CN115436307B - Method for nondestructively and rapidly detecting chlorophyll and carotenoid content in plant leaves - Google Patents
Method for nondestructively and rapidly detecting chlorophyll and carotenoid content in plant leaves Download PDFInfo
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Abstract
The invention discloses a method for nondestructively and rapidly detecting chlorophyll and carotenoid content in plant leaves, and relates to the technical field of photosynthetic pigment content detection. The method comprises the following steps: step 1, measuring absorbance value A of plant leaf at light source wavelength of 460+ -5 nm, 660+ -5 nm and 980+ -5 nm 1 、A 2 And A 3 The method comprises the steps of carrying out a first treatment on the surface of the Step 2, the A is carried out 1 、A 2 And A 3 Substituting the values into formulas (1), (2), (3) and (4) to obtain Cla, clb, clz and Clc values; cla represents the chlorophyll a content; clb represents the content of chlorophyll b; clz the total chlorophyll content; clc represents the carotenoid content. Compared with the prior art, the invention can realize nondestructive rapid detection of the contents of chlorophyll a, chlorophyll b, total chlorophyll and carotenoid in plant leaves, does not need to use any chemical reagent, and does not cause harm to the health of testers.
Description
Technical Field
The invention relates to the technical field of detection of photosynthetic pigment content of plant leaves, in particular to a method for nondestructively and rapidly detecting chlorophyll and carotenoid content in plant leaves.
Background
Photosynthetic pigments in plant leaves mainly consist of chlorophyll and carotenoid, and have the function of absorbing and converting light energy, and reflect the growth state, development stage and nitrogen condition of plants, so that the definition of the content of chlorophyll and carotenoid is of great significance for guiding the production of crops. Chlorophyll is composed of chlorophyll a and chlorophyll b. The existing research results show that the contents of chlorophyll a and chlorophyll b in different plant leaves or plant leaves in different growth stages are different, so that the absorption capacities of the plant leaves on red light and blue-violet light are different. Therefore, the content of chlorophyll a and chlorophyll b is clear, and the method has important significance for guiding the accurate light supplementing of greenhouse crops, improving the light supplementing efficiency, reducing the total energy consumption, saving energy and reducing emission. Carotenoids have the effects of absorbing and transmitting light energy, protecting chlorophyll, delaying rapid decomposition of chlorophyll in senescence leaves and the like, so that understanding the carotenoid content in plants is also of great significance for understanding the growth state and development condition of plants.
The Chinese agricultural industry standard NY/T3082-2007 prescribes that the detection method of chlorophyll and carotenoid content in plant leaves is a spectrophotometry method, and the detection result of the method is accurate and stable, but the method has the defects of complicated detection process, time consumption, higher cost, incapability of realizing on-site measurement and the like. In addition, because toxic acetone is required to be used in the detection process, potential risks exist for the health of operators, and the method is difficult to apply to actual production on a large scale. Therefore, research into methods for nondestructive rapid detection of chlorophyll and carotenoid content has been a dream of agricultural workers.
At present, a plurality of chlorophyll detectors are developed and designed by using 650nm and 940nm wavelengths according to the detection principle of an ultraviolet spectrophotometer, such as a SPAD-502 chlorophyll instrument, an FK-YL series chlorophyll instrument and the like. These instruments have the advantage of being simple and portable to operate, but they can only measure a relative value to chlorophyll content, such as SPAD (Soil-Plant Analyzer-Development), and cannot give the actual chlorophyll a, chlorophyll b, total chlorophyll and carotenoid content.
Chinese patent CN104777108A discloses a "detection device and method for chlorophyll content", which comprises irradiating chlorophyll solution with light having wavelengths of 645nm and 663nm, measuring intensity of transmitted light by photoelectric sensor to obtain absorbance value at corresponding wavelength, and calculating chlorophyll a, chlorophyll b and total chlorophyll content in the chlorophyll solution according to the relationship between the established absorbance value and chlorophyll concentration and the measured absorbance value. The method measures chlorophyll solution, so that nondestructive rapid detection of chlorophyll a, chlorophyll b and total chlorophyll in plant leaves cannot be realized.
Chinese patent CN208872661U discloses a three-wavelength-based plant leaf chlorophyll content detection device, which uses light emitting diodes at 460nm, 650nm and 940nm as light sources, and establishes a mathematical model for predicting chlorophyll content in plant leaves, thereby realizing nondestructive detection of chlorophyll content. However, the method in this patent can only detect the total chlorophyll content in the plant leaves, and cannot detect the contents of chlorophyll a, chlorophyll b and carotenoids.
Disclosure of Invention
The invention aims to provide a nondestructive rapid detection method for the contents of chlorophyll and carotenoid in plant leaves, which aims to solve the problems in the prior art, and can realize rapid detection for the contents of chlorophyll a, chlorophyll b, total chlorophyll and carotenoid in plant leaves under the condition of not damaging the plant leaves.
In order to achieve the above object, the present invention provides the following solutions:
according to one of the technical schemes, the method for constructing the model for nondestructive rapid detection of chlorophyll and carotenoid contents in plant leaves comprises the following steps:
testing absorbance A of plant leaf at light source wavelengths of 460+ -5 nm, 660+ -5 nm and 980+ -5 nm 1 、A 2 And A 3 ；
Spectrophotometry is used for measuring the contents of chlorophyll a, chlorophyll b, total chlorophyll and carotenoid in plant leaves, and the contents are respectively recorded as YLa, YLb, YLz and YLc;
the YLa, YLb, YLz and YLc are respectively used as dependent variables, and the A is used as the 1 、A 2 And A 3 And (3) establishing linear and nonlinear models for independent variables, and determining an optimal model according to the decision coefficients of the models and the complexity of the models.
The model with higher determination coefficient and simpler structure is used as the optimal model.
When spectrophotometry is used for measuring the content of photosynthetic pigments in plant leaves, detection is required within 20min after the plant leaves are picked.
The reason for choosing the absorbance values of the plant leaves tested at wavelengths of 460.+ -. 5nm, 660.+ -. 5nm and 980.+ -. 5nm is that: the absorbance of chlorophyll b is strongest near 460nm, the absorbance of chlorophyll a and the absorbance of chlorophyll b intersect near 660nm, and the absorbance of chlorophyll at 975-985 nm has little influence on the thickness of plant leaves, so that the absorbance values of plant leaves can be measured at the wavelengths of 460+ -5 nm, 660+ -5 nm and 980+ -5 nm to enable the established model to have better adaptability, and the chlorophyll a, chlorophyll b, total chlorophyll and carotenoid values calculated based on the model are closer to actual values.
Further, the methodThe A is 1 、A 2 And A 3 The test method of (1) comprises the following steps: the absorbance values of the plant leaves at each wavelength were calculated from the logarithm of the ratio of the intensity of light not transmitted and transmitted through the plant leaves at different wavelengths of 460.+ -.5 nm, 660.+ -.5 nm and 980.+ -.5 nm.
Further, the A 1 、A 2 And A 3 The test method of (1) comprises the following steps:
step a, setting up light intensity acquisition equipment by taking 3 light emitting elements with peak wavelengths or center wavelengths respectively at 460+/-5 nm, 660+/-5 nm and 980+/-5 nm as light sources and taking a photoelectric sensor as illumination acquisition equipment, wherein the 3 light emitting elements are opposite to the photoelectric sensor, the middle distance is 0.5-3 mm, and the original light intensities of the light emitting elements at the wavelengths of 460+/-5 nm, 660+/-5 nm and 980+/-5 nm are acquired and respectively marked as Y 1 、Y 2 And Y 3 ；
Step b, collecting plant leaves, and measuring the transmitted light intensities of 460+/-5 nm, 660+/-5 nm and 980+/-5 nm when the light intensity collecting device in the step 1 is used for transmitting the plant leaves, wherein the transmitted light intensities are respectively recorded as T 1 、T 2 And T 3 ；
Step c, calculating the absorbance of the plant leaf at each wavelength, specifically the absorbance A at the wavelength of 460+ -5 nm 1 ＝lg(Y 1 /T 1 ) Absorbance A at 660+ -5 nm wavelength 2 ＝lg(Y 2 /T 2 ) Absorbance A at 980.+ -. 5nm wavelength 3 ＝lg(Y 3 /T 3 )。
The light-emitting element is an LED light source.
According to a second technical scheme, the method for nondestructively and rapidly detecting the chlorophyll and carotenoid contents in plant leaves comprises the following steps:
step 1, measuring absorbance value A of plant leaf at light source wavelength of 460+ -5 nm, 660+ -5 nm and 980+ -5 nm 1 、A 2 And A 3 ；
Step 2, the A is carried out 1 、A 2 And A 3 Substituting the values into formulas (1), (2), (3) and (4) to obtain Cla, clb, clz and Clc values;
Cla＝-0.952A 1 +1.828A 2 -1.934A 3 +0.402 (1)
Clb＝-0.462A 1 +0.685A 2 -0.578A 3 +0.208 (2)
Clz＝-1.580A 1 +2.831A 2 -2.859A 3 +0.689 (3)
Clc＝-0.154A 1 +0.378A 2 -0.443A 3 +0.072 (4)
the Cla represents the content of chlorophyll a; the Clb represents the content of chlorophyll b; said Clz represents the total chlorophyll content; the Clc represents the carotenoid content.
The reason for choosing the absorbance values of the plant leaves tested at wavelengths of 460.+ -. 5nm, 660.+ -. 5nm and 980.+ -. 5nm is that: the absorbance of chlorophyll b is strongest near 460nm, the absorbance of chlorophyll a and the absorbance of chlorophyll b are intersected near 660nm, and the absorbance of chlorophyll at 975-985 nm has small influence on the light absorption and the thickness of plant leaves, so that the absorbance values of the plant leaves tested under the wavelengths of 460+/-5 nm, 660+/-5 nm and 980+/-5 nm are selected to be substituted into a formula, the established model has better adaptability, and the chlorophyll a, chlorophyll b, total chlorophyll and carotenoid values calculated based on the model are closer to actual values.
Further, in step 1, the absorbance value A 1 、A 2 And A 3 The test method specifically comprises the following steps: the absorbance values of the plant leaves at each wavelength were calculated from the logarithm of the ratio of the intensity of light not transmitted and transmitted through the plant leaves at different wavelengths of 460.+ -.5 nm, 660.+ -.5 nm and 980.+ -.5 nm.
Further, in step 1, the absorbance value A 1 、A 2 And A 3 The test method specifically comprises the following steps: the method comprises the steps of taking 3 light emitting elements with peak wavelengths or center wavelengths respectively at 460+/-5 nm, 660+/-5 nm and 980+/-5 nm as light sources, taking a photoelectric sensor as illumination collection equipment, enabling the 3 light emitting elements to be opposite to the photoelectric sensor, enabling the distance between the 3 light emitting elements to be 0.5-3 mm, and collecting the original light intensities of the light sources at the wavelengths of 460+/-5 nm, 660+/-5 nm and 980+/-5 nm respectively, wherein the original light intensities are marked as Y 1 、Y 2 And Y 3 ；
The light intensity collecting equipment is used for collecting the light intensity after the plant leaves are put into the light intensity collecting equipment, and the light intensity transmitted through the plant leaves under the wavelengths of 460+/-5 nm, 660+/-5 nm and 980+/-5 nm is respectively recorded as T 1 、T 2 And T 3 ；
According to said Y 1 、Y 2 、Y 3 And said T 1 、T 2 、T 3 Calculating absorbance values of plant leaves at wavelengths of 460+ -5 nm, 660+ -5 nm and 980+ -5 nm, specifically absorbance value A at wavelengths of 460+ -5 nm 1 ＝lg(Y 1 /T 1 ) Absorbance value A at 660+ -5 nm wavelength 2 ＝lg(Y 2 /T 2 ) Absorbance value A at 980.+ -. 5nm wavelength 3 ＝lg(Y 3 /T 3 )。
The invention discloses the following technical effects:
the invention realizes nondestructive rapid detection of chlorophyll a, chlorophyll b, total chlorophyllin and carotenoid content by establishing a relational expression based on absorbance of plant leaves at three wavelengths (460+/-5 nm, 660+/-5 nm and 980+/-5 nm) and the relation of actual chlorophyll a, chlorophyll b, total chlorophyll and carotenoid content measured according to spectrophotometry regulated by Chinese agricultural industry standard NY/T3082-2017. The method has the advantages of rapidness, simplicity, no damage, low cost and no need of any chemical reagent.
Compared with the prior art, the invention can realize nondestructive rapid detection of the contents of chlorophyll a, chlorophyll b, total chlorophyll and carotenoid in plant leaves, does not need to use any chemical reagent, and does not cause harm to the health of testers.
The device for carrying out nondestructive rapid detection on the chlorophyll and carotenoid contents by using the method can be prepared on the basis of the method, is beneficial to scientific researchers and agricultural growers to quickly know the chlorophyll a, chlorophyll b, total chlorophyll and carotenoid contents of plant leaves after popularization and use, and has important effects on guiding fertilization, light supplementing, variety cultivation and the like of crops.
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In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings can be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a schematic flow chart of nondestructive rapid detection of chlorophyll and carotenoid content in plant leaves according to the invention;
FIG. 2 is a comparison of chlorophyll-a content in vegetable leaves measured by the method of example 1 with actual chlorophyll-a content results measured by spectrophotometry;
FIG. 3 is a comparison of chlorophyll b content in vegetable leaves as determined by the method of example 1 with actual chlorophyll b content results as determined by spectrophotometry;
FIG. 4 is a comparison of the total chlorophyll content in vegetable leaves as determined by the method of example 1 with the actual total chlorophyll content results as determined by spectrophotometry;
FIG. 5 is a comparison of carotenoid content in vegetable leaves as determined by the method of example 1 with actual carotenoid content as determined by spectrophotometry.
Detailed Description
Various exemplary embodiments of the invention will now be described in detail, which should not be considered as limiting the invention, but rather as more detailed descriptions of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in this disclosure, it is also to be understood that each intervening value, between the upper and lower limit of that range is specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples of the present invention are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.
The invention provides a method for constructing a model for nondestructive rapid detection of chlorophyll and carotenoid content in plant leaves, which comprises the following steps:
testing absorbance A of plant leaf at light source wavelengths of 460+ -5 nm, 660+ -5 nm and 980+ -5 nm 1 、A 2 And A 3 ；
Spectrophotometry is used for measuring the contents of chlorophyll a, chlorophyll b, total chlorophyll and carotenoid in plant leaves, and the contents are respectively recorded as YLa, YLb, YLz and YLc;
the YLa, YLb, YLz and YLc are respectively used as dependent variables, and the A is used as the 1 、A 2 And A 3 And establishing linear and nonlinear models for independent variables, and taking the model with higher decision coefficient and simpler structure as the optimal model.
The reason why the absorbance values of the plant leaves are tested at the wavelengths of 460+ -5 nm, 660+ -5 nm and 980+ -5 nm is selected in the invention: the absorbance of chlorophyll b is strongest near 460nm, the absorbance of chlorophyll a and the absorbance of chlorophyll b are intersected near 660nm, and the absorbance of chlorophyll at 975-985 nm has small influence on the light absorption and the thickness of plant leaves, so that the calculated result can be more approximate to the actual values of chlorophyll a, chlorophyll b, total chlorophyll and carotenoid in the plant leaves by substituting the absorbance values of the tested plant leaves at wavelengths of 460+/-5 nm, 660+/-5 nm and 980+/-5 nm into the formula. The prior art describes the detection of total chlorophyll in plant leaves at wavelengths of 460nm, 650nm and 940nm, which, if selected at three wavelengths of 460nm, 650nm and 940nm, results in a greater impact of leaf thickness on the detection results.
Further, the A 1 、A 2 And A 3 The test method of (1) comprises the following steps: the absorbance values of the plant leaves at each wavelength were calculated from the logarithm of the ratio of the intensity of light not transmitted and transmitted through the plant leaves at different wavelengths of 460.+ -.5 nm, 660.+ -.5 nm and 980.+ -.5 nm.
Further, the A 1 、A 2 And A 3 The test method of (1) comprises the following steps:
step a, setting up light intensity acquisition equipment by taking 3 luminous elements with peak wavelengths or center wavelengths respectively at 460+/-5 nm, 660+/-5 nm and 980+/-5 nm as light sources, taking a photoelectric sensor as illumination acquisition equipment, setting up the light intensity acquisition equipment by taking the 3 luminous elements and the photoelectric sensor as opposite, setting the middle spacing to be 0.5-3 mm, and acquiring the original light intensities of the LED light sources at the wavelengths of 460+/-5 nm, 660+/-5 nm and 980+/-5 nm, which are respectively marked as Y 1 、Y 2 And Y 3 ；
Step b, collecting plant leaves, and measuring the transmitted light intensities of 460+/-5 nm, 660+/-5 nm and 980+/-5 nm when the light intensity collecting device in the step 1 is used for transmitting the plant leaves, wherein the transmitted light intensities are respectively recorded as T 1 、T 2 And T 3 ；
At measurement T 1 、T 2 And T 3 When the plant leaves are not picked, namely the transmitted light intensity can be collected in the growth state of the plant leaves, and the transmitted light intensity can be collected after the plant leaves are picked, so that the transmitted light intensity is required to be collected within 20 minutes after the plant leaves are picked.
Step c, calculating eachAbsorbance of plant leaf at each wavelength, specifically absorbance A at 460.+ -.5 nm 1 ＝lg(Y 1 /T 1 ) Absorbance A at 660+ -5 nm wavelength 2 ＝lg(Y 2 /T 2 ) Absorbance A at 980.+ -. 5nm wavelength 3 ＝lg(Y 3 /T 3 )。
The invention also provides a method for nondestructively and rapidly detecting the contents of chlorophyll and carotenoid in plant leaves, which comprises the following steps:
step 1, measuring absorbance value A of plant leaf at light source wavelength of 460+ -5 nm, 660+ -5 nm and 980+ -5 nm 1 、A 2 And A 3 ；
Step 2, the A is carried out 1 、A 2 And A 3 Substituting the values into formulas (1), (2), (3) and (4) to obtain Cla, clb, clz and Clc values;
Cla＝-0.952A 1 +1.828A 2 -1.934A 3 +0.402 (1)
Clb＝-0.462A 1 +0.685A 2 -0.578A 3 +0.208 (2)
Cl7＝-1.580A 1 +2.831A 2 -2.859A 3 +0.689 (3)
Clc＝-0.154A 1 +0.378A 2 -0.443A 3 +0.072 (4)
the Cla represents the content of chlorophyll a; the Clb represents the content of chlorophyll b; said Clz represents the total chlorophyll content; the Clc represents the carotenoid content.
Further, in step 1, the absorbance value A 1 、A 2 And A 3 The test method specifically comprises the following steps: the absorbance values of the plant leaves at each wavelength were calculated from the logarithm of the ratio of the intensity of light not transmitted and transmitted through the plant leaves at different wavelengths of 460.+ -.5 nm, 660.+ -.5 nm and 980.+ -.5 nm.
Further, in step 1, the absorbance value A 1 、A 2 And A 3 The test method specifically comprises the following steps: the light source is 3 light emitting elements with peak wavelength or center wavelength of 460+ -5 nm, 660+ -5 nm and 980+ -5 nm, and the photoelectric sensor is used for collecting illuminationThe light source is provided with 3 light emitting elements opposite to the photoelectric sensor, the distance is 0.5-3 mm, and the original light intensities of the light sources under the wavelengths of 460+ -5 nm, 660+ -5 nm and 980+ -5 nm are respectively collected and marked as Y 1 、Y 2 And Y 3 ；
The light intensity collecting equipment is used for collecting the light intensity after the plant leaves are put into the light intensity collecting equipment, and the light intensity transmitted through the plant leaves under the wavelengths of 460+/-5 nm, 660+/-5 nm and 980+/-5 nm is respectively recorded as T 1 、T 2 And T 3 ；
According to said Y 1 、Y 2 、Y 3 And said T 1 、T 2 、T 3 Calculating absorbance values of plant leaves at wavelengths of 460+ -5 nm, 660+ -5 nm and 980+ -5 nm, specifically absorbance value A at wavelengths of 460+ -5 nm 1 ＝lg(Y 1 /T 1 ) Absorbance value A at 660+ -5 nm wavelength 2 ＝lg(Y 2 /T 2 ) Absorbance value A at 980.+ -. 5nm wavelength 3 ＝lg(Y 3 /T 3 )。
Further, the formulas (1), (2), (3) and (4) in the step 2 are obtained by taking absorbance values of the plant leaves at wavelengths of 460+ -5 nm, 660+ -5 nm and 980+ -5 nm as independent variables, taking actual values of chlorophyll a, chlorophyll b, total chlorophyll and carotenoid contents in the plant leaves as independent variables, and establishing a relation between each independent variable and the independent variable.
Further, actual values of chlorophyll a, chlorophyll b, total chlorophyll content, and carotenoid content in the plant leaves were measured using spectrophotometry.
Further, the establishment of the formulas (1), (2), (3) and (4) in the step 2 is specifically:
measuring the chlorophyll a, chlorophyll b, total chlorophyll and carotenoid content of each plant leaf according to spectrophotometry specified in Chinese agricultural industry standard NY/T3082-2017, and taking the result as actual values of the chlorophyll a, chlorophyll b, total chlorophyll content and carotenoid content of each plant leaf, and respectively marking as YLa, YLb, YLz, YLc;
plant leaf suction at wavelengths of 460+ -5 nm, 660+ -5 nm and 980+ -5 nm with YLa, YLb, YLz, YLc as dependent variableLuminosity, i.e. A 1 、A 2 And A 3 And (3) respectively being independent variables, selecting linear models such as 'automatic linear modeling', 'linear', 'partial least squares', and the like under 'regression' under 'analysis' menu of IBM SPSS Statistics software, and nonlinear models such as 'multi-element logic', 'nonlinear', 'second-order least squares', and the like, and establishing a multi-element linear model and a nonlinear model of each independent variable and independent variable, wherein the models with larger determining coefficients and simpler relational expression are used as optimal models, namely the formulas (1), (2), (3) and (4) in the step (2).
The flow chart of the nondestructive rapid detection of chlorophyll and carotenoid content in plant leaves is shown in figure 1.
Example 1
(1) Light intensity acquisition equipment is built
Taking 3 Light Emitting Diodes (LEDs) with peak wavelengths (or center wavelengths) respectively at 460nm, 660nm and 980nm as light sources, taking a photoelectric sensor as illumination acquisition equipment, enabling the 3 light emitting diodes to be opposite to the photoelectric sensor, acquiring the original light intensity of the LED light sources at 3 wavelengths without plant leaves, and respectively marking as Y 1 、Y 2 And Y 3 。
(2) Plant leaf treatment
220 leaves of lettuce, green vegetables and spinach are collected, the leaves are tested within 20 minutes after picking, cut into square shapes with the length of 25mm multiplied by 25mm, and numbered, and then a plant leaf sample is obtained.
(3) Plant leaf light intensity acquisition and calculation
The built light intensity collection equipment is used for collecting the light intensity transmitted through the plant leaves, each plant leaf sample is collected for 3 times to obtain an average value, and data are recorded according to numbers. The intensity of light transmitted through the leaves of the plant at 460nm, 660nm and 980nm is respectively recorded as T 1 、T 2 And T 3 。
According to the original light intensity Y at each wavelength 1 、Y 2 And Y 3 And transmitted light intensity T after transmitted through plant leaves 1 、T 2 And T 3 Calculation of plant leaf absorptions at 460nm, 660nm and 980nmLuminosity A, in particular absorbance A at 460nm 1 ＝lg(Y 1 /T 1 ) Absorbance A at 660nm 2 ＝lg(Y 2 /T 2 ) Absorbance A at 980nm 3 ＝lg(Y 3 /T 3 )。
(4) Determination of actual chlorophyll a, chlorophyll b, total chlorophyll and carotenoid content of plant leaves
Actual values of chlorophyll a, chlorophyll b, total chlorophyll and carotenoid content in the plant leaves were measured according to spectrophotometry prescribed by chinese agricultural industry standard NY/T3082-2017, and are labeled YLa, YLb, YLz and YLc, respectively.
(5) Establishment of calculation formulas of chlorophyll a, chlorophyll b, total chlorophyll and carotenoid
The absorbance at 3 wavelengths is used as an independent variable, and chlorophyll a, chlorophyll b, total chlorophyll and carotenoid contents are respectively used as independent variables to establish a relation, wherein the relation is:
Cla＝-0.952A 1 +1.828A 2 -1.934A 3 +0.402 (1)
Clb＝-0.462A 1 +0.685A 2 -0.578A 3 +0.208 (2)
Clz＝-1.580A 1 +2.831A 2 -2.859A 3 +0.689 (3)
Clc＝-0.154A 1 +0.378A 2 -0.443A 3 +0.072 (4)
wherein Cla, clb, clz and CLc represent chlorophyll a, chlorophyll b, total chlorophyll and carotenoids, respectively. The decision coefficients of formulas (1), (2), (3) and (4) are 0.92, 0.88, 0.93 and 0.89, respectively.
(6) Verification of chlorophyll a, chlorophyll b, total chlorophyll and carotenoid calculation formulas
And (3) respectively taking 110 lettuce, green vegetables and spinach leaves, firstly utilizing the light intensity acquisition equipment constructed in the step (1) to acquire the original light intensity when the plant leaves are not put in, and then acquiring the transmitted light intensity when the plant leaves are transmitted after the plant leaves are put in. And then measuring the actual contents of chlorophyll a, chlorophyll b, total chlorophyll and carotenoid in the plant leaves according to a spectrophotometry specified by Chinese agricultural industry standard NY/T3082-2017. Substituting the calculated absorbance at each wavelength into the formula determined in the step (5), and calculating the contents of chlorophyll a, chlorophyll b, total chlorophyll and carotenoid. The calculated chlorophyll a, chlorophyll b, total chlorophyll and carotenoid content is compared with its actual content. The results are shown in FIGS. 2-5.
As can be seen from FIGS. 2-5, the root mean square error of the inventive method for chlorophyll a, chlorophyll b, total chlorophyll and carotenoid content was 0.068mg/g, 0.024mg/g, 0.085mg/g and 0.019mg/g, respectively, and the measurement time was less than 2s. The method can be used for nondestructively and rapidly detecting the contents of chlorophyll a, chlorophyll b, total chlorophyll and carotenoid in plant leaves.
Besides three kinds of lettuce, green vegetables and spinach listed in example 1, the method of the invention is also applicable to other plant leaves containing chlorophyll, and has wide application range.
The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.
Claims (3)
1. A method for nondestructive rapid detection of chlorophyll and carotenoid content in plant leaves, comprising the steps of:
step 1, measuring absorbance A of plant leaf at wavelengths of 460+ -5 nm, 660+ -5 nm and 980+ -5 nm 1 、A 2 And A 3 ；
Step 2, the A is carried out 1 、A 2 And A 3 Substituting the values into formulas (1), (2), (3) and (4) to obtain Cla, clb, clz and Clc values;
2. The method for the nondestructive rapid detection of chlorophyll and carotenoid content in plant leaves according to claim 1, wherein in step 1, the absorbance value a 1 、A 2 And A 3 The test method specifically comprises the following steps: the absorbance values of the plant leaves at each wavelength were calculated from the logarithm of the ratio of the intensity of light not transmitted and transmitted through the plant leaves at different wavelengths of 460.+ -.5 nm, 660.+ -.5 nm and 980.+ -.5 nm.
3. A method for the non-destructive rapid detection of chlorophyll and carotenoid content in plant leaves according to claim 2, wherein in step 1, said absorbance value a 1 、A 2 And A 3 The test method specifically comprises the following steps: taking 3 light emitting elements with peak wavelengths or center wavelengths of 460+/-5 nm, 660+/-5 nm and 980+/-5 nm as light sources, taking a photoelectric sensor as illumination collection equipment, enabling the 3 light emitting elements to be opposite to the photoelectric sensor, enabling the distance to be 0.5-3 mm, and collecting the original light intensities of the light sources with the wavelengths of 460+/-5 nm, 660+/-5 nm and 980+/-5 nm respectively, wherein the original light intensities are marked as Y 1 、Y 2 And Y 3 ；
The illumination collection equipment is used for collecting the light intensity after the plant leaves are put into the illumination collection equipment, and the light intensity transmitted through the plant leaves under the wavelengths of 460+/-5 nm, 660+/-5 nm and 980+/-5 nm is respectively recorded as T 1 、T 2 And T 3 ；
According to said Y 1 、Y 2 、Y 3 And said T 1 、T 2 、T 3 Calculating absorbance values of plant leaves at wavelengths of 460+ -5 nm, 660+ -5 nm and 980+ -5 nm, specifically absorbance value A at wavelengths of 460+ -5 nm 1 =lg（Y 1 /T 1 ) Absorbance value A at 660+ -5 nm wavelength 2 =lg（Y 2 /T 2 ) Absorbance value A at 980.+ -. 5nm wavelength 3 =lg（Y 3 /T 3 ）。
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