KR20030019863A - Method for discriminating the viability of seeds using near infrared spectroscopy - Google Patents
Method for discriminating the viability of seeds using near infrared spectroscopy Download PDFInfo
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- KR20030019863A KR20030019863A KR1020020047566A KR20020047566A KR20030019863A KR 20030019863 A KR20030019863 A KR 20030019863A KR 1020020047566 A KR1020020047566 A KR 1020020047566A KR 20020047566 A KR20020047566 A KR 20020047566A KR 20030019863 A KR20030019863 A KR 20030019863A
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- 238000000034 method Methods 0.000 title claims abstract description 49
- 238000004497 NIR spectroscopy Methods 0.000 title claims abstract description 5
- 230000035899 viability Effects 0.000 title 1
- 238000004458 analytical method Methods 0.000 claims abstract description 6
- 230000035784 germination Effects 0.000 claims description 48
- 238000012360 testing method Methods 0.000 claims description 14
- 238000001228 spectrum Methods 0.000 claims description 9
- 230000007226 seed germination Effects 0.000 claims description 6
- 238000010191 image analysis Methods 0.000 claims description 5
- 238000009331 sowing Methods 0.000 claims description 4
- 238000010899 nucleation Methods 0.000 claims description 3
- 238000000985 reflectance spectrum Methods 0.000 abstract description 5
- 230000002159 abnormal effect Effects 0.000 description 6
- 241000220259 Raphanus Species 0.000 description 5
- 235000006140 Raphanus sativus var sativus Nutrition 0.000 description 5
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 4
- 235000013339 cereals Nutrition 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 238000000513 principal component analysis Methods 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 241000201976 Polycarpon Species 0.000 description 2
- 230000002745 absorbent Effects 0.000 description 2
- 239000002250 absorbent Substances 0.000 description 2
- 230000001066 destructive effect Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000012850 discrimination method Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000003703 image analysis method Methods 0.000 description 2
- 238000002329 infrared spectrum Methods 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- 239000005355 lead glass Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- HUJXHFRXWWGYQH-UHFFFAOYSA-O sinapine Chemical compound COC1=CC(\C=C\C(=O)OCC[N+](C)(C)C)=CC(OC)=C1O HUJXHFRXWWGYQH-UHFFFAOYSA-O 0.000 description 2
- 230000003595 spectral effect Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 125000003831 tetrazolyl group Chemical group 0.000 description 2
- 238000012549 training Methods 0.000 description 2
- 235000007319 Avena orientalis Nutrition 0.000 description 1
- 244000075850 Avena orientalis Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 238000002306 biochemical method Methods 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 230000003412 degenerative effect Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 239000003925 fat Substances 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000012567 pattern recognition method Methods 0.000 description 1
- 239000003348 petrochemical agent Substances 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
- G01N21/359—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light using near infrared light
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
- G01N21/3563—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light for analysing solids; Preparation of samples therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/02—Food
Abstract
Description
본 발명은 근적외선을 이용하여 건전종자와 퇴화종자를 판별하는 방법에 관한 것이다. 더욱 상세하게는, 본 발명은 종자를 한 알 단위로 근적외선 분광광도계에서 1100 - 2500nm의 파장으로 조사(照射)하여 그 반사파장(스펙트럼)을 얻고 파장을 분석하여 건전하게 발아하는 종자와 그렇지 않는 종자를 판별하는 방법에 관한 것이다.The present invention relates to a method for discriminating healthy seeds and degenerated seeds using near infrared rays. More specifically, the present invention is a seed seed grain irradiated at a wavelength of 1100-2500nm in a near-infrared spectrophotometer to obtain its reflection wavelength (spectrum) and to analyze the wavelength and to germinate seeds that are not well germinated. It is about how to determine.
통상 근적외선을 이용하여 농산물, 약품, 석유화학, 토양 등에서 화학적인 조성을 분석하는 방법이 많이 이용되고 있다. 특히 많은 종자, 즉 밀, 귀리, 벼, 옥수수 및 조에서 단백질, 지방, 탄수화물 및 섬유소 등의 화학적인 조성을 분석하는 방법으로 근적외선이 많이 이용되고 있다. 그러나 종자품질을 평가하기 위하여 생리적인 특질, 특히 종자의 발아력을 측정하기 위한 방법으로 근적외선을 이용한 예는 없다.In general, a method of analyzing the chemical composition of agricultural products, drugs, petrochemicals, soil, etc. using near infrared rays is widely used. In particular, many seeds, such as wheat, oats, rice, corn, and crude, have been widely used as a method for analyzing the chemical composition of proteins, fats, carbohydrates, and fiber. However, there is no example of using near-infrared rays as a method for measuring the physiological characteristics, in particular, the germination power of seeds to evaluate seed quality.
지금까지 종자의 발아력을 검정하는 방법에는 생화학적인 방법으로서 테트라졸리움 테스트(tetrazolium test)와 전기전도도 측정(conductivity test) 등이 있으며, 비 파괴적인 방법으로는 퇴화종자에서 페놀물질인 시나핀(sinapine)이나 혹은 아미노산이 누출되는 원리를 이용하는 방법 등이 있다.To date, the germination of seeds has been tested by biochemical methods such as tetrazolium test and conductivity test. Non-destructive methods are phenolic sinapine in degenerative seeds. Or using the principle of leaking amino acids.
이때 테트라졸리움 테스트는 효소의 활성을 측정하는 방법으로서 종자를 절단하거나 침지하여야 하는 등 과정이 복잡하고 많은 시간을 요할 뿐 아니라 결과의 해석도 어려운 단점이 있다.At this time, the tetrazolium test is a method for measuring the activity of the enzyme, the process is complicated and requires a lot of time, such as cutting or immersing the seed has a disadvantage of difficult interpretation of the results.
또한 전기전도도 측정은 종자를 침지한 후 퇴화종자의 파괴된 세포막에서 나오는 전해질을 측정하는 방법인데 종자를 한 알 단위로 측정하기 어려워 주로 많은 종자를 침지하여 검정해야 하는 단점이 있다.In addition, the conductivity measurement is a method of measuring the electrolyte from the decomposed seed cell membrane after immersing the seed, it is difficult to measure the seed by a single unit, there is a disadvantage that mainly by soaking many seeds to test.
또한 퇴화종자에서 시나핀이나 단백질이 누출되는 원리를 이용하는 방법은종자코팅을 통하여 누출되는 물질을 흡착하여 발색하도록 처리한 후 분리하는 것으로서, 비 파괴적이긴 하나 여러 가지 전처리과정이 필요하다.In addition, the method of leaking cinnapine or protein from degenerated seeds is treated by separating and adsorbing the leaking material through seed coating, which is non-destructive but requires various pretreatment processes.
그러므로 본 발명은 상기한 단점을 해결하기 위한 것으로서, 종자를 파종하기 이전에 건전종자와 퇴화종자를 비 파괴적으로 판별하는 방법을 제공하는데 그 목적이 있다.Therefore, an object of the present invention is to provide a method for non-destructively discriminating healthy seeds and degenerated seeds before sowing seeds.
도 1은 본 발명의 방법에 의해 분광광도계에서 근적외선을 종자에 조사하는 것을 나타내는 모식도,BRIEF DESCRIPTION OF THE DRAWINGS The schematic diagram which shows that a seed irradiates a near-infrared ray with a spectrophotometer by the method of this invention,
도 2는 AOSA 규정에 의한 무 종자의 정상발아(A), 비정상발아(B, C) 그리고 불발아(죽은 종자, D)를 구별하여 나타내는 예시도,Figure 2 is an illustration showing the normal germination (A), abnormal germination (B, C) and non-germination (dead seed, D) of the seedless according to the AOSA regulations,
도 3은 삼차원 PCA 방법으로 분리된 발아종자(+)와 불발아종자( □)의 반사 파장의 차이를 나타내는 예시도,3 is an exemplary view showing the difference between the reflection wavelengths of germinated seeds (+) and ungerminated seeds (□) separated by a three-dimensional PCA method,
도 4는 삼차원 PCA 방법으로 분리된 정상발아종자(+)와 비정상발아종자( □)의 반사 파장의 차이를 나타내는 예시도이다.Figure 4 is an exemplary view showing the difference between the reflection wavelength of the normal germination seed (+) and abnormal germination seed (□) separated by a three-dimensional PCA method.
이러한 목적을 달성하기 위해 본 발명은 종자를 파종하기 전에 건전종자와 퇴화종자를 비 파괴적으로 판별하는 방법에 있어서, 근적외선 분광광도계(near infrared spectroscopy)에서 종자를 한 알 단위로 각 종자에 대한 반사파장(reflectance spectrum)을 얻는 단계; 반사파장을 얻은 종자를 파종하여 각각의 종자에 대한 발아력 검사 단계; 조사된 반사파장과 발아력을 비교하여 종자를 그룹으로 나누어 영상분석법으로 반사파장의 차이를 분석하는 단계; 및 반사파장의 차이가 확인되면 미지의 시료에 대하여 반사파장을 얻고 종자의 생리적 품질을 부분최소자승 2법 또는 소프트 인디펜던트 모델링 오브 클래스 아날로기법을 이용하여 판별하는 단계를 포함하여 이루어지는 것을 특징으로 한다.In order to achieve the above object, the present invention provides a method for nondestructively discriminating healthy seeds and degenerated seeds before seeding the seeds, and reflecting wavelength of each seed in a single grain unit in near infrared spectroscopy. obtaining a reflectance spectrum; Seeding the seed obtained the reflected wavelength germination power test step for each seed; Comparing the irradiated reflected wavelength with the germination force and dividing the seeds into groups to analyze the difference of the reflected wavelengths by image analysis; And when the difference in the reflected wavelengths is confirmed, obtaining the reflected wavelengths for the unknown sample and determining the physiological quality of the seeds using the partial least squares 2 method or the soft independent modeling of class analog technique. .
이와 같이 본 발명의 목적은 종자를 파종하기 전에 건전종자와 퇴화종자를 비 파괴적으로 판별하는 데 있어, 한 알 단위로 근적외선인 1100-2500nm의 파장을조사(照射)하고, 그 반사파장(reflectance spectrum)을 각 종자에서 얻은 다음, 건전발아종자와 그렇지 않은 종자에 대한 근적외선 반사파장을 PCA(principle component analysis) 등의 영상분석법을 이용하여 분석하고 반사파장의 차이가 확인되면 미지의 시료에 대하여 반사파장을 얻고 종자의 생리적 품질을 부분최소자승 2법 또는 소프트 인디펜던트 모델링 오브 클래스 아날로기법을 이용하여 판별함으로써 달성된다.As described above, an object of the present invention is to non-destructively discriminate healthy seeds and degenerated seeds before sowing seeds, and irradiate wavelengths of 1100-2500 nm, which are near infrared rays, on a grain basis, and reflect spectrum thereof. ) Is obtained from each seed, and then the near-infrared reflected wavelengths of the whole germinated seeds and the non-seed seeds are analyzed by image analysis such as PCA (principle component analysis). And the physiological quality of the seeds is determined by using the least-square 2 method or the soft independent modeling of class analog technique.
도 1은 본 발명의 방법에 의해 분광광도계에서 근적외선을 종자에 조사하는 것을 나타내는 모식도이다.BRIEF DESCRIPTION OF THE DRAWINGS It is a schematic diagram which shows that a seed is irradiated with near-infrared in a spectrophotometer by the method of this invention.
본 발명에서 모노크로메이터(monochromator)는 입사광(入射光) 속에 있는 임의의 파장의 단색광만을 추출하는데 사용되는 단색화장치로서 NIR 소스, 구체(integrating sphere) 등과 함께 사용된다. 이에 의한 근적외선 분광광도계는 NIRSystem 5000(Foss Co.) 또는 동등 이상의 기기가 적절하다. 근적외선은 하부방향에서 수정유리판을 통과하여 상부방향으로 조사(照射)되는데, 각 종자의 반사파장의 오차를 줄이면서 정확한 수치를 얻기 위해서 각 종자는 수정유리판 위에서 항상 동일한 위치에 고정되도록 한다. 즉, 동종의 종자에서 동일한 면이 조사(照射)되도록 하는 것이 중요하다.In the present invention, a monochromator is a monochromator used to extract only monochromatic light of any wavelength in incident light, and is used with an NIR source, an integrating sphere, or the like. NIRSystem 5000 (Foss Co.) or equivalent instrument is suitable for the near infrared spectrophotometer. Near-infrared rays are irradiated upward through the crystal glass plate in the downward direction, so that each seed is always fixed at the same position on the crystal glass plate in order to obtain an accurate value while reducing the error of the reflection wavelength of each seed. That is, it is important to make the same surface irradiate in the same kind of seed.
이때 종자를 고정할 수 있는 장치는 두께가 약 2-4 mm인 6각형 철판(예컨대 각변이 7.5 x 5.5 x 5.5 x 2.5 x 2.5 x 4.0cm)을 사용하고, 중간부에 종자 크기의 구멍을 형성하여 유리판 위에 고정한다.At this time, a device capable of fixing the seed is a hexagonal iron plate having a thickness of about 2-4 mm (eg, 7.5 x 5.5 x 5.5 x 2.5 x 2.5 x 4.0 cm on each side), and forms a seed-sized hole in the middle part. To be fixed on the glass plate.
그러나 종자를 고정할 수 있는 장치는 종자의 종류에 따라 여러 가지로 변형하여 사용할 수 있다. 근적외선 분광광도계 역시 종자의 반사파장을 얻을 수 있는 여러 종류의 기기를 사용할 수 있다.However, the device that can fix the seed can be used in various ways depending on the type of seed. Near-infrared spectrophotometers can also be used with a variety of instruments to obtain the reflected wavelength of the seed.
본 발명의 첫 번째 단계로서, 상기한 6각형 철판의 구멍에 종자를 한 알씩 동일한 방향으로 넣은 후 종자의 반사파장을 얻도록 한다.As a first step of the present invention, seed seeds are put in the same direction one by one in the hole of the hexagonal iron plate to obtain the reflection wavelength of the seeds.
본 발명의 다음 단계로서, 상기 첫 단계를 거친 종자를 파종하고 실내에서 발아력 검사를 실시한다. 이때, 발아 및 불발아, 발아한 종자 중에 비정상묘나 정상묘의 구별은 미국 공식종자검사자협회(Association of Official Seed Analysis: AOSA)의 발아검사 규정을 적용한다. 즉, 11x11x4cm 크기의 플라스틱 용기에 10x10cm의 흡습지 2겹을 깔고 밑에서 계속 수분을 흡습하도록 하여 흡습지 위에 파종한다. 발아검사는 매일 실시하며, 더 이상 발아하지 않을 때의 발아상태를 최종발아로 보았다.As a next step of the present invention, the seed having passed through the first step is sown and germination tests are performed indoors. At this time, germination, ungermination, and germination of the seed or abnormal seed germination of the seed germination test rules of the Association of Official Seed Analysis (AOSA). That is, two layers of 10x10cm absorbent paper are placed in a 11x11x4cm plastic container, and the moisture is continuously soaked from the bottom to be seeded on the absorbent paper. The germination test was carried out daily, and the germination state when no longer germinated was considered as the final germination.
본 발명의 다음 단계에서 발아가 조사된 종자와 그 종자 각각의 근적외선 반사파장과의 관계를 분석하기 위하여서는 판별을 목적으로 하는 다변량 영상분석법들을 이용하였는데, 특히 1100-2500nm의 영역의 근적외선 스펙트럼을 주성분 분석법(principle component analysis: PCA)으로 만들어진 세 개의 주성분(principle component)의 축을 이용하여 삼차원으로 구도를 구성하고 각 시료들이 가장 잘 분리되는 조건을 찾아 판별하는 방법을 사용한다. 그 외 부분최소자승 판별법(discriminant partial least squares: PLS) 등으로 판별할 수도 있다.In the next step of the present invention, in order to analyze the relationship between the seed irradiated and the near infrared reflection wavelength of each seed, multivariate image analysis methods for the purpose of discrimination were used, in particular, the near-infrared spectrum of the region of 1100-2500nm. Three-dimensional composition is constructed using the axes of three principal components (principal component analysis, PCA), and the method of finding and determining the conditions under which each sample is best separated is used. In addition, it may be determined by discriminant partial least squares (PLS).
본 발명의 마지막 단계에서 미지의 종자의 발아력을 검정하기 위해서는 상기한 첫 단계에서처럼 종자의 근적외선 반사파장을 얻은 후 이미 분석 완료된 자료와비교하여 반사파장이 속하는 그룹을 찾아 컴퓨터를 이용하여 판별함으로써 매우 쉽게 종자의 특성을 판별할 수 있다. 상기 판별은 부분최소자승 2(Partial Least Squares 2; PLS2)법 또는 소프트 인디펜던트 모델링 오브 클래스 아날로기(Soft Independent Modeling of Class Analogy; SIMCA)법을 이용하여 이루어질 수 있다.In order to test the germination power of unknown seeds in the last step of the present invention, it is very easy to obtain the near-infrared reflection wavelength of the seed as in the first step and compare it with the data already analyzed to find the group to which the reflection wavelength belongs, Seed characteristics can be determined. The determination may be made using the Partial Least Squares 2 (PLS2) method or the Soft Independent Modeling of Class Analogy (SIMCA) method.
이와 같이 본 발명의 종자 발아력 판별방법은 향후 미지의 종자를 기계적으로 발아력을 판별하여 자동 선별하기 위한 매우 중요한 기초기술에 속하며, 종자를 전처리 하거나 기타의 어떠한 처리도 필요하지 않아 매우 간편하고 효율적이며 경제적인 방법인 것이다.As described above, the seed germination force discrimination method of the present invention belongs to a very important basic technology for automatically sorting and automatically sorting unknown seeds in the future, and does not require pretreatment or any other treatment, which is very simple, efficient and economical. It is a way.
이하, 본 발명의 구체적인 방법을 실시예를 들어 상세히 설명하지만 본 발명의 권리범위는 이들 실시 예에만 한정되는 것은 아니다.Hereinafter, the specific method of the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited only to these Examples.
실시예 1 : 근적외선 분광광도계를 이용한 발아종자와 불발아종자의 스펙트럼 파장간의 차이 확인Example 1 Confirmation of the Difference Between Spectral Wavelengths of Germinated Seeds and Ungerminated Seeds Using Near Infrared Spectrophotometer
1993년에 수확한 국내 재배종인 청수궁중 품종의 무 종자를 시료로 사용하였다. 무 종자의 근적외선 반사파장을 얻기 위하여 NIRSystem 5000(Foss Co.)의 근적외선 분광광도계에서 1100-2500nm의 파장영역을 2nm 씩 파장을 증가하도록 하여 전체 반사파장을 각각의 종자에서 얻었다. 이들 종자를 AOSA 발아검사 규정에 의한 방법으로 파종하여 발아 및 불발아, 또 발아된 종자 중에서 정상발아와 비정상발아를 구별하였다.(도 2)Radish seeds of Cheongsu Palace, a Korean cultivar harvested in 1993, were used as samples. In order to obtain near-infrared reflected wavelength of seeds, the total reflected wavelength was obtained from each seed by increasing the wavelength of 1100-2500nm by 2nm in NIRSystem 5000 (Foss Co.) near-infrared spectrophotometer. These seeds were sown by the method according to the AOSA germination test to distinguish between normal and abnormal germination among germinated and ungerminated seeds.
그 결과 총 571개의 무 종자에서 발아된 종자는 325개이었고, 발아되지 않은종자는 246개이었다. 발아된 종자 중에서 정상발아가 283개, 비정상발아가 42개로 구분되었다. 이러한 결과를 가지고 다변량 영상분석법의 하나인 주성분분석법(principle component analysis)을 이용하여 발아된 종자와 불발아된 종자의 반사파장을 분석하였다.As a result, a total of 325 seeds germinated from 571 radish seeds and 246 non-germinated seeds. Among the germinated seeds, normal germination was 283 and abnormal germination was 42. With these results, the reflected wavelengths of germinated and ungerminated seeds were analyzed using principal component analysis, one of multivariate image analysis.
분석 결과는 도 3과 같으며 파장의 그룹은 뚜렷한 차이를 나타냈다. 분석은 WIN ISI II프로그램으로 컴퓨터상에서 수행되었다. 이에 따라 발아되는 종자와 발아되지 않는 종자는 서로 다른 근적외선 반사파장을 나타냄을 알 수 있었다.The analysis results are shown in FIG. 3, and the group of wavelengths showed distinct differences. The analysis was performed on a computer with the WIN ISI II program. Accordingly, the germinated seeds and the non-germinated seeds exhibited different near infrared reflection wavelengths.
또한 발아된 종자 중에서 정상발아와 비정상발아에 대한 근적외선 반사파장은 도 4와 같이 매우 뚜렷하게 파장의 차이를 나타내어 종자의 생리적인 품질인 발아력을 근적외선의 반사파장 만으로 판별할 수 있다는 것을 확인하였다.In addition, the near-infrared reflected wavelengths for normal and abnormal germination among the germinated seeds showed very different wavelengths as shown in FIG. 4, and it was confirmed that the germination force, which is the physiological quality of the seeds, could be determined only by the reflected wavelength of near infrared rays.
즉, 종자의 생리적인 상태에 따라서 분광광도계의 특정 파장에서 흡수, 배음의 크기가 변화되므로 건전종자와 퇴화종자가 용이하게 판별된다.That is, since the absorption and harmonic amplitudes are changed at specific wavelengths of the spectrophotometer according to the physiological state of the seeds, healthy seeds and degenerate seeds are easily distinguished.
실시예 2 : 미지의 종자에 대한 발아력 판별 시험Example 2 germination power determination test for unknown seeds
근적외선 분광광도계를 이용하여 종자의 스펙트럼을 얻고 이들 스펙트럼에 대하여 다변량 영상분석법의 하나인 주성분분석법(principle component analysis)에서 발아종자와 불발아종자가 뚜렷이 파장의 차이가 나타나는 것을 확인하고 미지의 무 종자에 대하여 발아종자인지 불발아종자인지를 판별하는 시험을 하였다.Seed spectra were obtained using a near-infrared spectrophotometer, and the principal component analysis, which is one of the multivariate image analysis methods, showed that the germinated and non-germinated seeds showed distinct wavelength differences. A test was conducted to determine whether the seeds were germinated or non-germinated.
판별방법으로서는 스펙트럼 페턴 인식법인 Partial Least Squares 2(PLS2) 방법과 Soft Independent Modeling of Class Analogy(SIMCA)법을 이용하였다.Partial Least Squares 2 (PLS2) method and Soft Independent Modeling of Class Analogy (SIMCA) method were used for discrimination.
PLS2 방법은 WinISI II 프로그램을 이용하였고, SIMCA 법은 Chemo HN1100 프로그램을 이용하였다. 먼저 미지의 종자를 판별하기 위해서는 기지의 발아종자와 불발아 종자에 대한 스펙트럼의 특성을 상기 판별식에 의하여 검량식(모델)을 만든 후 이 검량식을 이용하여 미지의 종자가 발아종자인지 불발아 종자인지를 판별하게 된다. 검량식을 위하여 300개의 무 종자에서 한 알 단위로 스펙트럼을 측정하고 종자는 각각 파종하여 발아여부를 조사하였다. 측정된 스펙트럼은 미세구조를 들어나게 하기위한 방법으로 1차와 2차 미분하였고 미분하지 않은 스펙트럼(raw spectrum)을 포함하여 3개의 모델을 만들었다. 모델을 만드는 과정에서 스펙트럼 데이터로부터 기준치를 크게 벗어나는 시료들을 수회에 걸쳐 제거하고 판별력이 우수한 모델(training set)을 선정하여 판별에 이용하였다.The PLS2 method used the WinISI II program and the SIMCA method used the Chemo HN1100 program. First, in order to discriminate unknown seeds, a calibration formula (model) is created by the above-described discriminant for the known germinated seeds and ungerminated seeds. Determine if it is a seed. For calibration, the spectra were measured in units of 300 radish seeds, and each seed was sown and examined for germination. The measured spectra were first and second derivatives and three models including the raw spectra as a means of eliciting the microstructure. In the model making process, samples that deviated significantly from the spectral data were removed several times, and a training set with excellent discriminating power was selected and used for discrimination.
표 1은 PLS2 방법으로 판별한 결과이다. 미지의 시료를 판별대상(testing set)으로 하여 판별한 결과 스펙트럼을 미분할 수록 판별력의 정확도가 증가하였으나 83% 까지가 가장 우수한 정확도 이었다.Table 1 shows the results determined by the PLS2 method. As a result of discriminating unknown sample as a test set, the accuracy of discrimination power increased with differentiation of spectrum, but the best accuracy was up to 83%.
또 표 2 에서와 같이 SIMCA방법에 의하여 판별한 결과 1차미분한 스펙트럼을 이용할 경우 발아종자를 91%, 불 발아종자를 90%까지 판별할 수 있었다. 따라서 위의 두가지 판별방법 중에서 SIMCA 방법이 판별능력이 더 우수하였다.As shown in Table 2, as a result of the SIMCA method, when the first differential spectrum was used, it was possible to distinguish germination seeds by 91% and fire germination seeds by 90%. Therefore, the SIMCA method outperformed the two discrimination methods.
이와 같이 미지의 종자에 대하여 근적외선 스펙트럼으로 페턴인식법을 이용하여 판별식의 모델을 이용하면 종자의 발아능을 신속, 정확하고 비파괴적으로 구분할 수 있다는 가능성을 확인하였다. 또한 앞으로 실제로 적용하기위해서 많은 시료 량으로써 정확한 판별식(training set)을 만들어 많은 변량을 수용하는 정성적인 모델을 확보한다면 더 정확한 정확도로서 판별할 수 있을 것이다.In this way, it was confirmed that the seed germination ability can be quickly, accurately and non-destructively classified using the pattern recognition method using the Peton recognition method in the near-infrared spectrum of unknown seeds. In addition, if a qualitative model that accommodates a large number of variables is made by making an accurate training set with a large amount of sample for practical application in the future, it may be determined with more accurate accuracy.
이상의 실시예를 통하여 설명한 바와 같이, 본 발명은 종자를 파종 전에 건전종자와 퇴화종자로 판별하는 방법에 관한 것으로서 근적외선 반사파장이 발아력에 따라 다르게 나타남을 이용하여 종자의 발아력을 성공적으로 판별할 수 있는 방법을 제공할 수 있는 효과가 있다.As described through the above embodiments, the present invention relates to a method for discriminating between healthy seeds and degenerated seeds before sowing seeds, and the near-infrared reflected wavelength is different depending on the germination power, and thus the seed germination power can be successfully determined. It has the effect of providing a method.
또한 종래의 어떠한 방법보다도 비 파괴적이고 간편하며 경제적으로 신속하게 종자의 생리적인 품질을 판별하는 방법이며 향후 기계적으로 종자를 자동 선별할 수 있도록 하는 기초기술을 제공할 수 있어 종자산업 상 매우 유용하게 응용될 수 있다.In addition, it is a non-destructive, simple, and economical method to determine the physiological quality of seeds faster than any conventional method, and it is very useful for the seed industry because it can provide the basic technology to automatically sort seeds in the future. Can be.
본 발명은 기재된 실시예에 한정되는 것은 아니고, 본 발명의 사상 및 범위를 벗어나지 않고 다양하게 수정 및 변형할 수 있음은 이 기술의 분야에서 통상의 지식을 가진 자에게 자명하다. 따라서 그러한 변형예 또는 수정예들은 본 발명의 특허청구범위에 속한다 해야 할 것이다.It is apparent to those skilled in the art that the present invention is not limited to the described embodiments, and that various modifications and variations can be made without departing from the spirit and scope of the present invention. Therefore, such modifications or variations will have to belong to the claims of the present invention.
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| US9513217B2 (en) | 2010-04-13 | 2016-12-06 | Kyungpook National University Industry-Academic Cooperation Foundation | Non-invasive method and apparatus for screening high-quality seeds |
| KR101033440B1 (en) * | 2010-12-14 | 2011-05-09 | 경북대학교 산학협력단 | Non invasive methods and apparatus for detection of cucumber seeds contaminated with virus |
| KR101341815B1 (en) * | 2012-12-04 | 2014-01-06 | 대한민국 | Screening devices of seeds using hyperspectral image processing |
| KR101509052B1 (en) * | 2013-05-02 | 2015-04-08 | 대한민국 | Method of predicting seed viability |
| WO2016204350A1 (en) * | 2015-06-16 | 2016-12-22 | Republic Of Korea(Management : Rural Development Administration) | Method for measurement of preharvest sprouting damaged grain using any one or more of the total dissolved solids and electric conductivity in wheat |
| KR20210119927A (en) * | 2019-04-30 | 2021-10-06 | 한국식품연구원 | Prediction System of Germination Rate of Wheat and Prediction Method of Germination Rate of Wheat using the same |
| KR102291771B1 (en) * | 2020-06-23 | 2021-08-19 | 강원대학교산학협력단 | Non-destructive Determination Method For Plant Seed Germination And Growth Using Near-Infrared Absorption Spectrum |
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