CN108226091A - A kind of NIRS model building methods for being used to predict cane sugar content in corn - Google Patents
A kind of NIRS model building methods for being used to predict cane sugar content in corn Download PDFInfo
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- CN108226091A CN108226091A CN201711262538.XA CN201711262538A CN108226091A CN 108226091 A CN108226091 A CN 108226091A CN 201711262538 A CN201711262538 A CN 201711262538A CN 108226091 A CN108226091 A CN 108226091A
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Classifications
-
- 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
- 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
- G01N21/3577—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light for analysing liquids, e.g. polluted water
Abstract
The invention discloses a kind of for predicting the NIRS model building methods of cane sugar content in corn, include the following steps：1) collection of sample near infrared spectrum：Obtain its near infrared light spectrum information；2) preparation of enzyme process sample liquid；3) measure of sample sucrose：Obtain sample chemical value；4) NIRS is modeled：The scatter correction and Mathematical treatment of spectrum are carried out to sucrose calibration model.The present invention uses the chemical score of infrared spectrum analysis combination enzymatic assays for cane sugar content in corn, pass through the scatter correction and Mathematical treatment of spectrum, the analysis statistics of Chemical Measurement has been carried out to full spectral band, build the NIRS models of cane sugar content, it can be used for the content of sucrose in fast prediction corn, and then provide valuable reference with quality breeding to accelerate the high-quality Screening of Germplasm of corn.
Description
Technical field
The present invention relates to field of plant breeding, the detection of the cane sugar content in more particularly to a kind of corn.
Background technology
Near-infrared spectral reflectance (NIRS) technology radiates sample by near-infrared light beam, collected light beam and responds back
Come reflectivity or transmissivity, basic principle be：Contain band hydrogen group (X-H) or carbon-carbon double bond in tested organic object
(C-C) etc. chemical bonds, under the irradiation of near-infrared light beam general frequency vibration or rotation can occur for these chemical bonds, then with diffusing reflection side
Formula obtains the absorption spectrum near infrared region.Different substance chemical compositions contains different hydric groups in sample, such as starch, sugarcane
Sugar, glucose, fructose and water-soluble polysaccharide etc. all have different hydric group.These different hydric groups are in near-infrared
Spectral regions can generate corresponding specific absorption spectrum characteristic strip, then by advanced mathematical method, be built using computer
The correlation models between sample spectrum and its related component content are played, so as to fulfill it is only necessary to be searched using near infrared spectrometer
Collection obtains relevant spectral information can just go out the constituent content of sample to be tested by model calculation.It is compared to traditional chemistry
Method, NIRS technologies are with finding speed is fast, sample preparation is simple, does not expend chemical reagent, is easy to operate, is at low cost, without dirt
The advantages that dye.NIRS technologies can not only analyze the ingredient related with hydric group such as protein, fat, starch, amino acid etc.,
Size of the density of substance, viscosity, granularity etc. can also be analyzed.With instrument hardware technology constantly improve and Chemical Measurement
The development of software, NIRS technologies are applied successfully to the quantitative and qualitative research in each field such as food, drug, agricultural product.In recent years
Application of the NIRS technologies in plant breeding and research of fruit germplasm resource has become an active research field.
Corn contains relatively rich sugar, food fiber, trace element and vitamin, have higher edible, nutrition,
Economy and value added.In the developed countries such as American-European, South Korea and Japan, corn is as one of main vegetables.China is to sweet tea jade
Rice consumption demand amount is also being continuously increased, and corn consumption market and development prospect possess huge potentiality.In recent years, China has been
Grown worldwide corn big country ranks are entered, China in 2015 becomes the second big country of world's planting of sweet corn, and corn is deep
Fabricated product is without exception various, and exports international market, and corn industry is gradually grown.
According to the difference of mutator, corn is divided into three basic forms of it：Super sweet tea type, general sweet tea type and reinforcement sweet tea type.It is general
Sweet tea type corn is about containing 10% soluble sugar and 35% water-soluble polysaccharide, but its quality decline is quickly, and shelf life is very short；It is super
Sweet tea type corn contains 25~30% high-content sugar, and content of starch is relatively low, can still be kept for a long time without water-soluble polysaccharide
Sweet taste, therefore it possesses longer harvest time and shelf life, can greatly reduce the loss of crop.And strengthen sweet tea type sweet tea jade
The sugar content of rice is higher by one times or more (20%~35%) than general sweet tea type, and water-soluble more contents are also very high, but quality
Also decline quickly as general sweet tea type.
Sugar content and composition are a key factors for influencing corn quality.Corn sugar includes soluble sugar
(fructose, glucose, sucrose), starch and water-soluble polysaccharide.The sugar amount containing solubility of seed is higher, correspondingly its sense organ sugariness
Will be stronger, often the Sweet corn variety of sweet tea more it can more be liked by masses.At present, the bottleneck of South China's breeding of sweet
Good kind is a lack of, the premise for solving quality breeding key is that the index of quality is quantified, and establishes quick and easy mirror
Determine appraisement system, then filter out good resource for genetic improvement.The method for measuring master of corn sugar content at present
If using chemical detection method for example high performance liquid chromatography, saccharometer method, anthrone colorimetry, 3,5- dinitrosalicylics acid system,
Enzymatic isolation method etc., but the shortcomings of these methods pollute environment there are of high cost, and operating technology requirement is high.Therefore it studies quick and easy
Accurate corn sugar detection method all has a very important significance Sweet Corn Germplasm Screening germplasm and breeding.
Invention content
It is an object of the invention in for the above-mentioned prior art for the deficiency of sugar content assay method in corn,
A kind of NIRS model building methods for being used to predict cane sugar content in corn are provided, it can be quickly to cane sugar content in corn
It is measured, and then reference is provided with quality breeding to accelerate the high-quality Screening of Germplasm of corn.
The technical solution used in the present invention is：A kind of NIRS model construction sides for being used to predict cane sugar content in corn
Method includes the following steps：
1) collection of sample near infrared spectrum：
Corn entire kernel 120~400 part works different and representative by choosing kind after pedigree method selection and breeding
For sample, sample is scanned and obtains its near infrared light spectrum information；
2) preparation of enzyme process sample liquid：
The milled 60 mesh sieve of corn sample, accurately weighs 0.5g dry sample powder and is put into 10mL centrifuge tubes, add in 4.5mL
Ultra-pure water, shake 30min, then 5000r/min centrifugal treating 30min using blending instrument, take 3mL supernatants in 10mL centrifuge tubes
And the abundant mixing of 7mL ultra-pure waters is added in, the rear accurate 0.1mL that measures adds in the dilution of 2.9mL ultra-pure waters, and the dilution is as enzyme process sample
Liquid；
3) measure of sample sucrose：
Enzyme process sample liquid obtained by 100 μ L steps 2) are added in 10mm quartz colorimetric utensils adds the fructosidase of 200 μ L, uses
Preservative film covers cuvette, is gently mixed, and places 5min at room temperature, sequentially adds 1900 μ L distilled water, 100 μ L imidazole buffers
The NADP of liquid, 100 μ L+With ATP mixed solutions in than ware, cuvette is covered with preservative film, is gently mixed, is placed at room temperature
3min reads absorbance A b at the 340nm of ultraviolet-uisible spectrophotometer1, the rear hexokinase and glucose for adding in 20 μ L
Dehydrogenase suspension covers cuvette with preservative film, is gently mixed, place 5min at room temperature, in UV, visible light point to cuvette
Absorbance A b is read at the photometric 340nm of light2, sucrose in each sample is calculated according to K-SUFRG_CALC data processing methods
Content, as sample chemical value；
4) NIRS is modeled：
Sample chemical value and step 1) gained sample near infrared light spectrum information according to obtained by step 3), it is fixed that each sample is divided into
Mark collects and verification collection, calibration collection establish calibration model using MPLS methods, verify the confidence level of set established model for verifying,
The scatter correction and Mathematical treatment of spectrum are carried out to calibration model, obtains cane sugar content calibration model.
As being further improved for said program, the scatter correction sum number of spectrum is carried out described in step 4) to calibration model
It is to combine to remove scatter correction of the scattering processing as spectrum using standard normalization to learn processing, using 3 order derivatives as mathematics at
Reason, derivative processing gap are 6, and smoothing processing wavelength points are 6, and cross validation standard deviation is 1.508, cross validation phase relation
Number is 0.820, and external certificate standard deviation is 1.880.
As being further improved for said program, the prediction related coefficient of cane sugar content calibration model described in step 4) is
0.891。
The beneficial effects of the invention are as follows：The present invention uses infrared spectrum analysis desmoenzyme for corn entire kernel sample
The chemical score that method measures, by the scatter correction and Mathematical treatment of spectrum, has carried out Chemical Measurement to full spectral band and has divided
Analysis statistics builds the NIRS models of cane sugar content, can be used for the content of sucrose in fast prediction corn, and then to accelerate
The high-quality Screening of Germplasm of corn provides valuable reference with quality breeding.
Specific embodiment
The present invention is specifically described with reference to embodiment, in order to technical field personnel to the present invention
Understand.It is necessary to it is emphasized that embodiment is only intended to, the present invention will be further described herein, it is impossible to be interpreted as to this
The limitation of invention protection domain, fields person skilled in the art, the non-intrinsically safe made according to foregoing invention content to the present invention
The modifications and adaptations of property, should still fall within protection scope of the present invention.Mentioned raw materials following simultaneously are unspecified, are
Commercial product；The processing step or preparation method not referred in detail be processing step known to a person skilled in the art or
Preparation method.
A kind of NIRS model building methods for being used to predict cane sugar content in corn, include the following steps：
1) collection of sample near infrared spectrum：
Corn entire kernel 120~400 part works different and representative by choosing kind after pedigree method selection and breeding
For sample, sample is scanned and obtains its near infrared light spectrum information.
2) preparation of enzyme process sample liquid：
The milled 60 mesh sieve of corn sample, accurately weighs 0.5g dry sample powder and is put into 10mL centrifuge tubes, add in 4.5mL
Ultra-pure water, shake 30min, then 5000r/min centrifugal treating 30min using blending instrument, take 3mL supernatants in 10mL centrifuge tubes
And the abundant mixing of 7mL ultra-pure waters is added in, the rear accurate 0.1mL that measures adds in the dilution of 2.9mL ultra-pure waters, and the dilution is as enzyme process sample
Liquid.
3) measure of sample sucrose：
Enzyme process sample liquid obtained by 100 μ L steps 2) are added in 10mm quartz colorimetric utensils adds the fructosidase of 200 μ L, uses
Preservative film covers cuvette, is gently mixed, and places 5min at room temperature, sequentially adds 1900 μ L distilled water, 100 μ L imidazole buffers
The NADP of liquid, 100 μ L+With ATP mixed solutions in than ware, cuvette is covered with preservative film, is gently mixed, is placed at room temperature
3min reads absorbance A b at the 340nm of ultraviolet-uisible spectrophotometer1, the rear hexokinase and glucose for adding in 20 μ L
Dehydrogenase suspension covers cuvette with preservative film, is gently mixed, place 5min at room temperature, in UV, visible light point to cuvette
Absorbance A b is read at the photometric 340nm of light2, sucrose in each sample is calculated according to K-SUFRG_CALC data processing methods
Content, as sample chemical value, cane sugar content calculation formula is as follows：
ΔAGlucose in urine=(Ab2-Ab1)Sample-(Ab2-Ab1)Blank；
Concentration calculates as follows：
The last volumes of V=(mL)
MW=detects the molecular weight (g/mol) of substance
Extinction coefficients of the ε=NADP at 340nm
=6300 (1 × mol-1×cm-1)
D=light paths (cm)
V=sample volumes (mL)
4) NIRS is modeled：
Sample chemical value and step 1) gained sample near infrared light spectrum information according to obtained by step 3), it is fixed that each sample is divided into
Mark collects and verification collection, calibration collection establish calibration model using MPLS methods, verify the confidence level of set established model for verifying,
The scatter correction and Mathematical treatment of spectrum are carried out to calibration model, obtains cane sugar content calibration model.
Preferred embodiment is further used as, carries out the scatter correction sum number of spectrum described in step 4) to calibration model
It is to combine to remove scatter correction of the scattering processing as spectrum using standard normalization to learn processing, using 3 order derivatives as mathematics at
Reason, derivative processing gap are 6, and smoothing processing wavelength points are 6, and cross validation standard deviation is 1.508, cross validation phase relation
Number is 0.820, and external certificate standard deviation is 1.880.
Preferred embodiment is further used as, the prediction related coefficient of cane sugar content calibration model described in step 4) is
0.891。
Above-described embodiment is the preferred embodiment of the present invention, all with similar technique of the invention and the equivalence changes made,
The protection category of the present invention should all be belonged to.
Claims (3)
1. a kind of NIRS model building methods for being used to predict cane sugar content in corn, which is characterized in that include the following steps：
1) collection of sample near infrared spectrum：
By being chosen after pedigree method selection and breeding, kind is different and 120~400 parts representative of corn entire kernel is used as sample
Product are scanned sample and obtain its near infrared light spectrum information；
2) preparation of enzyme process sample liquid：
The milled 60 mesh sieve of corn sample, accurately weighs 0.5g dry sample powder and is put into 10mL centrifuge tubes, add in the super of 4.5mL
Pure water shakes 30min, then 5000r/min centrifugal treating 30min using blending instrument, takes 3mL supernatants in 10mL centrifuge tubes and add
Enter the abundant mixing of 7mL ultra-pure waters, the rear accurate 0.1mL that measures adds in the dilution of 2.9mL ultra-pure waters, and the dilution is as enzyme process sample liquid；
3) measure of sample sucrose：
Enzyme process sample liquid obtained by 100 μ L steps 2) are added in 10mm quartz colorimetric utensils, adds the fructosidase of 200 μ L, and use is fresh-keeping
Membrane cover lives cuvette, is gently mixed, and places 5min at room temperature, sequentially adds 1900 μ L distilled water, 100 μ L imidazole buffers, 100
The NADP of μ L+With ATP mixed solutions in than ware, cuvette is covered with preservative film, is gently mixed, places 3min at room temperature, in
Absorbance A b is read at the 340nm of ultraviolet-uisible spectrophotometer1, the rear hexokinase for adding in 20 μ L and glucose dehydrogenase are hanged
Supernatant liquid covers cuvette with preservative film, is gently mixed, place 5min at room temperature, in ultraviolet-uisible spectrophotometer to cuvette
340nm at read absorbance A b2, cane sugar content in each sample is calculated according to K-SUFRG_CALC data processing methods, is made
For sample chemical value；
4) NIRS is modeled：
Sample chemical value and step 1) gained sample near infrared light spectrum information, are divided into calibration collection by each sample according to obtained by step 3)
Collect with verification, calibration collection establishes calibration model, the confidence level of verification set established model for verifying, to fixed using MPLS methods
Scatter correction and Mathematical treatment that model carries out spectrum are marked, obtains cane sugar content calibration model.
It is 2. according to claim 1 a kind of for predicting the NIRS model construction sides of glucose and fructose content in corn
Method, it is characterised in that：Described in step 4) to calibration model carry out spectrum scatter correction and Mathematical treatment be using standard just
Normalizing combines the scatter correction for going scattering processing as spectrum, and using 3 order derivatives as Mathematical treatment, derivative processing gap is
6, smoothing processing wavelength points are 6, and cross validation standard deviation is 1.508, and cross validation related coefficient is 0.820, external certificate
Standard deviation is 1.880.
It is 3. according to claim 1 a kind of for predicting the NIRS model construction sides of glucose and fructose content in corn
Method, it is characterised in that：The prediction related coefficient of cane sugar content calibration model described in step 4) is 0.891.
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| CN201711262538.XA CN108226091A (en) | 2017-12-04 | 2017-12-04 | A kind of NIRS model building methods for being used to predict cane sugar content in corn |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108593579A (en) * | 2018-08-01 | 2018-09-28 | 漳州傲农牧业科技有限公司 | The detection method of glycerol content in a kind of corn |
| CN111665217A (en) * | 2020-06-09 | 2020-09-15 | 吉林省农业科学院 | Near infrared spectrum detection method for sucrose content of soybean seeds |
| CN114018865A (en) * | 2021-11-18 | 2022-02-08 | 河北农业大学 | Method for constructing near-infrared model of peanut sucrose content with different seed coat colors |
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| CN101846617A (en) * | 2009-12-29 | 2010-09-29 | 中国科学院地球化学研究所 | Sterile detection method of cane sugar content in culture media based on spectrum analysis |
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| CN101846617A (en) * | 2009-12-29 | 2010-09-29 | 中国科学院地球化学研究所 | Sterile detection method of cane sugar content in culture media based on spectrum analysis |
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| 杨泉女等: "3,5-二硝基水杨酸法与酶法测定甜玉米还原糖和蔗糖含量的比较", 《中国农业科技导报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108593579A (en) * | 2018-08-01 | 2018-09-28 | 漳州傲农牧业科技有限公司 | The detection method of glycerol content in a kind of corn |
| CN111665217A (en) * | 2020-06-09 | 2020-09-15 | 吉林省农业科学院 | Near infrared spectrum detection method for sucrose content of soybean seeds |
| CN114018865A (en) * | 2021-11-18 | 2022-02-08 | 河北农业大学 | Method for constructing near-infrared model of peanut sucrose content with different seed coat colors |
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