Phototaxis measurement system and inference method

Abstract

To provide a phototaxis measurement system for measuring algal behavior data, and to provide a method for inferring algal phenotype, genotype, or gene expression based on the measured algal behavioral data.SOLUTION: Provided is a phototaxis measurement system that includes a culture chamber for accommodating algae and a plurality of light irradiating devices and enables measurement of behavioral data of the algae. The plurality of light irradiating devices are capable of each independently changing the wavelength or the wavelength and intensity of the irradiated light, and are configured to be installed at different positions around the culture chamber to irradiate the culture chamber with light. Provided is an inference method including: providing a model that relates behavioral data of the algae to one or more of a phenotype, a genotype, and a gene expression; measuring behavioral data of a target algae; and estimating one or more of a phenotype, a genotype, and a gene expression of the target algae based on the behavioral data and the model.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present disclosure relates to phototaxis measurement systems and estimation methods. [Background technology]

[0002] Phototaxis is the movement of an organism in response to light stimuli; the movement toward light is called "positive phototaxis," while the movement away from light is called "negative phototaxis."

[0003] Non-sessile algae are an example of an organism that can exhibit phototaxis. Algae have photoreceptors that absorb light of various wavelengths. Chlorophyll, used in photosynthesis, absorbs red and blue light well, channel rhodopsin, used in phototaxis, absorbs green light well, and cryptochrome, which acts as a switch to protect the photosynthetic apparatus from strong light, absorbs blue light well. Positive and negative phototaxis switches depending on various metabolic states, including photosynthetic activity, within the cell. The full mechanism of this switching has not yet been clarified, but it is known that the amount of reactive oxygen species within the cell acts as a signal, and when it is high, it exhibits positive phototaxis, and when it is low, it exhibits negative phototaxis (Non-Patent Document 1). Algae strains that are likely to exhibit positive phototaxis include those with high growth rates or high resistance to strong light (Non-Patent Document 2). Algae strains that are likely to exhibit negative phototaxis include those with high photosynthetic activity (Non-Patent Document 3).

[0004] Conventionally, for example, when trying to separate a specific algae using phototaxis, phototaxis of the algae was induced using a light source device emitting light from one direction at one wavelength (the wavelength of the absorption peak of the photoreceptor of that algae) (Non-Patent Document 4). [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Wakabayashi, K., Misawa, Y., Mochiji, S., Kamiya, R. (2011). Reduction-oxidation poise regulates the sign of phototaxis in Chlamydomonas reinhardtii. Proceedings of the National Academy of Sciences of the United States of America 108:11280-4 [Non-Patent Document 2] Morishita, J., Tokutsu, R., Minagawa, J., Hisabori, T., Wakabayashi, K. (2021)Characterization of Chlamydomonas reinhardtii mutants that exhibit strong positive phototaxisPlants 10: 1483 [Non-Patent Document 3] Kim, JYH, Kwak, HS, Sung, YJ, Choi, HI, Hong, ME, Lim, HS, Lee, JH, Lee SY, Sim, SJ Microfluidic high-throughput selection of microalgal strains with superior photosynthetic productivity using competitive phototaxis. Scientific Reports 6: 21155 [Non-Patent Document 4] Ueki, N. and Wakabayashi, K. (2017). Phototaxis Assay for Chlamydomonas reinhardtii. Bio-protocol 7(12): e2356. DOI: 10.21769 / BioProtoc.2356. Summary of the Invention [Problem to be solved by the invention]

[0006] However, the present inventors recognized that the conventional phototaxis measurement has a problem in that, for example, phototaxis induction cannot be performed under conditions in which the photosynthetic activity is adjusted to an arbitrary state. In addition, the present inventors recognized that such conventional techniques assume known algae whose photoreceptor absorption peaks are known, and therefore have a problem in that phototaxis assays cannot be performed on novel algae whose photoreceptors are unknown.

[0007] The present disclosure has been made to at least partially solve the above-mentioned problems, and aims to provide a phototaxis measurement system for measuring algal behavior data, and a method for estimating the phenotype, genotype, or gene expression of algae based on the measured algal behavior data. [Means for solving the problem]

[0008] One embodiment of the phototaxis measurement system of the present disclosure comprises a culture chamber for containing algae and a plurality of light irradiation devices, enabling measurement of behavioral data of the algae, wherein the plurality of light irradiation devices are capable of independently changing the wavelength, or the wavelength and intensity, of the irradiated light, and are configured to be installed at different positions around the culture chamber to irradiate light into the culture chamber.

[0009] A method for predicting one or more of an algal phenotype, genotype, and gene expression of one or more of an algae according to one embodiment of the present disclosure includes providing a model relating algal behavioral data to one or more of the phenotype, genotype, and gene expression, measuring behavioral data of a target algae, and predicting one or more of the phenotype, genotype, and gene expression of the target algae based on the behavioral data of the target algae and the model. Effect of the Invention

[0010] According to the present disclosure, a phototaxis measurement system for measuring behavior data of algae can be provided, and a method for inferring phenotype, genotype, or gene expression of algae based on the measured behavior data of algae can be provided. [Brief description of the drawings]

[0011] [Figure 1] FIG. 1 shows a configuration diagram of an apparatus according to an embodiment. [Diagram 2] FIG. 2 shows a schematic diagram of the device of the embodiment. [Diagram 3] FIG. 3 shows an example of algae isolation location data. [Figure 4] FIG. 4 shows an example of the analysis data. [Diagram 5] FIG. 5 shows a process flow of the embodiment. [Figure 6] FIG. 6 is a diagram showing the hardware configuration. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] Hereinafter, embodiments of the present disclosure will be described with reference to the drawings.

[0013] 1 is a diagram showing an example of a phototaxis measurement system according to an embodiment of the present invention, which includes a culture chamber 1, a plurality of light irradiation devices 2, and a measurement device 3, and further includes an isolation device 4, an analysis device 5, and a calculation device 6.

[0014] The culture chamber 1 is a culture vessel that contains the algae to be measured and analyzed. In the present disclosure, "culture" refers to maintaining the algae in a living state, and does not necessarily require the algae to grow. The algae in this embodiment are typically non-sessile algae. The algae may be, for example, unicellular algae. The culture chamber may have different shapes, including tank-shaped, well-shaped, tube-shaped, etc., depending on the type of algae of interest and the individual implementation purpose, and may contain different aqueous media, including seawater and freshwater. Embodiments in which organisms other than algae and / or substances other than aqueous media (e.g., soil) are included in the culture chamber are also contemplated. The culture chamber 1 may include a general configuration for culturing algae, such as an environmental control unit for controlling environmental conditions such as temperature, a liquid supply unit, a liquid drainage unit, etc., as necessary. The volume of the culture chamber 1 may also be changed depending on the type of algae and the individual implementation purpose, and is not particularly limited. For example, the culture chamber 1 may be an aquarium, a petri dish, or a microfluidic device.

[0015] Each of the light irradiation devices 2 can irradiate light to the culture chamber and thus to the algae in the culture chamber. The light irradiation devices 2 can change the wavelength of the irradiated light, or the wavelength and intensity. Thus, each of the light irradiation devices 2 can qualitatively and / or quantitatively adjust the irradiated light, and as a whole, the light irradiation state in the culture chamber can be adjusted. Changing the intensity of the light can also include turning off the light. The light irradiated by each of the light irradiation devices 2 at a given time can be light of a single color, wavelength, or wavelength range, or can include multiple colors, wavelengths, or wavelength ranges. The wavelength of the light irradiated by each of the light irradiation devices 2 at a given time can include visible light, ultraviolet light, infrared light, or a combination thereof. The light emitted by each light irradiation device 2 at a given time may include, but is not limited to, wavelengths in the range of blue (e.g., 430 nm or more and less than 490 nm), green (e.g., 490 nm or more and less than 550 nm), yellow (e.g., 550 nm or more and less than 590 nm), or red (e.g., 640 nm or more and less than 770 nm). Each light irradiation device 2 may emit, for example, any of four colors of light, the three primary colors of light plus white, with variable intensity. For example, in one embodiment, each light irradiation device 2 has three light sources that can emit red, green, and blue light, respectively, and can also emit white light as a mixture of the above three colors. That is, for example, a total of four lighting devices 2 (first to fourth) are installed around the culture chamber on all four sides, and when it is desired to emit red light from the first lighting device 2, only the red light source is turned on, and when it is desired to emit white light from the first lighting device 2, all of the red, green, and blue light sources are turned on.

[0016] As described above, the system of the embodiment includes a plurality of light irradiation devices 2. The number of light irradiation devices 2 may be two, three, four, five, six, or seven or more. It is preferable that there are three or more light irradiation devices 2, and it is more preferable that there are four or more light irradiation devices 2. The wavelength of the irradiation light, or the wavelength and intensity, of the plurality of light irradiation devices 2 can be set and changed by a user of the system or, for example, by an automatic program, independently of each other. That is, the wavelength of the irradiation light, or the wavelength and intensity, of the plurality of light irradiation devices 2 can be changed independently of each other.

[0017] In the system of the embodiment, the plurality of light irradiation devices 2 are installed at different positions around the culture chamber 1 and configured to irradiate the culture chamber 1 with light. In a preferred embodiment, the plurality of light irradiation devices 2 may be installed at substantially equal intervals around the culture chamber 1 (in this case, the light sources of the plurality of light irradiation devices may be installed at substantially the same height). For example, it is preferable that the system includes at least four light irradiation devices, and the four light irradiation devices are arranged on all four sides of the culture chamber 1. The first and second light irradiation devices may be installed facing each other across the culture chamber 1, and the third and fourth light irradiation devices may also be installed facing each other across the culture chamber 1, and the line connecting the first and second light irradiation devices and the line connecting the third and fourth light irradiation devices may be substantially perpendicular to each other. In some embodiments, not only two-dimensional behavior data but also behavior data of algae in a three-dimensional space may be measured. For example, the system may include light irradiation devices above and below the culture chamber in addition to the at least four light irradiation devices (FIG. 2).

[0018] The light irradiated by the light irradiation device 2 may be emitted by, for example, a light emitting diode (LED) as a light source. One light irradiation device may include one light source (e.g., an LED element) or may include multiple light sources (e.g., LED elements). The multiple light sources (e.g., LED elements) may have the same light irradiation characteristics or may have different light irradiation characteristics. The light irradiation device 2 may include a lens for diffusing or concentrating the irradiated light. The light irradiation device 2 may include a general control circuit required for using the light source (e.g., an LED element). For example, the light irradiation device 2 may include a control circuit for adjusting the output of the light source (e.g., an LED element).

[0019] The light irradiation device 2 may include a xenon lamp as a light source instead of or in addition to an LED element.

[0020] The light irradiation device 2 may include a light source that emits light having a wavelength range or polychromatic light, and optionally a monochromator, an optical filter, or both. The type of monochromator and optical filter is not limited as long as they can adjust the wavelength of the light emitted from the light source before it is irradiated onto the algae.

[0021] The phototaxis measurement system of the embodiment can provide several excellent properties by varying the wavelength or wavelength and intensity of the light emitted by the multiple light irradiation devices 2. Phototaxis as well as other photoresponsive reactivity and photosensitivity (including metabolic photoresponsiveness and photosensitivity) may differ or change among algae. In view of these properties, the system of the embodiment separates algae that exhibit positive or negative or other specific phototactic behavior to light of various light qualities (including wavelengths, intensities, or both) from other algae in a given algae population. In addition, the system of the embodiment may further perform genetic analysis, etc., of the separated algae in some cases. By the separation or genetic analysis, etc., of the algae performed by the system of the embodiment, algae with specific properties or algae lacking specific properties in some cases can be efficiently isolated. In addition, the photoreceptors of known or new algae whose photosensitive mechanism is unknown can be identified by performing various light irradiation to examine the behavior of the algae and comparing it with the behavior data of other algae.

[0022] For example, unlike conventional devices that use a light source device with one type of wavelength from one direction to induce phototaxis in algae, the phototaxis measurement system of this embodiment can induce phototaxis in algae with green light while adjusting photosynthetic activity to an arbitrary state with red and blue light. In addition, it is possible to efficiently isolate useful algae by utilizing the differences in phototaxis exhibited by algae to light of various wavelengths and intensities, for example, by isolating new algae or mutant strains that are likely to survive in wild environments or specific artificial environments, such as those that exhibit positive phototaxis even under strong red light that causes stress. Furthermore, it is possible to perform phototaxis assays on new algae that have unknown photoreceptors, and to explore the photoreception mechanism.

[0023] In this disclosure, the term behavior data is defined to include one or more of the position, movement distance, movement speed, movement direction, movement trajectory, and density in a specified area of ​​the measurement object (algae). Measuring behavior data may include detecting behavior data or changes therein. In the simplest embodiment, the measurement of behavior data of the algae in the culture chamber may be performed with the naked eye. If the phototaxis uniformity of the algae population in the culture chamber is relatively high, it can be seen with the naked eye that the density of algae is high in a particular area in the culture chamber. Alternatively, the system may include a measurement device that facilitates the measurement of behavior data of the algae. The measurement device 3 is an artificial device for detecting behavior data or changes therein, and is configured to measure behavior data of the algae contained in the culture chamber. These data may be measured in the entire area or any area in the culture chamber. The measurement device 3 may be, for example, a microscope such as a stereo microscope, a time-lapse image acquisition device, or a combination thereof. The measurement device is preferably configured to capture images of the algae contained in the culture chamber. When the measuring device 3 is a time-lapse image acquisition device, the measuring device 3 includes a camera for taking pictures, i.e., acquiring image data. The camera may be, for example, a CCD camera. When the measuring device 3 is a time-lapse image acquisition device, the measuring device 3 periodically acquires image data in the culture chamber 1. The image data acquired by the measuring device 3 may be image data of algae at any focal plane in the culture chamber 1. Software capable of tracking particles such as cells on the image data to provide behavior data is known to those skilled in the art and is described, for example, in Reference 4. The measuring device 3 may include a storage device for saving, and possibly accumulating, the measured behavior data. The accumulated behavior data can be used for model construction, which will be described later.

[0024] The system of the embodiment may include an isolation device 4. In a simple embodiment, the isolation device 4 may include a pipette or micropipette that manually or semi-manually collects algae that have moved or concentrated in a specific position in the culture chamber due to a specific phototaxis. Alternatively, the isolation device 4 may be a mechanical isolation device that is incorporated into the culture chamber and collects the culture medium containing the algae from a specific position in the culture chamber by sucking, discharging, scooping, or in other ways. The isolation device 4 may include a recording unit that records isolation position data corresponding to the collected sample when the algae are collected.

[0025] 3 shows an example of the configuration of isolation position data recorded by the isolation device 4. The isolation position data may be represented, for example, in a table format, and may have columns such as a sample identification number, recording date and time, and coordinates x, y, and z in the culture chamber.

[0026] The analysis device 5 may perform one or more of phenotypic analysis, genotypic analysis, and gene expression analysis of the algae sample obtained from the isolation device 4. The phenotype refers to what kind and / or amount of gene products and / or metabolites the collected algae possesses at that time, and may include morphological or physiological information of cells, subcellular structures, or tissues that may change depending on the kind and / or amount of gene products and / or metabolites. Examples of phenotypes include, but are not limited to, cell and tissue morphology and subcellular structure morphology, metabolic state, amount of reactive oxygen species, stress resistance such as high light resistance, growth rate, phototaxis, and photosynthetic activity. The genotype refers to what gene sequence the collected algae possesses in its genome. The gene expression analysis refers to what kind and / or amount of genes the collected algae are transcribing at that time. Methods and devices capable of analyzing phenotype, genotype, and gene expression from a given biological sample are known to those skilled in the art. These analyses may include, for example, but are not limited to, mass spectrometry, chromatography, electrophoresis, antibody-based detection, enzyme assays, nucleic acid sequencing, microarrays, reverse transcription PCR, or combinations thereof.

[0027] 4 shows an example of the structure of the analysis data obtained by the analysis device 5. The analysis data may be represented, for example, in a table format, and may have columns such as a sample identification number, a recording date and time, and an analysis value a, an analysis value b, and an analysis value c for identifying one or more of a phenotype analysis, a genotype, and a gene expression.

[0028] The calculation device 6 calculates and provides a model relating the algae behavior data measured by the measurement device to one or more of the phenotype, genotype, and gene expression identified by the analysis device 5. To this end, the calculation device 6 obtains the algae behavior data from the measurement device 3 and data relating to one or more of the algae phenotype, genotype, and gene expression from the analysis device 5.

[0029] The calculation device 6 can estimate one or more of the phenotype, genotype, and gene expression of the target algae based on a model relating behavioral data of the target algae to one or more of the phenotype, genotype, and gene expression of the algae.

[0030] The calculation device 6 may have a function of controlling one or more of the culture chamber 1, the light irradiation device 2, the measurement device 3, the isolation device 4, and the analysis device 5, and may include a control unit for that purpose. The calculation device 6 may control environmental parameters such as temperature in the culture chamber 1. The calculation device 6 may control the wavelength, intensity, or both of the irradiated light in the light irradiation device 2. The calculation device 6 may control exposure, frame rate, and other parameters related to camera measurement in the measurement device 3. The calculation device 6 may control parameters such as sample acquisition position, sample acquisition amount, and sample acquisition frequency in the isolation device 4. The calculation device 6 may also control phenotype, genotype, or gene expression analysis of the analysis device.

[0031] <Estimation method>

[0032] In one embodiment, a method for predicting one or more of an algal phenotype, genotype, and gene expression is provided, the method including providing a model relating algal behavior data and one or more of a phenotype, genotype, and gene expression, measuring the behavior data of a target alga, and predicting one or more of a phenotype, genotype, and gene expression of the target alga based on the behavior data of the target alga and the model.

[0033] Although some behavioral data may not necessarily be related to phototaxis (e.g., behavioral data related to chemotaxis), an example of an estimation process performed using the phototaxis measurement system of the above-mentioned embodiment will be described here with reference to the flowchart in Fig. 5. Regarding the components including the culture chamber 1, the light irradiation device 2, the measurement device 3, the isolation device 4, the analysis device 5, the calculation device 6, and the algae, and the manner of use thereof, the explanations provided in the context of the above-mentioned system embodiment may be applied to the embodiment of this estimation method, and vice versa.

[0034] In step S1, an algae culture solution is prepared in the culture chamber 1. The culture solution may contain components such as a buffer solution, nutrients, a carbon source, amino acids, vitamins, minerals, and rare metals. The algae may be used after they have grown to a predetermined density in the culture chamber 1, or the algae in the culture chamber 1 may be diluted so that the density is suitable for measurement. In addition, the algae may be cultured in a separate container in advance and placed in the culture chamber 1. The type of algae cultured in the culture chamber 1 is not limited. For example, any of the classifications such as Chlorella, Euglena, Botryococcus, Aurantiochytrium, Chlamydomonas, blue-green algae, diatoms, xanthophytes, dinoflagellates, and seaweeds may be used as the algae in the culture chamber 1. The algae may be a single species of algae or a plurality of species of algae. A genetically homogeneous (pure line) population of algae of the same species may be used, or a population of algae having genetic variation may be used.

[0035] In step S2, the culture chamber 1, and in turn the algae in the culture chamber, are irradiated with light from multiple directions at multiple wavelengths or a combination of multiple wavelengths and multiple intensities by multiple light irradiators. The wavelengths of the irradiated light may include wavelengths (e.g., 400-700 nm) that include green light that is not absorbed by chlorophyll, or may not include such wavelengths. By adjusting the intensity of light at wavelengths used or not used in photosynthesis, it is possible to perform measurements while adjusting the photosynthetic activity of the algae. Here, the algae in the culture chamber 1 behave in accordance with the phototaxis of each individual, and the algae population shows a specific distribution pattern within the culture chamber 1.

[0036] In step S3, the measurement device 3 measures behavior data of the algae in the culture chamber 1. The algae to be measured may be algae that have received light of multiple wavelengths, intensities, or combinations of wavelengths and intensities, each irradiated from a different direction. The phototaxis measurement system of the embodiment has multiple light irradiating devices and the wavelength and intensity of the irradiated light are variable, making it possible to measure the algae that have received light of multiple wavelengths, intensities, or combinations of wavelengths and intensities, irradiated from such different directions. The behavior data may be captured in time-lapse image data of the algae. The measured behavior data may be accumulated and used to build a model in step S6, or the behavior data measured for the target algae may be used to estimate at least one of the phenotype, genotype, and gene expression in step S7.

[0037] In step S4, the isolation device 4 isolates the algae in the culture chamber 1. Isolation of the algae may be performed for each specific position in the culture chamber 1. The isolated algae for each position in the culture chamber 1 may be collected, for example, in a tube or plate to which an identification number is assigned. The assigned number may be a number representing a specific position of the multi-tube or multi-well plate, or may include an identification number for each of the multi-tube or multi-well plate. The collected algae sample is sent to an analysis device and used for downstream analysis. At this time, the isolation device 4 may transmit the identification number for each of the multi-tube or multi-well plate together with the algae sample to the analysis device 5. Isolation position data (an example is shown in FIG. 3) in which the identification number and the associated coordinates in the culture chamber 1 where the algae were isolated are linked is transmitted to the calculation device 6.

[0038] In step S5, the algal sample obtained from the isolation device 4 is subjected to one or more of phenotypic analysis, genotypic analysis, and gene expression analysis.

[0039] In step S5, the phenotypic analysis performed by the analysis device 5 may be an analysis of one or more phenotypes of the algae, such as morphology, metabolic state, amount of reactive oxygen species, stress resistance such as strong light resistance, growth rate, phototaxis, photosynthetic activity, etc. These phenotypes may be analyzed by an analysis or assay method known to those skilled in the art, and analysis data may be obtained. Examples of specific methods of phenotypic analysis include, but are not limited to, metabolic analysis (metabolome analysis), protein analysis (proteome analysis), and cell surface analysis using an electron microscope.

[0040] In step S5, the genotype analysis performed by the analysis device 5 may be a whole genome analysis by next-generation sequence analysis. Alternatively, the genotype analysis performed by the analysis device 5 may be a DNA sequence analysis targeting a specific gene. The target gene of the DNA sequence analysis is not limited, and may be a gene encoding a photoreceptor protein or a gene encoding a chlorophyll synthesis protein. The photoreceptor protein may be, for example, a chlorophyll-containing protein. The chlorophyll-containing protein may be a photosystem complex protein. Alternatively, the photoreceptor protein may be a channel rhodopsin or a cryptochrome. These DNA sequence analyses may be performed by methods known to those skilled in the art.

[0041] In step S5, the gene expression analysis performed by the analysis device 5 may be next-generation sequence analysis or transcriptome analysis by microarray. The subject of the gene analysis may be the whole or partial transcriptome, or may be limited to individual target genes. The target genes of the transcriptome analysis or individual gene expression analysis are not limited, and may be, for example, the above-mentioned genes encoding photoreceptor proteins, genes encoding chlorophyll-containing proteins, or genes encoding chlorophyll synthesis proteins. These transcriptome analyses are performed by methods known to those skilled in the art.

[0042] The data obtained by the analysis in step S5 is linked to the identification number and sent to the calculation device 6 (FIG. 4).

[0043] In step S6, the calculation device 6 calculates and provides a model that correlates the behavior of the algae with one or more of the phenotype, genotype, and gene expression. For this purpose, the calculation device 6 obtains behavior data of the algae for constructing the model from the measurement device 3, and data on one or more of the phenotype, genotype, and gene expression of the algae for constructing the model from the analysis device 5. The model is not particularly limited as long as it correlates the behavior of the algae with one or more of the phenotype, genotype, and gene expression, and can be appropriately determined by a person skilled in the art based on ordinary knowledge. For example, the model may be a machine learning model. More specifically, models such as random forest, support vector machine, neural network, general linear model, regularized linear discriminant analysis, regularized logistic regression, and Lasso (least absolute shrinkage and selection operator) regression can be used. To construct the model, known algorithms used for constructing machine learning models or other models can be used. From the constructed models, a model in which the value output by inputting data for verification into the model most closely matches the actual measured value can be selected and used as the optimal model.

[0044] In step S7, the calculation device 6 estimates one or more of the phenotype, genotype, and gene expression of the target algae based on the behavior data of the target algae and the model. The behavior data of the algae may be obtained directly from the measurement device 3, or behavior data input to the calculation device 6 in another way may be used. The model may be calculated in advance by the calculation device 6 itself, or may be derived from a separate calculation and input to the calculation device 6. The estimation of one or more of the phenotype, genotype, and gene expression of the target algae can be performed using a known method depending on the type of each model exemplified in this specification. The estimated phenotype, genotype, and gene expression of the target algae may usually be one or more of the phenotype, genotype, and gene expression exemplified as the phenotype, genotype, and gene expression used to construct the model in step S6.

[0045] The calculation device 6 described above may include, for example, a general-purpose computer system as shown in Fig. 6. The illustrated computer system includes a CPU (Central Processing Unit, processor) 901, a memory 902, a storage 903 (HDD: Hard Disk Drive, SSD: Solid State Drive, etc.), a communication device 904, an input device 905, and an output device 906. The memory 902 and the storage 903 are storage devices. In this computer system, the CPU 901 executes a predetermined program loaded on the memory 902, thereby realizing the functions of the calculation device 6.

[0046] The computing device 6 may be implemented in one computer or in multiple computers. The computing device 6 may also be a virtual machine implemented in a computer. The program of the computing device 6 may be stored in a computer-readable recording medium such as a HDD, an SSD, a Universal Serial Bus (USB) memory, a Compact Disc (CD), or a Digital Versatile Disc (DVD), or may be distributed via a network. The computer-readable recording medium is, for example, a non-transitory recording medium.

[0047] The present invention is not limited to the above-described embodiment, and various modifications are possible within the scope of the present invention.

[0048] <Additional Notes> (Appendix 1) A phototaxis measurement system comprising a culture chamber for accommodating algae and a plurality of light irradiation devices, the phototaxis measurement system enabling measurement of behavior data of the algae, A phototaxis measurement system in which the multiple light irradiation devices are capable of independently changing the wavelength or wavelength and intensity of the irradiated light, and are configured to be installed at different positions around the culture chamber and to irradiate light into the culture chamber. (Appendix 2) The phototaxis measurement system of claim 1, further comprising a measurement device configured to photograph the algae contained in the culture chamber and measure behavior data of the algae. (Appendix 3) 3. The phototaxis measurement system according to claim 2, wherein the measurement device is a time-lapse image acquisition device. (Appendix 4) (i) the plurality of light irradiation devices include at least four light irradiation devices, and the four light irradiation devices are installed on four sides of the culture chamber; or (ii) In addition to the at least four light irradiation devices of (i), light irradiation devices are further provided above and below the culture chamber; 4. A phototaxis measurement system according to any one of claims 1 to 3. (Appendix 5) A method for predicting one or more of an algal phenotype, genotype, and gene expression, comprising: providing a model relating algal behavior data to one or more of phenotype, genotype, and gene expression; Measuring behavior data of the target algae; The method includes estimating one or more of a phenotype, a genotype, and a gene expression of the target algae based on behavior data of the target algae and the model. (Appendix 6) The estimation method according to claim 5, wherein the behavioral data is behavioral data of algae exposed to light of multiple wavelengths or combinations of multiple wavelengths and intensities, each irradiated from a different direction. (Appendix 7) The estimation method according to claim 5 or 6, wherein the behavior data of the target algae is measured using a phototaxis measurement system according to any one of claims 1 to 4. (Appendix 8) The provided model comprises: Measuring behavior data of model-constructing algae using a phototaxis measurement system according to any one of appendices 1 to 4; analyzing one or more of the phenotype, genotype, and gene expression of the model algae; A model constructed by a method including: constructing a model based on behavior data of the model-constructing algae; and constructing a model based on one or more of a phenotype, a genotype, and a gene expression of the model-constructing algae. The estimation method according to any one of appendix 5 to 7. [Explanation of symbols]

[0049] 1. Culture chamber 2 Light irradiation device 3. Measuring Equipment 4. Isolation device 5 Analysis device 6 Calculation device

Claims

1. A phototaxis measurement system comprising a culture chamber for accommodating algae and a plurality of light irradiation devices, the phototaxis measurement system enabling measurement of behavior data of the algae, The plurality of light irradiation devices are capable of independently changing the wavelength of the irradiation light or the wavelength and intensity of the irradiation light, and are configured to be installed at different positions around the culture chamber to irradiate the culture chamber with light. Phototaxis measurement system.

2. The phototaxis measurement system according to claim 1 , further comprising a measurement device configured to photograph the algae contained in the culture chamber and measure behavior data of the algae.

3. The phototaxis measurement system according to claim 2 , wherein the measurement device is a time-lapse image acquisition device.

4. (i) the plurality of light irradiation devices include at least four light irradiation devices, and the four light irradiation devices are installed on four sides of the culture chamber; or (ii) In addition to the at least four light irradiation devices of (i), light irradiation devices are further provided above and below the culture chamber; 3. A phototaxis measurement system according to claim 1 or 2.

5. A method for predicting one or more of an algal phenotype, genotype, and gene expression, comprising: Providing a model relating algal behavior data to one or more of phenotype, genotype, and gene expression; Measuring behavior data of the target algae; The method includes estimating one or more of a phenotype, a genotype, and a gene expression of the target algae based on behavior data of the target algae and the model.

6. The estimation method according to claim 5 , wherein the behavior data is behavior data of algae exposed to light of a plurality of wavelengths or a combination of a plurality of wavelengths and a plurality of intensities irradiated from different directions.

7. The estimation method according to claim 5, wherein the behavior data of the target algae is measured using a phototaxis measurement system according to any one of claims 1 to 3.

8. The provided model comprises: Measuring behavior data of model-constructing algae using the phototaxis measurement system according to any one of claims 1 to 3; analyzing one or more of the phenotype, genotype, and gene expression of the model algae; A model constructed by a method including: constructing a model based on behavior data of the model-constructing algae; and one or more of a phenotype, a genotype, and a gene expression of the model-constructing algae. The estimation method according to claim 5.

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