CN105136686B - The measuring method of Purple Leaf Plum anthocyanidin content - Google Patents
The measuring method of Purple Leaf Plum anthocyanidin content Download PDFInfo
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Abstract
The invention discloses a kind of measuring method of Purple Leaf Plum anthocyanidin content, belong to plant growth information lossless monitoring technical field in precision agriculture.This method measures the reflected spectrum data of Purple Leaf Plum using EO-1 hyperion radiation gauge, indoor measurement blade anthocyanidin content, then Purple Leaf Plum reflected spectrum data and anthocyanidin content data are blended, the sensitive wave length of anthocyanidin is determined by correlation analysis, construct the anthocyanidin content spectrum monitoring model based on characteristic wavelength, Purple Leaf Plum to be measured is measured in the reflected spectrum data of characteristic wavelength using spectral radiometer again, data are substituted into anthocyanidin content spectrum monitoring model, the anthocyanidin content of Purple Leaf Plum is calculated；This method is simple, efficient, practical, and accuracy is high, blade non-destructive testing is able to achieve, suitable for the different age of trees, the duplicate measurements in situ and big area monitoring of different colours sample.
Description
Technical field
The present invention relates to a kind of measuring method of Purple Leaf Plum anthocyanidin content, belongs to plant growth in precision agriculture and believe
Cease non-destructive monitoring technical field.
Background technique
Anthocyanidin is water-soluble flavone compound, it is third class primary pigments in pigment in plant leaf, is a kind of
Plant Secondary Metabolites with multiple physiological function.In general, anthocyanidin it is effective accumulation be as caused by many environment-stress,
Shortage, certain herbicides including strong light, ultraviolet irradiation, low temperature, arid, damage, bacterium and fungal infection, nitrogen and phosphorus
And pollutant etc., especially plant is immature and ageing leaves in rich content.Anthocyanidin is capable of the luminous environment of reblading, has
It potentially adjusts photosynthesis and limits the ability of Xanthophyll cycle and photobleaching, and the defence capability to photo damage.Cyanine
Element can be used as osmotic adjustment, improve the ability of Genes For Plant Tolerance frost and drought-resistant stress.In addition, anthocyanidin also has antioxygen
Change effect helps to repair the blade after damage.
Traditional anthocyanidin content measurement mainly uses wet chemistry method, including with anthocyanidin in solvent extraction blade, light splitting
The absorbance value of measurement is converted into anthocyanidin content by the absorbance of photometric determination anthocyanidin in a solvent.The party
Method the content of anthocyanidin can measure time-consuming, laborious but there are large labor intensity in Accurate Determining blade, need to blade
The problems such as being destroyed, not can be carried out duplicate measurements in situ and big area monitoring.Therefore, need it is a kind of simple, accurate, efficiently,
Practical anthocyanidin content measuring method.Existing research shows that phytochrome selectively can absorb or reflect specific wavelength
Light, using spectral absorption or reflection detection technique substitute destructive, time-consuming and laborious wet chemistry method, be beneficial to plant color
Quick, the non invasive estimation of element.Currently, the Purple Leaf Plum anthocyanidin measurement based on EO-1 hyperion monitoring model has not been reported.
Summary of the invention
The object of the present invention is to provide a kind of simple, accurate, efficient, practical Purple Leaf Plum anthocyanidin content measurement sides
Method.
In order to achieve the goal above, the technical scheme adopted by the invention is that:
The measuring method of Purple Leaf Plum anthocyanidin content, steps are as follows: measuring prunus cerasifera leaf using EO-1 hyperion radiation gauge
Piece substitutes into the reflected spectrum data measured in anthocyanidin content spectrum monitoring model in the reflected spectrum data of characteristic wavelength,
The anthocyanidin content of Purple Leaf Plum is calculated.
The foundation of the anthocyanidin content spectrum monitoring model the following steps are included:
1) sampling is acquired with data
The uniform multiple blades of color are formed a blade sample by the Purple Leaf Plum for acquiring the different age of trees, point
The anthocyanidin content and reflected spectrum data of blade in each sample are not measured；
2) data prediction
For a blade sample, 1nm first is resampled to reflected spectrum data, then calculates separately anthocyanidin content and again
The mean value of reflected spectrum data is sampled, and is denoted as one group of sample data, by obtained multiple groups sample data according to anthocyanidin content
Size sequence, forms a data set；
3) characteristic wavelength is determined
The anthocyanidin content and reflected spectrum data concentrate to data make correlation analysis, related coefficient maximum and two-tailed test
Reach extremely significant horizontal (correlation maximum between the two at this time)；
4) it models and tests mould
Using the reflected spectrum data fitting formula of anthocyanidin content and characteristic wavelength, anthocyanidin content spectrum monitoring is obtained
Model, test mould to get.
Conventional method in that art can be used in measurement anthocyanidin content in step 1), and preferably 0.1mol/L hydrochloric acid methanol low temperature soaks
Spectrophotometry is proposed, steps are as follows: broken Purple Leaf Plum being placed in 0.1mol/L hydrochloric acid acidified methanol and (is contained in every liter of methanol
Have 0.1mol HCl) in extract (extract 2 times such as at 45 DEG C, extract 2h every time), extract finish centrifuging and taking supernatant (or merging
Supernatant), spectrophotometric determination supernatant (or solution after supernatant constant volume) is calculated in the OD value of certain wave strong point
Into Purple Leaf Plum, (when specific wavelength takes 530nm, 620nm, 650nm, calculation method is referring to Xiong Qinge, leaf for anthocyanidin content
Treasure, Yang Shimin wait the Chengdu plant physiology experiment study course [M]: Sichuan science tech publishing house, 2003:94-95).
EO-1 hyperion radiation gauge can be used in measurement reflected spectrum data in step 1), as Spectra Vista company of the U.S. is raw
The SVC HR-1024i portable spectrometer of production, the spectral resolution of band value 350~2500nm, 350~1000nm wave band≤
Spectral resolution≤9.5nm of 3.5nm, 1000~1850nm wave band, the spectral resolution of 1850~2500nm wave band≤
6.5nm。
The reflected spectrum data of multiple groups sample data preferably 350~1000nm wave band in step 2), this is because anthocyanidin
Mainly ultraviolet, visible light wave range is responded, all band can be not considered.
Characteristic wavelength is 560nm in step 3).
Modeling in step 4) tests mould 2/3,1/3 data in data set is respectively adopted and carries out.
The softwares such as Excel, SPSS can be used in the fitting of the reflected spectrum data of anthocyanidin content and characteristic wavelength in step 4).
Multiple correlation coefficient (R is used in step 4)2) evaluation anthocyanidin content spectrum monitoring model, meet R2Model when > 0.8
Building is suitable.Meanwhile the Stability and veracity of verifying model is examined using independent sample, evaluation index selects multiple correlation coefficient
(R2), root-mean-square error (RMSE) and prediction residual deviation (RPD), meet R2When 0.5 μm of > 0.8, RMSE < ol/g, RPD > 2
Model construction success.
The calculation formula of above-mentioned evaluation index is as follows:
Formula 1:
Formula 2:
Formula 3:
In formula:For predicted value；For the mean value of observation；yiFor observation；N is sample number, and i is positive integer, i=1,
2,…,n；SDVFor the standard deviation of verifying collection；RMSEVFor the root-mean-square error of verifying collection.
Preferably, I representation model of following formula can be used in anthocyanidin content spectrum monitoring model.
Formula I: y=0.0189x-1.678, R2=0.823；
In formula: y: blade anthocyanidin content, μm ol/g；Reflected spectrum data at x:560nm wavelength；
The evaluation index of model verifying are as follows: coefficient of multiple correlation R2=0.876, RMSE=0.370 μm of ol/g of root-mean-square error,
Prediction residual deviation RPD=2.542.
Beneficial effects of the present invention:
The measuring method of Purple Leaf Plum anthocyanidin content is simple, efficient, practical in the present invention, and accuracy is high, is able to achieve
Blade non-destructive testing, suitable for the different age of trees, the duplicate measurements in situ and big area monitoring of different colours sample.This method uses
EO-1 hyperion radiation gauge measures the reflected spectrum data of Purple Leaf Plum, indoor measurement blade anthocyanidin content, then by prunus cerasifera
Blade reflected spectrum data is blended with anthocyanidin content data, and the sensitive wave length of anthocyanidin is determined by correlation analysis, building
Based on characteristic wavelength anthocyanidin content spectrum monitoring model (as shown in formula I, the model is simple, it is with good stability and
Adaptability, accuracy are high), then Purple Leaf Plum to be measured is measured in the reflected spectrum data of characteristic wavelength using spectral radiometer,
Data are substituted into anthocyanidin content spectrum monitoring model, the anthocyanidin content of Purple Leaf Plum is calculated, reaches monitoring
Purpose.
Detailed description of the invention
Fig. 1 is the building flow diagram of Purple Leaf Plum anthocyanidin content spectrum monitoring model in embodiment 1；
Fig. 2 is scatter plot and the matched curve of anthocyanidin content predicted value and measured value.
Specific embodiment
Only invention is further described in detail for following embodiments, but does not constitute any limitation of the invention.
Embodiment 1
The measuring method of prunus cerasifera (Prunus cerasifera) blade anthocyanidin content in the present embodiment, including it is following
Step:
One, establish Purple Leaf Plum anthocyanidin content spectrum monitoring model (flow diagram is shown in Fig. 1)
1) sampling is acquired with data
Acquire the Purple Leaf Plum of the different age of trees, blade is randomly choosed based on visualization feature, and color is from green, micro- red to complete
Quan Hong, and it is healthy, not damaged, uniform 3 blades of color are formed into a blade sample, measure each sample respectively
In the same blade anthocyanidin content and reflected spectrum data；
The measuring method of anthocyanidin content are as follows: take Purple Leaf Plum that impurity elimination is eluted with water, blots, be cut into 2 × 5mm strip
Fragment accurately weighs 0.15g, and 0.1mol/L hydrochloric acid acidified methanol 10mL is added, extracts 2 times, extraction time 2h at 45 DEG C,
It is centrifugated at 5000r/min after extraction, merges 2 centrifuged supernatants, filtered to get anthocyanidin extracting solution, by extracting solution
It is settled to 50mL, optical density of the anthocyanidin extracting solution under 530nm, 620nm, 650nm wavelength after spectrophotometric determination constant volume
Value, is calculated anthocyanidin content in Purple Leaf Plum；
Reflected spectrum data is adopted using the SVC HR-1024i portable spectrometer of U.S. Spectra Vista company production
Collection, spectral resolution≤3.5nm of band value 350~2500nm, 350~1000nm wave band, the light of 1000~1850nm wave band
Spectral resolution≤9.5nm, spectral resolution≤6.5nm of 1850~2500nm wave band；
2) data prediction
For a blade sample, 1nm first is resampled to reflected spectrum data, then calculates separately anthocyanidin content and again
The mean value of reflected spectrum data is sampled, and is denoted as one group of sample data, since anthocyanidin mainly rings ultraviolet, visible light wave range
It answers, the reflected spectrum data of 350~1000nm wave band is only chosen in pretreatment, and all band wouldn't be made to consider, then more by what is obtained
Group sample data forms a data set, as shown in table 1 below according to the ascending sequence of anthocyanidin content；
1 Purple Leaf Plum anthocyanidin content of table and spectroscopic data (part)
3) characteristic wavelength is determined
The anthocyanidin content and reflected spectrum data concentrate to data make Bivariate analysis (SPSS software), as a result show
Show that the related coefficient at 560nm wavelength is maximum (being equal to 0.80037), and two-tailed test reaches extremely significant horizontal (being equal to 0.000),
Therefore selection 560nm wavelength is characterized wavelength；
4) it models and tests mould
The data of selection 2/3 are made as calibration set (calibration set is modeling data herein), 1/3 data in data set
For verifying collection, in SPSS software, using the reflected spectrum data fitting formula of anthocyanidin content and characteristic wavelength, obtain as follows
Anthocyanidin content spectrum monitoring model shown in formula I, II, III, due to R in formula II, formula III2< 0.8, it is subsequent to use independent sample
The two models are not verified when examining verifying model accuracy and stability；
Formula I: y=0.0189x-1.678, R2=0.823；
In formula: y: blade anthocyanidin content, μm ol/g；Reflected spectrum data at x:560nm wavelength；
Formula II: y=350.1x2- 96.67x+6.877, R2=0.795；
In formula: y: blade anthocyanidin content, μm ol/g；Reflected spectrum data at x:560nm wavelength；
Formula III: y=-32.70x+4.317, R2=0.644；
In formula: y: blade anthocyanidin content, μm ol/g；Reflected spectrum data at x:560nm wavelength；
Using predict anthocyanidin content as abscissa, actual measurement anthocyanidin content be ordinate establish regression equation (see below formula a),
The scatter plot and matched curve of anthocyanidin content predicted value and measured value are shown in Fig. 2；
Formula a:y=1.018x+0.133, R2=0.876；
The intercept of regression equation and slope are comparatively close to 0 and 1 in formula, illustrate the prediction effect of the Single wavelength model compared with
It is good；
The evaluation index of model verifying are as follows: coefficient of multiple correlation R2=0.876, illustrate model accuracy up to 87.6%；It is square
RMSE=0.370 μm of ol/g of root error, illustrates that model predictive error is smaller；Prediction residual deviation RPD=2.542；Show the mould
Type is stable, accurate, can be used for the prediction of Purple Leaf Plum anthocyanidin content；
Two, Purple Leaf Plum anthocyanidin content measures
Prunus cerasifera to be measured is measured using the SVC HR-1024i portable spectrometer of Spectra Vista company of U.S. production
The reflected spectrum data measured is substituted into anthocyanidin content shown in formula I in the reflected spectrum data of 560nm characteristic wavelength by blade
In spectrum monitoring model, the anthocyanidin content of Purple Leaf Plum is calculated, see the table below 2.
Table 2 monitors the anthocyanidin content (part) of obtained reflectance spectrum and prediction and actual measurement
| Catalogue number(Cat.No.) | Reflectance spectrum at 560nm | Predict anthocyanidin content μm ol/g | Survey anthocyanidin content μm ol/g |
| 1 | 0.1728 | 0.3596 | 0.4055 |
| 2 | 0.0938 | 1.0025 | 0.9976 |
| 3 | 0.0861 | 1.1575 | 1.1643 |
| 4 | 0.0776 | 1.3781 | 1.4779 |
| 5 | 0.0575 | 2.2790 | 2.4386 |
| 6 | 0.0448 | 3.4643 | 3.3753 |
| 7 | 0.0408 | 4.0530 | 3.9595 |
Stability, the adaptability of Purple Leaf Plum anthocyanidin content spectrum monitoring model are good in the present embodiment, accuracy
Height, suitable for the different age of trees, the duplicate measurements in situ and big area monitoring of different colours sample.
Claims (6)
1. the measuring method of Purple Leaf Plum anthocyanidin content, it is characterised in that: steps are as follows: being measured using EO-1 hyperion radiation gauge
The reflected spectrum data measured is substituted into anthocyanidin content spectrum monitoring in the reflected spectrum data of characteristic wavelength by Purple Leaf Plum
In model, the anthocyanidin content of Purple Leaf Plum is calculated；
The foundation of the anthocyanidin content spectrum monitoring model the following steps are included:
1) sampling is acquired with data
The uniform multiple blades of color are formed a blade sample, surveyed respectively by the Purple Leaf Plum for acquiring the different age of trees
The anthocyanidin content and reflected spectrum data of blade in each fixed sample；
2) data prediction
For a blade sample, 1nm first is resampled to reflected spectrum data, then calculate separately anthocyanidin content and resampling
The mean value of reflected spectrum data, and it is denoted as one group of sample data, by obtained multiple groups sample data according to anthocyanidin content size
Sequence, forms a data set；
3) characteristic wavelength is determined
The anthocyanidin content and reflected spectrum data concentrate to data make correlation analysis, and related coefficient is maximum and two-tailed test reaches
The wavelength of extremely significant level is determined as characteristic wavelength；
4) it models and tests mould
Using the reflected spectrum data fitting formula of anthocyanidin content and characteristic wavelength, anthocyanidin content spectrum monitoring mould is obtained
Type, test mould to get；
Step 3) characteristic wavelength is 560nm；
The anthocyanidin content spectrum monitoring model is as shown in following formula I:
Formula I: y=0.0189x-1.678, R2=0.823；
In formula: y: blade anthocyanidin content, μm ol/g；Reflected spectrum data at x:560nm wavelength；
Test mould evaluation index are as follows: coefficient of multiple correlation R2=0.876, RMSE=0.370 μm of ol/g of root-mean-square error, prediction residual are inclined
Poor RPD=2.542.
2. measuring method according to claim 1, it is characterised in that: the method for measurement anthocyanidin content in step 1) are as follows:
Broken Purple Leaf Plum is placed in 0.1mol/L hydrochloric acid acidified methanol and is extracted, extracts and finishes centrifuging and taking supernatant, in measurement
Anthocyanidin content in Purple Leaf Plum is calculated in the OD value of certain wave strong point in clear liquid.
3. measuring method according to claim 1, it is characterised in that: measurement reflected spectrum data uses bloom in step 1)
Compose radiation gauge, spectral resolution≤3.5nm of band value 350~2500nm, 350~1000nm wave band, 1000~1850nm wave
Spectral resolution≤the 9.5nm, spectral resolution≤6.5nm of 1850~2500nm wave band of section.
4. measuring method according to claim 3, it is characterised in that: in step 2) multiple groups sample data selection 350~
The reflected spectrum data of 1000nm wave band.
5. measuring method according to claim 1, it is characterised in that: anthocyanidin content spectrum monitoring model is adopted in step 4)
Use coefficient of multiple correlation R2Evaluation, R2> 0.8.
6. measuring method according to claim 5, it is characterised in that: test mould in step 4) and examined using independent sample, commented
Valence index are as follows: coefficient of multiple correlation R2> 0.8, root-mean-square error RMSE < 0.5 μm of ol/g, prediction residual deviation RPD > 2.
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