Stoma observation system

Abstract

[Problem] To provide an integrated system suitable for the observation of a stoma. [Solution] This system includes: an imaging means 201 for capturing an image of a stoma on a plant leaf; a measuring means 203 for measuring the condition of the stoma on the basis of the captured image; a processing means 205 for processing data indicating the measured condition of the stoma and data associated with the measured stoma while associating these data with each other; and a display means 207 for selectively displaying a result of the processing performed in the processing means 205.

Description

Stomata Observation System

[0001] The present invention relates to a system for observing stomata in plant leaves.

[0002] The state of stomata in the leaves of cultivated crops is observed to verify the desired effects in crop cultivation, cultivation research, cultivar improvement, pesticide testing, etc. Stomata are gas exchange holes in plant leaves that open and close by changing the shape of cells.

[0003] To observe the state of stomata, images of the stomata are taken, and based on the images, the degree of opening, which indicates how open the holes are, and the opening area, which indicates the area of ​​the open part, are measured. The state of the stomata is then linked to the location of the crop being observed and environmental values ​​(temperature, humidity, PPFD (Photosynthetic Photon Flux Density), etc.).

[0004] ・Yusuke AIHARA, et al, “Identification and improvement of isothiocyanate-based inhibitors on stomatal opening to act as drought tolera nce-conferring agrochemicals”, [online], May 15, 2023, Nature, [Retrieved August 25, 2020], Internet <URL: https: / / www.nature.com / arti cles / s41467-023-38102-7>・Zhen Shang, et al, “Rapid non-destructive method to pheno type stomatal traits”, [online], March 31, 2023, Plant Methods, [Retrieved August 25, 2020], Internet <URL: https: / / plantmethods.biomed central.com / articles / 10.1186 / s13007-023-01016-y>

[0005] Each of the above steps for observing the state of pores is complicated, and in order to observe the state of pores, each of the above steps must be carried out individually, which causes a decrease in the throughput of pore observation.

[0006] For example, methods for capturing images of stomata include the SUMP method, in which nail polish or the like is applied directly to the leaves, a replica is taken, and images of the stomata are captured under a microscope, or a method of capturing images of the stomata using a handheld microscope.

[0007] However, the SUMP method requires applying nail polish to the leaf epidermis and obtaining a replica, which requires a long wait for the nail polish to dry and can damage or injure the leaf when peeling off the replica. Furthermore, the replica must be observed under a microscope, which makes it time-consuming to manage individual samples and makes it unsuitable for handling large amounts of data.

[0008] In addition, commercially available handheld microscopes are designed to be used for taking photos using standard apps. While stomatal observation data is sometimes saved separately for each plant variety or location, these microscopes are not designed for such purposes and are unable to save data suitable for stomatal observation.

[0009] Furthermore, the degree of stomatal opening and opening area are measured from the captured images by first visually identifying the location of the stomata, and then measuring the degree of stomatal opening and opening area. However, this measurement is carried out separately using a PC or other device, and it is not expected that measurements will be taken on the spot.

[0010] Furthermore, the linking with environmental values ​​obtained from the sensors is done manually or based on predetermined rules by accessing databases that store data on acquired stomatal aperture and open area, and databases that store data on environmental values ​​(temperature, humidity, PPFD, etc.), variety, photography location, etc. The data stored in each database is enormous, and similar data is stored, making management of each piece of data cumbersome.

[0011] The present invention has been made in view of the above circumstances, and has as its object to provide an integrated system suitable for observing stomata.

[0012] The stoma observation system of the present invention comprises an imaging means for capturing images of the stomata of plant leaves, a measuring means for measuring the state of the stomata based on the captured images, a processing means for processing data indicating the measured state of the stomata in association with data related to the measured stomata, and a display means for selectively displaying the results processed by the processing means.

[0013] With the above configuration, it is possible to seamlessly integrate a series of stoma observations, from taking images of the stomata, to measuring the state of the stomata based on the taken images, associating the measured stomata with data related to the stomata, and displaying the associated results.

[0014] In the above configuration, the photographing means includes an opening / closing section capable of clamping the plant leaf, a support section that supports the plant leaf while protecting the surface of the plant leaf, and a focusing section that performs focusing in accordance with the unevenness of the surface of the plant leaf.

[0015] According to the above configuration, it is possible to provide an imaging means that is configured to be suitable for imaging stomata in plant leaves.

[0016] The above configuration may further comprise an environmental value acquisition means for acquiring at least one environmental value of temperature, humidity, and PPFD.

[0017] According to the above configuration, it is possible to acquire environmental values ​​related to the measured pores.

[0018] In the above configuration, the processing means processes data indicating at least one of the pore opening degree and the pore opening area in association with the data indicating the environmental value, and further performs processing to identify the environmental value for forming pores having a desired pore opening degree and pore opening area based on the normality of the pore opening degree and pore opening area relative to a predetermined environmental value.

[0019] According to the above configuration, it is possible to identify the environmental values ​​necessary to form stomata with the desired stomatal aperture and stomatal aperture area based on the normality of the environmental values ​​and the stomatal aperture and stomatal aperture area. By cultivating plants in an environment with the identified environmental values, it is possible to cultivate plants with stomata with the desired aperture and aperture area, and to improve the reproducibility of the effects of pesticides and variety characteristics.

[0020] According to the present invention, it is possible to seamlessly integrate a series of stoma observations, from taking images of stomata, to measuring the state of the stomata based on the taken images, associating the measured stomata with data related to the stomata, and displaying the associated results.

[0021] Fig. 1 is a diagram showing a schematic configuration of a stomatal observation device used in the stomatal observation system according to the present invention. Fig. 2 is a block diagram showing the configuration of the stomatal observation system according to the present invention. Fig. 3 is a diagram showing the distribution of stomatal aperture. Fig. 4 is a diagram showing the distribution of stomatal open area. Fig. 4 is a diagram showing a state in which the open / closed state of stomata is specified and displayed on display means. Fig. 5 is a diagram showing a state displayed using a viewing application. Fig. 6 is a diagram showing the processing flow in the stomatal observation system.

[0022] Hereinafter, an embodiment of the present invention will be described with reference to the drawings.

[0023] 1 shows the schematic configuration of a stomatal observation device used in a stomatal observation system according to the present invention. The stomatal observation device 100 comprises an imaging unit 101 that takes images of the stomata of plant leaves, a measurement unit (not shown) that measures the state of the stomata based on the taken images, and a processing unit (not shown) that associates and processes data indicating the measured state of the stomata with data related to the measured stomata, and further comprises a display unit 103 that selectively displays at least one of the measured state of the stomata and the associated results.

[0024] The photographing unit 101 comprises a unit including a camera and a lens, and is equipped with an alligator-like opening and closing part 105 that can clamp the plant leaves, a support part (not shown) that supports the plant leaves while protecting the surface of the plant leaves, and a focusing part (not shown) that performs focusing in accordance with the unevenness of the surface of the plant leaves.

[0025] The opening and closing part 105 has a spring-type clip function, and holds the leaf in a pinched state by the spring elasticity. If the force clamping the leaf is too strong, the leaf surface will be damaged, and if the force clamping the leaf is too weak, it will be difficult to focus when photographing, so the elasticity is adjusted taking these into consideration.

[0026] The support portion is configured to support the leaves via a shock absorbing material such as a gel sheet in order to protect the surface of the leaves when they are clamped.

[0027] The photographing unit 101 supports a plurality of resolutions. The photographing unit 101 photographs the leaf by selecting the resolution necessary to measure the state of the stoma, such as the stomatal aperture and stomatal opening area. Furthermore, to ensure sufficient light during photographing or observation, the photographing unit 101 may be provided with a lighting function such as an LED, which may be turned on during observation or photographing.

[0028] The wavelength of the illumination light is not particularly limited as long as it is a wavelength that does not react with the stomata, and in addition to visible light, infrared light, or ultraviolet light can be selected. The intensity of the illumination light may also be changed. In addition to ensuring the required amount of light by the illumination function, a configuration may be adopted in which natural light is ensured, and a window 107 transparent to natural light may be provided to provide natural light to the subject.

[0029] The focus unit has a focus adjustment function that changes the distance to the leaf surface, which is the subject, by moving the unit including the camera and lens. The focus adjustment can be performed by moving the unit including the camera and lens mechanically or electronically.

[0030] The display unit 103 is a device equipped with a display function such as a tablet, smartphone, or PC, and is detachably supported by an attachment arm 111 attached to a gripping unit 109 extending from the imaging unit 101. The device and the main body of the device are connected by wire or wirelessly. Note that various processes may be performed by the device using a processing function installed in the tablet, smartphone, PC, or the like.

[0031] The stomatal observation device 100 can further be equipped with sensors for measuring environmental values ​​such as temperature, humidity, and PPFD, and can also be equipped with environmental value acquisition means (not shown) for acquiring various environmental values. The installation position of the sensor can be designed as appropriate, but for example, a sheet-shaped temperature sensor can be installed in the part that supports the leaves.

[0032] The stoma observation device 100 can further include a position information acquisition means (not shown) that acquires position information of the photographed area. Various sensors such as GPS, beacons, and Wi-Fi can be used to measure the position.

[0033] Fig. 2 is a block diagram showing the configuration of a stomatal observation system according to the present invention. As shown in Fig. 2, the stomatal observation system 200 comprises an imaging means 201 that captures images of the stomata of plant leaves, a measuring means 203 that measures the state of the stomata based on the captured images, a processing means 205 that associates and processes data indicating the measured state of the stomata with data related to the captured images, and a display means 207 that selectively displays at least one of the measured state of the stomata and the associated results.

[0034] The imaging means 201 has the functions of the imaging unit 101 shown in Fig. 1. The display means 207 selectively displays desired data by user operation on a screen provided on a device such as a smartphone, tablet, or PC as shown in Fig. 1.

[0035] The measuring means 203 measures at least one of the positions of stomata in the plant leaves, the open / closed state of stomata, the stomata opening degree, and the stomata opening area based on the captured image.

[0036] The processing means 205 processes data indicating the measured stomatal state in association with data related to the photographed image. The data indicating the stomatal state is data indicating the stomatal opening area and stomatal aperture. Processing the data indicating the stomatal state in association with environmental values ​​means processing the data in association with environmental values ​​such as temperature, humidity, and PPFD in association with the stomatal state such as the stomatal aperture and stomatal opening area.

[0037] 3 and 4 are histograms showing the distribution of stomatal aperture and stomatal open area for a total of 115 stomata from four photographed images.

[0038] Figure 3 shows the distribution of stomatal aperture. By observing the distribution of stomatal aperture for a given environmental value, the normality of stomatal aperture for the given environmental value can be determined. For example, if an environmental value can be identified that results in a symmetrical distribution of stomatal aperture, it can be determined that the normality of stomatal aperture for the identified environmental value is high.

[0039] 4 shows the distribution of stomatal aperture area, and by observing the distribution of stomatal aperture area with respect to a given environmental value, the normality of stomatal aperture area at the given environmental value can be determined. For example, if an environmental value can be identified that results in a symmetrical distribution of stomatal aperture area, it can be determined that the normality of stomatal aperture area for the identified environmental value is high.

[0040] As explained in FIGS. 3 and 4, the normality of the pore opening and pore area relative to a given environmental value can be considered, and environmental values ​​for forming pores having a desired pore opening and pore area can be identified.

[0041] The accuracy of histogram analysis is proportional to the number of samples. The histograms shown in Figures 3 and 4 are based on a total of 115 stomata from four captured images. However, to capture more stomata, it is necessary to increase the leaf area. Methods for increasing the leaf area include mechanically moving the subject using an XY stage, electronically moving the subject within the effective area of ​​the image sensor, and using a wide-angle lens.

[0042] Based on the knowledge that the state of stomata depends on the environmental values ​​approximately 0 to 60 minutes prior, the environmental value acquisition means 209 acquires data on environmental values ​​such as temperature, humidity, and PPFD, which are associated with the state of stomata, in advance of acquiring images of the stomata.

[0043] The position information acquisition means 210 acquires the position information of the location where the stomatal images were taken. By processing the acquired stomatal state including the position information of the stoma in association with the environmental values, it is possible to realize verification of the desired effects in crop cultivation, research on cultivation, improvement of cultivated varieties, testing of pesticides, etc., taking into account the geographical conditions of crop cultivation.

[0044] Fig. 5 shows the state of the image displayed on the display means 207. The state of the stomata and the results of association are arbitrarily displayed on the screen of the display means 207. Fig. 5 shows the state in which the stomata are identified and displayed based on the photographed image.

[0045] Stomata can be identified by visual inspection or by image recognition processing. It is also possible to display a numerical value indicating the stomatal aperture, as shown in the figure. It can be determined that stomata are open when the stomatal aperture exceeds a preset threshold.

[0046] The display mode on the display means 207 can be images, numerical values, or a combination of these, and the user can freely switch between graph display, gallery display, etc. Display data can be recorded in the stoma observation device main body, or can be recorded in storage external to the main body. The cloud can also be used as external storage.

[0047] In addition, the state of the stomata and related information can be displayed using a viewing app. Figure 6 shows the state displayed using the viewing app. The displayed content includes the information shown in Figures 3 to 5, the date and time of the photo, the variety, the average stomatal aperture, the number of stomata (pieces), the location of the photo, the temperature, humidity, weather, and detailed state of the stomata.

[0048] Figure 7 is a diagram showing the processing flow in the stomatal observation system, in which the processing required for stomatal observation is carried out using a photographing terminal 710 for photographing images of the stomata of crops, a viewing terminal 720 for viewing the state of the stomata and related information, and a server terminal 730 that performs various processes based on instructions from the photographing terminal 710 and the viewing terminal 720 and outputs the processing results to the photographing terminal 710 and the viewing terminal 720.

[0049] To achieve the above process, the server terminal 730 performs user registration processing (S731) based on user registration information such as an email address and password input from the viewing terminal 720. Next, the server terminal 730 performs authentication processing by comparing the user registration information including the email address and password input from the photographing terminal 710 with the user registration information input from the viewing terminal 730 (S732), and updates the database based on the authentication result (S733).

[0050] Next, the server terminal 730 performs an analysis process based on the data input from the photographing terminal 710 indicating the stomatal opening degree and stomatal opening area, environmental data such as temperature, humidity, and PPFD, data including user comments, and image data (S734), performs a judgment process based on the analysis results (S735), and notifies the photographing terminal 810 and the viewing terminal 830 that the judgment process has been performed (S735).

[0051] Next, server terminal 730 outputs data indicating stomatal aperture and stomatal open area, environmental data, data including comments, and data for displaying graphs to viewing terminal 720, and viewing terminal 720 displays this data so that it can be viewed (S721). Furthermore, server terminal 730 performs editing processing based on the data including environmental data and comments input from viewing terminal 720 (S737), and outputs the editing results to viewing terminal 720 as downloadable data. Viewing terminal 720 downloads this data (S722).

[0052] According to the process shown in FIG. 7, it is possible to seamlessly integrate and perform the processes of taking images of stomata, analyzing and determining based on the taken images, viewing the analysis and determination results, and saving the viewed data.

[0053] The present invention can be used to verify desired effects in crop cultivation, research on cultivation, improvement of cultivated varieties, testing of pesticides, and the like.

[0054] REFERENCE SIGNS LIST 100 Stomata observation device 101 Photography unit 103 Display unit 105 Opening / closing unit 107 Window 109 Holding unit 111 Arm 200 Stomata observation system 201 Photography means 203 Measuring means 205 Processing means 207 Display means 710 Photography terminal 720 Display terminal 730 Server terminal

Claims

1. A stoma observation system comprising: an imaging means for capturing images of stomata in plant leaves; a measuring means for measuring the state of the stomata based on the captured images; a processing means for processing data indicating the measured state of the stomata in association with data related to the measured stomata; and a display means for selectively displaying results processed by the processing means.

2. The stoma observation system of claim 1, wherein the photographing means comprises: an opening / closing section capable of clamping a plant leaf; a support section for supporting the plant leaf while protecting the surface of the plant leaf; and a focusing section for performing focusing in accordance with the unevenness of the surface of the plant leaf.

3. A stomatal observation system as described in claim 1 or 2, wherein the measuring means measures at least one of the stomatal opening degree and the stomatal opening area based on the captured image.

4. A stoma observation system according to any one of claims 1 to 3, comprising an environmental value acquisition means for acquiring at least one environmental value of temperature, humidity, and PPFD.

5. A stomatal observation system as described in claim 4, wherein said processing means processes data indicating at least one of the stomatal opening degree and the stomatal opening area in association with data indicating said environmental value.

6. A stomatal observation system as described in claim 5, wherein the processing means performs processing to identify environmental values ​​for forming pores having desired stomatal aperture and stomatal area, based on the normality of stomatal aperture and stomatal area relative to a predetermined environmental value.

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