JP2003023890A - Method for classifying plant individual by optical means and plant individual whose expressive character is classified by the optical means - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for classifying a plant individual by a non- destructive measurement method, and to provide the classified product. SOLUTION: This method for classifying the plant individual comprises applying a mating treatment, a radiation treatment, a chemical substance treatment or the like to a plant seed or vegetative organs to induce a genetic mutation, non-destructively measuring the seed or vegetative organs by an optical means for each individual, when a genetic character is expressed, and determining content ingredients from the analysis results of the optical spectra according to a computer program. Thus, the individual having the target specific ingredient can non-destructively be classified, and can be used as a kind and breeding raw material having the specific character.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for selecting plants, and more specifically to
The present invention relates to a method for detecting using a nondestructive measurement method and a selected plant body.

[0002]

2. Description of the Related Art Various methods have been adopted for large-scale selection of genetically mutated individuals of plants such as agricultural crops. As a method like this, it is possible to distinguish individuals with the naked eye, such as differences in color, height, shape, etc., and it is also possible to select the characteristics that change in association with the change. Targeted and widely used. However, these methods are limited to the properties that can be easily distinguished by the naked eye in the case of a large amount. For this reason, the content components of the plant body which are not apparently changed and the change thereof cannot be detected as they are, and it is the current situation that a large amount of samples cannot be selected except for the above properties. In particular, traits with a low frequency of appearance could not be considered as targets for breeding selection from the beginning. Furthermore, the precursors of the substances that are the criteria for content quality generated by processing and the like, or the target substances cannot be detected or quantified without complicated pretreatment and chemical analysis. The current situation is that it cannot be done.

Further, self-incompatible plants such as horseradish have a more difficult problem, it is difficult to produce a genetically fixed pure line, and genetically non-uniform seedlings obtained by natural mating are used. The method of using or using the vegetative propagation body of those seedlings is taken. Since there are changes in genetic traits for each individual or strain,
Individuals must be selected and the individuals evaluated for selection must be maintained in continuous growth. Therefore, when performing content component evaluation using chemical analysis and the like, the sample used for the analysis is quantitatively determined, it is difficult to maintain continuous growth of the individual, and complicated operations are required, At present, it is not possible to evaluate quickly and in large quantities, and it is not possible to make a selection based on the content components.

In order to solve the above problems, it is necessary to improve the efficiency so that a large amount can be selected.
However, if the target of breeding is seeds,
Since the genetic traits may change from seed to seed, it is necessary to select the seeds individually, and there is a problem that the seeds must be selected individually without changing the germination of the seeds. In addition, when the subject to be bred is a self-incompatible plant or its vegetative propagule, there is a change in genetic traits for each individual or line, so it is necessary to select them individually.
Further, there is a problem in that the plants must be individually sorted while maintaining a state in which they can be continuously grown.

Many studies have been carried out as a method of analyzing the content components of a plant by the nondestructive method. In particular, a method using light such as near-infrared rays is known, but a method for quantitatively selecting seeds and vegetative bodies at high speed for efficient breeding has not yet been solved.

[0006]

As described above, the conventional breeding methods for plant content components have their respective problems and are not satisfactory. In particular, in order to obtain an individual having a desired genetic trait by utilizing mutation,
It is necessary to target 100,000 to 10 million individuals. However, it was not possible to select a large number of individuals for mutation of plant content components unless there was an apparent change. For this reason, there has been a strong demand for a method of quickly and nondestructively and accurately selecting the difference in content component for each individual.

The present invention has been made from the above viewpoint, and an object of the present invention is to provide a method and a variety for efficiently selecting a genetic mutant, particularly a mutant strain caused by radiation.

[0008]

As a result of intensive studies to solve the above-mentioned problems, the present inventors have calculated the characteristics of reflected light, transmitted light, etc. obtained by irradiating a genetically-mutated individual with a computer. It was found that a variety having a desired content component and its change can be produced by applying a measurement method that analyzes the content at high speed by a program and reads quantitatively the content component and change accurately, and completed the present invention. It was

[0009] That is, the present invention is a breeding method relating to the content components of a plant in which genetically mutant individuals of a plant are selected for each individual from a nondestructive measurement method and a chemical quantification method using a computer program.

As a mode of the present invention, in the above-mentioned selection method, a nondestructive measurement method and a computer program are used to detect a difference in plant content components due to genetic mutation,
Provided is a method for producing a variety which is characterized by selecting a target individual. In the method of the present invention, seeds or trophozoites can be mentioned as the selection target of the genetically-mutated individuals. Further, the range to which the present invention is applied includes individual plants.

The method of inducing a genetic mutation in a plant used in the method of the present invention is not limited as long as it induces a genetic mutation, but radiation, preferably radiation,
Examples include X-rays and chemical substances. Of the radiation, γ rays, neutron rays, ion beams, and X rays are particularly preferable.

The optical measurement method used in the method of the present invention is
If a plant capable of non-destructive or germination and continuous growth is obtained, it is not particularly limited, but preferably visible light with little damage to the measurement object, near-infrared ray, and also for a short time, A measurement method using ultraviolet rays or fluorescent X-rays can be used. As a particularly preferable measuring method, it can be measured for each individual having a genetic mutation, and if it is a seed, the content component can be measured for each grain, a single grain component measuring method, and if it is a mutated vegetative body, the mutated part A two-dimensional measurement method capable of detecting can be exemplified.

By processing the optical information obtained by the optical measurement method of the present invention with a computer program, it is possible to provide a method for quantifying the content components contained in the plant body. The computer program is preferably a principal component analysis collectively referred to as a chemometrics method (chemometrics-new analytical chemistry, Aijima Zenro, Maruzen: 1992),
Cluster analysis, discriminant analysis, multiple regression analysis, principal component regression analysis, PLS (partial least squares) regression analysis, SIMCA
(Soft independent modeling of class analogy), KN
Examples include N (K-nearest neighbor method), neural network analysis, and the like, and a combination thereof.

In a computer program called data mining or Norwich Discovery System ([Special Issue] Knowledge Acquisition from Large Scale Database: Journal of Artificial Intelligence Vol. 12, No. 4), the target output is specified in advance. There are a goal-oriented type that refines the model by feeding back the calculation results, and a hypothesis discovery type that is suitable for extracting meaningful information from a large amount of discontinuous, multidimensional data. The hypothesis discovery type can be preferably used because it is easy to understand the basis of the output result.

[0015]

BEST MODE FOR CARRYING OUT THE INVENTION Embodiments of the present invention will be specifically described below.

(1) Plants to which the present invention can be applied In the method for selecting plants according to the present invention, in the case of seeds, the genetic mutation obtained by an operation such as radiation or mating is expressed as a gene after the second generation. Since the trait appears, it can be used in the second and subsequent generations. In the case of nutrition,
Mutants can be used in the first generation to obtain by vegetative propagation.

The plant that can be used as a seed is not particularly limited as long as it can be measured by an optical measurement method. The seeds whose content components can be quantified by near-infrared and computer spectrum analysis programs can be used particularly preferably. Many seeds wear seed coats, but these are
If germination is not affected, it can be removed and measured.

Specific examples of plants to be selected include seeds of cereals such as rice, wheat, beans, corn, rapeseed, and their vegetative bodies. Examples of fruits such as tomato and melon include germinated seeds and vegetative bodies. As for vegetables such as cabbage, Chinese cabbage, Japanese radish, broccoli, and cauliflower, germinated seeds and vegetative bodies can be mentioned. Further, regarding carnations, roses, margarets, chrysanthemums and other flowers, nutritional substances can be mentioned.

For example, in horseradish, content quality such as spiciness and flavor is more important than appearance quality such as size, color and shape. In particular, the precursor sinigrin, isothiocyanates changed by the enzyme tyrosinase in the process of processing, etc. are the most important pungency and flavor components, and the purpose is to select varieties and strains of higher quality depending on the content of these substances. And However, with wasabi, isothiocyanates are produced after processing such as grinding,
Isothiocyanates cannot be used as an index in selection by component analysis of nutrients and the like. Therefore, in the present invention, a correlation was found between the content of sinigrin as a precursor and the content of isothiocyanates generated after treatment such as grinding. Furthermore, a correlation was found between sinigrin in the wasabi leaf blade and the content of isothiocyanates in the rhizome after treatment such as grinding. Using near-infrared spectroscopy, measuring the sinigrin of rhizomes or leaf blades, found a method of estimating the pungency component of the rhizome, that is, isothiocyanates, from a mutant individual of wasabi obtained by treatment such as radiation and mating, An inventor has invented a method for efficiently selecting individuals containing a large amount of spicy ingredients.

(2) Kind of light used in the present invention and measuring method Next, an optical measuring method will be described. The light used in the present invention is preferably visible light, near infrared light, and infrared light, and for a short time, ultraviolet light, fluorescent X-rays, and the like are used for measurement. Since these lights have no or little damage to plants, they have little adverse effect on plant activities such as germination and can be used. In particular, near infrared rays are nondestructive and excellent in measurement, and can be preferably used for this purpose.

Further, the genetic mutation can be detected most efficiently by a device capable of measuring at a high speed for each seed in the case of seeds and for each individual in the case of trophozoites. Especially nutrient stems, leaves,
For the detection of genetically-mutated individuals in tissue culture, more efficient detection is expected by providing a device that can measure each individual continuously and at high speed. Particularly preferably, a method of continuously selecting rice, wheat grains, and beans one by one can be exemplified. For example, in horseradish, the part to be selected is a nutrient, that is, a rhizome,
Stems and leaves can be used, and the target component can be measured by using a horseradish nutrient itself, a rhizome slice collected to the extent that continuous growth is not impaired, a leaf blade separated from the rhizome, and a petiole.

In addition, particularly in the selection of vegetative bodies, a genetic mutation can be detected most efficiently without disturbing continuous growth by a device that can measure each individual in a field in a short time. Especially in the case of horseradish, a small and lightweight device (for example, the device shown in FIG. 5 or Fruit Nondestructive Quality Research Institute FT-
20) etc., the genetically-mutated individual can be preferably selected. In addition, in nutrient foliage and tissue culture,
In the detection of genetically-mutated individuals, by arranging them in a plane or in a line, by providing a device that can detect them,
More efficient detection is expected. In FIG. 5, reference numeral 1 is a light source, 2 is a spectroscope, 3 is a detector and an amplifier, and 4 is A.
D converter, 5 is a calculation and aggregation device, 6 is an optical fiber, and 7 is a sample to be measured.

(3) Computer program used for quantification Since the optically measured data contains information on the content components of the plant body, the optical data and multiple regression analysis method, principal component analysis method, cluster analysis method are used. , Discriminant analysis method,
By clarifying the relationship with the analysis results by the principal component regression analysis, PLS method, etc., which compensates for the drawbacks of the multiple regression analysis method,
A quantification method of the content component can be exemplified. Furthermore, even if seeds with the same trait are used for cultivated plants depending on soil conditions, weather conditions, irrigation water, cultivation conditions, fertilization conditions, etc., the components of the seeds and trophozoites make a large difference. , In order to read genetic variation, in addition to the conventional methods of linear statistical model, non-linear model, neural network model considering discontinuous and multi-dimensional effects, regularity is found in a large amount of data. Therefore, an estimation method using a hypothesis discovery type program that can improve measurement accuracy can be preferably used.

(4) Components and Reactions of Plants to be Measured In the measurement using near infrared rays, it is not limited as long as they are substances and reactions characterized by absorption and reflection characteristics of near infrared rays. Using the above computer program,
Sugars, acids, vitamins, amino acids, tannins, fatty acids, starch, nitrogen, phosphorus, potassium, sodium, magnesium and the like can be preferably measured. Similarly, with respect to horseradish, isothiocyanates such as allyl isothiocyanate, which is a substance that affects the content quality, and its precursor, sinigrin, can be preferably measured. With visible light and ultraviolet light, the turbidity inside the seed can be measured by absorption of transmitted light, so that it is possible to determine the stickiness and stickiness. Also,
It is possible to quantify or detect with high accuracy by treating a reagent that reacts with a specific substance that does not affect germination and measuring the specific absorption wavelength of the reagent by reflected light or transmitted light. It is expected that the measurement using fluorescent X-rays can measure metals higher than beryllium, and can measure the difference in reaction based on the product type. Further, for example, the antibody can be efficiently obtained by synthesizing the allergen substance based on the result of the amino acid sequence and injecting it into the blood of the animal. The purified antibody can be used for a quantitative assay with high accuracy.

(5) Method for selecting plants and method for producing varieties of the present invention In the method of the present invention, seeds or vegetative bodies used for producing varieties are treated with radiation or X-rays, mutagens such as chemical substances, Induces a genetic mutation in a plant. The amount of the mutagen to be treated may be such that a genetic mutation is induced, but it is preferably treated so that the germination rate or growth is 60 to 70% or more as compared with the untreated one.

The plant in which the genetic mutation is induced can be selected after the second generation since the seeds express the genetic characteristics in their progeny. For self-pollinating seeds, the second generation (M
2nd generation) and later can be preferably used. For selection, it is expected that the same genetic operation will be repeated after the third generation (M3 generation) and the target genetic mutation can be reliably obtained. Further, in the trophozoite, after the mutation treatment, the grown portion is selected. Also in this case, it is expected that the chimera can be eliminated early by repeating the selection.

Especially. For plants whose purpose is to produce fruits,
The culturable part that accompanies the target mutant individual obtained by selecting fruits that have undergone mutation treatment such as radiation or naturally occurring mutation branches (branch change) is used for tissue culture or cuttings. Can be preferably used to obtain a complete plant having the desired trait. Regarding the selection method using near infrared rays, for seeds having a low water content, reflected or transmitted light of near infrared rays in the vicinity of 1000 to 2300 nm can be preferably used. Elements such as lead sulfide and gallium arsenide can be preferably used as the light receiving element. An array in which gallium arsenide elements are laminated can be particularly preferably used because all wavelengths can be measured quickly.
In the selection method using visible light, the turbidity is measured using reflected light and transmitted light with wavelengths near 400 to 600 nm,
The degree of stickiness or stickiness of the seed can be determined. Further, in the case of brewed rice, the degree of occurrence of soothing white can be measured by the same method. When using fluorescent X-rays, 1 mm of the seed surface
It is expected that it will be possible to measure without obstructing the germination function of seeds, because it will be possible to measure each individual by carving out more than four sides.

The plant to which the present invention can be applied is not particularly limited as long as it is a plant that can be selected using the above-mentioned method, but as a target of the selection method using near infrared rays and fluorescent X-rays, rice is used. Dry seeds of wheat, barley, corn and soybean can be preferably used because their shapes are suitable for mechanical operations. As the vegetative body, leaves, stems and tissue cultures of tea, grapes, citrus fruits, pears and the like can be preferably used.

As described above, by analyzing the content components and reactions measured by various lights of plant individuals genetically mutated by the treatment with radiation using a computer program, the genetically mutated individuals can be efficiently treated. You can select well. As a result, varieties with the desired performance can be produced with good reproducibility.

Further, in the original seed cultivation, the purity of the variety is the most important. By comparing the variation due to the occurrence of mutation, the contamination of other varieties and the crossing with the undesired pollen with the characteristics of the spectrum stipulated in the variety. Can be eliminated. Similar sequence analysis (with IBM Intelligent Minor) is an excellent method for efficiently applying this method. Thus, in the present invention, in addition to utilizing the selection of plants for breeding, by detecting the optical spectrum showing the characteristics of the variety such as the original species management, prevent the mixed species, it is possible to obtain a highly pure variety. it can.

[0031]

EXAMPLES The principle of the present invention will be described prior to the description of specific examples. When a plant is irradiated with near infrared rays, light is scattered, reflected, and absorbed. Due to the absorption of light, a molecular normal vibration appears in a specific spectral range. For example, as the absorption wavelength of rice protein, 1656, 167
2, 2176, 1776, 2116 nm and the like are known. For these, a calibration curve can be created by clarifying the correlation with the actual measurement value by PLS regression analysis, multiple regression analysis, etc., and can be quantified accurately.
In the case of single grain analysis, the shape and weight are different, but by using the optical measurement array shown in the configuration specifications, which can measure the transmitted light of rice grains and measure them instantly, the accuracy is about ± 0.35%. It is possible to measure one grain at a speed of 2 seconds or less. The importance of the present invention is that even if there are very few mutations, mutations due to radiation or mutations that occur naturally have an extremely small probability, if the calibration curve is established, accurate selection is possible. There is a target point. Specific examples will be given below, but the present invention is not limited to these.

[0032]

[Example 1] (1) Name of measuring device Brown rice high-speed continuous component analyzer CTC-3 type (i) Outline of device This device irradiates brown rice single grain with near-infrared light and determines the brown rice from the transmission or reflection spectrum. It is a component analyzer that finds proteins.

(Ii) Block diagram The brown rice continuously supplied grain by grain from the sample supply unit is irradiated with light from a light source. Reflected light or transmitted light from the sample is collected by an optical fiber and introduced into a spectroscope. Inside the spectroscope, the incident light is separated into each wavelength by the spectroscope, and is incident on the linear array detector arranged at the emission part. The detection is 400 to 70 in the short wavelength region.
0 nm and 650 to 1050 nm are Si-CCD, and 1000 to 1600 nm and 1500 to 2300 in the long wavelength region
nm with four detectors of InGaAs respectively. The analog output from the detector is digitally converted by the A / D converter and read by the computer. The protein of the sample is measured based on the calibration curve stored in the database from the spectrum converted into the reflectance or the transmittance by comparison with the reference plate. The measured brown rice is sorted according to the hierarchy according to the protein measurement value and the preset rank setting, and stored in the tank. Figure 1 shows a block diagram of the brown rice high-speed continuous component analyzer.

[0034] (Iii) Configuration specifications   This case (sample supply unit / measurement unit / arithmetic control unit / sorting / storage unit)   Appearance: Desktop type power supply AC100V   Sample supply unit; within 1 grain / second 4kg storage                 If multiple grains are supplied, separate the samples or                 It should have a structure that returns to the supply section.   Measuring part; within 1 second / grain                 With a structure that allows continuous operation, correction of wavelength accuracy etc. can be done in a week                 Not more than once.                 The measurement can be either transmission or reflection.   Measurement wavelength: 400 to 700 nm / 650 to 1050 nm / 1000 to 16                 00nm / 1500-2300nm   Wavelength resolution: 5-20 nm   Data interval: 1 nm   Light source: halogen lamp   Condensing optical system; Optical fiber optical system   Spectrometer ； Diffraction grating spectrometer                 Spectrometer 1 Wavelength range 400-700nm                 Spectrometer 2 Wavelength range 650 to 1050 nm                 Spectrometer 3 Wavelength range 1000-1600nm                 Spectrometer 4 Wavelength range 1500-2300 nm   Detector: Detection element Si-CCD and InGaAs linear array                 Detector 1 wavelength range 400-700 nm 0.14 nm /                                       pixel                 Detector 2 wavelength range 650 to 1050 nm 0.19 nm                                       /pixel                   (Spectroscopes 1, 2; 2100 pixels, no electronic cooling)                 Detector 3 wavelength range 1000-1600 nm 2.34n                                       m / pixel                 Detector 4 wavelength range 1500-2300 nm 3.13n                                       m / pixel                   (Spectroscope 3, 4; 256 pixels, electronic cooling)   Arithmetic control unit; within 1 second / grain including measurement unit                 Able to easily set the rank of brown rice sorting based on the calibration curve.                 Be able to display the number of grains sorted by rank.                 Measured individual brown rice data (absorption spectrum, component value, classification                 Link) is stored in the storage device (HDD) and at the same time as measurement.                 Can be output to the outside.                 If the sample runs out, or if there is an abnormality such as a light source outage or a power outage,                 The system can be stopped automatically.   Sorting unit; within 1 grain / second                 Select from 5 or more levels determined in advance by protein value                 Possible.                 The sorting method and the sorting section have little damage to brown rice and impair germination.                 No structure.   Reservoir: Max. 4 kg   Overall sorting speed: within 2 seconds (all processes including sample supply, measurement and sorting)

(Iv) Software PLS regression analysis software for protein measurement (v) General specifications type; CTC-3 type target grain; Brown rice (quality; Hitomebore, Yamada Nishiki, Koshihikari) Measurement time: Within 2 seconds / grain (supply / Measurement, selection and storage) Measurement range: Protein 5-10% (dry matter) Measurement accuracy: Protein SEP ≤ 0.5% (when water content is 12-18%)

[0036]

[Example 2] Selection of wheat varieties with low allergens using near infrared rays (1) Test materials and methods Wheat varieties (normal, single grain, macaroni, club, cultivation emmer, Poland, rivet, sperta type) The allergen content of 370 samples in total, 3 to 6 grains per variety, was measured. The relative value (ELISA value) calculated from the colorimetric value due to the antigen-antibody reaction or the segment based on it is used as the target variable, and the analysis software uses Cleme.
Nine 5.2.1 (manufactured by SPSS) was used to predict the allergen content by the near infrared absorption spectrum. (2) The test result data was examined separately for the calibration curve and the test. In the prediction of the ELISA value using the neural network analysis, there was a tendency that the prediction was lower than the actual measurement value (see FIG. 2). In the segment prediction of the ELISA value by C5.0 (decision tree) analysis, the accuracy rate was 63 to 68%, but C5.
In the prediction combining 0 and the neural network analysis, it was possible to predict with a high probability in the category with a low allergen content (see the tables in FIGS. 3 and 4).

[0037]

[Example 3] Method for estimating allyl isothiocyanate content from sinigrin content in horseradish (1) Method (i) Test material Wasabi "Mazuma", "Shizu system No. 17" cultivated at wasabi branch of Shizuoka Agricultural Experiment Station , "D-52" and irradiated seeds M1 of these varieties. (ii) Analysis Method and Timing of Analysis Both the sinigrin content and the allyl isothiocyanate content were measured using HPLC. (2) Summary of results A strong correlation (correlation coefficient r = 0.961, 1% significant) was observed between the allyl isothiocyanate content after crushing wasabi rhizome and its precursor sinigrin content (see FIG. 6), It was clarified that the amount of allyl isothiocyanate can be estimated by measuring the content of sinigrin. Furthermore, there is a strong correlation between the content of sinigrin in leaf blades and the content of allyl isothiocyanate in rhizomes (correlation coefficient r = 0.7.
79 (1% significant) was observed (see FIG. 7), and it was clarified that the amount of allyl isothiocyanate after crushing rhizomes can be estimated from leaf blade sinigrin.

[0038]

[Example 4] Selection method of wasabi varieties with high sinigrin content using near infrared rays Generation of wasabi mutant individuals (1) Method (i) Test material 47 horseradish plants cultivated at Wasabi branch of Shizuoka Agricultural Experiment Station (“Mazuma”, “Amagi Midori”, “Static Line 13” and irradiated seeds M1 of these varieties) were harvested in January and August 1999 and used for the test. (ii) Measurement of near-infrared spectrum 5 mm below the wasabi rhizome to the extent that continuous growth is not hindered.
It was sliced into thick pieces, and the center part was hollowed out with a cork borer having a diameter of 17 mm. Place the wasabi rhizome in a reflective sample cell and use a near-infrared spectrophotometer (NIRSystems650
0) using the transmission reflection spectrum (400-2500n
m) was measured (integration number: 50 times). (iii) Measurement of sinigrin content Horseradish rhizomes were homogenized and then analyzed by high performance liquid chromatography (Shimadzu LC10-ATVP system). (iv) Preparation and evaluation of calibration curve: Using multiple regression analysis and PLS regression analysis as analytical methods, the calibration curve with the sinigrin content of the rhizome as the target variable and the absorbance at each wavelength of the second derivative spectrum as the explanatory variable is prepared. did. About 1/3 of all samples were used as unknown samples to evaluate the measurement accuracy of the calibration curve. (2) Summary of results (i) When 7 varieties were used as samples, as shown in Table 1, rhizome sinigrin content was 7.41 to 17.69.
mg / gF. W (0.74 to 1.77%) was a relatively wide range width.

[0039]

[Table 1]

(Ii) Multiple regression analysis In the original spectrum, large peaks at 1450 nm and 1950 nm, which are considered to be water absorption, were observed. The wavelength range where the correlation between the second derivative spectrum and the component concentration is high is 10
Wavelength exceeded. Therefore, when each wavelength was manually selected as the first wavelength and a calibration curve was created and evaluated, the highest accuracy in the calibration curve evaluation was 1628 at the first wavelength.
In the calibration curve using nm, the measurement error (hereinafter referred to as SEP) when evaluating the calibration curve was 1.35 (see Table 2 and FIG. 8A). 1628 nm is in the vicinity of 1626 nm, which is the attribution wavelength of a pure product of sinigrin, and SC = N or C in wasabi.
It was considered that the functional group of ═N was captured.

[0041]

[Table 2]

(Iii) PLS regression analysis SEP1.29 and a calibration curve with higher accuracy than the multiple regression analysis were obtained in the state where all wavelengths were analyzed with the same weighting (Weight). Furthermore, when the weighting of each wavelength was considered while considering the wavelengths attributed to sinigrin, 2222 nm
The calibration curve obtained by weighting 8 times in the vicinity and 2 times in the vicinity of 1650 nm had the highest accuracy, and the SEP was 1.03 (see Table 3 and FIG. 8B).

[0043]

[Table 3]

From the above results, it was clarified that the sinigrin content of horseradish can be measured by using near-infrared spectroscopy, and that it can be used for selection of wasabi having a lot of spicy components. In addition, multiple regression analysis and PLS regression analysis could be used as analysis methods.

[0045]

The characteristics of reflected light and transmitted light obtained by irradiating a genetically mutated individual with light are analyzed at high speed by a computer program, and by applying a measuring method for precisely reading the content components and changes, It was found that varieties with the intended content components and their changes can be produced.

[Brief description of drawings]

FIG. 1 is an explanatory diagram showing a block diagram of a brown rice high-speed continuous component analyzer.

[Fig. 2] ELISA by neural network analysis
It is a graph which shows the relationship between the predicted value of a value, and a measured value.

FIG. 3 is a table showing the prediction accuracy of ELISA values by C5.0 analysis.

FIG. 4 is a table showing prediction accuracy when a neural network analysis is performed after C5.0.

FIG. 5 is an explanatory view showing various embodiments of a measuring machine capable of measuring an individual in a field so as not to hinder the growth of the individual plant.

FIG. 6 is a graph showing the relationship between the content of sinigrin in the wasabi rhizome and the content of allyl isothiocyanate after crushing.

FIG. 7 is a graph showing the relationship between the sinigrin content of wasabi leaf blades and the content of allyl isothiocyanate after crushing rhizomes.

FIG. 8 is a graph (a) showing a relationship between an estimated value and a measured value of wasabi rhizome sinigrin content by multiple regression analysis, and P
It is a graph (b) which shows the relationship between the estimated value and the actual measurement value of the wasabi rhizome sinigrin content by LS regression analysis.

[Explanation of symbols]

1 light source 2 spectroscope 3 Detector and amplifier 4 AD converter 5 Calculation and aggregation device 6 optical fiber 7 Sample to be measured

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) G01N 21/33 G01N 21/33 21/35 21/35 Z 23/223 23/223 33/48 33/48 N (72) Inventor Hideyuki Matsuura 678-1 Tomioka, Toyota-cho, Iwata-gun, Shizuoka Inside Shizuoka Agricultural Experiment Station (72) Inventor Hiroshi Arakawa 678-1 Tomioka, Toyota-cho, Iwata-gun Shizuoka Prefecture Inside the Agricultural Experiment Station (72) Ina Takehiro 2860-25 Amagi Yugashima-cho, Takata-gun, Shizuoka Prefecture Wasabi Branch of Shizuoka Prefectural Agricultural Experiment Station (72) Inventor Takahiro Makino 678-1 Tomioka, Toyota-cho, Iwata-gun, Shizuoka Prefecture F-term (reference) 2B030 AA02 AB03 AD09 CA09 CA28 2G001 AA01 BA04 CA01 KA01 LA01 2G045 AA31 CB20 FA11 JA01 2G059 AA01 AA10 BB12 CC16 EE01 EE02 EE12 GG10 HH01 HH02 HH03 JJ05 JJ17 MM01 MM02 MM09 MM12 PP04

Claims (12)

[Claims] 1. A difference in content components caused by a genetic mutation and its change are measured by an optical method and quantified by a spectrum analysis program using a computer so that a desired genetic mutation can be obtained. A method for selecting plant individuals using an optical method, which is characterized by conducting a large amount of high-speed survey and selecting each plant individual. 2. The individual plant to be selected is a plant whose content component largely affects quality, or a plant whose precursor substance is changed to another substance at the time of use and whose substance amount largely affects quality. Alternatively, such as vegetatively propagating plants, self-incompatible plants, or plants obtained by natural crossing thereof, or vegetative propagules thereof, there is a change in genetic traits from individual to individual or lineage. The method for selecting plant individuals using the optical method according to claim 1, wherein the plant is a plant that needs to be selected based on its components and that must maintain continuous growth of the selected individuals. 3. An optical method for measuring the content components of the plant individual and changes thereof is a technique that extremely suppresses adverse effects on the activity of the plant such as germination, and the plant individual is quantitatively analyzed. The method for selecting plant individuals using the optical method according to claim 1 or 2, wherein the selection is carried out hierarchically based on information. 4. The optical method for measuring the content component of the plant individual and its change is near infrared, visible light, ultraviolet ray, X-ray.
A method for selecting plant individuals using the optical method according to claim 1, 2 or 3, which is an optical measurement method using one or a plurality of X-rays and fluorescent X-rays in combination. 5. The genetic mutation induced in the plant individual,
5. The optical method according to claim 1, 2, 3 or 4, wherein at least one of radiation, X-rays, chemical substances, naturally occurring mutations and mutations due to mating is used as a mutagen. How to select individual plants. 6. The chemometrics method capable of creating an accurate quantitative calibration curve is applied to a computer program for analyzing the content component of the plant individual and its change. A method for selecting plant individuals using the optical method described in 3, 4, or 5. 7. When the plant individual is cultivated under discontinuous conditions with different cultivation environments such as variety, region, place, meteorological factor, fertilization condition, cultivation year, etc., and is sample data having multidimensional information, , The hypothesis-discovery type data mining and Norwich Discovery system, which can detect the regularity from a large amount of data and measure it with higher accuracy, are applied to the computer program that analyzes the content components of plant individuals and their changes. 7. A method for selecting plant individuals using the optical method according to claim 6. 8. When selecting a large amount of said plant individuals, information obtained by a non-destructive measurement method such as an optical method is used to utilize lateral buds of vegetatively propagating plants or the bottom of rhizomes. The test individual is selected by utilizing a part of the edible portion by a method of continuously growing the test individual as it is, according to claim 1, 2, 3, 4, 5, 6 or 7. A method for selecting plant individuals using an optical method. 9. When quantifying the content component of the plant individual and its change, a quantitative determination of a gene product based on the DNA analysis result and a substance synthesized based on the result of the amino acid sequence, such as by spectroscopic method, 9. The plant individual selection using the optical method according to claim 1, characterized in that genetic information of the plant individual based on a qualitative method is used. Method. 10. In selecting the plant individual, a precursor substance in the plant is changed to another substance during processing and utilization, and the amount of the substance is a target substance having a great influence on the quality.
Measured in the form before processing and utilization, and estimated from the amount of the substance that has a high correlation with the production amount of the precursor substance or the target substance,
The method for selecting individual plants using the optical method according to claim 1, 2, 3, 4, 5, 6, 7, 8 or 9, wherein the individual is examined and selected. 11. In selecting the plant individual, the amount of substance produced at a target site for breeding is estimated from the amount of substance in a part of the tissue of a plant that has a high correlation with this,
A method for selecting a plant individual using the optical method according to claim 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, wherein the individual is investigated and selected. 12. A method for measuring differences and changes in content components caused by genetic mutation by an optical method and quantifying by a spectrum analysis program using a computer,
The method according to any one of claims 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11 so that the desired genetic mutation can be obtained.
A plant individual whose phenotypic trait has been selected by an optical method characterized by being selected by the selection method described.