阅读说明：本技术 一种植物叶绿素荧光参数的校正方法和装置 (Method and device for correcting fluorescence parameters of plant chlorophyll ) 是由 翁海勇 刘浪 田雅 杨林 何城城 黄逸平 崔蕴涵 黄俊昆 叶大鹏 于 2019-09-30 设计创作，主要内容包括：本发明提出一种植物叶绿素荧光参数的校正方法和装置,包括以下步骤；A1、对植物暗适应处理；A2、获取植物的最小叶绿素荧光图像,再获取植物的最大叶绿素荧光图像；A3、使植物处于光化光照射环境下,获取植物此时的初始反射图像；A4、以光化光对植物照射并持续t时长,使植物达到光适应状态后,获取植物反射图像和动态瞬时荧光图像；A5、使植物被紫外光光照t时长后,获取植物稳态的蓝色光谱荧光图像、绿色光谱荧光图像、红色光谱荧光图像和远红外光谱荧光图像；A6、计算叶绿素荧光产量偏差系数；A7、进行校正计算,得到光照条件下实际的叶绿素荧光产量参数；A8、经运算得到实际叶绿素荧光参数图像以进行分析；本发明能更加准确检测光合作用效率。(The invention provides a method and a device for correcting a plant chlorophyll fluorescence parameter, which comprises the following steps; a1, dark adaptation treatment of plants; a2, acquiring a minimum chlorophyll fluorescence image of a plant, and acquiring a maximum chlorophyll fluorescence image of the plant; a3, placing the plant in an environment of actinic light irradiation, and acquiring an initial reflection image of the plant at the moment; a4, irradiating the plants with actinic light for t duration to enable the plants to reach a light adaptation state, and then acquiring a plant reflection image and a dynamic instantaneous fluorescence image; a5, obtaining a blue spectrum fluorescence image, a green spectrum fluorescence image, a red spectrum fluorescence image and a far infrared spectrum fluorescence image of the plant in a stable state after the plant is illuminated by ultraviolet light for a time period t; a6, calculating a chlorophyll fluorescence yield deviation coefficient; a7, carrying out correction calculation to obtain actual chlorophyll fluorescence yield parameters under the illumination condition; a8, obtaining an actual chlorophyll fluorescence parameter image through calculation for analysis; the invention can detect the photosynthesis efficiency more accurately.)

1. a correction method of plant chlorophyll fluorescence parameters is used for measuring the plant chlorophyll fluorescence parameters, and is characterized in that: the method comprises the following steps;

a1, placing the plant in a dark environment, and carrying out dark adaptation treatment on the plant to reset the photosynthetic system of the plant to an initial state;

A2, in a dark environment, measuring light illumination, acquiring a minimum chlorophyll fluorescence image of the plant after dark adaptation treatment by a camera through a filter element, and acquiring a maximum chlorophyll fluorescence image of the plant after dark adaptation treatment by the camera through the filter element by oversaturated light illumination;

a3, exposing the plant to the environment of actinic light, and immediately acquiring an initial reflection image of the plant exposed to the environment of actinic light through a camera;

a4, irradiating the plants with actinic light for t time, and acquiring reflected images of the plants irradiated with the actinic light for t time by a camera after the plants reach a light adaptation state; then a camera is used for obtaining the plant dynamic instantaneous fluorescence image at the moment through a light filtering element;

A5, after the plant is in an ultraviolet light illumination environment for a time t, a camera is used for obtaining a blue spectrum fluorescence image, a green spectrum fluorescence image, a red spectrum fluorescence image and a far infrared spectrum fluorescence image of the plant steady state after the plant is irradiated by the ultraviolet light for the time t through a light filtering element;

A6, obtaining the chlorophyll yield Ro at the moment according to the initial reflection image of the plant in the actinic light irradiation environment; obtaining the chlorophyll yield Rt at the time according to the reflection image of the plant irradiated by the long-time actinic light at t, and calculating the deviation coefficient of the chlorophyll fluorescence yield caused by chloroplast movement under the illumination condition according to the formula c =1+ ((Rt-Ro)) ⁄ Ro;

A7, correcting the dynamic and steady chlorophyll fluorescence yield to obtain actual chlorophyll fluorescence yield parameters under illumination conditions;

And A8, performing mathematical operation on the corrected chlorophyll fluorescence yield parameters to obtain a chlorophyll fluorescence parameter image capable of reflecting the actual photosynthesis of the plant for analysis.

2. The method according to claim 1, wherein the dark adaptation treatment is performed on the plant for not less than 25 minutes in step A1, and the plant is irradiated with 740nm far-infrared light having an intensity of about 10 μmol-m ^-2 -s ^-1 during the dark adaptation treatment.

3. the method for correcting the fluorescence parameters of plant chlorophyll according to claim 1, wherein the method comprises the following steps: in step a3, the actinic light irradiation environment simulates a natural environment with actinic light of varying light intensity; the pattern of light intensity variations comprises one or more of a constant light intensity, a sin variation, a cos variation or a step variation.

4. the method for correcting the fluorescence parameters of plant chlorophyll according to claim 1, wherein the method comprises the following steps: the chlorophyll fluorescence parameters comprise non-actinic light quenching coefficients, minimum fluorescence variable fluorescence in a light adaptation state and steady-state fluorescence parameters in the light adaptation state.

5. a correction device of plant chlorophyll fluorescence parameter is characterized in that: the correction device is a device used by the method of claim 1;

The correcting device comprises a control module and a box body, wherein a reflector is arranged on the side wall of the box body, and the box body can contain plants to be measured; a light source component and a camera are arranged on the top plate of the box body; the photographing direction of the camera points to the plants in the box; a lens is arranged in front of the camera, and a filter element is arranged between the camera and the lens; the light source assembly can change the illumination environment of the plant under the control of the control module; the light filtering element can filter the light rays emitted to the light inlet end of the camera under the control of the control module.

6. The device for correcting the fluorescence parameters of plant chlorophyll according to claim 5, wherein: a conveying belt is arranged in the box body; the transmission belt is provided with a hole tray for loading plants; the transmission band can be sent into or send the box with the plant.

7. the device for correcting the fluorescence parameters of plant chlorophyll according to claim 5, wherein: the camera is a monochrome CCD camera.

8. The device for correcting the fluorescence parameters of plant chlorophyll according to claim 7, wherein: the filtering element is a filtering wheel; six working positions are arranged on the filtering wheel; the working positions comprise five filtering positions and a zero position; the zero position does not filter light; each light filtering position is provided with a light filter for filtering light; the filters of each filtering position are respectively a 680nm band-pass filter, a 440nm band-pass filter, a 520nm band-pass filter, a 690nm band-pass filter and a 740nm band-pass filter.

9. The device for correcting the fluorescence parameters of plant chlorophyll according to claim 7, wherein: the light source assembly comprises a plurality of LED light sources which are arranged in an annular shape; the camera is arranged at the center of the ring where the LED light sources are arranged; the plant is located under the camera when measuring.

10. the device for correcting the fluorescence parameters of plant chlorophyll according to claim 7, wherein: the LED light source comprises an actinic light source with the central wavelength of 620nm and an ultraviolet light source with the central wavelength of 400 nm; the actinic light source is used for exciting dynamic chlorophyll fluorescence and providing a light source for acquiring a reflection image; the ultraviolet light source is used for exciting the steady chlorophyll fluorescence.

Technical Field

the invention relates to the technical field of plant phenotype analysis, in particular to a method and a device for correcting plant chlorophyll fluorescence parameters.

Background

Photosynthetic efficiency is an important trait in plant phenotype and is one of the key points of research. The chloroplast is used as a mechanism of photosynthesis, and the rapid nondestructive monitoring of the physiological state of the chloroplast is an important link for realizing high-flux photosynthesis analysis. Chlorophyll fluorescence is a "probe" of plant photosynthesis, and can reflect the real-time plant photosynthesis efficiency. However, when measuring chlorophyll fluorescence signals of plant leaves, the behavior (aggregation and movement) that chloroplasts can move under illumination is often ignored, so that the measured fluorescence yield is deviated, the accuracy of measuring the photosynthesis efficiency is reduced, and the selection of varieties with excellent photosynthesis efficiency is not facilitated.

Disclosure of Invention

the invention provides a method and a device for correcting plant chlorophyll fluorescence parameters, which can be used for correcting an original chlorophyll fluorescence image to obtain a real chlorophyll fluorescence image and can realize the purpose of more accurately detecting the photosynthesis efficiency.

The invention adopts the following technical scheme.

A method for correcting a plant chlorophyll fluorescence parameter, which is used for measuring a plant chlorophyll fluorescence parameter, and comprises the following steps;

A1, placing the plant in a dark environment, and carrying out dark adaptation treatment on the plant to reset the photosynthetic system of the plant to an initial state;

A2, in a dark environment, measuring light illumination, acquiring a minimum chlorophyll fluorescence image of the plant after dark adaptation treatment by a camera through a filter element, and acquiring a maximum chlorophyll fluorescence image of the plant after dark adaptation treatment by the camera through the filter element by oversaturated light illumination;

A3, exposing the plant to the environment of actinic light, and immediately acquiring an initial reflection image of the plant exposed to the environment of actinic light through a camera;

A4, irradiating the plants with actinic light for t time, and acquiring reflected images of the plants irradiated with the actinic light for t time by a camera after the plants reach a light adaptation state; then a camera is used for obtaining the plant dynamic instantaneous fluorescence image at the moment through a light filtering element;

A5, after the plant is in an ultraviolet light illumination environment for a time t, a camera is used for obtaining a blue spectrum fluorescence image, a green spectrum fluorescence image, a red spectrum fluorescence image and a far infrared spectrum fluorescence image of the plant steady state after the plant is irradiated by the ultraviolet light for the time t through a light filtering element;

a6, obtaining the chlorophyll yield Ro at the moment according to the initial reflection image of the plant in the actinic light irradiation environment; obtaining the chlorophyll yield Rt at the time according to the reflection image of the plant irradiated by the long-time actinic light at t, and calculating the deviation coefficient of the chlorophyll fluorescence yield caused by chloroplast movement under the illumination condition according to the formula c =1+ ((Rt-Ro)) ⁄ Ro;

a7, correcting the dynamic and steady chlorophyll fluorescence yield to obtain actual chlorophyll fluorescence yield parameters under illumination conditions;

And A8, performing mathematical operation on the corrected chlorophyll fluorescence yield parameters to obtain a chlorophyll fluorescence parameter image capable of reflecting the actual photosynthesis of the plant for analysis.

In the step A1, the dark adaptation treatment is performed on the plant for not less than 25 minutes, and in the dark adaptation treatment, 740nm far-infrared light having an intensity of about 10. mu. mol · m ^-2 · s ^-1 may be used to irradiate the plant.

In step a3, the actinic light irradiation environment simulates a natural environment with actinic light of varying light intensity; the pattern of light intensity variations comprises one or more of a constant light intensity, a sin variation, a cos variation or a step variation.

The chlorophyll fluorescence parameters comprise non-actinic light quenching coefficients, minimum fluorescence variable fluorescence in a light adaptation state and steady-state fluorescence parameters in the light adaptation state.

a correction device for plant chlorophyll fluorescence parameters, wherein the correction device is a device used in the method;

the correcting device comprises a control module and a box body, wherein a reflector is arranged on the side wall of the box body, and the box body can contain plants to be measured; a light source component and a camera are arranged on the top plate of the box body; the photographing direction of the camera points to the plants in the box; a lens is arranged in front of the camera, and a filter element is arranged between the camera and the lens; the light source assembly can change the illumination environment of the plant under the control of the control module; the light filtering element can filter the light rays emitted to the light inlet end of the camera under the control of the control module.

a conveying belt is arranged in the box body; the transmission belt is provided with a hole tray for loading plants; the transmission band can be sent into or send the box with the plant.

the camera is a monochrome CCD camera.

The filtering element is a filtering wheel; six working positions are arranged on the filtering wheel; the working positions comprise five filtering positions and a zero position; the zero position does not filter light; each light filtering position is provided with a light filter for filtering light; the filters of each filtering position are respectively a 680nm band-pass filter, a 440nm band-pass filter, a 520nm band-pass filter, a 690nm band-pass filter and a 740nm band-pass filter.

The light source assembly comprises a plurality of LED light sources which are arranged in an annular shape; the camera is arranged at the center of the ring where the LED light sources are arranged; the plant is located under the camera when measuring.

the LED light source comprises an actinic light source with the central wavelength of 620nm and an ultraviolet light source with the central wavelength of 400 nm; the actinic light source is used for exciting dynamic chlorophyll fluorescence and providing a light source for acquiring a reflection image; the ultraviolet light source is used for exciting the steady chlorophyll fluorescence.

Compared with the prior art, the invention has the beneficial effects that:

(1) The method and the device can overcome the deviation of chlorophyll fluorescence signal measurement caused by the movement of chloroplast caused by the response to light, and can more accurately acquire the photosynthesis information of plants

(2) The method and the device set different illumination modes through programs, simulate the light intensity change of natural conditions, strengthen the difference of the plant light response to different illumination and realize the accurate detection of the plant photosynthesis efficiency.

(3) the obtained corrected fluorescence signals comprise dynamic fluorescence and steady-state fluorescence, and the photosynthesis difference of plants with different genotypes can be explained from different photosynthesis scales.

The invention provides a method and a device for correcting chlorophyll fluorescence parameters, which are used for calculating a deviation coefficient by respectively obtaining fluorescent images of plants under dark adaptation and illumination conditions, collecting red light reflection images corresponding to an initial illumination time and a t time, and correcting an original chlorophyll fluorescence image to finally obtain a real chlorophyll fluorescence image, thereby realizing the purpose of more accurately detecting the photosynthesis efficiency.

Drawings

The invention is described in further detail below with reference to the following figures and detailed description:

FIG. 1 is a schematic view of the apparatus of the present invention;

FIG. 2 is a schematic view of the present invention at the top panel of the cabinet;

FIG. 3 is a schematic flow diagram of the process of the present invention;

in the figure: 1-a camera; 2-a plant; 3-plug tray; 4, conveying the belt; 5-a box body; 6-source of actinic light; 7-ultraviolet light source; 14-a filter wheel; 15-a control module; 201-lens; 202-working bit.

Detailed Description

1-3, a method for correcting a fluorescence parameter of chlorophyll in a plant for measuring a fluorescence parameter of chlorophyll in a plant, said method comprising the steps of;

A1, placing the plant in a dark environment, and carrying out dark adaptation treatment on the plant to reset the photosynthetic system of the plant to an initial state;

A2, in a dark environment, measuring light illumination, acquiring a minimum chlorophyll fluorescence image of the plant after dark adaptation treatment by a camera through a filter element, and acquiring a maximum chlorophyll fluorescence image of the plant after dark adaptation treatment by the camera through the filter element by oversaturated light illumination;

A3, exposing the plant to the environment of actinic light, and immediately acquiring an initial reflection image of the plant exposed to the environment of actinic light through a camera;

A4, irradiating the plants with actinic light for t time, and acquiring reflected images of the plants irradiated with the actinic light for t time by a camera after the plants reach a light adaptation state; then a camera is used for obtaining the plant dynamic instantaneous fluorescence image at the moment through a light filtering element;

A5, after the plant is in an ultraviolet light illumination environment for a time t, a camera is used for obtaining a blue spectrum fluorescence image, a green spectrum fluorescence image, a red spectrum fluorescence image and a far infrared spectrum fluorescence image of the plant steady state after the plant is irradiated by the ultraviolet light for the time t through a light filtering element;

a6, obtaining the chlorophyll yield Ro at the moment according to the initial reflection image of the plant in the actinic light irradiation environment; obtaining the chlorophyll yield Rt at the time according to the reflection image of the plant irradiated by the long-time actinic light at t, and calculating the deviation coefficient of the chlorophyll fluorescence yield caused by chloroplast movement under the illumination condition according to the formula c =1+ ((Rt-Ro)) ⁄ Ro;

a7, correcting the dynamic and steady chlorophyll fluorescence yield to obtain actual chlorophyll fluorescence yield parameters under illumination conditions;

And A8, performing mathematical operation on the corrected chlorophyll fluorescence yield parameters to obtain a chlorophyll fluorescence parameter image capable of reflecting the actual photosynthesis of the plant for analysis.

in the step A1, the dark adaptation treatment is performed on the plant for not less than 25 minutes, and in the dark adaptation treatment, 740nm far-infrared light having an intensity of about 10. mu. mol · m ^-2 · s ^-1 may be used to irradiate the plant.

In step a3, the actinic light irradiation environment simulates a natural environment with actinic light of varying light intensity; the pattern of light intensity variations comprises one or more of a constant light intensity, a sin variation, a cos variation or a step variation.

the chlorophyll fluorescence parameters comprise non-actinic light quenching coefficients, minimum fluorescence variable fluorescence in a light adaptation state and steady-state fluorescence parameters in the light adaptation state.

A correction device for plant chlorophyll fluorescence parameters, wherein the correction device is a device used in the method;

The correcting device comprises a control module 15 and a box body, wherein a reflector is arranged on the side wall of the box body, and the box body can contain the plant 2 to be measured; a light source component and a camera 1 are arranged on the top plate of the box body; the photographing direction of the camera points to the plants in the box; a lens 201 is arranged in front of the camera, and a filter element is arranged between the camera and the lens; the light source assembly can change the illumination environment of the plant under the control of the control module; the light filtering element can filter the light rays emitted to the light inlet end of the camera under the control of the control module.

A conveying belt 4 is arranged in the box body; the transmission belt is provided with a hole tray 3 for loading plants; the transmission band can be sent into or send the box with the plant.

The camera is a monochrome CCD camera.

the filter element is a filter wheel 14; six working positions 202 are arranged on the filter wheel; the working positions comprise five filtering positions and a zero position; the zero position does not filter light; each light filtering position is provided with a light filter for filtering light; the filters of each filtering position are respectively a 680nm band-pass filter, a 440nm band-pass filter, a 520nm band-pass filter, a 690nm band-pass filter and a 740nm band-pass filter.

the light source assembly comprises a plurality of LED light sources which are arranged in an annular shape; the camera is arranged at the center of the ring where the LED light sources are arranged; the plant is located under the camera when measuring.

the LED light source comprises an actinic light source 6 with the central wavelength of 620nm and an ultraviolet light source 7 with the central wavelength of 400 nm; the actinic light source is used for exciting dynamic chlorophyll fluorescence and providing a light source for acquiring a reflection image; the ultraviolet light source is used for exciting the steady chlorophyll fluorescence.