CN102426161A - Method for evaluating senescence degree of plant leaves based on photon radiation - Google Patents

Abstract

The invention discloses a method for evaluating the senescence degree of plant leaves based on photon radiation. According to the method, a leaf to be measured is cleaned with distilled water and put in dark for a plurality of minutes, and spontaneous photon radiation ISL per unit time of the leaf is measured by using a measuring system based on single photon detection; the leaf to be measured is irradiated for a certain period of time T by using foreign unsaturated exciting light, delayed luminescence of the leaf in the certain period of time T after optical excitation is measured by using the measuring system based on single photon detection, and delayed luminescence integrated intensity I(T) in the period of time T is obtained through mathematical fitting; the state parameter Q of the leaf and the relative change rate K of the Q are calculated, and the senescence degree of the leaf is evaluated according to K. The invention brings forward the method for describing, analyzing and measuring senescence of plant leaves based on photon radiation, proves reliability and maneuverability of the method through experiments and enables nondestructive test of the senescence degree of plant leaves to be realized.

Description

Estimate the method for plant leaf blade aging degree based on photon radiation

Technical field

The invention belongs to biological technical field, relate to a kind of method of estimating the plant leaf blade aging degree, be specifically related to a kind of method of estimating the plant leaf blade aging degree based on photon radiation.

Background technology

The plant leaf blade aging is that (Programmed cell death, PCD), it not only receives the expression regulation of internal gene, receives the influence of plant growth external environment condition simultaneously in a kind of procedural cell death.Research to plant leaf blade aging degree evaluation method is one of key problem of plant life scientific research, also has great economic benefit and social benefit.

At present, the research old and feeble to plant leaf blade has been deep into gene level, and still, how Nondestructive Evaluation is a basic problem that does not have solution for a long time with detecting the plant leaf blade aging.Since during plant senescence the result of chlorophyll degradation cause leaf color from green flavescence, redden or orange, thereby often with former the process and old and feeble sign of chlorophyll content as aging.In addition, protein content, mda content, SCAVENGING SYSTEM OF ACTIVATED OXYGEN (SOD, POD, CAT) content, and hormone-content, plasmalemma permeability and carboxylase activity or the like also are used as old and feeble index.But; Because the plant leaf blade aging relates to the complex physical process; Directly judge that through the leaf look aging degree can not accurately reflect old and feeble real conditions, and the acquisition of weighing the physiological and biochemical index of leaf senile needs destructive the measurement, can not accomplish Non-Destructive Testing.

Plant leaf blade self can send photon radiation; This bio-photon radiation is relevant closely with many life processes such as physiological metabolism, photosynthesis, cell divisions; Be the active electromagnetic signal that sends in the plant leaf blade cell activities, this signal can be measured.If can set up a cover plant leaf blade aging degree is described the method with quantitative test, just might carry out Non-Destructive Testing the plant leaf blade aging degree based on the bio-photon radiation.

Summary of the invention

The purpose of this invention is to provide and a kind ofly estimate the method for plant leaf blade aging degree, solve that the existing method that detects and estimate the plant leaf blade aging degree has destructiveness, measuring period is long and detect inaccurate problem based on photon radiation.

The technical scheme that the present invention adopts is based on the method for photon radiation evaluation plant leaf blade aging degree, specifically to implement according to following steps:

Step 1: clean blade to be measured with distilled water, it was placed several minutes in the dark, use the spontaneous photon radiation I that the measuring system that detects based on single photon is measured the blade unit interval _SL

Step 2: with blade to be measured with external unsaturation excitation light irradiation certain time interval T; Use the measuring system that detects based on single photon to measure it and receive the delayed luminescence in certain time interval T after the optical excitation, obtain the delayed luminescence integrated intensity I (T) in time T through the mathematics match;

Step 3: calculate the state parameter Q value of blade and the relative change rate K of Q value:

$Q=\frac{I\left(T\right)}{I_{\mathrm{SL}}T},$

$K=1-\frac{Q}{Q_0},$

Wherein, Q ₀State parameter for the blade initial state; Q is blade certain state parameter constantly in aging course, and according to the aging degree of this blade of size evaluation of K value, evaluation criterion is following: the size of K value is between 0～1; The K value of blade approaches 0 more, and the aging degree of this blade is low more; The K value approaches 1 more, and the aging degree of blade is big more.

Characteristics of the present invention also are,

Unsaturation exciting light in the step 2 wherein adopts a kind of in LED, laser, daylight lamp or the natural light.

Process mathematics match in the step 2 wherein obtains the delayed luminescence integrated intensity I (T) in time T, specifically implements according to following steps:

Step a. is expressed as blade delayed luminescence kinetics equation:

$I\left(t\right)=\frac{I_0}{{(1+\frac{t}{\mathrm{\τ}})}^{\mathrm{\β}}},$

Wherein, β, τ and I ₀Be constant, t is delayed luminescence die-away time;

Following formula is obtained the expression formula of delayed luminescence integrated intensity I (T) a measurement period T integrates:

$I\left(T\right)=\frac{\mathrm{\τ}{I}_0}{\mathrm{\β}-1}[1-\frac{1}{{(1+T/\mathrm{\τ})}^{\mathrm{\β}-1}}],$

After step b. cleans blade to be measured set by step 1 measure the blade unit interval spontaneous photon radiation I _SL

Step c with blade to be measured with external unsaturation excitation light irradiation after, use the measuring system that detects based on single photon to measure blade and receive after the optical excitation delayed luminescence change curve of t in time;

Steps d. with the delayed luminescence that obtains among the step c over time curve carry out the mathematics match, obtain delayed luminescence characteristic parameter: I ₀, τ and β;

Step f. is with the Measuring Time T and the delayed luminescence characteristic parameter I of delayed luminescence ₀, τ and β substitution delayed luminescence integrated intensity I (T) expression formula, obtain delayed luminescence integrated intensity I (T).

Process mathematics match in the step 2 wherein obtains the delayed luminescence integrated intensity I (T) in time T, specifically implements according to following steps:

Step a: blade delayed luminescence kinetics equation is expressed as:

$I\left(t\right)=I_{\mathrm{SL}}+\frac{I_0}{{(1+\frac{t}{\mathrm{\τ}})}^{\mathrm{\β}}},$

Wherein, I _SLBe the spontaneous photon radiation of blade unit interval, β, τ and I ₀Be constant, t is delayed luminescence die-away time, and T is measuring period;

Following formula is obtained the expression formula of delayed luminescence integrated intensity I (T) a measurement period T integrates:

$I\left(T\right)=I_{\mathrm{SL}}T+\frac{\mathrm{\τ}{I}_0}{\mathrm{\β}-1}[1-\frac{1}{{(1+T/\mathrm{\τ})}^{\mathrm{\β}-1}}],$

After step b. cleans blade to be measured set by step 1 measure the blade unit interval spontaneous photon radiation I _SL

Step c with blade to be measured with external unsaturation excitation light irradiation after, use the measuring system that detects based on single photon to measure blade and receive after the optical excitation delayed luminescence change curve of t in time;

Steps d. will go up the delayed luminescence that obtains of step over time curve carry out the mathematics match, obtain delayed luminescence characteristic parameter: I ₀, τ and β;

Step f. is with the above-mentioned I that obtains _SL, T, I ₀, τ and β substitution delayed luminescence integrated intensity I (T) expression formula, obtain delayed luminescence integrated intensity I (T).

Process mathematics match in the step 2 wherein obtains the delayed luminescence integrated intensity I (T) in time T, specifically implements according to following steps:

Step a. is expressed as the delayed luminescence kinetics equation:

$I\left(t\right)=I_0+I_1{e}^{-t/{\mathrm{\τ}}_1}+I_2{e}^{-t/{\mathrm{\τ}}_2},$

Wherein, I ₀, I ₁, I ₂And τ ₁, τ ₂Be constant, following formula obtained the expression formula of I (T) a measurement period T integrates:

$I\left(T\right)=I_{0}T+{\mathrm{<figure-callout\; id="1"\; label="I"\; filenames="HDA0000101331560000031.png"\; state="\{\{state\}\}">I</figure-callout>}}_1{\mathrm{\&tau;}}_1[1-\exp (-\frac{T}{{\mathrm{\&tau;}}_1})]+{\mathrm{<figure-callout\; id="2"\; label="I"\; filenames="HDA0000101331560000031.png"\; state="\{\{state\}\}">I</figure-callout>}}_2{\mathrm{\&tau;}}_2[1-\exp (-\frac{T}{{\mathrm{\&tau;}}_2})],$

Step b. uses external excitation light irradiation with blade to be measured, uses the measuring system that detects based on single photon to measure this blade and receives the delayed luminescence change curve of t in time after the optical excitation;

Step c will go up the delayed luminescence that obtains of step and carry out the mathematics match over time, obtain delayed luminescence characteristic parameter: I ₀, I ₁, I ₂, τ ₁, τ ₂

Steps d. with the above-mentioned T that obtains, I ₀, I ₁, I ₂, τ ₁, τ ₂The expression formula of substitution delayed luminescence integrated intensity I (T) obtains delayed luminescence integrated intensity I (T).

Q in the step 3 wherein ₀Acquisition, specifically implement according to following steps:

Step 1: the fresh blade when will be old and feeble is cleaned with distilled water, dries surface moisture, places in the dark, uses the spontaneous photon radiation I that the measuring system that detects based on single photon is measured the blade unit interval _SL

Step 2: the fresh blade when inciting somebody to action aging is with external unsaturation excitation light irradiation certain hour; Use the measuring system that detects based on single photon to measure it and receive the delayed luminescence in certain time interval T after the optical excitation, obtain the delayed luminescence integrated intensity I (T) in time T through the mathematics match;

Step 3: the spontaneous photon radiation I of unit interval when old and feeble with the above-mentioned blade that obtains _SL, delayed luminescence integrated intensity I (T) and delayed luminescence Measuring Time T bring into

Obtain the state parameter Q of blade when old and feeble ₀

The invention has the beneficial effects as follows, propose, and proved through experiment and the reliability and the operability of this method to have realized Non-Destructive Testing the plant leaf blade aging degree based on the plant leaf blade of photon radiation old and feeble description, analysis and measuring method.

Description of drawings

Fig. 1 is the variation of chlorophyll content in the spinach blade aging course in the embodiment of the invention;

Fig. 2 is the variation of spontaneous luminescence intensity in the spinach blade aging course in the embodiment of the invention;

Fig. 3 is the variation of delayed luminescence die-away curve in the spinach blade aging course in the embodiment of the invention;

Fig. 4 is the variation of Q and K value in the spinach blade aging course in the embodiment of the invention;

Fig. 5 is the relation that chlorophyll content and Q and K change in the spinach blade aging course in the embodiment of the invention.

Embodiment

Below in conjunction with accompanying drawing and embodiment the present invention is elaborated.

The plant leaf blade cell is a life system, and the state parameter (representing with Q) of describing the blade cell state is defined as:

$Q=\frac{E}{\mathrm{\&epsiv;}}---\left(1\right)$

Wherein, E is the energy of blade cell storage, and ε is the energy that cell dissipates away in vital movement.The Q value is big more, and cell state is good more.In the leaf senile process, the energy of blade cell storage is fewer and feweri, and meanwhile, the energy that dissipates away is more and more, and its Q value is more and more littler.

Plant leaf blade is excited with the unsaturation ambient light,, show that its energy storage capability is strong more if the photon number that the certain time interval T intra vane after exciting light stops to send is many more.Therefore; The summation of all photon number that plant leaf blade is sent in certain time interval T under external optical excitation is called the delayed luminescence integrated intensity; With I (T) expression, then can calculate the energy storage capability of blade cell through the integrated intensity I (T) that measures the interior delayed luminescence of section T sometime; And the photon radiation I that the energy that blade cell dissipates away can send in the unit interval through blade cell in the dark _SLMeasure.Like this, formula (1) can obtain through following formula:

$Q=\frac{I\left(T\right)}{I_{\mathrm{SL}}T}---\left(2\right)$

Wherein, Q is a nondimensional cardinar number.

On mechanism, the research that bio-photon is learned confirms that delayed luminescence integrated intensity I (T) is relevant with cellular metabolism intensity, photon radiation I _SLIt is the sign of cell peroxidating degree; The order state of cell is kept in cellular metabolism, and the cell peroxidating impels cell ageing, and cell state depends on the balance of cellular metabolism and cell levels of peroxide; Therefore, the state with formula (2) description blade cell is rational.

Because in the leaf senile process, along with the dissipation of blade cell self-energy and the raising of levels of peroxide, the Q value of blade cell will be more and more littler, therefore, can measure the aging degree of blade with the size of the Q value of blade cell.But for the cell of different plants, the Q value is different.In order to eliminate the deviation in the leaf senile degree evaluation that causes owing to individual difference, can be defined as the aging degree of blade with the relative change rate of blade cell Q value in the aging course, represent with K, promptly

$K=1-\frac{Q}{Q_0}---\left(3\right)$

Wherein, Q ₀Be certain blade state parameter of (initial state) under normal condition, the state parameter of Q is this blade in aging course some day.Because the Q during initial state ₀Maximum, along with the blade cell dissipation of energy, Q diminishes in aging course, and it is big that the K value becomes, and the aging degree of blade is also just big more.

Advantage with K value representation aging degree is that the K value of each blade is the cardinar number of a no unit, and all between 0～1, the K value is more little, shows that the leaf senile degree is more little in its variation; K=0 shows that blade does not have aging; K approaches 1 more, shows that the aging degree of blade is big more, thereby has eliminated the individual difference in the different leaf senile degree evaluations, makes the evaluation of leaf senile degree that unified standard arranged.

The present invention is based on photon radiation and estimate the method for plant leaf blade aging degree, specifically implement according to following steps:

Step 1: clean blade to be measured with distilled water, it was placed several minutes in the dark, use the spontaneous photon radiation I that the measuring system that detects based on single photon is measured the blade unit interval _SL

Step 2: blade is shone certain hour with external unsaturation exciting light (LED, laser, daylight lamp, natural light or other various external light sources); Use the measuring system that detects based on single photon to measure it and receive the delayed luminescence in certain time interval T after the optical excitation, obtain the delayed luminescence integrated intensity I (T) in this time T through the mathematics match; Specifically implement according to following method:

A kind of method is:

A. blade delayed luminescence kinetics equation is expressed as

$I\left(t\right)=\frac{I_0}{{(1+\frac{t}{\mathrm{\&tau;}})}^{\mathrm{\&beta;}}}---\left(4\right)$

Wherein, β, τ and I ₀Be constant, t is delayed luminescence die-away time.

Formula (4) is obtained the expression formula of delayed luminescence integrated intensity I (T) a measurement period T integrates:

$I\left(T\right)=\frac{\mathrm{\&tau;}{I}_0}{\mathrm{\&beta;}-1}[1-\frac{1}{{(1+T/\mathrm{\&tau;})}^{\mathrm{\&beta;}-1}}]---\left(5\right)$

B. after blade to be measured being cleaned set by step 1 measure the blade unit interval spontaneous photon radiation I _SL

C. with blade to be measured with after external unsaturation exciting light (LED, laser, daylight lamp, natural light or other the various external light sources) irradiation, receive after the optical excitation delayed luminescence change curve of t in time with the measuring system measurement blade that detects based on single photon in the step 1;

D. with the delayed luminescence that obtains by surveying instrument among the step c over time curve set by step the formula (4) among a carry out the mathematics match, obtain the delayed luminescence characteristic parameter in the formula (4): I ₀, τ and β;

F. with the Measuring Time T and the delayed luminescence characteristic parameter I of delayed luminescence ₀, τ and β substitution formula (5), obtain delayed luminescence integrated intensity I (T).

Another kind method is:

A. blade delayed luminescence kinetics equation is expressed as

$I\left(t\right)=I_{\mathrm{SL}}+\frac{I_0}{{(1+\frac{t}{\mathrm{\&tau;}})}^{\mathrm{\&beta;}}}---\left(6\right)$

Wherein, I _SLBe the spontaneous photon radiation of blade unit interval, β, τ and I ₀Be constant, t is delayed luminescence die-away time, and T is measuring period.

Formula (6) is obtained the expression formula of delayed luminescence integrated intensity I (T) a measurement period T integrates:

$I\left(T\right)=I_{\mathrm{SL}}T+\frac{\mathrm{\&tau;}{I}_0}{\mathrm{\&beta;}-1}[1-\frac{1}{{(1+T/\mathrm{\&tau;})}^{\mathrm{\&beta;}-1}}]---\left(7\right)$

B. after blade to be measured being cleaned set by step 1 measure the blade unit interval spontaneous photon radiation I _SL

C. with blade to be measured with after external unsaturation exciting light (LED, laser, daylight lamp, natural light or other the various external light sources) irradiation, receive after the optical excitation delayed luminescence change curve of t in time with the measuring system measurement blade that detects based on single photon in the step 1;

D. will go up delayed luminescence that the step obtains by surveying instrument over time curve set by step the formula (6) among a carry out the mathematics match, obtain the delayed luminescence characteristic parameter in the formula (6): I ₀, τ and β;

F. with the above-mentioned I that obtains _SL, T, I ₀, τ and β substitution formula (7), obtain delayed luminescence integrated intensity I (T).

The third method is:

A. the delayed luminescence kinetics equation is expressed as

$I\left(t\right)=I_0+I_1{e}^{-t/{\mathrm{\&tau;}}_1}+I_2{e}^{-t/{\mathrm{\&tau;}}_2}---\left(8\right)$

Wherein, I ₀, I ₁, I ₂And τ ₁, τ ₂Be constant, formula (8) obtained the expression formula of I (T) a measurement period T integrates:

$I\left(T\right)=I_{0}T+I_1{\mathrm{\&tau;}}_1[1-\exp (-\frac{T}{{\mathrm{\&tau;}}_1})]+I_2{\mathrm{\&tau;}}_2[1-\exp (-\frac{T}{{\mathrm{\&tau;}}_2})]---\left(9\right)$

B. blade to be measured is used external excitation light irradiation, measure this blade with the measuring system that detects based on single photon in the step 1 and receive the delayed luminescence change curve of t in time after the optical excitation;

C. will go up the delayed luminescence that obtains of step and carry out the mathematics match by formula (8) over time, obtain delayed luminescence characteristic parameter: I ₀, I ₁, I ₂, τ ₁, τ ₂

D. with the above-mentioned T that obtains, I ₀, I ₁, I ₂, τ ₁, τ ₂Substitution formula (9) obtains delayed luminescence integrated intensity I (T).

Step 3: the state parameter Q value of definition blade and the relative change rate K of Q value are:

$Q=\frac{I\left(T\right)}{I_{\mathrm{SL}}T}---\left(10\right)$

$K=1-\frac{Q}{Q_0}---\left(11\right)$

Wherein, Q ₀Be the state parameter of blade initial state (promptly old and feeble time), Q is blade certain state parameter constantly in aging course.Estimate the aging degree of this blade according to the size of K value, evaluation criterion is following: the size of K value is between 0～1, and the K value of blade approaches 0 more, and the aging degree of this blade is low more; The K value approaches 1 more, and the aging degree of blade is big more.

Q ₀Acquisition specifically implement according to following steps:

Step 1: the fresh blade when inciting somebody to action aging is cleaned with distilled water, dries surface moisture.Place a few minutes in the dark, use the spontaneous photon radiation I that the measuring system that detects based on single photon is measured the blade unit interval _SL

Step 2: this blade is shone certain hour with external unsaturation exciting light (LED, laser, daylight lamp, natural light or other various external light sources); Measure it with above-mentioned measuring system and receive the delayed luminescence in certain time interval T after the optical excitation, obtain the delayed luminescence integrated intensity I (T) in this time T through the mathematics match; Specifically implement according to following steps:

A kind of method is:

A. will be old and feeble blade with external unsaturation exciting light (LED, laser, daylight lamp, natural light or other various external light sources) irradiation after, receive after the optical excitation delayed luminescence change curve of t in time with aforesaid measuring system measurement blade based on the single photon detection;

B. will go up delayed luminescence that the step obtains by surveying instrument over time curve carry out the mathematics match by formula (4), obtain the delayed luminescence characteristic parameter in the formula (4): I ₀, τ and β;

C. with the Measuring Time T and the above-mentioned I that obtains of delayed luminescence ₀, τ and β substitution formula (5), obtain delayed luminescence integrated intensity I (T).

Another kind method is:

A. will be old and feeble blade use external excitation light irradiation, receive the delayed luminescence change curve of t in time after the optical excitation with aforesaid this blade of measuring system measurement based on the single photon detection;

B. will go up the delayed luminescence that obtains of step and carry out the mathematics match by formula (6) over time, obtain delayed luminescence characteristic parameter: I ₀, τ and β;

C. with the Measuring Time T and the above-mentioned I that obtains of delayed luminescence ₀, τ and β substitution formula (7), obtain delayed luminescence integrated intensity I (T).

The third method is:

A. will use external excitation light irradiation for the blade of aging, and measure this blade with the aforesaid measuring system that detects based on single photon and receive the delayed luminescence change curve of t in time after the optical excitation;

B. will go up the delayed luminescence that obtains of step and carry out the mathematics match by formula (8) over time, obtain the I in the formula (8) ₀, I ₁, I ₂, τ ₁, τ ₂

C. with the Measuring Time T and the above-mentioned I that obtains of delayed luminescence ₀, I ₁, I ₂, τ ₁, τ ₂Substitution formula (9) obtains delayed luminescence integrated intensity I (T).

Step 3: the spontaneous photon radiation I of unit interval when old and feeble with the above-mentioned blade that obtains _SL, delayed luminescence integrated intensity I (T) and delayed luminescence Measuring Time T bring formula (10) into and obtain the state parameter Q of blade when old and feeble ₀

Embodiment

Choose the close fresh spinach blade of growing way; With cleaning surperficial dirt in the deionized water, filter paper blots the moisture on surface, is cut into the disk (avoiding the blade sustainer during punching) of Φ 10mm then with card punch; Mixing is placed in the double dish that is equipped with middling speed filter paper, and use concentration is 2.5% H ₂O ₂Carry out aging and handle H ₂O ₂Treating fluid added once in per 8 hours, continued to handle 50 hours.

At H ₂O ₂When handling back 0h, 12h, 24h, 36h and 48h, take out 10 spinach blades respectively, blot surface liquid, shred behind the weighing fresh weight with filter paper, mixing, with conventional colourimetry measurement chlorophyll content, triplicate when at every turn measuring is averaged.Simultaneously measure spontaneous luminescence and delayed luminescence in the superweak photon radiation of spinach respectively with step according to the method described above; The measurement of spontaneous luminescence adopts the BPCL Weak-luminescence measuring instrument of Institute of Biophysics, Academia Sinica's development to measure, and representes spontaneous photon steradiance (counts/s) with the photon number of unit interval.Get 10 spinach blades when measuring spontaneous luminescence at every turn, blot surface liquid, put into the spontaneous luminescence of working sample behind the reason 5min of dark place, measuring instrument sample chamber with filter paper.Each 50s that measures, image data is spaced apart 1s, WV-1000V.The darkroom measures ambient temperature is made as 25 ℃, respectively surveys background 1 time before and after measuring, and deducts background.Each measurement respectively repeats 3 times, averages.

Delayed luminescence is measured after 1 minute with blue unsaturation LED irradiation, and Measuring Time is 50s, and image data is spaced apart 1s, WV-1000V, and the darkroom measures ambient temperature is made as 25 ℃.Respectively survey background 1 time before and after measuring, and deduct background.Each measurement respectively repeats 3 times, averages.

At H ₂O ₂Coerce in the aging course, the variation of spinach chlorophyll content in leaf blades is as shown in Figure 1.Because the general at present index that adopts chlorophyll content as the leaf senile degree, this result shows, along with H ₂O ₂The prolongation in processing time, blade are old and feeble gradually.

The variation of the spontaneous luminescence in the leaf senile process is as shown in Figure 2, and the variation of delayed luminescence is as shown in Figure 3.By formula (4) match, each fitting parameter that obtains is as shown in table 1 with each curve among Fig. 3.Each data substitution formula (5) in the table 1 is obtained the variation of delayed luminescence integrated intensity I (T), itself and Fig. 2 data substitution formula (10) and formula (11) are obtained the variation of Q and K value in the leaf senile process shown in Figure 4.Fig. 4 and Fig. 1 are relatively obtained the mutual relationship that Q and K value and chlorophyll content change in the leaf senile process shown in Figure 5.

Delayed luminescence fitting parameter in the table 1 spinach blade aging course

Visible by Fig. 5, in spinach blade aging course, the variation of blade Q value and K value all presents good linear relationship (R with the variation of chlorophyll content ²=0.91441,0.91443).Because chlorophyll content is the physiologic index of present evaluation plant leaf blade aging degree of generally acknowledging, this mutual relationship has explained in the leaf senile process that the aging degree of blade can characterize with the Q value or the K value of the present invention's definition.Different with chlorophyll content evaluation leaf senile degree methods in the measurement blade; The Q value that the present invention proposes or the preparation method of K value need not destroy plant leaf blade; Directly get final product with its photon radiation of apparatus measures; Have the advantage that not damaged detects, thereby the inventive method is useful with respect to existing method.

Claims (9)

1. estimate the method for plant leaf blade aging degree based on photon radiation, it is characterized in that, specifically implement according to following steps:

Step 1: clean blade to be measured with distilled water, it was placed several minutes in the dark, use the spontaneous photon radiation I that the measuring system that detects based on single photon is measured the blade unit interval _SL

Step 2: with blade to be measured with external unsaturation excitation light irradiation certain time interval T; Use the measuring system that detects based on single photon to measure it and receive the delayed luminescence in certain time interval T after the optical excitation, obtain the delayed luminescence integrated intensity I (T) in time T through the mathematics match;

Step 3: calculate the state parameter Q value of blade and the relative change rate K of Q value:

$Q=\frac{I\left(T\right)}{I_{\mathrm{SL}}T},$

$K=1-\frac{Q}{Q_0},$

Wherein, Q ₀State parameter for the blade initial state; Q is blade certain state parameter constantly in aging course, and according to the aging degree of this blade of size evaluation of K value, evaluation criterion is following: the size of K value is between 0～1; The K value of blade approaches 0 more, and the aging degree of this blade is low more; The K value approaches 1 more, and the aging degree of blade is big more.

2. according to claim 1ly estimate the method for plant leaf blade aging degree based on photon radiation, it is characterized in that, the unsaturation exciting light in the described step 2 adopts a kind of in LED, laser, daylight lamp or the natural light.

3. the method based on photon radiation evaluation plant leaf blade aging degree according to claim 1 is characterized in that, the process mathematics match in the described step 2 obtains the delayed luminescence integrated intensity I (T) in time T, specifically implements according to following steps:

Step a. is expressed as blade delayed luminescence kinetics equation:

$I\left(t\right)=\frac{I_0}{{(1+\frac{t}{\mathrm{\&tau;}})}^{\mathrm{\&beta;}}},$

Wherein, β, τ and I ₀Be constant, t is delayed luminescence die-away time;

Following formula is obtained the expression formula of delayed luminescence integrated intensity I (T) a measurement period T integrates:

$I\left(T\right)=\frac{\mathrm{\&tau;}{I}_0}{\mathrm{\&beta;}-1}[1-\frac{1}{{(1+T/\mathrm{\&tau;})}^{\mathrm{\&beta;}-1}}],$

After step b. cleans blade to be measured set by step 1 measure the blade unit interval spontaneous photon radiation I _SL

Step c with blade to be measured with external unsaturation excitation light irradiation after, use the measuring system that detects based on single photon to measure blade and receive after the optical excitation delayed luminescence change curve of t in time;

Steps d. with the delayed luminescence that obtains among the step c over time curve carry out the mathematics match, obtain delayed luminescence characteristic parameter: I ₀, τ and β;

Step f. is with the Measuring Time T and the delayed luminescence characteristic parameter I of delayed luminescence ₀, τ and β substitution delayed luminescence integrated intensity I (T) expression formula, obtain delayed luminescence integrated intensity I (T).

4. the method based on photon radiation evaluation plant leaf blade aging degree according to claim 1 is characterized in that, the process mathematics match in the described step 2 obtains the delayed luminescence integrated intensity I (T) in time T, specifically implements according to following steps:

Step a: blade delayed luminescence kinetics equation is expressed as:

$I\left(t\right)=I_{\mathrm{SL}}+\frac{I_0}{{(1+\frac{t}{\mathrm{\&tau;}})}^{\mathrm{\&beta;}}},$

Wherein, I _SLBe the spontaneous photon radiation of blade unit interval, β, τ and I ₀Be constant, t is delayed luminescence die-away time, and T is measuring period;

Following formula is obtained the expression formula of delayed luminescence integrated intensity I (T) a measurement period T integrates:

$I\left(T\right)=I_{\mathrm{SL}}T+\frac{\mathrm{\&tau;}{I}_0}{\mathrm{\&beta;}-1}[1-\frac{1}{{(1+T/\mathrm{\&tau;})}^{\mathrm{\&beta;}-1}}],$

After step b. cleans blade to be measured set by step 1 measure the blade unit interval spontaneous photon radiation I _SL

Step c with blade to be measured with external unsaturation excitation light irradiation after, use the measuring system that detects based on single photon to measure blade and receive after the optical excitation delayed luminescence change curve of t in time;

Steps d. will go up the delayed luminescence that obtains of step over time curve carry out the mathematics match, obtain delayed luminescence characteristic parameter: I ₀, τ and β;

Step f. is with the above-mentioned I that obtains _SL, T, I ₀, τ and β substitution delayed luminescence integrated intensity I (T) expression formula, obtain delayed luminescence integrated intensity I (T).

5. the method based on photon radiation evaluation plant leaf blade aging degree according to claim 1 is characterized in that, the process mathematics match in the described step 2 obtains the delayed luminescence integrated intensity I (T) in time T, specifically implements according to following steps:

Step a. is expressed as the delayed luminescence kinetics equation:

$I\left(t\right)=I_0+I_1{e}^{-t/{\mathrm{\&tau;}}_1}+I_2{e}^{-t/{\mathrm{\&tau;}}_2},$

Wherein, I ₀, I ₁, I ₂And τ ₁, τ ₂Be constant, following formula obtained the expression formula of I (T) a measurement period T integrates:

$I\left(T\right)=I_{0}T+I_1{\mathrm{\&tau;}}_1[1-\exp (-\frac{T}{{\mathrm{\&tau;}}_1})]+I_2{\mathrm{\&tau;}}_2[1-\exp (-\frac{T}{{\mathrm{\&tau;}}_2})],$

Step b. uses external excitation light irradiation with blade to be measured, uses the measuring system that detects based on single photon to measure this blade and receives the delayed luminescence change curve of t in time after the optical excitation;

Step c will go up the delayed luminescence that obtains of step and carry out the mathematics match over time, obtain delayed luminescence characteristic parameter: I ₀, I ₁, I ₂, τ ₁, τ ₂

Steps d. with the above-mentioned T that obtains, I ₀, I ₁, I ₂, τ ₁, τ ₂The expression formula of substitution delayed luminescence integrated intensity I (T) obtains delayed luminescence integrated intensity I (T).

6. the method based on photon radiation evaluation plant leaf blade aging degree according to claim 1 is characterized in that Q in the described step 3 ₀Acquisition, specifically implement according to following steps:

Step 1: the fresh blade when will be old and feeble is cleaned with distilled water, dries surface moisture, places in the dark, uses the spontaneous photon radiation I that the measuring system that detects based on single photon is measured the blade unit interval _SL

Step 2: the fresh blade when inciting somebody to action aging is with external unsaturation excitation light irradiation certain hour; Use the measuring system that detects based on single photon to measure it and receive the delayed luminescence in certain time interval T after the optical excitation, obtain the delayed luminescence integrated intensity I (T) in time T through the mathematics match;

Step 3: the spontaneous photon radiation I of unit interval when old and feeble with the above-mentioned blade that obtains _SL, delayed luminescence integrated intensity I (T) and delayed luminescence Measuring Time T bring into

Obtain the state parameter Q of blade when old and feeble ₀

7. the method based on photon radiation evaluation plant leaf blade aging degree according to claim 6 is characterized in that, the process mathematics match in the described step 2 obtains the delayed luminescence integrated intensity I (T) in time T, specifically implements according to following steps:

Step a. is expressed as blade delayed luminescence kinetics equation:

$I\left(t\right)=\frac{I_0}{{(1+\frac{t}{\mathrm{\&tau;}})}^{\mathrm{\&beta;}}},$

Wherein, β, τ and I ₀Be constant, t is delayed luminescence die-away time;

Following formula is obtained the expression formula of delayed luminescence integrated intensity I (T) a measurement period T integrates:

$I\left(T\right)=\frac{\mathrm{\&tau;}{I}_0}{\mathrm{\&beta;}-1}[1-\frac{1}{{(1+T/\mathrm{\&tau;})}^{\mathrm{\&beta;}-1}}],$

After step b. cleans blade to be measured set by step 1 measure the blade unit interval spontaneous photon radiation I _SL

Step c with blade to be measured with external unsaturation excitation light irradiation after, use the measuring system that detects based on single photon to measure blade and receive after the optical excitation delayed luminescence change curve of t in time;

Steps d. with the delayed luminescence that obtains among the step c over time curve carry out the mathematics match, obtain delayed luminescence characteristic parameter: I ₀, τ and β;

Step f. is with the Measuring Time T and the delayed luminescence characteristic parameter I of delayed luminescence ₀, τ and β substitution delayed luminescence integrated intensity I (T) expression formula, obtain delayed luminescence integrated intensity I (T).

8. the method based on photon radiation evaluation plant leaf blade aging degree according to claim 6 is characterized in that, the process mathematics match in the described step 2 obtains the delayed luminescence integrated intensity I (T) in time T, specifically implements according to following steps:

Step a. is expressed as blade delayed luminescence kinetics equation

$I\left(t\right)=I_{\mathrm{SL}}+\frac{I_0}{{(1+\frac{t}{\mathrm{\&tau;}})}^{\mathrm{\&beta;}}},$

Wherein, I _SLBe the spontaneous photon radiation of blade unit interval, β, τ and I ₀Be constant, t is delayed luminescence die-away time, and T is measuring period;

Following formula is obtained the expression formula of delayed luminescence integrated intensity I (T) a measurement period T integrates:

$I\left(T\right)=I_{\mathrm{SL}}T+\frac{\mathrm{\&tau;}{I}_0}{\mathrm{\&beta;}-1}[1-\frac{1}{{(1+T/\mathrm{\&tau;})}^{\mathrm{\&beta;}-1}}],$

After step b. cleans blade to be measured set by step 1 measure the blade unit interval spontaneous photon radiation I _SL

Step c with blade to be measured with external unsaturation excitation light irradiation after, use the measuring system that detects based on single photon to measure blade and receive after the optical excitation delayed luminescence change curve of t in time;

Steps d. will go up the delayed luminescence that obtains of step over time curve carry out the mathematics match, obtain delayed luminescence characteristic parameter: I ₀, τ and β;

Step f. is with the above-mentioned I that obtains _SL, T, I ₀, τ and β substitution delayed luminescence integrated intensity I (T) expression formula, obtain delayed luminescence integrated intensity I (T).

9. the method based on photon radiation evaluation plant leaf blade aging degree according to claim 6 is characterized in that, the process mathematics match in the described step 2 obtains the delayed luminescence integrated intensity I (T) in time T, specifically implements according to following steps:

Step a. is expressed as the delayed luminescence kinetics equation:

$I\left(t\right)=I_0+I_1{e}^{-t/{\mathrm{\&tau;}}_1}+I_2{e}^{-t/{\mathrm{\&tau;}}_2},$

Wherein, I ₀, I ₁, I ₂And τ ₁, τ ₂Be constant, following formula obtained the expression formula of I (T) a measurement period T integrates:

$I\left(T\right)=I_{0}T+I_1{\mathrm{\&tau;}}_1[1-\exp (-\frac{T}{{\mathrm{\&tau;}}_1})]+I_2{\mathrm{\&tau;}}_2[1-\exp (-\frac{T}{{\mathrm{\&tau;}}_2})],$

Step b. uses external excitation light irradiation with blade to be measured, uses the measuring system that detects based on single photon to measure this blade and receives the delayed luminescence change curve of t in time after the optical excitation;

C. will go up the delayed luminescence that obtains of step and carry out the mathematics match over time, obtain delayed luminescence characteristic parameter: I ₀, I ₁, I ₂, τ ₁, τ ₂

D. with the above-mentioned T that obtains, I ₀, I ₁, I ₂, τ ₁, τ ₂The expression formula of substitution delayed luminescence integrated intensity I (T) obtains delayed luminescence integrated intensity I (T).

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