JP7329834B2 - Nondestructive measurement device and nondestructive measurement method for plant metabolites, and plant cultivation system and cultivation method using the same - Google Patents

Description

TECHNICAL FIELD The present invention relates to a nondestructive measuring device and measuring method for nondestructively measuring components in tissues of humans, animals or plants, and particularly relates to metabolites such as sugars produced by photosynthesis of growing plants or their metabolites. The present invention relates to a nondestructive measuring device and a nondestructive measuring method for metabolites of a growing plant that nondestructively measures migration. The present invention also relates to a plant cultivation system and a plant cultivation method for growing plants by feedback-controlling the growth environment of growing plants based on measurement data obtained by the measuring device.

In horticultural cultivation such as so-called vinyl greenhouses and hydroponic cultivation, an environment considered to be optimal for target plants is artificially created to promote the growth of the plants. For this reason, in greenhouses or hydroponics equipment, various factors that are considered to be closely related to plant growth, such as temperature, humidity, water supply, light irradiation (insolation), fertilizer supply, etc. It is managed as strictly as possible according to the conditions considered to be optimal for
However, the optimum environment for plants is one in which the three elements of sunlight, temperature, moisture, carbon dioxide and fertilizer, nitrogen, phosphoric acid, potassium, and many other elements are intricately and mutually related. , but it is not yet fully understood. Therefore, in the conventional method, as exemplified in Patent Documents 1 to 3, environmental conditions for cultivation that are considered to be the optimum environment for growth derived from past cultivation results and empirical measurements are set, and the optimum environment is set. Plant growth environment is managed by controlling temperature, water, fertilizer and other factors (cultivation environment factors) to meet the conditions. Further, Patent Literature 4 discloses a technique of observing the growth state by photographing the stem of a plant with a television camera.

On the other hand, as a device for non-destructively measuring the components inside the ecology of plants and animals, for example, the sugar content in fruits such as apples, melons, tomatoes, etc., and the sugar content in human blood are non-destructive (or non-invasive). There is prior art that measures at . This method uses light to measure the sugar content of fruits, vegetables, other plants, and human blood (hereinafter referred to as "organisms"), like spectroscopic analysis. The measurement of sugar content by light is to measure the sugar content by utilizing the fact that the absorbance of light changes depending on the concentration of sugars contained in the tissue of an organism. are doing. These prior arts irradiate harvested fruits, agricultural crops, or human measurement sites with light of a predetermined frequency of near-infrared light through an optical element such as a lens, and the light reflected from the measurement target or Light that has passed through the object to be measured is received by an imaging device and measured. By measuring changes in the intensity of the reflected light and transmitted light of the irradiated light and comparing this with a calibration curve previously collected from many samples, the sugar content of the material to be measured is calculated according to the absorbance. Further, Patent Literature 10 filed by the present patent applicant discloses a noninvasive blood glucose level measuring device that measures the pedigree concentration in human blood by irradiating light onto a person's earlobe or the like.

JP-A-64-51023 JP 2018-121527 A JP 2019-118284 A JP 2015-202056 A JP-A-2-197940 JP-A-4-208842 JP-A-6-186159 JP-A-6-213804 Japanese Patent No. 3903147 JP 2018-159663 A

In the prior art, as described above, although there are empirically elucidated optimal growth environmental conditions for plants, there are a great many factors (factors or cultivation environment factors) related to the growth conditions of plants. We have not been able to identify exactly how factors contribute or affect growth. Furthermore, since the optimal growth environment differs depending on the type of plant and the growth process from seedling to harvest, whether or not the optimal environment is provided during the growth process of the plant depends on the metabolic situation during the actual growth process of the plant. It is difficult to understand without real-time understanding. This is the same in both horticultural cultivation and hydroponics.

All of the prior arts disclosed in Patent Documents 1 to 3 related to vinyl greenhouses and water grain cultivation are suitable for optimal environments based on empirically acquired knowledge in horticultural cultivation and cultivation. It controls the growth of plants by controlling the cultivation environment (growth environment) of plants such as amount and fertilizer, and directly observes the metabolism of plants during growth and controls the growth environment based on that. isn't it. Further, Patent Document 4 is for observing the growing condition of a plant by imaging it with a camera, and is intended to grasp the growing condition of a growing plant. However, the camera only observes external changes in the plant, and does not directly observe the metabolic state inside the plant tissue.

In order to observe the metabolic state of growing plants, it is necessary to continuously measure metabolites such as sugars and ammonium ions produced by photosynthesis in real time without damaging the plants. Patent Documents 5 to 9 disclose techniques capable of nondestructively measuring saccharides. However, these techniques measure harvested fruits and the like, and do not measure metabolites of growing plants. In addition, the measurement method is to irradiate the object to be measured with light, measure the reflected light or transmitted light, and compare the measured absorbance with a so-called "calibration curve" showing the average intensity distribution of the object to be measured. is used to calculate the sugar content of the non-measured material. Therefore, in order to measure metabolites using these techniques, a "calibration curve" against which to compare is required. However, in the prior art, there was no information about the calibration curve of the metabolites of the growing plant, nor was there any idea of directly measuring the metabolites of the growing plant.

Patent document 10 is a patent application filed by the present patent applicant. The applicant of this patent filed patent application 10 to provide a measuring device for non-invasively and accurately measuring blood glucose concentration in human or animal blood without using a calibration curve. At the time of filing of Patent Document 10, there was no recognition of the problem of measuring the metabolism of growing plants in real time. Nothing was disclosed or suggested.

As is clear from the above, in the prior art including Patent Documents 1 to 10, changes in metabolites of living plants are directly and non-destructively continuously measured to determine the growth environment and metabolism of growing plants. There is no disclosure about clarifying the relationship between the two, and there is no recognition of such a problem. Furthermore, there has been no concept of measuring the movement of plant metabolites during growth and feedback-controlling the growth environment of plants in horticulture or hydroponics based on the measured data. The applicant of the present patent has conducted intensive research and testing, and by directly measuring the movement of metabolites of growing plants to each site in horticultural cultivation or hydroponics, the relationship between cultivation environmental factors and plant growth In addition to being able to analyze plant growth in more detail, we came up with the idea that plant cultivation could be feedback-controlled almost in real time. The inventors have arrived at the invention of the present application.

If it were possible to measure the tissue components (especially metabolic components) of plants under cultivation without damaging the plants, it would be possible to inspect and observe the growth conditions of plants and the health conditions of animals in real time. Real-time observation of the movement of plant metabolites enables us to study the metabolism of plants during each growth process and the factors necessary for plant growth and fruit ripening, such as solar radiation, temperature, water, saturation, and fertilizer. By measuring and observing both factors (cultivation or growth environment factors) at the same time, detailed comparative analysis of how the cultivation environment factors affect the metabolism of plants in a short time. becomes possible. By controlling the cultivation environmental factors in real time while observing actual plant metabolism, it becomes possible to more accurately control plant growth and fruit ripening. That is, if the metabolic state of a plant and the data of various cultivation environmental factors at that time are simultaneously known, it is possible to directly know how the various cultivation environmental factors affect the metabolism of the plant. As a result, it is possible to control plant growth by identifying environmental factors that are thought to affect plant growth and the relationship between their amounts and changes, and by adjusting the cultivation environment to achieve optimal conditions for each growth process. It becomes possible. In addition, if the metabolic status of plants can be known in real time during plant cultivation, it will be possible to control various environmental factors related to harvest timing and quality, such as plant growth and fruit ripening, in accordance with the growth process of plants. becomes possible.

The present invention has been made on the basis of the above idea, and provides a non-destructive measuring apparatus for metabolites of a plant and a measuring method for measuring the metabolites of a growing plant. An object of the present invention is to provide a plant growing system and a plant growing method using a nondestructive measuring device.

In order to achieve the above object, the nondestructive measurement device for metabolites according to the present invention comprises a fixing part for fixing a peduncle or petiole of a plant; and a nondestructive measuring unit that nondestructively measures the metabolites of the plant by the transmitted light or the reflected light of the light,
The non-destructive measurement unit includes: a light emitting unit that separately emits light of a first wavelength as reference light and a second wavelength of measurement light; and an optical element that irradiates the portion to be measured, receives the reflected light or transmitted light from the portion to be measured of the irradiated light of the first wavelength and the second wavelength, and outputs a detection signal corresponding to the amount of received light. and the amount of light emitted from each of the light emitting units, and the amount of reflected light or transmitted light of the first wavelength and the second wavelength received by the light receiving unit, respectively, are the absorption coefficient of the first wavelength and an absorptance calculation unit that obtains the absorptivity of the second wavelength and outputs as measurement data a value obtained by correcting the absorptance of the second wavelength with the absorptance of the first wavelength; A storage unit for storing data, an output unit for outputting the stored measurement data to the outside, and a control unit for controlling each of the above units to continuously measure changes in the movement of metabolites in plants. and

This configuration allows continuous measurement of metabolites (sucrose, fructose, glucose, etc.) in petioles or flower stalks of growing plants. For example, the growth state of plants, particularly the growth state of plants intended for fruit harvesting, is strongly influenced by sugars produced by photosynthesis. For example, changes in the concentration of sugars in petioles or flower stalks indicate the movement of sugars produced by photosynthesis in leaves, and by measuring this, it is possible to understand the state of photosynthesis in growing plants. Become. It is also possible to detect rapid changes in the movement of sugars and rapid changes in pedigree values. In particular, in plants, continuous measurement of the amount of saccharide movement in petioles and flower stalks over time makes it possible to control fruit growth and predict future fruit growth conditions.

In another embodiment of the nondestructive measurement device for plant metabolites according to the present invention, the lights of the first wavelength and the second wavelength are generated by the light emitting unit, the optical element, and the measurement control unit, It is characterized in that the light beams pass through the same optical axis and are alternately irradiated to the same place of the part to be measured. By alternately irradiating reference light beams with different wavelengths along the same optical axis, it is possible to accurately irradiate the target site with light beams of different wavelengths, enabling accurate irradiation of the measurement light and highly accurate correction processing. becomes.

Further, in another embodiment of the nondestructive measuring device for plant metabolites according to the present invention, the control unit controls the light emitting unit, the light receiving unit, and the absorbance calculation unit to perform the first measurement. The measured value calculated at the time is stored as the first measured value, the same measurement and calculation are performed at the time of the second measurement after a certain period of time, and the third measured value is stored after a certain period of time. Similar measurements and calculations are performed at the time of the measurement of , and stored as a third measured value, and the difference between the third measured value and the first measured value is calculated to express the concentration change of the substance to be measured in the part to be measured. A measurement control unit that outputs a calculated value of 1, divides the difference between the second measured value and the first measured value by time, and outputs a second calculated value representing the rate of change over time. Characterized by As a result, it is possible to measure the amount of change after a certain period of time has elapsed, and to measure the time rate of change in sugars during a part of the measurement time of the amount of change. The storage unit can store a first measured value, a second measured value, a first calculated value, and a second calculated value in addition to the measurement data.

Furthermore, by providing a communication function for outputting the concentration change value and time change rate to the outside, the growth of plants can be constantly monitored in real time by a PC or the like, and the growth environment of the plants can be controlled. becomes possible.

INDUSTRIAL APPLICABILITY The present invention is suitable for grasping the state of photosynthesis in plants in real time. Sugars are preferred. It is also possible to configure such that the measurement control unit repeats similar measurements and calculations at regular time intervals to continuously output the concentration change value and time change rate.
It is also possible to measure the blood sugar level of mammals such as humans as the substance to be measured. In this case, it is possible to easily measure a sudden increase in blood sugar level.

The correction of the second absorption coefficient by the first absorption coefficient can obtain the measured value by subtracting the first absorption coefficient from the second absorption coefficient. Further, the correction of the second absorption coefficient by the first absorption coefficient may be obtained by dividing the second absorption coefficient by the first absorption coefficient to obtain the measured value. Furthermore, it is possible to configure so as to have a mechanism for correcting the irradiation position and angle of the light irradiated to the part to be measured.

INDUSTRIAL APPLICABILITY According to the present invention, it becomes possible to measure the metabolic state in a plant in real time without damaging the growing plant. As a result, it is possible to measure actual data in real time about how growth environment factors affect plant metabolism, and to directly observe the effects of environmental factors on plant metabolism. This makes it possible to continuously observe the state of photosynthesis in real time and to feedback-control environmental factors for plant growth according to the metabolic state of the plant. That is, it is possible to more accurately control plant metabolism, such as promoting plant growth and delaying the harvest time, while observing the actual metabolic state of the plant. In addition, even in living animals including humans, it is possible to continuously measure the metabolic state according to the purpose and collect data.

Graphs predicting changes in glucose transport through petioles and flower stalks with changes in solar radiation and temperature in plants. The figure which shows the example which attached the nondestructive ecological measuring device (measuring device) of this invention to a petiole or a flower peduncle. The figure which shows the structure of the part which fixes the to-be-measured part of a measuring device. A typical graph for explaining a measuring procedure of the present invention. FIG. 2 is a schematic diagram for explaining the basic structure of a transmitted light type measuring device; FIG. 2 is a schematic diagram for explaining the basic structure of a reflected light type measuring device; FIG. 4A is a diagram illustrating adjustment of the optical axis using an actuator, and FIG. 4A is a schematic diagram illustrating tilt adjustment as viewed from the side of a measurement site (petiole or flower stalk) fixed to the device, and FIG. 10A is a schematic diagram illustrating shift adjustment in the axial direction of a measurement site viewed from the top, and FIG. 11C is a schematic diagram illustrating tilt adjustment and shift adjustment viewed from the front. FIG. 2 is a schematic diagram illustrating basic optical components of a transmitted light type measuring device using two light sources. FIG. 2 is a schematic diagram illustrating basic optical components of a reflected light type measuring device using two light sources; FIG. 4 is a schematic diagram showing an example of a clip structure for an optical portion of a transmission-type measuring device. 1 is a block diagram of an electric circuit showing a first embodiment of a measuring device of the present invention; FIG. FIG. 2 is a block diagram of an electric circuit showing a second embodiment of the biological measuring device of the present invention; FIG. 4 is a diagram for explaining an example of switching timing of LD1 and LD2; The figure which shows an example of the graph for calculating|requiring a final judgment value. Graph exemplified for explaining the relationship between satiety control and plant metabolites in strawberry horticultural cultivation. 4 is a flowchart illustrating a control procedure of saturation control; The graph illustrated for demonstrating the relationship of irrigation and the metabolism of a plant. Flowchart illustrating the procedure for controlling irrigation

Hereinafter, a nondestructive measuring device and a nondestructive measuring method for plant metabolites, a plant cultivation system and a plant cultivation method using the same according to the present invention will be described with reference to the drawings. In the following description, an example of examining the growth state of a plant by measuring plant sugars produced mainly by photosynthesis using light of a predetermined wavelength will be used. Target substances (metabolites, etc.) are not limited to these.

Since plants grow through photosynthesis, if it is possible to measure the amount of substances (metabolic products) produced by photosynthesis that contribute to growth in real time during the plant growth process, it will be possible to predict the growth status of plants and their future. Not only can it be predicted, but it is also possible to control the cultivation environment and promote growth. For example, photosynthesis in plants produces various sugars such as glucose, and the amount of these sugars produced is closely related to the plant growth. It is possible to grasp the growth of the plant and the growth of the fruit from when the fruit is still small, and to adjust the maturity degree and timing of the growth of the plant and the fruit.

In addition, sudden changes in the supply of sugars and water during the growth process of plants can adversely affect fruit growth, and in animals, rapid changes in blood sugar levels can damage blood vessels. and other adverse effects. For example, during the growth process of tomatoes, a sudden increase in water is considered to be the cause of tomato cracks, and a sudden rise in human blood sugar levels is known as a blood sugar level spike, which is known to cause great damage to blood vessels and the like. Therefore, not only the magnitude of the measured value but also the amount of change over time is important information for knowing the growth and health condition of plants and animals. In particular, by measuring not only short-term changes, but also medium- and long-term changes in metabolic organisms, it is possible to directly observe the effects of changes in plant growth and growth environment on plant growth. By feedback-controlling the growth environment factor based on the measurement data, it is possible to provide a more appropriate plant growth environment. In the present invention, by continuously measuring the movement of metabolites that occur within the tissue of a growing plant, it is possible to grasp more precise and accurate information that cannot be obtained only from empirically obtained knowledge about the cultivation environment. In addition, it becomes possible to control cultivation environmental factors in real time according to the actual metabolic situation of growing plants.

In the following, by non-destructively measuring the absorbance and diffusivity of light, instead of measuring the sugar content of harvested fruits, we measure changes in the amount of sugars and ammonium ions produced by photosynthesis in living plants. Then, an example of grasping the metabolic state of a plant will be explained. Metabolites such as sugars and ammonium ions have a wavelength of light that exhibits their own absorbance. By measuring the concentration of the metabolite, it is possible to measure the concentration of the metabolite. As for the degree of diffusion, the amount of diffusion of light increases as the molecular weight increases, as in the case of sugars. Therefore, the degree of diffusion of reflected light and transmitted light when light is irradiated is proportional to the concentration of sugars. Therefore, when there are many metabolites such as sugars, the amount of absorption and diffusion increases, and the amount of transmitted light and reflected light decreases according to the concentration.

Embodiments of a measuring device and method for measuring the concentration of metabolites (sugars, etc.) in plants, and how to attach the measuring device and measuring method according to the present invention will be described below.
First, an apparatus for measuring glucose, which is one of sugars produced by photosynthesis, will be described. Glucose produced by photosynthesis in plants moves to fruits through petioles (stems at the base of leaves) and peduncles (stems at the base of flowers). As the glucose produced in the leaf blade moves to the flower or fruit side, the glucose in the whole leaf decreases and the sugar in the fruit side increases. Using near-infrared light, it is possible to estimate the amount of sugars produced by photosynthesis that contribute to fruit growth by measuring the amount of change in sugars that pass through the petiole or flower stalk. Become.

Photosynthesis is performed by taking in light, water, carbon dioxide, etc., and is also affected by temperature. Substances produced by photosynthesis are supplied to flowers and fruits by moving through petioles and floral stalks. Among the metabolites and other substances that pass through the petiole and peduncle of the plant, it is thought that the glucose, water content, and the growth of the plant itself may vary greatly during the day.

FIG. 1 is a graph that predicts changes in glucose migration in petioles and flower stalks with changes in solar radiation and temperature in plants. The horizontal axis is the time axis, and the vertical axis represents the temperature, the concentration of glucose at each measurement site, the amount of solar radiation, the temperature, and the like. The size of the notation on the vertical axis differs depending on each factor, and the unit and scale of the size of each factor are adjusted so that mutual correlation can be understood. In the figure, 5 indicates temperature, 6 indicates change in solar radiation, 7 indicates change in glucose in petioles (predicted), and 8 indicates changes in glucose in flower stalks (predicted). The movement (translocation) of sugars in plants is said to be initiated by changes in temperature. As shown in FIG. 1, the movement of glucose in the peduncle seems to be affected by a decrease in temperature and a decrease in the amount of solar radiation. For plant growth, especially for harvesting fruits, it is important to increase the amount of sugars that pass through the flower stalks, and for this purpose, it is important to promote photosynthesis. By measuring the displacement of sugars in petioles or flower stalks, the state of photosynthesis can be inferred. In other words, in plants, metabolism and the movement of metabolites occur in a daily cycle. can be grasped.

Since plants repeat a certain metabolism on a daily basis, measuring the amount of metabolites passing through petioles or flower stalks on a daily basis enables us to understand the movement patterns of metabolites in the plant. It is possible to accurately grasp the actual metabolic situation of plants in individual plants. Therefore, by using a measuring device that can non-destructively measure the metabolites (mainly sugars) of growing plants, various factors (temperature, humidity, water) that affect plant growth on a daily basis or on a several-day basis can be analyzed. By measuring changes in the amount of metabolite transfer while changing cultivation environmental factors such as , solar radiation, fertilizer, etc., the relationship between cultivation environmental factors that affect plant growth and growth in each growth process of plants. can be grasped. In addition, based on the findings obtained from these measurements, it becomes possible to control the environment so as to provide the optimum environment for plant growth. For example, it becomes possible to grasp the actual metabolic state of a plant based on measurement data, and to feedback-control environmental factors in the growth environment so as to promote or suppress the metabolism of the plant.

FIG. 2 is a diagram illustrating a state in which the nondestructive metabolite measuring device of the present invention (hereinafter simply referred to as "measuring device") is attached to a petiole or flower peduncle. The measuring device of the present invention measures metabolites by irradiating a site to be measured with light and measuring the amount of transmitted light or reflected light. By continuously measuring the pathway of metabolites, it is possible to measure changes in the amount of migration. Details of the measuring device will be described later. A measuring device 16 or 17 is attached and fixed to either one or both of the petiole 18a or the peduncle 18b, which is the passageway of plant metabolites, and moves in the petiole 18a or the peduncle 18b (commutation 18' or 18'') It is preferable to measure the change in saccharide movement in the translocation component. In the case of plants such as strawberries and watermelons that grow like crawling on the ground or on walls, it is often not necessary. or 17 is preferred. As a result, it is possible to minimize the influence of the position of the measurement site (measured part) shifting due to wind, rain, or vibration. Since photosynthesis is carried out by the leaf blade and translocated from the vein to the petiole, the state of photosynthesis can be determined by measuring the translocation 18' at the petiole 18a. It is possible to know whether the product (glucose) is being sent to the fruit.

For continuous measurement, it is important to ensure that measurement data does not fluctuate due to displacement of the measurement point. It is preferable to firmly fix the petiole or flower peduncle of the measurement site to the measuring device so that the optical axis of the light emitted from the measuring device does not shift due to vibrations such as wind and rain and so as not to hinder the growth of the plant. With reference to FIG. 3, an example of a device and a method for fixing a measured portion 21 (for example, a petiole or a flower stalk) of a plant to a measuring device will be described. FIG. 3 is a schematic diagram showing only the structure for fixing the measurement site 21 of the plant to be measured. FIG. 3 exemplifies a thin stem-shaped measurement site 21 such as a petiole or a flower stalk, but the structure of the fixing part and housing can be appropriately changed and adjusted according to the measurement site. 3, (a) and (c) are side views, (b) is a plan view, (d) is a front view, and (e) is a cross-sectional view along the line AA' shown in (b) ( 72b is a cross-sectional view of the fixing piece 72b.

A lower housing 70b of the measuring device 16 or 17 is provided with a V-shaped groove 71 extending in the longitudinal direction (vertical direction in FIG. 1B) for arranging the measurement site 21 such as the petiole 18a or the floral stalk 18b. there is In addition, two fixing pieces 72a and 72b which cross the V-shaped groove 71 at regular intervals and fix the measurement site 21 therebetween are provided substantially in the center of the lower housing 70b in order to fix and hold the measurement site 21. A U-shaped fixture 72 is provided. The fixture 72 is fixed so as to be embedded in a substantially flat hollow surface provided in a U-shape provided in the lower housing 70b (see FIGS. 7B and 7E). The fixture 72 is made of an elastic member (plate spring or the like), and the root portions of two integrally formed fixing pieces 72a and 72b are fixed to the housing 70b by screws, pins or the like 74. As shown in FIG.

It is preferable that the elastic force of the fixing pieces 72a and 72b of the fixture 72 is such that the growth of the measurement site 21 is not hindered. Moreover, the surfaces of the fixing pieces 72a and 72b that come into contact with the measurement site 21 are preferably covered with a sponge, cloth, or other soft material. The fixture 72 is rotatably fixed by a hinge, and the fixing pieces 72a and 72b of the fixing portion 72 press the measurement site 21 by an elastic member such as a torsion ring, a plate spring, or a coil spring, thereby fixing the measurement site 21. It is good also as a structure which urges so that it may carry out.

<Nondestructive measuring device>
In an embodiment of the present invention, near-infrared light is irradiated to substances passing through petioles or flower stalks, and metabolites are measured from the difference between the transmitted or reflected light and the irradiated light. However, in addition to glucose and ammonium ions, there are multiple substances other than glucose and ammonium ions that absorb light in the vicinity of near-infrared light. be. Regarding this point, the measurement accuracy can be improved as follows.
Among substances that pass through petioles and flower stalks, substances whose amount changes in a short period of time are predominantly sugars such as glucose, ammonium ions, and water. It is believed that it is these substances that greatly affect the data. Therefore, if the influence of water can be removed, glucose and ammonium ions can be measured.

For example, if light with a wavelength of about 1500 nm to 1600 nm is used as measurement light, this measurement light absorbs sugars, ammonium ions, and water. On the other hand, light with a wavelength around 1300 nm has the same degree of water absorption as the measurement light. Therefore, by irradiating light with a wavelength near 1300 nm as reference light and subtracting the measurement data of the reference light from the measurement data of the measurement light, water can be separated from the measurement data.
That is, measurements are performed using two light beams (measurement light) and a reference light having an absorption wavelength of glucose or the like, and the measurement data of the measurement light is corrected by the measurement data of the reference light to obtain metabolites such as glucose. and water can be separated. In addition, the tissue change (physical change) due to the growth of the plant itself is a displacement of the diffuser that does not depend much on the wavelength, and acts in the same way on the wavelengths of both the measurement light and the reference light. can be offset. Correction is not essential, and it is possible to use the measured data as it is. Since the measurement data of the reference light is the measurement data of moisture, the reference light can be used not only as correction data but also as measurement data of moisture movement alone.

Also, in order to accurately measure absorbance and reflected light, it is important to configure the part to be measured so that light of each wavelength passes coaxially. In addition, originally, in order to identify substances other than glucose, glucose cannot be measured without creating a calibration curve based on a large amount of data measured in a wide spectrum. For example, the measurement state itself at a certain point in time acts as a calibration curve, and after all, the calibration curve is not necessary. As described above, the apparatus of this time is characterized by basically measuring the amount of change over time in the amount of rise and fall in glucose. In addition, when measuring the amount of displacement as in this measurement, it is possible to improve the measurement accuracy and reproducibility because errors due to individual variations in plant types and sizes can be offset.

A photodiode (hereinafter referred to as "PD") is used to measure the amount of reflected light or transmitted light. Determine the diffusion range in consideration of the diameter and degree of diffusion. The amount of light detected by PD is reduced by absorption by glucose and at the same time is diffused by tissue (diffuser) and glucose. When the amount of diffused light increases in this way, the amount of light detected by the PD decreases, and it acts as if the absorbance of glucose has increased, so diffusion increases the detection sensitivity of changes in glucose. In the present invention, the measured value obtained by superimposing the absorbance and the diffusivity is used as the basic detected amount of received light, and the absorbance or the absorbance of the object to be measured is obtained from the detected amount in the sensitized state. Also, since it is known that light absorption varies with temperature, the temperature of the measurement site is measured, the detected amount is corrected according to the temperature value, and the final light absorbance is obtained. This correction is based on the known temperature coefficient of absorption of sugar.

In the present invention, the amount of metabolites passing through the same site is continuously measured at time intervals, and the amount of change in glucose that moves through the petiole or flower stalk is measured based on the difference in measurement data at regular time intervals. be. In order to clearly detect this state of change, in the present invention, as a basic configuration of measurement, measurement is performed three times at timings t1, t2, and t3 at regular time intervals, for example, 10-minute intervals. A measurement of , a second measurement, and a third measurement are obtained. From this, the amount of change in absorbance is calculated based on the difference between the measured values at time t3 and time t1 (third measured value - first measured value) to obtain the first calculated value (ds). Based on the measured values of t2 and t1 (the second measured value and the first measured value), a short time change amount or an instantaneous change amount is calculated to obtain a second calculated value (dts). The present invention comprehensively evaluates the growth state of plants based on these two calculated values ds and dts.

FIG. 4 is a schematic graph for explaining the measuring method of the present invention. Time intervals between the first measurement time t1, the second measurement time t2, and the third measurement time t3 are set constant (for example, 10 minutes). This time interval can be appropriately set according to the change speed of the plant or substance to be measured. In other words, it is possible to make the measurement easier by shortening the time interval for plants with large changes and by setting the time intervals wider for plants with small changes.

The basic concept of calculating the first calculated value ds and the second calculated value dts is as follows. In the measurement (measurement 12) at the first measurement time t1, the difference between the amount of light received by the light receiving element and the amount of light emitted from the light source, or the ratio of the two (absorbance) Let the first measured value (S1) represent the rate). In the measurement (measurement 13) at the second measurement time t2, the difference or ratio (absorbance) between the amount of light received by the light receiving element and the amount of light emitted from the light source is used as the absorbance or absorbance of the object to be measured. 2 measured value (S2). In the measurement (measurement 13) at the third measurement time t3, the difference between the amount of light received by the light receiving element and the amount of light emitted from the light source, or the ratio of the two (absorbance), represents the amount of absorption or the absorbance of the non-measurement. This is the third measured value (S3). Preferably, such measurements are repeated sequentially to level the measured data.

As explained in the example of glucose and water, the first to third measured values (S1 to S3) are desirably corrected by using the reference light and the measurement light, which are lights of different wavelengths. For example, if the reference light is light of a first wavelength and the measurement light is light of a second wavelength, by subtracting the light reception amount or absorption rate of the reference light from the light reception amount or absorption rate of the measurement light, Correction is possible by removing the influence. Alternatively, correction can be made by dividing the amount of light received or the absorptivity of the measurement light by the amount of light received or the absorptance of the reference light. In this case, the corrected values become the first to third measured values (S1 to S3), respectively.

Next, (S3-S1), which is a value obtained by subtracting the first measured value (S1) from the third measured value (S3), is obtained and used as the first calculated value ds. Further, (S2-S1), which is a value obtained by subtracting the first measured value (S1) from the second measured value (S2), is obtained, and this value is defined as the first calculated value dts. That is, the first calculated value is the difference in the tenth measurement period, and the second calculated value is the difference in the tenth measurement period.

5 and 6 illustrate the basic structure of the measuring device 16 or 17 attached and fixed to the flower stalk and leaf stalk shown in FIG. 2, and FIG. 6 is a diagram for explaining the basic structure for measurement using reflected light. In the case of the transmitted light method shown in FIG. 5, light 19′ is irradiated from a light source 19 through an objective lens 20 to a measurement site 21, which is a portion to be measured such as a petiole or flower stalk, and the transmitted light 19'' passing through the measurement site 21 is The amount of light is detected by a photodiode (PD) 23 as the magnitude of voltage or current. In the case of the reflected light method shown in FIG. 5, the light from the light source passes through the quarter-wave plate 24 and is irradiated onto the measurement site with the polarization shifted by 45 degrees. A portion of the irradiated light is reflected in the measurement site and returns. Since this returning light (reflected light) passes through the 1/4 wavelength plate again, the polarized light is shifted by 90 degrees. be. Although one light source 19 is illustrated in FIGS. 4 and 5, reference light and measurement light can be output by using a two-wavelength laser.

The fixture 72 is used to fix the measurement site, and the V-shaped groove on the lower side in the figure is used as a support, and the upper side is a pressure fixture 72 that presses with a weak force that does not hinder the growth of the measurement site. It is preferable to reduce the pressing load of the toe 21 . This is because, especially when the object to be measured is a plant, its physical shape changes as it grows, so that if a large load continues for a long period of time, the growth will not be hindered. When the measurement is performed on the petiole or flower stalk, measurement errors occur due to changes in the optical path, resulting in a decrease in measurement accuracy. Therefore, the optical path is fixed so that it does not change, but if it moves for some reason, there is a possibility that the underlying tissue at the measurement site will change, resulting in a decrease in measurement accuracy. In addition, when the incident state of light changes due to external factors such as wind, rain and vibration, the measurement accuracy is lowered. Therefore, first, it is important to fix the measuring device to the measurement site so that the measuring device and the measurement site do not shift.

By adopting a structure in which the petiole or flower stalk is inserted into the measuring device, it is possible to keep the optical path length almost constant. Its physical displacement may not be unidirectional. In addition, it is assumed that the measurement site is displaced due to vibration due to the influence of wind, rain, etc., and these factors reduce the measurement accuracy. Therefore, we used a high-speed response actuator with a small light path (actuator with a structure similar to the drive system capable of high-speed response, such as that used in optical pickups of CD and DVD readers) to adjust the angle and position of the lens. It is desirable to provide a mechanism for adjusting the irradiation position to the measurement site and adjusting so that the detected light is maximized.

This mechanism has a real-time adjustment mechanism (dynamic control mechanism) to ensure measurement accuracy. FIG. 7 is a diagram for explaining the situation in which the lens 20 is moved by the high-speed response actuator to adjust the optical axis. The lens 20 can be tilted or shifted by a high-speed response actuator that combines a plurality of coils and permanent magnets that oppose the coils provided in a support portion that supports the lens. FIG. 7(a) is a side view of the measurement site 21 of the petiole or flower stalk, (b) is a schematic plan view, and (c) is a front view. By changing the tilt angle of the objective lens 20 with an actuator as shown in FIGS. By shifting the two objective lenses (shift correction 27), it is possible to suppress vibrations due to physical displacement of the measurement site and wind swaying, etc., and stabilize the measurement accuracy (actuator Operation control will be described later).

Positional deviation and angle change caused by vibrations due to wind and rain can be adjusted by actuators. However, there are things such as plant growth and other physical changes that occur over time that cannot be corrected by the adjustment mechanism using actuators. In this regard, for example, with respect to changes accompanying plant growth, etc., reference light and measurement light having different wavelengths from the measurement light are irradiated to the measurement site at different timings so as to pass exactly the same optical path, and the respective By measuring the amount of received light of the wavelength, it is possible to adjust the measured value of the measurement light using the measured value of the reference light. The absorption rate of light due to physical changes such as plant growth is considered to be approximately the same as the wavelength of the measurement light used for measuring glucose and the wavelength of the reference light, so the difference between the detected values at both wavelengths is taken. This is because physical changes can be offset.

FIG. 8 is a schematic diagram illustrating an optical configuration of a transmitted light type measuring device. As a feature, a plurality of different wavelengths are used as near-infrared light sources (in this configuration, two semiconductor laser diodes 19a and 19b are used), and the plurality of light sources are coaxially emitted. As wavelengths, a second wavelength light source LD2 (19b) that outputs light with a wavelength that exhibits large absorption in sugar, for example, around 1500 nm as measurement light, and a first wavelength light source LD1 (19a) that outputs light with a wavelength of 1300 nm as reference light are used. do. 7, 8 and 9, the positions of LD1 (19a) and LD2 (19b) may be interchanged.

Laser light is desirable as the light source. The reason is that the emission wavelength range is very narrow and can be treated as a single wavelength. Naturally, the light source may have emission characteristics with a deviation of about 10 nm as the range considered to be a single wavelength. In addition, it is also possible to indirectly measure the amount of nitrogen by using another wavelength for the LD2, for example near-infrared light of 1660 nm as a wavelength having strong absorption characteristics of ammonia. It can be used as a judgment whether nitrogen, which is a compulsory substance for growth, is appropriate. Also in this case, a wavelength around 1300 nm is selected as the wavelength of the reference light LD1.

This combination of wavelengths is determined by the substance to be measured, and it is possible to change the measurement target of the change. The basis of the combination is a combination of light having a wavelength that exhibits light absorption characteristics with respect to the substance to be measured and wavelengths that do not have light absorption characteristics. The reason why 1310 nm is used as the reference light LD1 is that this wavelength exhibits high absorbance to moisture, but does not exhibit large absorbance to glucose. By combining the wavelength of the reference light LD1 and the light of the wavelength of the measurement light LD2, it is possible to perform correction to remove the water content from the detection amount of the measurement light (LD2) based on the detection amount of LD2. This correction method may be a method of obtaining the difference between the amounts detected by LD2 and LD1, or a method of obtaining the ratio of the two, whereby values other than the displacement of glucose contained in the amount detected by LD2 are offset.

Further, the amount detected by the reference light LD1 can be used as a control amount for controlling the actuator for correcting the vibration of the measurement site and the incident light state, and avoiding obstacles on the optical path. An actuator, which will be described later in detail, is electrically controlled so that the amount of light detected by this reference light (LD1) is maximized. FIG. 7 shows a configuration example of a transmitted light method with respect to a measurement site, and FIG. 8 is a schematic diagram illustrating an optical configuration of a reflected light method measuring apparatus.

Light from the light sources 19a and 19b is condensed into small diameter beams by lenses 28a and 28b and the like to become collimated light 19'. By narrowing the beam to a small diameter, it is possible to ensure brightness without using a light source with a large output, and it is possible to reduce power consumption and costs. Also, if there is an obstacle on the optical path, it can be avoided. This beam is made coaxial by the PBS 25a. However, the two light sources are controlled so as not to emit light at the same time. The irradiation light can correct the irradiation position on the measurement site 21 by the objective lens 20 having an actuator function. The operation of this actuator has a shift movement function (parallel movement function) and a tilt function (tilt angle adjustment function), and performs adjustment by real-time control so that the detected value by the reference light is maximized. Since this adjustment corresponds to relatively slow changes in the movement of the measurement site due to wind and rain, this actuator does not require a high-speed response performance up to a CD optical pickup.

A measuring device for measuring plant metabolites is configured to be foldable by connecting one end of two structures with a hinge or the like. Sometimes, it is desirable to have a configuration (sandwich structure) in which the measurement site 21 is fixed at a predetermined measurement position. This makes it easier to fix the measurement site 21 so that it does not move at the measurement position. In addition, the simple structure makes it possible to shorten the distance from the light source and the light receiving section to the measurement site 21 while always maintaining a constant distance. FIG. 10 shows a schematic diagram illustrating an outline of an optical configuration of a transmission-type measurement device fixed to a measurement site of a folded structure. A side view of a state in which a measuring portion 21 of a growing petiole or flower stalk is fixed to the measuring device 16 is drawn as a transparent drawing so that the layout of the main components of the measuring portion can be understood. In the folding measuring device 16, the upper portion 70a of the housing 70 can rotate counterclockwise in FIG. 10 around the rotating shaft 29c. FIG. 10 illustrates a transmitted light type measuring device 16 .

In FIG. 10, as the light source 19, it is desirable to use a two-wavelength laser capable of emitting two wavelengths of reference light and measurement light. In FIG. 10, the illumination light 19' from the light source 19 is focused by the lens 28, reflected by the PBS 25a, and guided to the objective lens 20. However, it is also possible to use an optical fiber or the like to guide the light to the objective lens 20. . The irradiation light 19 ′ is applied to the measurement site 21 , and the transmitted light 19 ″ is collected by the condenser lens 30 , reflected by the mirror 31 and detected by the PD 23 . In addition, although FIG. 10 shows a folding structure using a hinge, a clip structure may be employed in which two housings are sandwiched using an elastic member such as a spring.

Although FIG. 10 shows the structure of the transmitted light system, a similar folding structure or a spring-type clip mechanism can also be used in the reflection system. In this case, the optical structure shown in FIG. built in. With such a folding structure or clip structure, the measuring device 16 can be easily attached to the petiole or flower stalk of the plant. In addition, since a narrow beam is used in equipment such as an ordinary optical reader, strict adjustment of the optical axis is required for alignment. However, in this method, the irradiation light 19′ irradiated to the measurement site is diffused at the measurement site and has a relatively large beam diameter. Therefore, it is necessary to strictly adjust the detection of the transmitted light 19'' after passing through the measurement site. disappears. Therefore, it is possible to realize the desired measurement accuracy even by mounting with the clip structure as in this case. This makes it possible to reduce the cost of the mechanism of the device.
<First embodiment of non-destructive measuring device>

FIG. 11 is a block diagram of an electric circuit showing the first embodiment of the nondestructive inspection apparatus for living organisms according to the present invention. Although FIG. 11 shows a configuration using transmitted light, the same electrical circuit configuration may be used even when reflected light is used. Oscillator 1 (OSC1: 32a) is a signal used for measurement, for example, a signal for AC-modulating an optical output at 1 Khz. The measured value is a value detected by the PD 23 as transmitted light attenuated by absorption and diffusion of the irradiated light modulated by the OSC 1 and attenuated by the measurement site.

11 and 12, the lenses of the optical system are partially omitted. Also, the arrangement positions of the LD1 (19a) and the LD2 (19b) are reversed from those in FIGS. can also In addition, as a PD that is a light receiving element such as an image sensor, it is equipped with a main PD 23 that measures the amount of received light and sub PDs 23s and 23b that are used as optical axis adjustment signals according to the adjustment and vibration of the optical axis and lens. there is Functions and roles of the sub PDs 23s and 23b will be described later.

The oscillator 2 (OSC2: 32b) is for switching between the light source 1 (LD1: 19a) and the light source 2 (LD2: 19b) by means of a switch 33. When the LD1 emits light, the LD2 is inactive, and when the LD2 emits light. alternately switches the light source from which LD1 emits light as if at rest. For example, when the output of OSC2 (32b) is high (H), LD1 emits light, and when it is low (L), LD2 emits light. Also in this case, LD1 is used as the reference light, and LD2 is used as the measurement light. The output of a light receiving element (main PD) 23 (shared for reference light and measurement light) is converted by a current/voltage element 37 and amplified by a synchronous amplifier 38 .

The light source driving circuits 1 and 2 (34a, 34b: hereinafter referred to as LDD1 and LDD2) have high-frequency superposition functions 36a, 36b in their respective laser diodes, and in order to avoid unstable laser light emission due to reflected light, Used for single-mode to multi-mode oscillation, the optical output is kept constant by an APC circuit (not shown) such as a front monitor or a back monitor. A temperature sensor is also installed to compensate for changes due to temperature.

The RMS circuit 39 outputs the effective value of the detected signal and inputs it to the servo amplifiers 40 and 42 . A circuit 43b for holding the output of the RMS circuit 39 when the LD1 emits light and a reference voltage 41 (corresponding to the reference light quantity) are input to the LD1 servo AMP to calculate the difference, and the calculation result is held by the LD1LD1 control amount hold circuit 43a. to form a servo loop for automatically controlling the amount of light emitted by the LD1. By this operation, the light amount of the reference light received by the main PD 23 becomes constant by eliminating the influence of the basic transmission amount.

When the output of the LDI servo amplifier 40 is large, it indicates that the amount of light attenuation in the object to be measured is large. becomes a reference value for controlling By obtaining this reference value, the output intensity of the light source LD2 required for measuring the object to be measured is automatically obtained. In addition, a tissue with physical displacement of the measurement site (physical variation of the object to be measured due to plant growth, etc.) has the same attenuation characteristics for light of both wavelengths LD1 and LD2 ( It does not affect the absorption characteristics and diffusivity characteristics), so the amount detected by LD1 reflects the amount of physical displacement and the correction amount of absorbance due to moisture that may change over time. Become.

Further, by calculating the difference between the output of the circuit 43c that holds the output of the RMS circuit 39 when the LD2 emits light and the output of the circuit 43a that holds the control amount of the LD1, and using it as the control output of the LD2, It becomes possible to keep the output of LD2 constant (the optimum ratio of the light emission amount of LD1 and the light emission amount of LD2 is obtained in advance, and the gain of LDD is determined according to the ratio). The circuit 43a for holding the control amount of the LD1 holds the output from the RMS circuit 39 of the LD1 when the output of the oscillator 2 (32b: OSC2) is high (H), for example. When the output of is low (L), the LD2 detection hold circuit 43c holds. The output of the measured value correction circuit 44, which calculates the difference between the controlled variable hold circuit 43a and the controlled variable of the LD2, is finally the measured value obtained by correcting the physical displacement and the moisture displacement from the LD2 detected quantity.

Next, position adjustment of the objective lens 20 will be described. The objective lens 20 is adjusted by the emission period 51 of the reference light LD1 (see FIG. 13(a)). When light is emitted for the first time, the light intensity distribution detection circuit 45 calculates the difference between the outputs of the sub PDs 23s and 23b provided next to the main PD 23, thereby detecting which side the center of the beam is on. This detection operation adjusts the center of the intensity of light detected by the main PD 23 to be the center of the main PD. In the configuration of FIG. 11, the output signal S (37S) and the output signal B (37b) of the I/V (current/voltage) conversion circuit 37 are input to the light intensity distribution detection circuit 45. FIG. When the output signal S is large, the output of the light intensity distribution detection circuit 45 appears on the (+) side of the reference voltage, and the shift drive circuit 46b is driven in the direction in which this output decreases. When the output signal B (37b) is large, the (-) output appears from the reference voltage, so the shift drive circuit 46b is driven in the opposite manner to the output signal S (35S) so that this output becomes small. .

In the measurement, the light source LD1 of the reference light and the shift drive mechanism 49 are driven at the same time, and after obtaining the amount of reference light detected by the LD1, detection by the LD2 is performed during the light emission period 52 of the LD2 (see FIG. 12). to get the final measurements. In order to improve the SNR (signal-to-noise ratio) of the measured value, it is preferable that the measured value is an average value (superposed value) of a plurality of measured values. FIG. 13 is a diagram for explaining an example of the switching timing of driving the light source LD1 for the reference light and the light source LD2 for the measurement light. In the example of this figure, a continuous signal is used as the modulation signal 32c as shown in (b), but a pulse signal with a low duty ratio as shown in (c) may also be used.

If the objective lens 20 (FIGS. 8, 9, etc.) for irradiation light has a tilt function (see FIG. 7), tilt adjustment is performed using the reference light LD1 before measurement. Therefore, the light emission period 51 of the LD1 is used a plurality of times to measure the reference light from the LDI. At that time, the tilt drive mechanism 50 is driven by changing the output from the tilt drive reference voltage generation circuit 48 (which can be configured with a very small MPU or the like) each time the measurement is performed so that the LD1 control amount is minimized. ask for a good condition. After that, the shift direction is adjusted by the shift drive mechanism, and the measurement cycle starts. Adjustments by the tilt drive mechanism 50 and the shift drive mechanism 49 are performed in real time (dynamic control) in order to eliminate the influence of the tissue structure of the measurement site and to correct deviations due to vibration and the like. adjustment) is desirable. The signal output by this circuit is an analog signal. In practice, this value is A/D converted and then input to a personal computer (PC) or the like to obtain the final measurement result. It is the measured value S1 at one measurement 12 (t1 in FIG. 4), and the final measured value is obtained from the subsequent two measurements 13, 14 (S2, S3).
<Second embodiment of non-destructive measuring device>

FIG. 12 shows a block diagram of an electric circuit showing a second embodiment of the nondestructive inspection apparatus for living organisms according to the present invention. Although FIG. 11 shows an analog servo loop configuration for the measurement portion as an electric circuit, FIG. In this case, the output from FIG. 12 (the output equivalent to the output to the display device 55) can be captured and processed by a PC or the like. In this case, data transmission/reception to/from a PC or the like can be performed not only by wire but also by wireless communication means.

In the case of an analog servo loop, the light emission of LD1 and LD2 is normally emitted with a waveform 32c from the oscillator as shown in FIG. Pulsed light emission 32d, for example, pulsed light emission of about 30 ns to 1 μs can also be realized. In this case, the output of the main PD 23 appears in response to the pulsed light emission, but the absorbance can be measured by measuring the peak value output (voltage). can be improved.

In addition, this pulse emission makes it possible to avoid the temperature rise of the measurement site due to the energy of the light. By suppressing the temperature rise, an improvement in measurement accuracy can also be expected. Moreover, when the site to be measured is continuously irradiated with very strong light, it is possible to avoid the possibility of burning due to the light. Furthermore, the synchronous amplifier 37 can also be realized by digital signal processing. The control amounts of LD1 and LD2 of this servo loop themselves result in detection amounts corresponding to absorbance and diffusion.

The operation of FIG. 12 will be briefly described. First, the value 38a at the time of light emission of the LD1 (the light emission control amount is a predetermined amount) is inputted to the AD terminal several times, and the driving amount of the tilt driving circuit 47b is changed so that the detection amount of the LD1 is minimized. Amount is detected to determine the optimum state of tilt. In this state, the signals 35S and 35b from the sub PD are input to the AD terminal of the MPU 54 for adjustment by the shift drive mechanism 49, and calculation (light intensity distribution detection Calculation corresponding to the circuit 45) to drive the shift drive circuit 46b. This series of tilt control and shift control is performed before measurement by LD1 and LD2.

When driving the LD1 and LD2, the MPU 54 drives the LD1 and LD2, and the ON/OFF signal 34d of the LD1 and the ON/OFF signal 34e of the LD2 are controlled so as to correspond to the modulation by the OSC1 (32a). In the measurement, first, the output of the light emission control amount 34c from the MPU to the LD1 is adjusted by a constant amount so that the value 38a input from the AD terminal becomes a predetermined value (corresponding to the LD1 reference voltage generation circuit 41). The detected amount is obtained and used as the detected value of LD1. Subsequently, similarly, the LD2 light emission control amount 34f is adjusted so that the input value 38a inputted to the MPU from the AD terminal becomes a constant amount based on the amount detected by the LD1. The LD2 light emission control amount 34f at this time is set as the detection amount by the LD2.

Next, the MPU 54 subtracts the amount detected by LD1 from the amount detected by LD2, and the value is corrected by the signal 35a from the temperature correction sensor 35 (the amount of correction is obtained experimentally based on the value obtained from the absorbance characteristics depending on temperature). is the final measured value. With this configuration, it is assumed that the measured glucose concentration ranges from 50 mg/dl to 200 mg/dl.

Next, the treatment of the three measured values and the output of the final measured value, which are one of the features of this measuring apparatus, will be specifically described. Measurement is started when the power is turned on to the device or after the operation switch is operated, and the value at that time is measured as the reference value. Let the measured value at this time be the value (t1, S1) of the measurement 12 in FIG. Measurements are taken at fixed time intervals, and the values are sequentially recorded in the storage device in the MPU 54. The value measured at t2 (measurement 12) after a fixed time, for example, 10 minutes, is defined as the measured value (t2, S2). Furthermore, let the measured value of t3 (measurement 13) after 10 minutes pass be (t3, S3) (the time intervals of t1, t2, and t3 are set to be the same).

ds=S3-S1 is obtained from the measured values at these three points. This value is the basic measured quantity for this device. Next, dts=S2-S1 is obtained. This value is a time differential value indicating how much it has changed in a short period of time, whereas the ds value is a time differential value in a long period of time compared to dts. The measuring device of the present invention outputs the value of ds and the value of dts, and can make complex judgments based on these measured values ds and dts. When the value of dts is large, it is understood that there was a large displacement in a short period of time, and since ds is a displacement in a relatively long period of time, it indicates continuous displacement. By combining these values, it becomes possible to observe the state of photosynthesis and the state of translocation.

Specifically, when these values (ds, dts) are large, it can be evaluated that photosynthesis was performed well and growth was promoted, and conversely, when these values were small, photosynthesis was not performed well Probability is high. In this case, if it is judged that the water content is low based on the monitoring data of the environmental conditions, automatic control such as increasing the amount of irrigation water becomes possible. On the other hand, even if it suddenly mutates, it can be imagined that something is wrong, and it is possible to avoid a big accident such as withering. In addition, when it is determined that photosynthesis does not function well, it is possible to take measures such as changing the growth environment in greenhouse cultivation.

When performing non-destructive measurement using light, changes in external factors related to light reflectance and absorbance greatly affect measurement precision. However, according to the measuring method of the present invention, since the difference between measured values obtained by measuring a plurality of times at relatively short time intervals is calculated compared to the conventional method, compared to the case of measuring changes at long time intervals, , the error caused by the displacement of the measurement site and the growth of the object to be inspected are reduced, and the deviation against the accuracy is offset, thereby improving the measurement accuracy and reproducibility. This is not limited to plant measurements. For example, in the measurement of blood sugar level, components other than blood sugar hardly change in a short period within two hours after a meal. In addition, since the condition of the skin, which greatly affects transmitted light and reflected light, hardly changes in a short period of time, it is almost necessary to consider errors due to external factors when obtaining changes based on the first measured value. Therefore, it is possible to improve the accuracy of measurement using light. When measuring the blood glucose level of an animal or human, it is desirable to select a measurement site with good light transmission such as an earlobe.

FIG. 14 illustrates a graph for judging the state of photosynthesis and the like based on the measured values. The horizontal axis 58 represents the value of ds=S3-S1, and the vertical axis 59 represents the evaluation value "dds" for the final judgment indicating the progress of photosynthesis, for example. A plurality of straight lines shown in this graph represent the evaluation value dds by the following function with ds as a variable, and are straight lines with a slope of 1/(1+dts).
dds=(1/(1+dts))ds+dts

Knowing the measured value of dts establishes each straight line. By obtaining this straight line from the measured dts and substituting the measured ds into the straight line, the evaluation value dds can be obtained. Here, dts=S2-S1 and ds=S3-S1. When dts=0, the slope of the straight line is "1", and the straight line passes through the origin and has a slope of 45°. The ds, dts, and dds characteristics show that even when the value of ds(58) is the same, the value of dds(57) increases when the value of dts(57) increases. Since the value at is emphasized, it is possible to clarify the point of change. For example, in the case of plants, it changes very slowly, so it was difficult to determine what factors (temperature, amount of insolation/time, irrigation, etc.) depend on the change. Since the point of change is emphasized, the point of change is easy to understand, and it becomes easy to specify the factor by checking the environmental data at that time. When a display device 53 is connected to the device, the display device can display different colors according to the magnitude of the numerical value of this dds, or gradually change to a darker or lighter color as the numerical value increases. By adopting such a display configuration, the change in dds can be sensuously grasped as a change in color, and the change and state of the evaluation value dds can be easily recognized.

In vinyl greenhouses and hydroponics, the status of plant metabolism can be measured in real time by placing one or more at important measurement points using the above-described nondestructive measurement device for plant metabolites according to the present invention. It is possible to control the cultivation environment by feeding it back.
At that time, in order to analyze the causal relationship between the measurement data of the metabolites and the cultivation environment factors, and to control the cultivation environment, temperature control, humidity control, water supply control, fertilizer supply control, etc. that can manage the cultivation environment Equipment that can control the environment (automatic control is preferable, but manual control is also acceptable) and various sensors (air, soil, etc. temperature sensors, humidity sensors, solar radiation measurement sensors, EC sensors, etc.) It is preferable to have

In the above description, the present invention has been described as an invention relating to an apparatus. A non-destructive measurement method of a living organism for measuring a substance to be measured, comprising the step of individually irradiating a part to be measured with light of a first wavelength as reference light and a second wavelength as measurement light;
a step of receiving reflected light or transmitted light from the portion to be measured of the irradiated light of the first wavelength and the second wavelength and outputting a detection signal corresponding to the amount of received light;
The ratio of the light amount of the irradiation light irradiated to the part to be measured and the light amount of the reflected light or the transmitted light of the first wavelength and the second wavelength received by the light receiving part is the absorbance of the first wavelength and A step of determining the absorbance of the second wavelength and outputting a value obtained by correcting the absorbance of the second wavelength with the absorbance of the first wavelength as a measured value;
The measured value calculated at the time of the first measurement is stored as the first measured value, and the same measurement and calculation are performed at the time of the second measurement after a certain period of time has passed, and the measured value is stored as the second measured value. Similar measurements and calculations are performed at the time of the third measurement later, and the result is stored as a third measured value. outputting as a first calculated value representing the change, and dividing the difference between the second measured value and the first measured value by time to output a second calculated value representing the rate of change over time;
It also discloses a non-destructive measurement method for living organisms, characterized by comprising:

The measurement data obtained by measuring the floral pattern of strawberries in the horticultural cultivation of strawberries by the nondestructive measurement device for metabolites according to the present invention is shown below. In horticultural cultivation in which plants are grown while controlling the growth environment of the plants using a greenhouse or the like, it is extremely important to control saturation and irrigation water. will be explained. Saturation indicates the ratio of the saturated water vapor pressure to the actual water vapor pressure, and is expressed by the following formula with temperature and humidity as factors.
Saturation = saturated water vapor pressure x (1 - relative humidity/100)
Irrigation usually refers to watering plants, but in horticultural cultivation, liquid fertilizer, which is a mixture of water and fertilizer, is often used. The following measurement data also shows an example of measurement under a cultivation environment in which water mixed with fertilizer (liquid fertilizer) is irrigated.
15 (a) and (b) show the measurement data for a day from 0:00 am on February 7, 2019 to 0:00 am the next day in the horticultural cultivation of strawberries at approximately every 30 minutes, and FIG. shows the measurement data for 7 days from April 7th to April 15th, 2019 every 3 hours and 40 minutes. FIG. 15 illustrates the relationship between satiation control and plant metabolites, and FIG. 17 illustrates the relationship between irrigation and plant metabolism.

In FIGS. 15(a) and 15(b), the measurement data (Agri-1 and Aguri-2) indicated by symbols (1) and (2) show the measurement results of the nondestructive measuring device for metabolites of the present invention. there is 15A and 15B are divided into two graphs because it is difficult to see if all the data measured at the same time under the same environment are written in one graph. Agricultural 1 (1) and Agricultural 2 (2) are shown in two graphs (a) and (b) so that the relationship with other indices such as saturation (4) and solar radiation (8) can be easily compared. The same measurement data is displayed redundantly. Agri 1 is the measurement data of the reference light (for example, 1310 nm wavelength), and Agri 2 is the measurement data of the measurement light (for example, 1550 nm wavelength). In either case, the intensity of the received light is used as the measurement data, so when the intensity of the received light is high, that is, when the absorbance is low, the measured value is large. Therefore, in the measurement data of Aguri 1 and 2 in FIGS. 15 and 16, the larger the value of the measurement data, the less the metabolites. In FIGS. 15 and 17, the measurement data of the reference light and the measurement light without correcting the measurement light with the reference light are displayed as AGR1 and AGR2, respectively, with the intensity of the received light as they are. Since the amount of water movement and the degree of plant growth can be known from the light reception data of the reference light, the light reception data of the reference light and the measurement light are displayed as raw measurement data. Of course, in addition to this, it is also possible to graphically display data obtained by correcting the measurement light with the reference light.

In addition to Agricultural 1 and Agricultural 2, the graph in FIG. Measured data (8) is shown. The EC value (5) represents the concentration of the fertilizer in the soil, and the EC value x20(3) represents the amount of change of the EC value (5) multiplied by 20.
Observation of FIG. 15(a) reveals that the EC value x20 (soil fertilizer concentration) (3) varies randomly throughout the day, suggesting that the plant takes some reaction such as absorbing the fertilizer. However, from 0:00 to 19:30, there is no significant change between Aguri 1 and Aguri 2, so there is no migration of metabolites in the peduncle. On the other hand, after the amount of solar radiation (8) disappeared, it was observed that agriculture 2 was activated in synchronization with saturation (4) from around 19:30. In the first place, the output of the EC value (5) is small and difficult to understand, but it can be observed that the EC value x20 (2) is changing relatively actively.

From these changes, it can be seen that the movement of metabolites in the peduncle can be activated by stimulating by varying the saturation value after the daytime insolation has disappeared. This is the first knowledge obtained by observing the metabolism of growing plants using the metabolite measuring device according to the present invention. Appropriate timing of metabolite migration in the peduncle during growth has a significant impact on fruit growth and sugar content. In addition, it is empirically known that sudden and large fluctuations in the saturation value are not good for plants. Therefore, by controlling the temperature and humidity, it is preferable to change the variation range of the saturation value within an empirically known appropriate range or within an appropriate range found by the present invention. FIG. 16 shows a flow chart illustrating a control procedure for such saturation control.

For example, the temperature and humidity are controlled to increase the saturation value from around 19:30, when a certain period of time has passed after sunset indicating that the agriculture 2 indicated by reference numeral (2) in FIG. 15 is activated (S1). . After that, for example, at intervals of 5 to 10 minutes, the metabolite migration state in the flower stalk or petiole is measured at regular time intervals using the nondestructive measuring device for metabolites according to the present invention (S2). Next, for example, changes in measured data (amount of displacement, rate of change, rate of mutation, etc.; displacement rate is illustrated in FIG. 16) are calculated every 30 minutes (S3). If the displacement rate of the metabolite is smaller than the predetermined reference value (S4: No), it is checked whether the range of the saturation value is within the proper range, and if it is within the proper range (S9: Yes), The measurement (S3) and checks (S4, S9) by the measuring device are repeated while maintaining the setting for increasing the saturation difference. Here, the predetermined reference value of the displacement amount and the appropriate range of the saturation value can be determined based on knowledge based on experience, but the value obtained from the result of observation with the measuring device according to the present invention is used as the reference value or the appropriate range. You can decide.

If the displacement is greater than a predetermined reference value (S4: Yes), or if the saturation value is out of the proper range (S9: No), change the temperature and humidity settings to lower the saturation value. (S5), the state of change in metabolites is measured at regular time intervals in the same manner as in S2 and S3 (S6), the rate of change is calculated (S7), and the value is compared with a predetermined reference value. (S8). If the mutation rate is smaller than the predetermined reference value (S8: No), it is checked whether the saturation value is within the proper range. Measurement by the device (S6), calculation of rate of change (S7) and checking (S8, S10) are repeated. If the displacement is smaller than a predetermined reference value (S8: Yes) or if the saturation value is out of the proper range (S10: No), change the temperature and humidity settings to increase the saturation value. (S1), and the same processing is repeated.
If the measurement data of metabolites does not fluctuate due to changes in saturation (temperature and humidity), the reason may be that the fruit is in a mature state. Continued observation is preferred based on management.

Next, with reference to FIG. 17, the relationship between the input of irrigation water (liquid fertilizer) and the metabolism of plants will be described. FIG. 17 is also a graph showing measurement data of the plant metabolite measuring device of the present invention and other cultivation environment factors. As the measurement data of the cultivation environment factor, the water content (liquid fertilizer) indicated by the symbol (3) of the EC value in FIG. It is
FIG. 17 shows measurement data for 9 days, and the liquid fertilizer is applied four times every other day at the timing indicated by vertically long circles a1 to a4. Despite the fact that liquid fertilizer was applied twice (a1, a2) during the first five days of FIG. not Rather, during the first five days, the measured data for Aguri 1 and Aguri 2 show a gentle upward trend. The large measured values of Aguri 1 and 2 indicate that the amount of received light is large, indicating that there are few metabolites in the peduncle, which means that they do not contribute much to the growth of the fruit. ing. Therefore, even if liquid fertilizer is added in such a situation, it is ineffective. Therefore, in such a case, it is better to irrigate with water instead of liquid fertilizer. This can prevent damage due to excessive fertilizer application.

On the other hand, from the 6th day to the 9th day, both Aguri 1 and Aguri 2 showed a gradual decrease in the amount of received light, indicating that the movement of metabolites was active. From this measurement data, the amount of metabolite movement in the peduncle of metabolites is low during the daytime and increases at night, which agrees with empirical findings. Since the plants are in a growing state during such metabolism, it is desirable to apply liquid fertilizer. In this case, it is preferable not to apply the liquid fertilizer all at once, but to apply it dispersedly so that the osmotic pressure within the plant does not change abruptly.

A control procedure for controlling irrigation (liquid fertilizer) will be described below with reference to the flowchart illustrated in FIG. First, 1/n of the planned amount of liquid fertilizer (irrigation) is irrigated (S11), and the movement of metabolites in the flower stalk or petiole is measured at regular time intervals by the nondestructive measuring device for metabolites according to the present invention. (S12). Next, based on the measured data, changes in the measured data (displacement amount, rate of change, mutation rate, etc.: displacement rate is exemplified here) are calculated (S13). When the displacement rate is larger than the predetermined reference value (S14: Y), the movement of metabolites is active, so liquid fertilizer is irrigated (S15), and when it is smaller (S14: N), water is irrigated. (S17). Next, it is confirmed whether liquid fertilizer or water has been watered n times, and if watering has not been done n times (S16: N), the steps from S12 to S16 are repeated. If watering has been performed n times (S16; Y), the watering is terminated. In this way, the metabolic situation during actual growth is observed in real time, and liquid fertilizer is irrigated when the movement of metabolites is active, and water is irrigated when the movement is not active.

In the above explanation, we explained the measurement of sugars, but since the wavelength of light of 1550 nm also reacts with the absorption of ammonium ions, we can measure not only sugars but also ammonium ions depending on the measurement timing even if the wavelength is the same. can. Whether it is sugars or ammonium ions can be determined based on empirical knowledge of cultivation based on the amount of sunlight at the time of measurement and a combination of other factors (whether the time of measurement is during the day or at night, etc.). can. In addition, by using light of different wavelengths as measurement light, it is possible to measure the movement of metabolites other than sugars and ammonium ions.
Furthermore, the measurement data from the reference light can be used not only to measure physical displacement such as vibration to control actuators and to correct water content, but also to observe water movement in petioles or flower stalks. It can also be used as data for observing the growth of flower stalks or petioles.
Further, the correction by the measurement light using the reference light is not essential, and as described with reference to FIGS. 15 to 18, it is possible to measure the reference light and the measurement light as two independent measurement lights.
Furthermore, in the above description, only an example in which two types of light with different wavelengths are irradiated and an object to be measured is measured based on the difference between them is shown. It may be configured to irradiate the object so as to pass through the same optical axis and measure the object using the measurement results of each wavelength. In addition, the measurement site for measuring the movement of metabolites is not limited to floral stalks or petioles, and it is also included in the scope of the present invention to set the measurement site at a desired location where it is desired to know the movement of metabolites. At that time, the fixing structure of the measurement site can be changed as appropriate.

By using this measuring device, the state of photosynthesis and the state of translocation of plants can be grasped in real time. The present invention is applicable not only to plants. For example, it can also be used for continuous observation of metabolic organisms of animals, insects and other organisms. The measuring device and measuring method of the present invention can also be used for new health management of animals including human beings who continuously measure changes in blood sugar levels. It can also be applied as a diagnostic device for early detection of so-called latent diabetes.

For example, it is known that the pedigree level of a person generally rises in about 30 minutes after eating, and returns to the pre-meal blood sugar level after about 2 hours. Therefore, the first measurement time t1 in this measurement method shown in the graph of FIG. 4 is about 30 minutes before the meal and the second measurement time t2 after the meal. do. The third measurement time t3 is about two hours after the first measurement time t1. These 30 minutes and 2 hours are examples, and in the case of an actual device, the time is measured by the clock function in the device and used. The state of occurrence of a so-called blood sugar level spike can be grasped from the time differential values at times t1 and t2 before and after meals of the user. Also, when the rate of change of t1 and t3 is observed, it is possible to grasp the state of sugar metabolism in the body. Generally, if the blood sugar level returns to a normal level after about two hours have passed after a meal, the amount of the blood sugar level is judged to be normal. However, even if the blood sugar level returns to normal, an event in which the blood sugar level rises sharply in the process is judged to be abnormal.
According to the present invention, the rate of increase in blood sugar level and the state of sugar metabolism can be evaluated in a composite manner, and can be used as a comprehensive index for the increase in blood sugar level. After a meal, blood sugar levels at multiple points in time, such as changes in actual blood sugar levels at 10-minute intervals, are continuously observed, and a glucose tolerance test is performed to diagnose diabetes from the state of the changes by measuring 3 points. will perform the same function as

In addition, if the metabolite to be measured changes with time, the change affects the health, growth, etc. of the organism, and the absorbance or diffusivity of light of the object to be measured varies depending on the wavelength. , can be similarly applied to components other than sugar.

5 temperature 6 solar radiation 7 concentration of glucose in petiole 8 change in glucose in flower stalk 16, 17 metabolite non-destructive measuring device (measuring device)
19 light source 19a LD1 (light source 1: reference light source)
19b LD2 (light source 2: light source of measurement light)
19′ irradiated light 19″ transmitted light 20 objective lens 21 measurement site (object to be measured)
23 light receiving element (main PD)
23s, 23b Sub-PD
70 housing 71 V-shaped groove 72 fixture

Claims (14)

a fixing part for fixing the peduncle or petiole of the plant;
a non-destructive measurement unit that irradiates light on a portion of the fixed petiole or the floral stalk and non-destructively measures metabolites of the plant by transmitted light or reflected light of the light;
A non-destructive measurement device for plant metabolites comprising:
The fixing part has a groove part in which the stem part, which is the flower stalk or the petiole of the growing plant, can be embedded in the nondestructive measurement device,
The nondestructive measurement unit
A first wavelength, which is a laser beam in the near-infrared region and serves as a reference light that is absorptive to water and has a low absorptivity to metabolites of plants, and the absorptivity to water is the first wavelength. a light-emitting unit that individually emits light of a second wavelength that serves as measurement light that is nearby and has a high absorption rate for metabolites of the plant ;
an optical element that focuses the light of the first wavelength and the second wavelength from the light emitting unit into a small diameter beam and irradiates the measured part of the plant ;
a light-receiving unit that receives reflected light or transmitted light from the measured part of the irradiated light of the first wavelength and the second wavelength and outputs a detection signal according to the amount of received light;
The amount of light emitted from each of the light emitting units and the amount of reflected light or transmitted light of the first wavelength and the second wavelength received by the light receiving unit are each represented by a first absorption coefficient of the first wavelength. and a second absorption coefficient that is the absorption coefficient of the second wavelength , and the second absorption coefficient is corrected with the first absorption coefficient , thereby removing at least the amount of water absorbed from the plant metabolism an absorptance calculator that outputs a value including a value indicating the amount of change in the product as measurement data;
a storage unit that sequentially stores the measurement data;
an output unit that outputs the measured data to the outside;
a control unit that controls the light-emitting unit, the optical element, the light-receiving unit, the absorbance calculation unit, the storage unit, and the output unit to continuously measure changes in the movement of metabolites in plants ;
comprising a
A non-destructive measurement device for plant metabolites. The light of the first wavelength and the light of the second wavelength are configured to alternately irradiate the same location of the measured part through the same optical axis by the light emitting unit, the optical element, and the control unit. The non-destructive measurement device for plant metabolites according to claim 1. The control unit stores the measurement data calculated during the first measurement by controlling the light emitting unit, the light receiving unit, and the light absorption coefficient calculation unit as a first measurement value, and stores the measurement data as a first measurement value after a lapse of a certain time. Perform the same measurement and calculation at the time of measurement and store it as the second measured value. Further, at the time of the third measurement after a certain period of time has passed, perform the same measurement and calculation and store it as the third measured value. calculating a difference between the measured value and the first measured value and outputting it as a first calculated value representing a concentration change of the substance to be measured in the part to be measured, and obtaining the second measured value and the first measured value; Non-destruction of plant metabolites according to claim 1 or 2, characterized by comprising a measurement control unit that controls to output a second calculated value representing the rate of change over time by dividing the difference between measuring device. 4. The non-destructive measurement device for plant metabolites according to claim 3, further comprising a communication function for outputting said concentration change and said rate of change with time to the outside. 5. The nondestructive measurement device for plant metabolites according to claim 1, wherein the metabolites are sugars produced by photosynthesis. 4. The plant metabolite according to claim 3 , wherein the measurement control unit repeats similar measurements and calculations at regular time intervals to continuously output the concentration change and the time rate of change. Nondestructive measurement device. 7. The nondestructive measurement device for plant metabolites according to any one of claims 1 to 3 or 5 to 6, further comprising a communication function for outputting the measurement data to the outside. 8. The measurement data is obtained by subtracting the first absorption coefficient from the second absorption coefficient in the correction of the second absorption coefficient using the first absorption coefficient. A non-destructive measuring device for the plant metabolite according to any one of Claims 1 to 3. 8. Correction of said second absorption coefficient by said first absorption coefficient is characterized in that said measurement data is obtained by dividing said second absorption coefficient by said first absorption coefficient. A non-destructive measuring device for the plant metabolite according to any one of Claims 1 to 3. 10. The non-destructive measurement device for plant metabolites according to any one of claims 1 to 9, further comprising a mechanism for dynamically adjusting the irradiation position and angle of the light irradiated to the measured portion. . Having a two- dimensional data table centered on the first calculated value representing the density change and the second calculated value representing the rate of change over time, and measuring the measured A calculation is performed to convert the values preset in the data table of the first calculated value and the second calculated value representing the measured time change rate into a color , and the output shown in claim 1 is performed. 11. The non-destructive measurement device for plant metabolites according to any one of claims 3 to 10, wherein the device has a function of displaying the converted color. Light is applied to the stem, which is the petiole or flower stalk of a growing plant placed and fixed in a groove in a non-destructive inspection device , and metabolites of the plant are measured from the transmitted light or reflected light of the light. A nondestructive measurement method for
A first wavelength, which is a laser beam in the near-infrared region and serves as a reference light that is absorptive to water and has a low absorptivity to metabolites of plants, and the absorptivity to water is the first wavelength. A step of individually irradiating the measurement target part with light of a second wavelength, which is a measurement light that is close and has a high absorption rate to the metabolites of the plant, focused into a small diameter beam ;
a step of receiving reflected light or transmitted light from the measured part of the irradiated light of the first wavelength and the second wavelength with a light receiving part and outputting a detection signal corresponding to the amount of received light;
The ratio of the light amount of the irradiation light irradiated to the measured part and the light amount of the reflected light or transmitted light of the first wavelength and the second wavelength received by the light receiving part is the absorbance of the first wavelength. A first absorption coefficient and a second absorption coefficient that is the absorption coefficient of the second wavelength are obtained, and at least the absorption amount of water is removed by correcting the second absorption coefficient with the first absorption coefficient. a step of outputting a value including a value indicating the amount of change in the metabolite of the plant as measurement data ;
A method for nondestructive measurement of plant metabolites, comprising: Furthermore , the measured data calculated during the first measurement is stored as the first measured value, and the same measurement and calculation are performed during the second measurement after a certain period of time has elapsed, and stored as the second measured value. Similar measurements and calculations are performed at the time of the third measurement after the lapse of time, and stored as a third measured value, and the difference between the third measured value and the first measured value is calculated to calculate the measured value at the measured portion. outputting as a first calculated value representing the change in concentration of the substance to be measured , and dividing the difference between the second measured value and the first measured value by time to output a second calculated value representing the time rate of change; 13. The method for nondestructive measurement of plant metabolites according to claim 12 , comprising the steps of: A non-destructive measuring device for metabolites according to any one of claims 1 to 10;
At least one of a temperature control device equipped with a temperature sensor to control at least one of the temperatures of air, soil, and water, a humidity control device equipped with a humidity sensor, a water supply control device, a solar radiation control device, and a fertilizer supply device. a growing environment control unit comprising one facility;
an output unit that receives and outputs measurement data from the nondestructive measurement device and detection data from the growth environment control unit;
a control unit that controls the growth environment control unit;
A plant cultivation system comprising: