytes can be rhizobia, and may be bacteria selected from, for example, the genera Rhizobium, Bradyrhizobium, Sinorhizobium, or Mesorhizobium. In this specification, bacterial endophytes include bacteria of the genus Rugamonas. Bacteria of the genus Rugamonas are endogenous bacteria of barley (roots) and may exert growth-promoting effects in infected plants. Fungal or filamentous endophytes may be mycorrhizal fungi. Mycorrhizal fungi may be, for example, arbuscular mycorrhizal fungi (AM fungi or VA fungi) or ericoid mycorrhizal fungi. Arbuscular mycorrhizal fungi may be selected from the genera Gigaspora, Glomus, and Rhizophagus, for example. Erycoid mycorrhizal fungi may be selected from the genera Neotyphodium and Epicloe, for example.

[0032] Plants infected with endophytes may have enhanced immunity, accelerated growth, and / or increased resistance to environmental stresses such as strong light, high temperatures, and / or drought, which could result in increased yields. Therefore, in embodiments where the microorganism is an endophyte, the plant treatment method of this disclosure can be said to be a method for increasing plant yield. Therefore, this disclosure is, Light in the wavelength range of 300-325 nm is applied to plants, especially plant roots, at an irradiation dose of 4000 μmol / m³. 2 More than 50000μmol / m 2 The irradiation process is as follows, and is performed such that no light in the wavelength range of 290 nm or less is irradiated, or the amount of such light is less than 20% of the amount of light irradiated in the wavelength range of 300 to 325 nm. An infection process in which endophytes are infect plants, particularly the roots of plants, that have been irradiated with the aforementioned light, The present invention provides a method for increasing plant yields, including [specific method / feature].

[0033] Furthermore, in embodiments where the microorganism is an endophyte, the method for producing microbially infected plants according to this disclosure can be described as a method for producing endophyte-infected plants. Therefore, this disclosure is, The plants were exposed to light in the wavelength range of 300-325 nm, with an irradiation dose of 4000 μmol / m³. 2 More than 50000μmol / m 2 The irradiation process is as follows, and is performed such that no light in the wavelength range of 290 nm or less is irradiated, or the amount of such light is less than 20% of the amount of light irradiated in the wavelength range of 300 to 325 nm. An infection process in which endophytes are infected into plants irradiated with the aforementioned light, This invention provides a method for producing endophyte-infected plants, including those containing endophytes.

[0034] In some embodiments, the microorganisms are microbial pesticides. These microbial pesticides are used as insecticides, nematicides, or fungicides to control plant diseases and pests. Examples of bacterial microbial pesticides include Bacillus thuringiensis, Pasteuria penetrans, Agrobacterium radiobacter (or Agrobacterium tumefaciens), Pseudomonas fluorescens, Pseudomonas rhodesiae, Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus simplex, Variovorax paradoxus, Lactiplantibacillus plantarum, and non-pathogenic Erwinia carotovora. Examples of filamentous fungal microbial pesticides include Verticillium lecanii, Paecilomyces tenuipes, Paecilomyces fumosoroseus, Beauveria bassiana, Beauveria brongniartii, Metarhizium anisopliae, Coniothyrium minitans, Talaromyces flavus, and Tricoderma atroviride.Examples of microbial pesticides that are viruses include nuclear polyhedrosis viruses (NVPs) and granulosis viruses (GVs) of the genus Baculovirus, as well as cytoplasmic polyhedrosis viruses of the genus Cypovirus. Specifically, these include baculovirus tea tortrix granulosis virus, apple tortrix granulosis virus, and beet armyworm nuclear polyhedrosis virus.

[0035] In the plant treatment method of the present invention, by using microbial pesticides as microorganisms, it becomes possible to efficiently apply microbial pesticides to plants. Specifically, it is possible to shorten the time it takes for microbial pesticides to establish themselves on plants and / or reduce the amount of microbial pesticides used. Furthermore, plants infected with pesticide microorganisms have increased resistance to harmful organisms, and as a result, their yield may increase.

[0036] Therefore, in embodiments where the microorganism is a microbial pesticide, the plant treatment method of the present invention can be said to be a method for increasing plant yield and / or controlling pests and diseases in plants. Therefore, this disclosure is, The plants were exposed to light in the wavelength range of 300-325 nm, with an irradiation dose of 4000 μmol / m³. 2 More than 50000μmol / m 2 The irradiation process is as follows, and is performed such that no light in the wavelength range of 290 nm or less is irradiated, or the amount of such light is less than 20% of the amount of light irradiated in the wavelength range of 300 to 325 nm. An infection step is to infect the plant that has been irradiated with the aforementioned light with microorganisms that act as microbial pesticides in the plant, The present invention provides a method for increasing plant yields and / or controlling plant diseases and pests, including the following:

[0037] Furthermore, in embodiments in which the microorganism is a microbial pesticide, the method for producing microbially infected plants according to this disclosure can be described as a method for producing plants with increased resistance to diseases and pests. Therefore, this disclosure is, The plants were exposed to light in the wavelength range of 300-325 nm, with an irradiation dose of 4000 μmol / m³. 2 More than 50000μmol / m 2 The irradiation process is as follows, and is performed such that no light in the wavelength range of 290 nm or less is irradiated, or the amount of such light is less than 20% of the amount of light irradiated in the wavelength range of 300 to 325 nm. An infection step is to infect the plant that has been irradiated with the aforementioned light with microorganisms that act as microbial pesticides in the plant, This invention provides a method for producing plants with increased resistance to diseases and pests, including [specific element].

[0038] Examples of pests and diseases include tortrix moths, beet armyworms, lice, thrips, whiteflies, aphids, spider mites, diamondback moths, cabbage worms, tobacco budworms, mites, scale insects, longhorn beetles, corn borers, armyworms, diamondback moths, onion moths, white-spotted armyworms, and root-knot nematodes. Plant diseases caused by pests and diseases include powdery mildew, cucumber mosaic, wilt, sclerotinia rot, black rot, gray mold, leaf mold, sooty mold, anthracnose, gall, bacterial blight, bacterial seedling blight, brown stripe, bakanae disease, rice blast, seedling blight, purple root rot, white mold, crown gall, flower bud rot, soft rot, black rot, bacterial leaf spot, bacterial black spot, bacterial soft rot, bacterial stem necrosis, bacterial spot, canker, bacterial branch blight, white spot, leaf blight, white spot leaf blight, black blight, leaf spot, gray mold, white rust, black spot, bacterial wilt, and bacterial wilt / root rot. Increasing resistance to pests and diseases can reduce the occurrence of these plant diseases.

[0039] In some embodiments, the bacteria have the ability to transfer at least a portion of their internal DNA, such as plasmids, into the genome of an infected plant, such as the plant's genome. In certain embodiments, the microorganism is Agrobacterium. Preferably, the Agrobacterium is tumorigenic Agrobacterium (Rhizobium radiobacter or Agrobacterium tumefaciens) or rhizogenic Agrobacterium (Rhizobium rhizogenes or Agrobacterium rhizogenes).

[0040] In some embodiments, the microorganism or infectious agent is a plant virus vector. A plant virus vector is a vector based on a plant virus. Here, the plant virus is not particularly limited, as long as it can infect plant cells used in the method of this disclosure that have been irradiated with light in the wavelength range of 300-325 nm, and therefore cells whose resistance to microbial infection has been reduced compared to before irradiation. The plant virus may be, for example, a virus selected from the genera Tobamovirus, Potexvirus, Potivirus, Tobulavirus, Tombusvirus, Cucumovirus, Bromovirus, Alphamovirus, Comovirus, Calmovirus, Chelavirus, Weikavirus, and Kalimovirus. The plant virus is preferably a plant single-stranded positive-sense RNA virus. Specific examples of plant virus vectors include cauliflower mosaic virus (CaMV) vector, cucumber mosaic virus (CMV) vector, tobacco mosaic virus (TMV) vector, potato X virus (PVX) vector, tomato mosaic virus (ToMV) vector, plum poxvirus (PPV) vector, alfalfa mosaic virus (AIMV) vector, cowpea mosaic virus (CPMV) vector, and zucchini yellow mosaic virus (ZYMV) vector.

[0041] The microorganisms may be wild-type or genetically modified. In some embodiments, the microorganism has a gene encoding a target polypeptide, including a protein, to be expressed in the plant used in the method of the present disclosure. Herein, the gene is foreign to the plant. In the infection step, the foreign gene can be introduced into the plant by infecting the plant with the microorganism containing the target foreign gene. In this case, the “infection step” may also be called the “foreign gene introduction step.” Plants into which foreign genes have been introduced (genetically modified plants) may transiently express the target polypeptide (transient transformant or transient transfectant) or stably express it (transgenic plant or genetically modified plant in the narrow sense).

[0042] Therefore, in embodiments in which the microorganism contains foreign genes, the plant treatment method of this disclosure can be said to be a method for introducing foreign genes into plants. Therefore, this disclosure is, The plants were exposed to light in the wavelength range of 300-325 nm, with an irradiation dose of 4000 μmol / m³. 2 More than 50000μmol / m 2 The irradiation process is as follows, and is performed such that no light in the wavelength range of 290 nm or less is irradiated, or the amount of such light is less than 20% of the amount of light irradiated in the wavelength range of 300 to 325 nm. An infection process in which a plant irradiated with the aforementioned light is infected with a microorganism containing an exogenous gene, This invention provides a method for introducing foreign genes into plants, including [specific example].

[0043] Furthermore, in embodiments in which the microorganism contains foreign genes, the method for producing microbially infected plants according to this disclosure can be said to be a method for producing genetically modified plants. Therefore, this disclosure is, The plants were exposed to light in the wavelength range of 300-325 nm, with an irradiation dose of 4000 μmol / m³. 2 More than 50000μmol / m 2The irradiation process is as follows, and is performed such that no light in the wavelength range of 290 nm or less is irradiated, or the amount of such light is less than 20% of the amount of light irradiated in the wavelength range of 300 to 325 nm. An infection process in which a plant irradiated with the aforementioned light is infected with a microorganism containing an exogenous gene, The present invention provides a method for producing genetically modified plants, including those mentioned above.

[0044] The polypeptide to be expressed in a plant can be any polypeptide, as long as it can be expressed in the plant used, and may, for example, be a polypeptide of medical or industrial use. More specifically, the polypeptide of interest may be, for example, an immunogenic or antigenic polypeptide derived from a pathogen such as a virus, an antibody or its binding fragment, a cytokine, an enzyme, a growth factor or other physiologically active protein or its functional fragment. It is particularly preferable that the immunogenic or antigenic polypeptide can induce neutralizing antibodies, for example, one that can be used as a vaccine, including a VLP vaccine. The foreign gene may be included in the expression cassette. In the expression cassette, the foreign gene may be activatably linked upstream to a promoter appropriate for the plant being used. Examples of promoters include, but are not limited to, the 35S promoter for cauliflower mosaic virus and the ubiquitin promoter for maize. The expression cassette may also include other expression regulatory elements such as terminators.

[0045] Microbial infection of plants can be achieved by bringing a suspension of microorganisms (hereinafter also referred to as "microbial suspension") into contact with the plant. The liquid in which the microorganisms are suspended may be a culture medium suitable for the microorganisms and may contain a surfactant. When the microbial suspension comes into contact with a plant, the microorganisms infect the plant cells by penetrating into the plant tissue. More specifically, contact between the microbial suspension and the plant can be achieved by applying or spraying the microbial suspension onto the plant or by immersing the plant in the microbial suspension. Alternatively, the microbial suspension may be infiltrated into the plant tissue, for example, by injection using a syringe. Alternatively, infection can be brought into contact with plants using a dried powder obtained by freeze-drying or spray-drying a microbial suspension, such as a wettable powder, driftless (DL) powder, flow dust (FD) agent, or powder. In this case, contact can be achieved, for example, by spraying the powder or by using a blower mechanism installed in a greenhouse to disperse the powder.

[0046] Contact with the microbial suspension does not have to be with the entire plant, but may be with a part of the plant, such as all or some of the leaves of the plant, or all or part of a single leaf, the entire root or part of the root, all or part of the above-ground parts, or all or part of the flowers and / or buds. In this specification, “above-ground parts” may be leaves and / or stems, and may optionally include flowers and / or buds.

[0047] When infection is carried out by immersion of plants in a microbial suspension, a reduced pressure treatment may be performed during immersion to promote the penetration of the microbial suspension into the plant tissue. The target pressure during reduced pressure may be, for example, 0.005 to 0.5 atmospheres, more specifically 0.005 to 0.3 atmospheres, more specifically 0.01 to 0.3 atmospheres, more specifically 0.01 to 0.2 atmospheres, more specifically 0.01 to 0.1 atmospheres, or more specifically 0.02 to 0.1 atmospheres. Alternatively, the target pressure may be, for example, 0.5 to 50 kPa, more specifically 0.5 to 30 kPa, more specifically 1 to 30 kPa, more specifically 1 to 20 kPa, more specifically 1 to 10 kPa, or more specifically 2 to 10 kPa. The duration of the depressurization process could be, for example, 10 seconds to 10 minutes, more specifically 10 seconds to 5 minutes, more specifically 20 seconds to 5 minutes, more specifically 20 seconds to 3 minutes, more specifically 30 seconds to 3 minutes, or more specifically 30 seconds to 2 minutes. After the depressurization process, the pressure is restored to near atmospheric pressure or ambient pressure. The time required for restoration is not particularly limited and may be, for example, 10 seconds or less, more specifically 5 seconds or less, more specifically 3 seconds or less, or more specifically 1 second or less.

[0048] Specific examples of methods for infecting plants with microorganisms include the Agrobacterium method (also known as the agro-infiltration method; including the vacuum infiltration method), the floral spray method, the floral dip method, and the plant virus vector method, all of which are well known in this field.

[0049] The infection process is preferably carried out in the dark. By keeping the plants in the dark during the infection process, that is, by preventing photosynthesis in the cells of the plants, the reduction in microbial infection resistance induced in the irradiation process can be sustained throughout the infection process, thereby achieving efficient infection. More preferably, the plants are kept in the dark from the start of the irradiation process at the latest until the end of the infection process at the earliest. After the infection process, any microorganisms remaining on the plant surface may be removed using water or sterile / sterilized water such as a 70% ethanol aqueous solution.

[0050] (Storage in a dark place process) Plants that have been infected with microorganisms during the infection process may then be placed in a dark place. By placing the plants in the dark after the infection process, the reduction in microbial infection resistance induced during the irradiation process can be sustained, allowing a larger proportion of the microorganisms that invaded the plant tissue during the infection process to infect the plant cells. The upper limit of the period during which plants are placed in the dark after contact with the microbial suspension or its dried powder is not particularly limited and may be, for example, 48 hours, 36 hours, 24 hours, 18 hours, or 12 hours. The lower limit of the said period is not particularly limited and may be, for example, 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, or 6 hours. The range of the dark placement period may be any combination of the above lower and upper limits. Specific examples of periods of placement in darkness include, but are not limited to, 5 minutes to 48 hours, 15 minutes to 48 hours, 30 minutes to 48 hours, 1 hour to 48 hours, 2 hours to 48 hours, 3 hours to 48 hours, 6 to 48 hours, 6 to 36 hours, 6 to 24 hours, 6 to 18 hours, or 6 to 12 hours. The plants are preferably placed in the dark from the start of the infection process at the latest, and more preferably from the start of the irradiation process at the latest.

[0051] (Plant growth process or cultivation process) The plants infected with microorganisms in the aforementioned infection process may be allowed to grow thereafter. Plant growth can be carried out by any cultivation method known in the field, and therefore, the plant growth process can also be called the cultivation process. Cultivation may be soil cultivation, more specifically open-field cultivation or greenhouse cultivation, or hydroponic cultivation, more specifically hydroponics, spray cultivation or solid substrate cultivation. Hydroponic cultivation can be carried out under sterile conditions.

[0052] Cultivation, particularly hydroponics, may be carried out under controlled conditions. Controlled environmental conditions include, for example, light-dark cycle, temperature, humidity, the amount of natural and / or artificial light, and carbon dioxide concentration. These conditions are not particularly limited as long as they are suitable for the cultivation / growth of the plants used. The light-dark cycle can be appropriately selected according to the plants being cultivated and their growth stage. The light-dark cycle may be, for example, a long-day condition with 14-18 hours of light, or a short-day condition with 6-10 hours of light. The artificial light source is not particularly limited as long as it is suitable for cultivating the plants being grown. For example, incandescent lamps, fluorescent lamps, white lamps, high-intensity discharge (HID) lamps, such as high-pressure sodium lamps, metal halide lamps, high-pressure mercury lamps, LEDs, such as white LEDs, cold cathode fluorescent lamps (CCFLs), and organic EL lamps can be used. The artificial light is irradiated at a photosynthetically active photosynthetic photon flux density (PFS) that is appropriately set according to the plants being cultivated and their growth stage. The PFS is, for example, 50-600 μmol / m³. 2 / s, more specifically 100-500 μmol / m³ 2 It could be / s.

[0053] The temperature could be, for example, 20-30°C, and the humidity could be, for example, approximately 40-100%, more specifically approximately 50-95%, and more specifically approximately 50-80%. The carbon dioxide concentration can be, for example, approximately 300 to 5000 ppm, more specifically, approximately 500 to 3000 ppm, and more specifically, approximately 1000 to 1500 ppm. Fertilizers / liquid fertilizers can be appropriately selected depending on the plants being cultivated. Generally, fertilizers / liquid fertilizers contain nitrogen, phosphorus, and potassium. Cultivation may be carried out in an open-type plant factory or a (completely) closed-type plant factory. If the plant is a genetically modified plant, the target polypeptide can be extracted from the microbially infected part of the plant, for example, from the infected leaves, after approximately 3 to 7 days of cultivation.

[0054] <Method for producing fermented plant products> The method for producing plant fermented products disclosed herein is: The plants were exposed to light in the wavelength range of 300-325 nm, with an irradiation dose of 4000 μmol / m³. 2 More than 50000μmol / m 2 The irradiation process is as follows, and is performed such that no light in the wavelength range of 290 nm or less is irradiated, or the amount of such light is less than 20% of the amount of light irradiated in the wavelength range of 300 to 325 nm. An infection process in which microorganisms are infected into plants that have been irradiated with the aforementioned light, The process of fermenting the infected plants, Includes.

[0055] In other words, the method for producing a plant fermentation product according to the present disclosure includes a step of fermenting a plant treated by the plant treatment method according to the present disclosure.

[0056] The plant fermentation method of the plant fermentation method of the present disclosure is obtained by fermenting plant raw materials, preferably edible plant raw materials, with microorganisms, and may be, for example, brewed alcoholic beverages including fruit wine, brewed vinegar, pickles including pickles, miso, soy sauce, fermented tea, etc. Examples of fermented alcoholic beverages include wine (grape wine) and cider (apple wine). Examples of brewed vinegars include grain vinegars such as rice vinegar, barley vinegar, wheat vinegar, malt vinegar, Job's tears vinegar, soybean vinegar, and corn vinegar, as well as fruit vinegars such as apple cider vinegar, grape vinegar, wine vinegar, balsamic vinegar, and persimmon vinegar. Examples of miso include barley miso, rice miso, and soybean miso.

[0057] The plants that can be used in this method are plants that can serve as raw materials for the plant fermentation product to be produced, such as grains, legumes, vegetables, and fruits. Specific examples include rice, barley, wheat, corn, soybeans, adzuki beans, peas, Chinese cabbage, Hiroshima greens, Takana, Nozawana, Komatsuna, Rapeseed, Mizuna, radish, turnip, cucumber, eggplant, cabbage, perilla, potato, sweet potato, taro, bamboo shoots, and tea plants.

[0058] The irradiation step in the method for producing plant fermented products is as described in the section above, "<Plant Treatment Methods and Methods for Producing Microbially Infected Plants>". The infection process in the method for producing plant fermented products is as described in the section above, <Plant Treatment Methods and Methods for Producing Microbially Infected Plants>, excluding the use of microorganisms. The microorganisms used in the method for producing plant fermented products may be those commonly used in the production of fermented foods, including beverages made from plants, and may be bacteria, yeasts, or filamentous fungi.

[0059] Bacteria include Bacillus subtilis subsp. natto; plant-derived lactic acid bacteria: for example, Lactobacillus (e.g., Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus casei, Lactobacillus paracasei), Leuconostoc (e.g., Leuconostoc mesenteroides), Streptococcus, Enterococcus (e.g., Enterococcus faecalis, Enterococcus faecium), or Tetragenococcus (e.g., Tetragenococcus halophilus). Lactic acid bacteria belonging to the halophilus genus; acetic acid bacteria: for example, acetic acid bacteria belonging to the genus Acetobacter (e.g., Acetobacter aceti) or the genus Gluconacetobacter (e.g., Gluconacetobacter xylinus). Specific examples of yeast include those belonging to the genera Saccharomyces or Zygosaccharomyces, more specifically Saccharomyces cerevisiae, Saccharomyces ellipsoideus, Saccharomyces bayanus, Saccharomyces pastorianus, and Zygosaccharomyces rouxii. Specific examples of filamentous fungi include those belonging to the genus Aspergillus, more specifically Aspergillus oryzae, Aspergillus sojae, Aspergillus luchuensis, and Aspergillus niger.

[0060] Microbial infection of plants can be carried out by bringing a microbial suspension or microbial culture, also known as a "starter," into contact with the plant. The process of fermenting plants infected with microorganisms (fermentation process) can be carried out by methods known in the relevant field, depending on the fermented food being manufactured. According to the method for producing plant fermented products of this disclosure, it is possible to infect the raw plant material with more microorganisms and shorten the required fermentation time by promoting fermentation, thereby improving the production efficiency of plant fermented products.

[0061] <Plant treatment device> The plant processing apparatus disclosed herein is A plant holding section configured to hold plants, A UV light irradiation unit is configured to irradiate at least a portion of the plant held in the plant holding unit with UV light in the wavelength range of 300 to 325 nm, such that light in the wavelength range of 290 nm or less is not irradiated or the amount of such light is less than 20% of the amount of light in the wavelength range of 300 to 325 nm. A plant immersion unit comprising a storage tank configured to contain a liquid containing microorganisms in a liquid-containing area partitioned within its internal cavity, wherein the portion of the plant irradiated with ultraviolet light by the ultraviolet light irradiation unit is positioned within the liquid-containing area, or a liquid spraying unit configured to spray the liquid containing microorganisms toward the portion of the plant irradiated with ultraviolet light by the ultraviolet light irradiation unit, It is equipped with.

[0062] The plant treatment apparatus of this disclosure is suitable for carrying out the plant treatment method and the method for producing microbially infected plants of this disclosure, and is particularly suitable for carrying out the plant treatment method and the method for producing microbially infected plants of this disclosure in which the infection step is carried out by the agro-infiltration method.

[0063] (Plant holding part) The plant holding unit is capable of holding a plant and can be positioned, at least temporarily, within the irradiation area of ​​the ultraviolet light irradiation unit, for example, when the plant holding unit is in a predetermined position ("ultraviolet light irradiation position"), to irradiate the portion of the held plant (the portion to be irradiated with ultraviolet light in the wavelength range of 300 to 325 nm). The irradiated portion may be the entire plant, or one or more portions. The plant holding part may directly hold the plant, or it may indirectly hold the plant by holding a container that directly holds the plant, such as a container for hydroponics, or more specifically, a pot. The manner in which the plant-holding part holds the plant is not particularly limited and may include, for example, placing, housing, clamping, gripping, or fitting.

[0064] In the plant processing apparatus of the present disclosure, which includes a plant immersion section, the plant holding section is preferably configured to hold the plant in an upright position, i.e., with the above-ground part facing upward and the underground part facing downward, and / or in an inverted position. The plant holding section may also be configured to be able to move the plant being held between the upright and inverted positions, i.e., to be able to invert it upside down. This configuration makes it easy, for example, to immerse all or part of the above-ground part of a plant in a liquid containing microorganisms ("microbial suspension") contained in the storage tank of the plant immersion section. The plant holding unit may be equipped with reflective members around the plant being held, configured to reflect light from the ultraviolet light irradiation unit back towards the plant. This configuration allows for irradiation of parts of the plant that would be shaded by direct light from the ultraviolet light irradiation unit, resulting in irradiation of a wider area and thus achieving energetically and / or temporally efficient irradiation. In a particular embodiment, the plant holding section includes a member that shields light in the wavelength range of 300 to 325 nm, and is configured such that, with respect to direct light from the ultraviolet light irradiation section, the portion of the plant held by the plant holding section that is behind the member is not irradiated with light from the ultraviolet light irradiation section.

[0065] (Ultraviolet light irradiation section) The ultraviolet light irradiation unit is configured to irradiate a plant (hereinafter also referred to as the "irradiated plant") held in a predetermined position (ultraviolet light irradiation position; first position) by the plant holding unit with ultraviolet light in the wavelength range of 300 to 325 nm (hereinafter also referred to as the "predetermined ultraviolet light"), such that light in the wavelength range of 290 nm or less is not irradiated, or the amount of irradiation or irradiance of such light is less than 20%, preferably less than 10%, more preferably less than 5%, and more preferably less than 1% of the amount of irradiation or irradiance of light in the wavelength range of 300 to 325 nm. The ultraviolet light irradiation section is equipped with a light source capable of emitting ultraviolet light in the wavelength range of 300 to 325 nm. The ultraviolet light in the wavelength range of 300 to 325 nm has a main peak wavelength preferably within 310 ± 7 nm, more preferably within 310 ± 5 nm, and more preferably within 310 ± 3 nm. From the viewpoint of energy efficiency, it is preferable that the ultraviolet light irradiation section does not emit light in the wavelength range of 330 nm or higher, or emits light in the wavelength range of 300 to 325 nm at a dose or irradiance of, for example, less than 50%, preferably less than 30%, more preferably less than 20%, more preferably less than 10%, more preferably less than 5%, and more preferably less than 1%.

[0066] Examples of light sources for the ultraviolet light irradiation section include, for example, light-emitting diodes (LEDs), laser diodes (LDs), and xenon lamps, fluorescent lamps, incandescent lamps, metal halide lamps, and high-pressure mercury lamps having the necessary optical filters. The optical filter may be an optical filter whose transmittance to light in the wavelength range of 300 to 325 nm is greater than its transmittance to light in the wavelength range of 290 nm or less, or an optical filter whose transmittance to light in the wavelength range of 300 to 325 nm is greater than its transmittance to light in the wavelength range of 290 nm or less and light in the wavelength range of 330 nm or more. Light-emitting diodes (LEDs) or laser diodes (LDs) are particularly preferred as the light source. When using LEDs or LDs, it becomes easy to irradiate plants with wavelengths of light useful for reducing microbial infection resistance, while avoiding irradiation with wavelengths of light that may be unhelpful or harmful to reducing the plant's resistance to microbial infection. Furthermore, the use of LEDs or LDs is also preferable from the viewpoint of energy efficiency and economics due to their energy-intensive nature, low heat generation, low power consumption, and long lifespan. In addition, it becomes easier to control or manage the illuminance or irradiation amount.

[0067] Suitable LEDs or LDs as light sources are, specifically, LEDs or LDs having a peak wavelength of 310±7nm and a spectrum with a full width at half maximum of 1-15nm, more specifically 5-15nm, or more specifically 5-10nm; more specifically, LEDs or LDs having a peak wavelength of 310±5nm and a spectrum with a full width at half maximum of 1-15nm, more specifically 5-15nm, or more specifically 5-10nm; and more specifically, LEDs or LDs having a peak wavelength of 310±3nm and a spectrum with a full width at half maximum of 1-15nm, more specifically 5-15nm, or more specifically 5-10nm. LDs or LEDs may be provided in the form of an array, matrix, or cluster.

[0068] The ultraviolet light irradiation unit may irradiate the plant to be irradiated with predetermined ultraviolet light directly from any direction around the plant, for example, from above and / or from the side, or from one or more directions. The ultraviolet light irradiation unit does not necessarily have to irradiate the entire plant to be irradiated with the predetermined ultraviolet light. The ultraviolet light irradiation unit may be configured to irradiate only specific parts of the plant to be irradiated with the predetermined ultraviolet light, either by itself or together with the plant holding unit, more specifically with a shielding member that the plant holding unit may have. The ultraviolet light irradiation unit may be configured to irradiate the predetermined ultraviolet light as directional light. In the ultraviolet light irradiation area, when irradiated with ultraviolet light in the wavelength range of 300-325 nm, the photosynthetic photon flux density is 10 μmol / m³. 2 It is preferable that the value is less than or equal to 5 μmol / m². 2More preferably, it is below / s, and more preferably 2 μmol / m 2 More preferably, it is below / s, and more preferably 1 μmol / m 2 More preferably, it is below / s. Thus, in a specific embodiment, the ultraviolet light irradiation unit is disposed in an irradiation chamber that shields photosynthetically active radiation from the outside. As used herein, "shielding photosynthetically active radiation from the outside" means that when the illumination provided inside is turned off, the photosynthetically active photon flux density inside is 10 μmol / m 2 / s or less, more specifically 5 μmol / m 2 / s or less, more specifically 2 μmol / m 2 / s or less, more specifically 1 μmol / m 2 / s or less, which means that it can be achieved. According to this embodiment, since the plant can be kept in the dark, photosynthesis in the plant can be substantially prevented, and a decrease in the microbial infection resistance of the plant associated with irradiation of predetermined ultraviolet light by the ultraviolet light irradiation unit can be induced more efficiently.

[0069] The ultraviolet light irradiation unit may include an optical system composed of one or more optical system components known in the art, such as a lens, a mirror, an optical filter, a mask, a diffusion plate, etc. Additionally or alternatively, the ultraviolet light irradiation unit may include a control unit that controls the light source and / or the optical system. The control unit may control the dimming of the light source and / or the timing of lighting and extinguishing. The control unit may control whether the ultraviolet light irradiation unit emits continuous light, intermittent light, or a combination thereof. When the ultraviolet light irradiation unit can emit intermittent light, the control unit may control the pulse width and / or duty ratio of the intermittent light. Such a control unit may be, for example, a pulse width modulation circuit, or a pulse width modulation circuit and a timer, and may be composed of, for example, a microcomputer, a relay, and / or a switching element. The control unit may be configured to specify, based on information from a sensor, which of the plurality of light sources included in the ultraviolet light irradiation unit should emit light. The plurality of light sources may be in the form of, for example, an array, a matrix, or a cluster.

[0070] In certain embodiments, the ultraviolet light irradiation unit is configured to irradiate a specific part of a plant held in a plant holding unit with predetermined ultraviolet light (preferably, predetermined directional ultraviolet light). In this case, the specific part of the plant is detected or identified in advance by a sensor. The control unit may control the ultraviolet light irradiation unit based on information from the sensor so that the predetermined ultraviolet light is directed towards the specific part. The sensor may be configured as part of the ultraviolet light irradiation unit, or it may be located in a sensor unit outside the ultraviolet light irradiation unit.

[0071] (Plant soaking section) The plant immersion unit includes a storage tank configured to contain a liquid in a liquid-containing area partitioned within its internal cavity, and is configured such that the portion of the plant held in the plant holding unit at a predetermined position that is irradiated with ultraviolet light by the ultraviolet light irradiating unit is located within the liquid-containing area. Here, the predetermined position, which can also be called the immersion position, may be the position in which the plant held in the plant holding unit is irradiated with ultraviolet light by the ultraviolet light irradiating unit (ultraviolet light irradiation position; first position), or it may be a position different from the ultraviolet light irradiation position (second position). In the former case, the plant is immersed in the microbial suspension at the position irradiated with the predetermined ultraviolet light. When liquid is placed in the liquid-containing area, the portion of the plant held in the plant-holding section at the immersion position that is exposed to ultraviolet light is immersed in the liquid contained in the liquid-containing area. Specifically, the liquid is a liquid containing microorganisms, i.e., a microbial suspension.

[0072] The storage tank may have one or two openings for moving plants held in the plant holding section into and out of its internal cavity. The openings may, for example, open to the top of the storage tank or to the side. The openings may have a rim along their outer circumference. The rim is configured to be in close contact with a portion of the lid member described later, thereby sealing the internal cavity of the storage tank. In certain embodiments, the plant immersion section further includes a lid member configured to close the opening of the storage tank and seal its internal cavity. According to this embodiment, while the plants are immersed in the microbial suspension within the internal cavity, the infiltration of the microbial suspension into the plant tissue can be promoted by adjusting the air pressure within the internal cavity. The air pressure within the internal cavity may be adjusted by a pressure adjustment unit. In this embodiment, the storage tank may be configured to function as a depressurization chamber. The lid member may be provided with a tightly fitting member or mechanism, such as a sealing material, on the periphery of the opening, or on the outer rim if the opening has an outer rim.

[0073] (Pressure regulating section) The pressure regulating unit includes a pressure reduction mechanism configured to reduce the pressure in the internal cavity of the storage tank, which is sealed by a lid member. The pressure reduction mechanism may be, for example, a vacuum pump. The pressure reduction mechanism is connected to the internal cavity of the storage tank via a valve, enabling fluid communication. The pressure reduction mechanism may be configured to reduce the pressure in the internal cavity of the storage tank to, for example, 0.5 to 50 kPa, more specifically 0.5 to 30 kPa, more specifically 1 to 30 kPa, more specifically 1 to 20 kPa, more specifically 1 to 10 kPa, or more specifically 2 to 10 kPa. The pressure regulating section may further include a mechanism, such as a pressure release valve, for restoring the pressure inside the internal cavity to the ambient pressure, for example, a pressure near atmospheric pressure.

[0074] (Liquid injection part) The liquid spraying unit is configured to spray liquid toward the portion of the plant held in the plant holding unit that is exposed to ultraviolet light. Here, the predetermined position, which can also be called the spraying position, may be the ultraviolet light-irradiated position (first position) or a position different from the ultraviolet light-irradiated position (second position). In the former case, the plant is sprayed with a microbial suspension at the position irradiated with the predetermined ultraviolet light. The liquid may specifically be a microbial suspension. The liquid spraying unit can employ any configuration that allows it to spray liquid towards the plant held in the plant holding unit. The liquid spray from the liquid spraying unit may be directed from any direction around the plant held by the plant holding unit, for example, from above and / or from the side, or from two or more directions. The injection may utilize water pressure and / or air pressure. The injection pattern or spray pattern may be of any shape, for example, linear, fan-shaped (flat), or full-cone.

[0075] The liquid injection unit has, for example, one or more nozzles that spray liquid toward a plant held by the plant holding unit. The nozzles are configured to spray liquid or a mixture of liquid and gas. The nozzles are, for example, one-fluid nozzles or two-fluid nozzles. The liquid spraying unit may spray the liquid in a mist form. In certain embodiments, the liquid spray unit is configured to spray liquid as a stream directed at a specific part of a plant held in the plant holding unit. In this case, the specific part of the plant is detected or identified in advance by a sensor. A control unit in the liquid spray unit may control the direction of the nozzle based on information from the sensor so that the liquid is sprayed toward the specific part. The sensor may be located in the ultraviolet light irradiation unit or in a sensor unit outside the ultraviolet light irradiation unit.

[0076] (Sensor unit) In certain embodiments, the plant processing apparatus of the present disclosure includes a sensor unit configured to identify a specific part of a plant held in a plant holding unit based on color and / or shape, and to transmit its positional information to a control unit in an ultraviolet light irradiation unit. This enables efficient and reliable identification of plant parts whose resistance to microbial infection should be reduced and irradiation of those parts with predetermined ultraviolet light by the plant processing apparatus of the present disclosure. The specific part may be, for example, a specific organ, such as a leaf, flower and bud, or a root. The sensor unit may be configured to transmit positional information to a control unit located in the liquid ejection unit. This allows for efficient and reliable spraying of the microbial suspension by the plant processing apparatus of this disclosure onto the site to be infected with microorganisms. The sensor unit comprises one or more sensors capable of discriminating color and / or shape. The sensors may be, for example, two-dimensional sensors, preferably two-dimensional color sensors. Specific examples of sensors include, but are not limited to, monochrome or color image discrimination sensors, and may include CCD sensors, CMOS sensors, etc. The sensors may be arranged so that their sensing area covers the irradiation area of ​​the ultraviolet light irradiation unit.

[0077] (Photosynthesis-effective radiation irradiation section) The plant processing apparatus of this disclosure may further include a photosynthetically active radiation irradiation unit. The photosynthetically active radiation irradiation unit is configured to irradiate photosynthetically active radiation toward a plant ("irradiated plant") that is held in a predetermined position (photosynthetically active radiation irradiation position; third position) by the plant holding unit. The photosynthetically active radiation irradiation unit is equipped with a light source capable of emitting light in the wavelength range of 400 to 700 nm. The light source is not particularly limited as long as it can emit light suitable for the growth of the irradiated plants, and any light source available in the field can be used, such as incandescent lamps, fluorescent lamps, white lamps, high-intensity discharge (HID) lamps, such as high-pressure sodium lamps, metal halide lamps, high-pressure mercury lamps, LEDs, such as white LEDs, cold cathode fluorescent lamps (CCFLs), and organic EL lamps. Of the aforementioned light sources, LEDs or LDs are particularly preferred from the viewpoint of energy efficiency, economy, and ease of control and management. LDs or LEDs may be provided in the form of arrays, matrices, or clusters.

[0078] The photosynthetically active radiation irradiation section preferably irradiates the entire plant to be irradiated with photosynthetically active radiation. The photosynthetically active photon flux density is, for example, 50 to 600 μmol / m³. 2 / s, more specifically 100-500 μmol / m³ 2 It can be / s. The photosynthetically active radiation irradiation unit may irradiate the plant to be irradiated from any direction around the plant, but it is preferable to irradiate from above. The photosynthetically active radiation irradiation unit may include a control unit that controls the emission pattern of photosynthetically active radiation. The emission pattern may include a light-dark cycle. The photosynthetically active radiation irradiation unit may be incorporated into a cultivation rack (e.g., a multi-tiered rack) configured to hold the plant holding unit. In this case, the cultivation rack may incorporate a nutrient solution supply system as a nutrient solution supply unit that supplies nutrient solution to the plants held by the plant holding unit held by the cultivation rack.

[0079] (Plant transport mechanism) In some embodiments, the plant processing apparatus of the present disclosure includes a first transport mechanism configured to transport a plant holder from at least an ultraviolet light irradiation section to a plant immersion section or a liquid spray section. More specifically, the first transport mechanism may be configured to transport at least from an ultraviolet light irradiation position (first position) within the ultraviolet light irradiation section to an immersion position within the plant immersion section or a spray position (second position) within the liquid spray section. In a particular embodiment that includes a plant immersion section, the first transport mechanism may be configured to include an inversion mechanism that inverts the plant holding section between the ultraviolet light irradiation position and the immersion position. This configuration makes it easier to immerse, for example, all or part of the above-ground portion of a plant in a microbial suspension contained in the storage tank of the plant immersion section. Preferably, the first transport mechanism is configured to shield photosynthetically active radiation from the plant holding section on the transport path from at least a first position to a second position, and more specifically from the plants held in the plant holding section. For example, the transport path may be provided within a passage that shields from photosynthetically active radiation from the outside.

[0080] In some embodiments, the plant processing apparatus of the present disclosure additionally or alternatively comprises a second transport mechanism configured to transport the plant holding section at least from the plant immersion section or liquid injection section to the photosynthetically active radiation irradiation section. More specifically, the second transport mechanism may be configured to transport at least from a second position to a third position. Preferably, the second transport mechanism is configured to shield photosynthetically active radiation from the plant holding section on the transport path from at least the second to the third position, and more specifically from the plants held in the plant holding section. For example, the transport path may be provided within a passage that shields from photosynthetically active radiation from the outside.

[0081] The transport mechanism may be configured, for example, with a transport stage, a transport robot, or a robotic arm. The second transport mechanism may be configured integrally with the first transport mechanism. A part of the first and / or second transport mechanism may constitute a plant holding section.

[0082] The plant processing apparatus of this disclosure will be described below with reference to Figures 1 and 2, which are schematic diagrams showing specific examples of embodiments of the plant processing apparatus of this disclosure.

[0083] (Embodiment 1) The plant processing apparatus 100 of the present disclosure, schematically shown in Figure 1, comprises a plant holding unit 110, an ultraviolet light irradiation unit 120, a liquid spray unit 130, and a control unit 140 that controls the ultraviolet light irradiation unit 120 and the liquid spray unit 130. The plant holding unit 110, the ultraviolet light irradiation unit 120, and the liquid spray unit 130 may be contained within a housing 180. The plant holding unit 110 is capable of holding the plant P. The plant processing device 100 may have multiple plant holding units 110. The ultraviolet light irradiation unit 120 includes a light source 122 capable of emitting ultraviolet light in the wavelength range of 300 to 325 nm, and a control unit 124 that controls the light source 122, and can irradiate the plant P with ultraviolet light in the wavelength range of 300 to 325 nm. The light source 122 may be in the form of a light source array such as an LD array or an LED array. The liquid injection unit 130 is capable of spraying liquid toward the plant P held in the plant holding unit 110. The liquid injection unit 130 is controlled by the control unit 140 to spray liquid toward the plant P after it has been irradiated with ultraviolet light in the wavelength range of 300 to 325 nm by the ultraviolet light irradiation unit 120. Here, the liquid may more specifically be a microbial suspension. The enclosure 180 may be entirely made of light-shielding material, or it may be a darkroom. The plant processing device 100 may also be provided with a reflective material 112 capable of reflecting ultraviolet light in the wavelength range of 300 to 325 nm. The reflective material 112 may be provided in the plant holding unit 110, or it may be arranged on at least a part of the inner surface of the housing 180. The reflective material 112 can reflect ultraviolet light in the wavelength range of 300 to 325 nm emitted from the ultraviolet light irradiation unit 120 toward the plant P held in the plant holding unit 110.

[0084] (Embodiment 2) The plant processing apparatus 200 of this disclosure, schematically shown in Figure 2, comprises a plant holding unit 210, an ultraviolet light irradiation unit 220, and a plant immersion unit 240. The plant processing apparatus 200 further comprises an optional pressure adjustment unit 250. The plant holding section 210 is capable of holding plants P. The plant processing device 200 may have multiple plant holding sections 210. The plant holding section 210 may be equipped with a reflective material that can reflect ultraviolet light in the wavelength range of 300 to 325 nm toward the plant P being held. The plant holding section 210 may be transported by a transport mechanism from within the irradiation area of ​​the ultraviolet light irradiation section 220, more specifically from the ultraviolet light irradiation position, to within the liquid storage area partitioned within the internal cavity of the storage tank 242 of the liquid immersion section 240, more specifically to the immersion position. The plant holding section 210 may be inverted during transport or at the immersion position to facilitate immersion.

[0085] The ultraviolet light irradiation unit 220 includes a light source 222 capable of emitting ultraviolet light in the wavelength range of 300 to 325 nm, and a control unit 224 that controls the light source 222, and can irradiate the plant P with ultraviolet light in the wavelength range of 300 to 325 nm. The light source 222 may be in the form of a light source array such as an LD array or an LED array. The ultraviolet light irradiation unit 220 may be located inside the irradiation chamber 280. In Figure 2, the ultraviolet light irradiation unit 220 is positioned to irradiate the plant P from above, but it may be additionally or alternatively positioned to irradiate the plant P from the side and / or below. It is preferable that the irradiation chamber 280 can shield against photosynthetically active radiation from the outside. The liquid immersion section 240 includes a storage tank 242 in which a liquid storage area 244 is partitioned within its internal cavity. The liquid immersion section 240 further includes, optionally, an opening 246 and a lid member 248 that closes the opening. The liquid containment area 244 can contain a liquid in which the plant P is immersed, more specifically a microbial suspension. The opening 246 is provided for moving the plant P, held in the plant holding section 210, into and out of the internal cavity of the storage tank 242. In Figure 2, the opening 246 opens upward, but it may also open to the side. The lid member 248 can close the opening 246 and seal the internal cavity.

[0086] The pressure regulating unit 250 includes a vacuum pump 252 capable of reducing the pressure in the internal cavity of the storage tank 242 sealed by the lid member 248, and a pressure release valve 254 for returning the pressure in the internal cavity to a pressure near atmospheric pressure. The vacuum pump 252 and the pressure release valve 254 are connected to the internal cavity of the storage tank 242 via the valve so as to be able to communicate with fluids. The plant P held in the plant holding section 210 is transported into the internal cavity of the storage tank 242 through the opening 246. After being irradiated with ultraviolet light in the wavelength range of 300-325 nm by the ultraviolet light irradiation section 220, the plant P held in the plant holding section 210 is immersed in the liquid contained in the liquid storage area 244 at the immersion position within the internal cavity. After transport, the lid member 248 seals the opening 246 and seals the internal cavity of the storage tank 242. After sealing, the internal cavity of the storage tank 242 is depressurized to a predetermined pressure by the vacuum pump 252. After a predetermined time has elapsed, the pressure release valve 254 is opened, and the pressure in the internal cavity of the storage tank 242 returns to a pressure near atmospheric pressure. At this time, the immersion of the liquid into the plant P is promoted. Subsequently, the plant P held in the plant holding section 210 is transported out of the internal cavity of the storage tank 242 through the opening 246. [Examples]

[0087] (light source) In the following examples, LEDs emitting light with peak wavelength ± full width at half maximum of 280±10 nm, 310±10 nm, and 340±10 nm, respectively (hereinafter also referred to as "280 nm-LED light," "310 nm-LED light," and "340 nm-LED light," respectively) were used. The emission spectra of the LEDs used are shown in Figure 3. 310nm-LED light has most of its wavelength components in the 300-325nm range. On the other hand, 280nm-LED light and 340nm-LED light have almost no wavelength components in the 300-325nm range.

[0088] Example 1: Agro-infiltration test 1 (method) <Cultivation of Benthamia tobacco> Tobacco benthamiana was cultivated in soil for approximately four weeks (using "Yosaku" special growing medium for fruit and vegetable crops; J-Cam Agri Co., Ltd.). The cultivation conditions were as follows: temperature 24°C, 14 hours of light, white light illumination (100 μmol / m²). 2 ( / s). <Preparation of Agrobacterium suspension> Agrobacterium (strain LBA4404) introduced with intron-inserted firefly luciferase was cultured with shaking at 28 °C and 200 rpm in LB medium. When the OD600 reached around 0.5, the bacteria were collected by centrifugation at 3,000 rpm for 15 minutes at room temperature. The precipitated bacterial cells were suspended in infiltration medium (10 mM MgCl2, 10 mM MES, 150 μM acetosyringone, pH 5.7) to an OD600 of 0.5 and left standing at room temperature in the dark for about 24 hours.

[0089] <Ultraviolet LED light irradiation> Nicotiana benthamiana plants at about 4 weeks old were used in their potted state. For each plant, treatment sections with and without LED light irradiation were set up for 1 to 3 leaves per plant. Half of each leaf was covered with aluminum foil to block light, serving as the non-irradiated section (hereinafter also simply referred to as the "non-irradiated part"), and the remaining half was the irradiated section (hereinafter also simply referred to as the "irradiated part") (Figure 4). One of the above three types of LED lights was irradiated onto the leaves of the test section at 2.5 μmol / m 2 / s for 45 minutes. The irradiation dose was 6750 μmol / m 2 . Immediately after irradiation, the aluminum foil covering the leaves was removed. Thereafter, Nicotiana benthamiana was stored in the dark for 24 hours. <Infection treatment (agroinfiltration) and infection verification> After 24 hours of storage in the dark, the above-mentioned Agrobacterium suspension was applied to the irradiated and non-irradiated sections of the leaves of Nicotiana benthamiana irradiated with 310 nm-LED light or 340 nm-LED light using a needleless syringe. For each leaf, the same number (2 - 6) of applications were made in each treatment section of the irradiated and non-irradiated parts. Immediately after application, a 0.2 mM D-luciferin solution was sprayed over the entire leaf. After spraying, it was left standing at 22 °C in the dark for 24 hours. After 24 hours of storage in the dark, the non-irradiated and irradiated sections were imaged with a CCD color camera, and the luminescence amounts of firefly luciferase in both treatment sections were compared on the image. The luminescence amount of luciferase in the leaf reflects the infection amount of luciferase-introducing Agrobacterium into the leaf cells.

[0090] (result) In the area irradiated with 280nm-LED light, significant wilting occurred 24 hours after irradiation, resulting in decreased moisture content and thinner leaves (Figure 5). This trend was more pronounced in leaves that were not irradiated, i.e., leaves that were entirely irradiated with 280nm-LED light. It is thought that the significant wilting occurred due to ultraviolet light stress associated with 280nm-LED light irradiation, as 4-week-old tobacco plants have low resistance to ultraviolet light. Since the 280nm-LED light irradiation area was not in a state where agro-infiltration could be performed, 280nm-LED light was not used in subsequent embodiments. Figures 6 and 7 show the luminescence images of tobacco leaves irradiated with 310nm-LED light or 340nm-LED light, respectively. The areas enclosed by frames are the unirradiated areas. The upper panel shows an image of tobacco leaves superimposed with the luminescence image of firefly luciferase, and the lower panel shows the luminescence image of firefly luciferase. In the upper image, areas with high luminescence are shown in black. The area irradiated with 310nm-LED light shows significantly more light emission compared to the unirradiated area, which is shown as a framed region in the figure (Figure 6). Therefore, it is thought that 310nm-LED light irradiation improved the infection efficiency of Agrobacterium, and it is presumed that this is due to a decrease in the plant's resistance to microbial infection. On the other hand, in leaves irradiated with 340nm-LED light, no difference in luminescence was observed between the irradiated and unirradiated areas (Figure 7). This is thought to be because 340nm-LED light did not affect the plant's resistance to microbial infection.

[0091] Figures 8 and 9 show the luminescence levels in the un-irradiated and irradiated areas of each leaf of Tobacco benthamiana, irradiated with 310nm-LED light and 340nm-LED light, respectively. A paired t-test (two-tailed test) was performed between the un-irradiated and irradiated areas. The results showed that the luminescence level was significantly increased in the 310nm-LED irradiated area compared to the un-irradiated area (Figure 8; p<0.05). This indicates that more Agrobacterium infects plant cells in the 310nm-LED irradiated area compared to the un-irradiated area. On the other hand, there was no significant difference in luminescence between the 340nm-LED irradiated area and the unirradiated area (Figure 9; p≧0.05). This indicates that 340nm-LED light irradiation does not affect Agrobacterium infection of plant cells. Based on these results, it is considered that 310nm-LED light induces a decrease in microbial infection resistance in plant cells, while 340nm-LED light does not have such an effect.

[0092] Example 2: Agro-infiltration test 2 (method) Tobacco benthamiana leaves were exposed to 310nm-LED light at a concentration of 2.5 μmol / m². 2 The same tests as in Example 1 were performed, except that the irradiation was performed at a rate of 4.5 minutes, 90 minutes, 225 minutes, or 360 minutes. The irradiation doses were 675, 13,500, 33,750, or 54,000 μmol / m², respectively. 2 That was the case.

[0093] (result) Figures 10 to 13 show 310nm-LED light at concentrations of 675, 13,500, 33,750, and 54,000 μmol / m², respectively. 2 The luminescence levels in the unirradiated and irradiated areas of each leaf of tobacco benthamiana irradiated with the specified dose are shown. A paired t-test (two-tailed test) was performed between the unirradiated and irradiated areas, yielding 13,500 and 33,750 μmol / m³, respectively. 2 In the light-irradiated area, the amount of light emitted was significantly increased compared to the unirradiated area (Figure 11, p<0.01; Figure 12, p<0.05). On the other hand, at 675 μmol / m³2 In the light-irradiated area, there was no significant difference in luminescence between the irradiated and unirradiated areas (Figure 10; p≧0.05). Also, 54,000 μmol / m 2 In the light-irradiated areas, the amount of light emitted tended to increase compared to the unirradiated areas, but wilting was also observed. Figure 14 shows the irradiation dose of 13,500 μmol / m². 2 The image shows the luminescence of a tobacco leaf used in a 310nm-LED light irradiation test. The area enclosed by the frame is the unirradiated area. The upper panel shows an image of tobacco leaves superimposed with the luminescence image of firefly luciferase, and the lower panel shows the luminescence image of firefly luciferase. In the upper image, areas with high luminescence are shown in black. From the figure, it can be seen that the 310nm-LED light-irradiated area emits significantly more light than the unirradiated area, which is shown as the framed area in the figure. Combining these results with those of Example 1, the 310nm-LED light has a molecular weight of approximately 1,000 to 4,000 μmol / m³. 2 At the above irradiation doses, a decrease in microbial infection resistance can be induced in plant cells, but 54,000 μmol / m³ 2 It is thought that the above irradiation doses may damage plant cells.

[0094] Example 3: Agro-infiltration test 3 (method) The same test as in Example 1 was performed, except that only 310nm-LED light was used and storage in the dark after irradiation was omitted. (result) Figure 15 shows the 310nm-LED light at 6,750 μmol / m². 2 The luminescence levels in the unirradiated and irradiated areas of each leaf of tobacco plants irradiated with 310nm-LED light were shown. The luminescence level in the 310nm-LED irradiated area was significantly increased compared to the unirradiated area (paired t-test (two-tailed), p<0.05). From this, it can be understood that plants irradiated with 310nm-LED light show a significant decrease in resistance to microbial infection immediately afterward.

[0095] Example 4: Floral Inoculation Test (method) Following the method of M. Narusaka et al. (Plant Biotechnology 27, 349-351 (2010)), the flower buds of Arabidopsis thaliana were irradiated with 310 nm-LED light at a dose of 6,750 μmol / m². 2 (=2.5 μmol / m³) 2 After irradiation at 45 minutes ( / s), a suspension containing Agrobacterium with a kanamycin resistance gene introduced into the culture medium was inoculated using a micropipette (floral inoculation). Arabidopsis thaliana was grown according to conventional methods until the flower buds bloomed and seeds were formed, and then the seeds were harvested as dried seeds. For control, dried seeds were harvested from flower buds of another Arabidopsis thaliana plant that underwent floral inoculation without 310 nm-LED light irradiation. 264 dried seeds obtained from flower buds irradiated with 310nm-LED light and 1000 dried seeds obtained from unirradiated flower buds were sown on kanamycin-containing agar medium (MS medium) and kanamycin selection was performed.

[0096] (result) Seeds obtained from unirradiated flower buds did not grow on kanamycin-containing agar medium. This suggests that genetic recombination by the floral inoculation method did not occur in flower buds subjected to floral inoculation without 310 nm-LED light irradiation. On the other hand, of the 264 seeds obtained from flower buds that underwent floral inoculation after irradiation with 310nm-LED light, two grew on kanamycin-containing agar medium. This suggests that the efficiency of genetic recombination by the floral inoculation method was increased in flower buds that underwent floral inoculation after irradiation with 310nm-LED light. These results indicate that irradiation with 310nm-LED light improves the efficiency of genetic modification by the floral inoculation method. Considering the results of Examples 1-3, this is thought to be a result of the 310nm-LED light irradiation reducing the resistance of flower buds to microbial infection.

[0097] Example 5: Gene expression analysis of Arabidopsis thaliana irradiated with 310nm-LED light. 310nm-LED light irradiation amount 6,750μmol / m 2 (=2.5 μmol / m³) 2 Gene expression analysis was performed on Arabidopsis thaliana immediately after irradiation at 45 minutes ( / s). (result) In Arabidopsis thaliana irradiated with 310nm-LED light, transcription factors related to defense against bacteria and filamentous fungi (WRKY60, MYB28, MYB29, MYB45) and genes associated with salicylic acid response (UGT1, PARN) were significantly downregulated. Furthermore, we were able to confirm the downregulation of genes (GAD1, ALDH2B7) related to GABA biosynthesis, which are known to inhibit gene transfer into plants by Agrobacterium. Therefore, it can be understood that 310nm-LED light weakens the defense mechanisms of plant cells against bacteria and filamentous fungi (in other words, reduces resistance to microbial infection), and further promotes microbial infection of plant cells by suppressing GABA biosynthesis in cells.

[0098] This disclosure includes the methods and apparatus described in the following sections. Section 1: The plants were exposed to light in the wavelength range of 300-325 nm, with an irradiation dose of 4000 μmol / m³. 2 More than 50000μmol / m 2 The irradiation process is as follows, and is performed such that no light in the wavelength range of 290 nm or less is irradiated, or the amount of such light is less than 20% of the amount of light irradiated in the wavelength range of 300 to 325 nm. An infection process in which microorganisms are infected into plants that have been irradiated with the aforementioned light, Methods for processing plants, including those mentioned above. Section 2: The method according to item 1, wherein, in the infection step, the foreign gene is introduced into the plant by infecting it with a microorganism containing the target foreign gene. Section 3: The method according to item 1 or 2, wherein the irradiation step is performed in the dark. Section 4: The method according to any one of claims 1 to 3, further comprising the step of placing the plant infected with microorganisms in the aforementioned infection step in the dark for 6 to 48 hours. Section 5: The method according to any one of claims 1 to 4, further comprising the step of growing a plant infected with microorganisms in the aforementioned infection step. Item 6: In the irradiation step, the light within the wavelength range of 300 to 325 nm is 0.05 to 300 μmol / m³ 2 The method described in any one of items 1 to 5, irradiated with a photon flux density of / s. Section 7: The method according to any one of claims 1 to 6, wherein in the irradiation step, no light in the wavelength range of 330 nm or more is irradiated, or the amount of such irradiation is less than 30% of the amount of light irradiated in the wavelength range of 300 to 325 nm. Section 8: The method according to any one of claims 1 to 7, wherein the infection in the infection step is by immersion of the plant in a liquid containing microorganisms or by spraying or injecting the liquid into the plant. Section 9: The infection in the aforementioned infection step is carried out by the agro-infiltration method or the plant virus vector method, according to any one of items 1 to 8. Section 10: The method according to any one of items 1 to 9, wherein the light in the 300 to 325 nm wavelength range is light having a wavelength spectrum with a peak wavelength of 310 ± 5 nm and a full width at half maximum of 5 to 15 nm. Section 11: The plants were exposed to light in the wavelength range of 300-325 nm, with an irradiation dose of 4000 μmol / m³. 2 More than 50000μmol / m 2The irradiation process is as follows, and is performed such that no light in the wavelength range of 290 nm or less is irradiated, or the amount of such light is less than 20% of the amount of light irradiated in the wavelength range of 300 to 325 nm. An infection process in which microorganisms are infected into plants that have been irradiated with the aforementioned light, A method for producing plants infected with microorganisms, including [specific microorganisms]. Section 12: The irradiation process and infection process described in item 11, The process of fermenting the infected plants, A method for producing plant fermented products containing Section 13: A plant holding section configured to hold plants, A UV light irradiation unit is configured to irradiate at least a portion of the plant held in the plant holding unit with UV light in the wavelength range of 300 to 325 nm, such that light in the wavelength range of 290 nm or less is not irradiated or the amount of such light is less than 20% of the amount of light in the wavelength range of 300 to 325 nm. A plant immersion unit comprising a storage tank configured to contain a liquid containing microorganisms in a liquid-containing area partitioned within its internal cavity, wherein the portion of the plant irradiated with ultraviolet light by the ultraviolet light irradiation unit is positioned within the liquid-containing area, or a liquid spraying unit configured to spray the liquid containing microorganisms toward the portion of the plant irradiated with ultraviolet light by the ultraviolet light irradiation unit, A plant processing device equipped with the following features. Section 14: The first transport mechanism is further configured to transport the plant holding section from the ultraviolet light irradiation section to the plant immersion section or the liquid spray section, The plant processing apparatus according to claim 13, wherein the first transport mechanism is configured to shield photosynthetically active radiation to the plants on the transport path. Section 15: A plant processing apparatus according to item 13 or 14, further comprising a pressure adjustment unit, The storage tank has an opening, The plant immersion section further comprises a lid member configured to close the opening and seal the internal cavity of the storage tank, The plant processing apparatus comprises a pressure adjustment unit equipped with a pressure reduction mechanism configured to reduce the pressure of the internal cavity of the storage tank when it is sealed by the lid member. Section 16: The plant processing apparatus according to any one of claims 13 to 15, wherein the ultraviolet light irradiation unit, or the ultraviolet light irradiation unit and the plant holding unit, are configured to irradiate ultraviolet light only to specific parts of the plant held in the plant holding unit. Section 17: A plant processing apparatus according to any one of claims 13 to 16, further comprising a sensor unit configured to detect a specific part of a plant, The ultraviolet light irradiation unit is configured to irradiate the ultraviolet light toward the portion detected by the sensor unit, The plant processing apparatus is configured such that the liquid injection unit is capable of spraying liquid toward the part detected by the sensor unit. Section 18: The plant processing apparatus according to any one of claims 13 to 17, wherein the ultraviolet light irradiation unit comprises a light source that emits light having a wavelength spectrum with a peak wavelength of 310 ± 5 nm and a full width at half maximum of 5 to 15 nm. [Industrial applicability]

[0099] The methods of this disclosure can be used in several embodiments to produce recombinant proteins that serve as raw materials for biopharmaceuticals (e.g., interferons, immunogens for vaccines, antibodies, antigens, growth factors), industrial enzymes (e.g., cellulases, proteases, amylases), and foods and feeds (e.g., protein supplements, nutritional supplements). The method disclosed herein can be used in several embodiments for the production of transgenic plants. The method disclosed herein is particularly effective for plants with low gene transfer efficiency due to high disease resistance or the small number of species obtained at one time. The method disclosed herein can be used in some embodiments for the efficient production of plant fermentations. The method disclosed herein can be used in some embodiments to produce plants that can grow under various environmental stresses. [Explanation of Symbols]

[0100] 100, 200... Plant treatment device, 110, 210... Plant holding section, 112... Reflective material, 120, 220... Ultraviolet light irradiation section, 122, 222... Light source, 124, 224... Control section, 130... Liquid injection section, 140... Control section, 180... Housing, 240... Liquid immersion section, 242... Storage tank, 244... Liquid storage area, 246... Opening, 248... Lid member, 250... Pressure adjustment section, 252... Vacuum pump, 254... Pressure release valve, 280... Irradiation chamber, P... Plant

Claims

1. The plants were exposed to light in the wavelength range of 300-325 nm, with an irradiation dose of 4000 μmol / m³. 2 More than 50000μmol / m 2 The irradiation process is as follows, and is performed such that no light in the wavelength range of 290 nm or less is irradiated, or the amount of such light is less than 20% of the amount of light irradiated in the wavelength range of 300 to 325 nm. An infection process in which microorganisms are infected into plants that have been irradiated with the aforementioned light, Methods for processing plants, including those mentioned above.

2. The method according to claim 1, wherein, in the infection step, the foreign gene is introduced into the plant by infecting it with a microorganism containing the target foreign gene.

3. The method according to claim 1 or 2, wherein the irradiation step is performed in the dark.

4. The method according to claim 1 or 2, further comprising the step of placing the plant infected with microorganisms in the infection step in a dark place for 6 to 48 hours.

5. The method according to claim 1 or 2, further comprising the step of growing a plant infected with microorganisms in the infection step.

6. In the irradiation step, the light within the wavelength range of 300 to 325 nm is 0.05 to 300 μmol / m³ 2 The method according to claim 1 or 2, wherein irradiation is performed at a photon flux density of / s.

7. The method according to claim 1 or 2, wherein in the irradiation step, light in the wavelength range of 330 nm or more is not irradiated, or the amount of such irradiation is less than 30% of the amount of light irradiated in the wavelength range of 300 to 325 nm.

8. The method according to claim 1 or 2, wherein the infection in the infection step is performed by immersing the plant in a liquid containing microorganisms or by spraying or injecting the liquid into the plant.

9. The method according to claim 1 or 2, wherein the infection in the infection step is performed by an agro-infiltration method or a plant virus vector method.

10. The method according to claim 1 or 2, wherein the light in the 300-325 nm wavelength range is light having a wavelength spectrum with a peak wavelength of 310 ± 5 nm and a full width at half maximum of 5-15 nm.

11. The plants were exposed to light in the wavelength range of 300-325 nm, with an irradiation dose of 4000 μmol / m³. 2 More than 50000μmol / m 2 The irradiation process is as follows, and is performed such that no light in the wavelength range of 290 nm or less is irradiated, or the amount of such light is less than 20% of the amount of light irradiated in the wavelength range of 300 to 325 nm. An infection process in which microorganisms are infected into plants that have been irradiated with the aforementioned light, A method for producing plants infected with microorganisms, including [specific microorganisms].

12. The irradiation step and infection step according to claim 11, The process of fermenting the infected plants, A method for producing plant fermented products containing

13. A plant holding section configured to hold plants, A UV light irradiation unit is configured to irradiate at least a portion of the plant held in the plant holding unit with UV light in the wavelength range of 300 to 325 nm, such that light in the wavelength range of 290 nm or less is not irradiated or the amount of such light is less than 20% of the amount of light in the wavelength range of 300 to 325 nm. A plant immersion unit comprising a storage tank configured to contain a liquid containing microorganisms in a liquid-containing area partitioned within its internal cavity, wherein the portion of the plant irradiated with ultraviolet light by the ultraviolet light irradiation unit is positioned within the liquid-containing area, or a liquid spraying unit configured to spray the liquid containing microorganisms toward the portion of the plant irradiated with ultraviolet light by the ultraviolet light irradiation unit, A plant processing device equipped with the following features.

14. The first transport mechanism is further configured to transport the plant holding section from the ultraviolet light irradiation section to the plant immersion section or the liquid spray section, The plant processing apparatus according to claim 13, wherein the first transport mechanism is configured to shield photosynthetically active radiation to the plants on the transport path.

15. A plant processing apparatus according to claim 13 or 14, further comprising a pressure adjustment unit, The storage tank has an opening, The plant immersion section further comprises a lid member configured to close the opening and seal the internal cavity of the storage tank, The plant processing apparatus comprises a pressure adjustment unit equipped with a pressure reduction mechanism configured to reduce the pressure of the internal cavity of the storage tank when it is sealed by the lid member.

16. The plant processing apparatus according to claim 13 or 14, wherein the ultraviolet light irradiation unit, or the ultraviolet light irradiation unit and the plant holding unit, are configured to irradiate ultraviolet light only to specific parts of the plant held in the plant holding unit.

17. A plant processing apparatus according to claim 13 or 14, further comprising a sensor unit configured to detect a specific part of a plant, The ultraviolet light irradiation unit is configured to irradiate the ultraviolet light toward the portion detected by the sensor unit, The plant processing apparatus is configured such that the liquid injection unit is capable of spraying liquid toward the part detected by the sensor unit.

18. The plant processing apparatus according to claim 13 or 14, wherein the ultraviolet light irradiation unit comprises a light source that emits light having a wavelength spectrum with a peak wavelength of 310 ± 5 nm and a full width at half maximum of 5 to 15 nm.

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