CN101900679A - Method for rapidly detecting proline content of rape leaf - Google Patents
Method for rapidly detecting proline content of rape leaf Download PDFInfo
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- CN101900679A CN101900679A CN 201010219036 CN201010219036A CN101900679A CN 101900679 A CN101900679 A CN 101900679A CN 201010219036 CN201010219036 CN 201010219036 CN 201010219036 A CN201010219036 A CN 201010219036A CN 101900679 A CN101900679 A CN 101900679A
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Abstract
The invention discloses a method for rapidly detecting proline content of a rape leaf. The method comprises the following steps of: fixing, drying and crushing the rape leaf to obtain rape leaf powder; acquiring rape leaf reflectivities corresponding to light waves with wave lengths of 2,308nm, 1,444nm, 1,418nm, 1,994nm, 2,432nm, 2,366nm, 2,020nm, 1,100nm, 1,942nm, 2,058nm, 1,338nm, 1,730nm, 2,294nm, and 2,186nm respectively; and substituting each reflectivity into a multiple linear regression equation and calculating the proline content of the rape leaf. Through the method, the multiple linear regression analysis is performed through a few characteristic wavelengths and the proline content of the rape leaf is rapidly and accurately detected, so that the detection time is greatly shortened, the environmental pollution is reduced, and the detection cost is lowered.
Description
Technical field
The present invention relates to the rape detection range, relate in particular to a kind of method for rapidly detecting proline content of rape leaf.
Background technology
Rape is one of the main oil crops of China and nectar source crop, and the cultivated area of rape accounts for more than 40% of the national oil crops total area, and output accounts for more than 60% of national edible vegetable oil total production, occupies first place in the world.The upgrowth situation of rape different times is related to the output and the quality of rapeseed.Amino acid content in the rape leaf is very important physical signs in the growth of rape process, and is significant to physiology courses such as rape leaf physiology and organic matter accumulations.Therefore can obtain the upgrowth situation of rape by detecting the amino acid content in the rape leaf.In recent years, because a large amount of herbicides has been used in rural laborer's minimizing in the rapeseed cultivation.Some herbicide is a kind of growth inhibitor, and for example propyl-ester nitorfen can stop the amino acid whose biosynthesizing of straight chain, has rape safety, kills that the grass spectrum is wide, activity of weeding is high, the lasting period is long, and low toxicity, low-residual, with characteristics such as environmentally compatible.Be in adverse circumstances when rape and coerce down, the accumulation of the proline that causes jointly by the inhibition of proline biosynthesizing activation and proline degraded in the plant, and the utilization that after adverse circumstance pressure is removed, is decomposed rapidly.Generally believe that a large amount of accumulation of proline may have very great help to the recovery of plant after pressure is removed under the environment stress, because proline can be used as carbon source and nitrogenous source, and can provide lot of energy for cellular-restoring.Therefore, its content can be used as one of physical signs of stress resistance of plant, proline amount height, and plant resistance to environment stress is strong.To the detection of proline content in the rape leaf, can reflect of the influence of some herbicide to rape physiology and environment stress, the land for growing field crops of realizing the growth of rape situation is detected and continuous monitoring, the output and the quality that improve rapeseed are significant.
Traditional proline content of rape leaf detection method is according to GB/T5009.124-2003, adopt ten thousand/balance accurately to take by weighing the ampere bottle that powder sample 0.05g places the 10ml volume, add 5ml6M HCL, vacuumize and on alcohol blast burner, seal to be placed in 110 ℃ of baking ovens with the circulating vacuum pump and clear up 24h; Cooled digestion solution is transferred to constant volume in the 50ml volumetric flask fully, shaken up; Draw the 5ml supernatant and place round bottom ground conical flask, (60～70 ℃) vacuumize 15min left and right sides evaporate to dryness hydrochloric acid on Rotary Evaporators; Again add 0.02M HCl (pH 2.2) 10ml, ultrasound wave fully dissolves back one-shot injector sucking-off, disposable filter through 0.22 μ m is filtered in the special-purpose sample bottle of amino acid analysis, 4 ℃ of dark preservations are with the L-8900 of Hitachi automatic amino acid analyser (Japan) assay determination proline content.This method sample preprocessing process and measuring process is time-consuming, effort, chemical depletion product expensive, and environment is caused certain pollution.Therefore, be badly in need of research a kind of simple, quick, harmless proline content of rape leaf detection method and technology.
As seen/near infrared spectrum can reflect that the frequency multiplication of groups such as the organic principle of material inside, particularly C-H, O-H, N-H and sum of fundamental frequencies absorb, and can be used for the content of quantitative measurement material organic substance.Spectral technique has been widely used in industries such as agricultural, food, petrochemical complex, pharmacy, feed.In the research of plant life information fast detecting, has application potential widely
Summary of the invention
The invention provides a kind of method for rapidly detecting proline content of rape leaf, solved problems such as existing detection method is time-consuming, effort, complicated operation, contaminated environment.
A kind of method for rapidly detecting proline content of rape leaf may further comprise the steps:
(1) rape leaf completed, dries the back and pulverize, obtain the blade powder;
(2) gather the reflectivity of wavelength 2308nm, 1444nm, 1418nm, 1994nm, 2432nm, 2366nm, 2020nm, 1100nm, 1942nm, 2058nm, 1338nm, 1730nm, 2294nm and the pairing rape leaf of 2186nm light wave respectively;
(3) with each reflectivity difference substitution multiple linear regression equations: y=-151770 λ
1-179.233 λ
2-156.080 λ
3+ 90.881 λ
4-61.178 λ
5-85.245 λ
6-97.499 λ
7+ 14.884 λ
8-47.067 λ
9+ 39.998 λ
10-35.203 λ
11-1.957 λ
12-7.455 λ
13+ 60.409 λ
14+ 1.8470, calculate proline content in the rape leaf;
Y is a proline content of rape leaf in the equation, and unit is mg/100mg DW; λ
1～λ
14Be respectively the reflectivity of wavelength 2308nm, 1444nm, 1418nm, 1994nm, 2432nm, 2366nm, 2020nm, 1100nm, 1942nm, 2058nm, 1338nm, 1730nm, 2294nm and the pairing rape leaf Powder samples of 2186nm light wave.
In the step (1), the blade powder is 40～80 orders, and the temperature that completes is 95～115 ℃, and fixation time is 20～40min.This complete temperature and fixation time are suitable, help the reservation of major component in the rape leaf, particularly proline.In the step (1), bake out temperature is 70～90 ℃, and blade is dried to constant weight, and this bake out temperature is suitable, can burned blade.
Because the rape leaf Powder samples is a non-transparent medium, when light-wave irradiation in its when surface, except that part is absorbed, all the other all reflect, and what (perhaps reflectivity) that light wave absorbs are to be correlated with the inner content of chemical substances of rape leaf, and only the reflectivity with some characteristic wavelength light wave of minority is relevant at the concrete material of this class of proline content.
Use the successive projection algorithm and in 1100～2500nm wavelength coverage, carry out choosing of characteristic wavelength, obtain these characteristic wavelengths and be respectively 2308nm, 1444nm, 1418nm, 1994nm, 2432nm, 2366nm, 2020nm, 1100nm, 1942nm, 2058nm, 1338nm, 1730nm, 2294nm and 2186nm.With the reflectance value of the above-mentioned wavelength input variable as multiple linear regression model, the proline content of actual measurement calculates above-mentioned multiple linear regression equations as output variable.
The present invention carries out multiple linear regression analysis by a few features wavelength, quick and precisely detects proline content in the rape leaf, has shortened the time of detecting greatly, has reduced environmental pollution, has reduced the detection cost.
Description of drawings
Fig. 1 is a diffusing some distribution plan for modeling collection sample of the present invention predicts the outcome;
Fig. 2 is a diffusing some distribution plan for forecast set sample of the present invention predicts the outcome.
Embodiment
248 rape leafs of random collecting, the rape leaf sample of gathering for making has more representativeness and diversity, not only comprise the rape leaf (totally 62) under the normal growth state in 248 blades being gathered, also comprise and sprayed variable concentrations (100,500,1000mg/L) rape leaf of propyl-ester nitorfen herbicide is 186 (100,500, each concentration gradient of 1000mg/L is gathered 62 of rape leafs), spraying time is one heart stages of five leaves, spray pattern is a foliage-spray.The propyl-ester nitorfen herbicide is a kind of proline inhibitor, and is inhibited to the rape leaf proline, can make the proline content in the rape leaf have bigger scope, makes institute's established model have better adaptability and robustness.
248 rape leafs are placed under 80 ℃, dry after completing 30 minutes in 105 ℃ the baking oven to constant weight.Rape leaf sample mill powdered after will drying with mortar again, and cross 60 mesh sieves.At room temperature measure the reflectance value (Foss NIR Systems 5000) of rape leaf Powder samples, utilize the mentioned classic method of background technology to measure the proline content of all blades simultaneously at wavelength 2308nm, 1444nm, 1418nm, 1994nm, 2432nm, 2366nm, 2020nm, 1100nm, 1942nm, 2058nm, 1338nm, 1730nm, 2294nm and 2186nm place.Select 186 samples as modeling collection sample then at random, all the other 62 as the forecast set sample, with the reflectance value of each characteristic wavelength of 186 rape leafs of modeling collection sample as input variable, with the proline content of corresponding measurement as output variable, utilize multiple linear regression method (Multiple linearregression MLR) obtains following multiple linear regression equations:
Y=-151770 λ
1-179.233 λ
2-156.080 λ
3+ 90.881 λ
4-61.178 λ
5-85.245 λ
6-97.499 λ
7+ 14.884 λ
8-47.067 λ
9+ 39.998 λ
10-35.203 λ
11-1.957 λ
12-7.455 λ
13+ 60.409 λ
14+ 1.8470, calculate proline content of rape leaf.
Y in the equation (mg/100mg DW) is a proline content of rape leaf, and unit is mg/100mgDW; λ
1～λ
14Be respectively the reflectivity of wavelength 2308nm, 1444nm, 1418nm, 1994nm, 2432nm, 2366nm, 2020nm, 1100nm, 1942nm, 2058nm, 1338nm, 1730nm, 2294nm and the pairing rape leaf Powder samples of 2186nm light wave.
Utilize above-mentioned multiple linear regression equations that the proline content of modeling collection and forecast set rape leaf sample is predicted then, and estimate predicting the outcome.Related coefficient and slope approach 1 more in the evaluation index, and the absolute value of root-mean-square error, deviation and the absolute value of intercept are more little, illustrate that the model prediction performance is good more.It is as shown in the table to predict the outcome:
Modeling collection and forecast set are predicted the facies relationship number average between the predicted value of proline content of rape leaf and the true chemical method measured value has obtained satisfied precision of prediction greater than 0.95.The diffusing some distribution plan that modeling collection and forecast set sample are predicted the outcome as depicted in figs. 1 and 2, sample has good linear prediction effect near being distributed in the regression straight line both sides among the figure.The detection that method of the present invention can realize proline content of rape leaf is rapidly and accurately used in the The above results explanation.
Claims (4)
1. method for rapidly detecting proline content of rape leaf may further comprise the steps:
(1) rape leaf completed, dries the back and pulverize, obtain the blade powder;
(2) gather the reflectivity of wavelength 2308nm, 1444nm, 1418nm, 1994nm, 2432nm, 2366nm, 2020nm, 1100nm, 1942nm, 2058nm, 1338nm, 1730nm, 2294nm and the pairing rape leaf of 2186nm light wave respectively;
(3) with each reflectivity difference substitution multiple linear regression equations: y=-151770 λ
1-179.233 λ
2-156.080 λ
3+ 90.881 λ
4-61.178 λ
5-85.245 λ
6-97.499 λ
7+ 14.884 λ
8-47.067 λ
9+ 39.998 λ
10-35.203 λ
11-1.957 λ
12-7.455 λ
13+ 60.409 λ
14+ 1.8470, calculate proline content in the rape leaf;
Y is a proline content of rape leaf in the equation; λ
1～λ
14Be respectively the reflectivity of wavelength 2308nm, 1444nm, 1418nm, 1994nm, 2432nm, 2366nm, 2020nm, 1100nm, 1942nm, 2058nm, 1338nm, 1730nm, 2294nm and the pairing rape leaf Powder samples of 2186nm light wave.
2. method for rapidly detecting proline content of rape leaf according to claim 1 is characterized in that: described blade powder is 40～80 orders.
3. method for rapidly detecting proline content of rape leaf according to claim 1 is characterized in that: in the step (1), the temperature that completes is 95～115 ℃, and fixation time is 20～40min.
4. method for rapidly detecting proline content of rape leaf according to claim 1 is characterized in that: in the step (1), bake out temperature is 70～90 ℃.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106248468A (en) * | 2016-10-09 | 2016-12-21 | 塔里木大学 | A kind of detection method of humic acids from different soils content |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| JP2008151760A (en) * | 2006-11-21 | 2008-07-03 | Nippon Telegr & Teleph Corp <Ntt> | Method and apparatus for qualitatively and quantitatively analyzing substance having weak interaction |
| CN101871893A (en) * | 2009-04-21 | 2010-10-27 | 陈延霞 | Colorimetric method for determining free L-hydroxyproline (L-HYP) in milk powder |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008151760A (en) * | 2006-11-21 | 2008-07-03 | Nippon Telegr & Teleph Corp <Ntt> | Method and apparatus for qualitatively and quantitatively analyzing substance having weak interaction |
| CN101871893A (en) * | 2009-04-21 | 2010-10-27 | 陈延霞 | Colorimetric method for determining free L-hydroxyproline (L-HYP) in milk powder |
Non-Patent Citations (3)
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| 《光谱学与光谱分析》 20091130 刘飞等 连续投影算法在油菜叶片氨基酸总量无损检测中的应用 3079－3083 1-4 , 第11期 2 * |
| 《农业机械学报》 20070525 牛智有等 鱼粉中氨基酸近红外光谱定量分析 114-117 1-4 第38卷, 第05期 2 * |
| 《四川轻化工学院学报》 19980630 付大友等 多元线性回归在吸光光度法测量氨基酸组分中的应用 6-10 1-4 第11卷, 第02期 2 * |
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| CN106248468A (en) * | 2016-10-09 | 2016-12-21 | 塔里木大学 | A kind of detection method of humic acids from different soils content |
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