CN105486669A - Chlorophyll fluorescence diagnostic method for deficiency of indispensable plant nutritive elements - Google Patents
Chlorophyll fluorescence diagnostic method for deficiency of indispensable plant nutritive elements Download PDFInfo
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Abstract
The invention discloses a chlorophyll fluorescence diagnostic method for deficiency of indispensable plant nutritive elements. The method includes the steps that a, a plurality of nutrient solution formulas are made; b, plant seeds to be tested are sterilized; c, accelerating germination is conducted on the sterilized seeds; d, the seeds are cultured in culture cups after the seeds sprout; e, 10-30 plant culture cups are arranged for each nutrient solution in step a, namely, 10-30 times of repetition are set for each nutrient solution, then plants are placed into an illumination incubator, and illumination intensity is set to be 0-4000 lux, the temperature is 4 DEG C-30 DEG C, parameters are continuous and adjustable, and the optimal culture temperature of the plants is set, and culture is conducted for 4-8 weeks; f, fluorescent information collection is conducted on lacking elements and chlorophyll of leaves of plants growing normally; g, chlorophyll fluorescence diagnostic calculation is conducted on the lacking elements of the plants. Aiming at adverse factors of existing plant nutrient element deficiency diagnosis, the method starts from rapid and damage-free diagnosis, the convenient, fast, damage-free and accurate indispensable plant nutritive element diagnosis method based on plant chlorophyll fluorescence is developed, and the method has a quite high technical advantage and is a significant part of agriculture intellectualization.
Description
Technical field
The present invention relates to a kind of chlorophyll fluorescence diagnostic method, be specifically related to the chlorophyll fluorescence diagnostic method that a Plants essential nutrient element wanes.
Background technology
The 1860's, Sachs and knop established plant nutrient theory, determined the mineral element that plant growth is necessary, a great number of elements, moderate-element and trace element.Wherein macronutrient comprises: carbon, hydrogen, oxygen, nitrogen, phosphorus, potassium; Middle amount nutrient comprises: calcium, magnesium, sulphur; Micronutrient element comprises: iron, boron, manganese, copper, zinc, molybdenum, chlorine.Outside carbon, hydrogen, oxygen three kinds of elements, other elements can be added by the mode of fertilising.When lacking any one element in growing process and all can cause the exception of plant growth, this is that plant needs nutrient " Bucket Principle ".Therefore, the type of elements in Soil and content can, along with changes such as soil types, dimension, longitudes, in order to make the normal, healthy and strong of plant length on producing, need in soil, apply dissimilar fertilizer, and once lack after any one lacks element, plant all can show symptom.
In production, the diagnostic method that plant lacks nutrient comprises morphological diagnosis, chemical diagnosis and fertilization diagnosis.Plant forms, leaf morphology also color etc. is mainly observed in morphological diagnosis, needs very strong practical experience, subjectivity strong and be easy to produce erroneous judgement; The constituent content that chemical diagnosis mainly detects in plant or soil differentiates, this method needs to carry out sample analysis to soil and plant, and needs the instrument and equipment of costliness, specialty for plant elements analysis, consuming time, effort, detection costliness; Fertilization diagnosis applies fertilizer to the plant of performance nutritional deficiency symptom, and judged by the change of plant symptom, this kind of method correctness is very low, and too time-consuming, the easily treatment of delay plant symptom.
Summary of the invention
Technical matters to be solved by this invention is the correctness technical matters that is low, too time-consuming that in prior art, diagnostic method exists.
Solving the problems of the technologies described above adopted technical scheme is the chlorophyll fluorescence diagnostic method that a Plants essential nutrient element wanes, and comprises the steps:
The configuration of the some nutrient solution prescriptions of a:
B, by test plants seed distilled water immersion 2-3h, then with 5-10% liquor natrii hypochloritis or 3-5% hydrogen peroxide solution sterilization 10-20min, outwells thimerosal, cleans seed 5-8 time with distilled water;
Seed after sterilization is placed in the double dish being covered with layer 2-3 filter paper by c, is placed in vernalization under 25-30 DEG C of condition;
After d germination, consistent for germinating energy seed is placed on the meshed flat board of band, its flat board is allowed to swim in continued growth in ultrapure water, wherein mesh diameter is less than seed particle size, prevent seed from falling in water, intensity of illumination is set by illumination box: 0-4000lux, temperature 4-30 DEG C, continuous parameters is adjustable, arrange plant optimum cultivation temperature to cultivate 1 week, after axis minister becomes, the root system of consistent for growing way plant is taken out carefully from mesh, plant stem is put on the spot with sponge, then it is fixed on in the hole of KT plate central authorities, this KT plate is fixed on and cultivates on cup, be cultivation plant cup of the present invention, the volume cultivating cup is 400-600ml,
In step e a, often kind of nutrient solution arranges 10-30 and cultivates plant cup, namely often kind of nutrient solution arranges 10-30 repetition, then plant is placed in illumination box and intensity of illumination is set: 0-4000lux, temperature 4-30 DEG C, continuous parameters is adjustable, arranges plant optimum cultivation temperature and cultivates, and is finished pancebrin 0-4 week and cultivates, grow up to after seedling until plant, each nutrient solution prescription of 4-8 week step a is cultivated;
F nutritional deficiency and the information acquisition of normal growth plant leaf blade chlorophyll fluorescence;
The chlorophyll fluorescence diagnosis of g plant nutritional deficiency calculates.
Lack the unfavorable factor of diagnosis for existing plant nutrient, the present invention diagnoses from quick, not damaged start with convenient, quick, the not damaged, accurately plant essential nutrient element diagnostic method developed based on plant chlorophyll fluorescence.Having very strong technical advantage, is the pith of intelligent agriculture.
Accompanying drawing explanation
Fig. 1 is chlorophyll from maize leaf fluorescence parameter Fv/Fm, Fv '/Fm ', Φ PSII value;
Fig. 2 is that corn indispensable element N, P, K, Fe wane diagnosis space figure;
Fig. 3 is soybean leaves chlorophyll fluorescence parameters Fv/Fm, Fv '/Fm ', Φ PSII value;
Fig. 4 is that soybean indispensable element N, P, K, Fe wane diagnosis space figure.
Embodiment
Below in conjunction with embodiment, the present invention is described in more detail, but the invention is not restricted to these embodiments.
The present invention discloses the chlorophyll fluorescence diagnostic method that a Plants essential nutrient element wanes, and comprises the steps:
The configuration of the some nutrient solution prescriptions of a:
B, by test plants seed distilled water immersion 2-3h, then with 5-10% liquor natrii hypochloritis or 3-5% hydrogen peroxide solution sterilization 10-20min, outwells thimerosal, cleans seed 5-8 time with distilled water;
Seed after sterilization is placed in the double dish being covered with layer 2-3 filter paper by c, is placed in vernalization under 25-30 DEG C of condition;
After d germination, consistent for germinating energy seed is placed on the meshed flat board of band, its flat board is allowed to swim in continued growth in ultrapure water, wherein mesh diameter is less than seed particle size, prevent seed from falling in water, intensity of illumination is set by illumination box: 0-4000lux, temperature 4-30 DEG C, continuous parameters is adjustable, arrange plant optimum cultivation temperature to cultivate 1 week, after axis minister becomes, the root system of consistent for growing way plant is taken out carefully from mesh, plant stem is put on the spot with sponge, then it is fixed on in the hole of KT plate central authorities, this KT plate is fixed on and cultivates on cup, be cultivation plant cup of the present invention, the volume cultivating cup is 400-600ml,
In step e a, often kind of nutrient solution arranges 10-30 and cultivates plant cup, namely often kind of nutrient solution arranges 10-30 repetition, then plant is placed in illumination box and intensity of illumination is set: 0-4000lux, temperature 4-30 DEG C, continuous parameters is adjustable, arranges plant optimum cultivation temperature and cultivates, and is finished pancebrin 0-4 week and cultivates, grow up to after seedling until plant, each nutrient solution prescription of 4-8 week step a is cultivated;
F nutritional deficiency and the information acquisition of normal growth plant leaf blade chlorophyll fluorescence;
The chlorophyll fluorescence diagnosis of g plant nutritional deficiency calculates.
Being configured to of some nutrient solutions in step a:
(1) with ultrapure water (18.2M Ω) configuration: 7.45mg/LNa
2– EDTA, 5.57mg/LFeSO
47H
2o, 820.7mg/LCa (NO
3)
2, 502.6mg/LKNO
3, 616.2mg/LMgSO
47H
2o, 272.2mg/LKH
2pO
4, 2.860mg/LH
3bO
3, 1.015mg/LMnSO
4, 0.079mg/LCuSO
45H
2o, 0.220mg/LZnSO
47H
2o, 0.090mg/LH
2moO
4solution adjust ph 5.5-6.5, obtains complete nutrition liquid;
(2) with ultrapure water (18.2M Ω) configuration: 616.2mg/LMgSO
47H
2o, 272.2mg/LKH
2pO
4, 7.45mg/LNa
2– EDTA, 5.57mg/LFeSO
47H
2o, 555mg/LCaCl
2, 372.8mg/LKCl, 2.860mg/LH
3bO
3, 1.015mg/LMnSO
4, 0.079mg/LCuSO
45H
2o, 0.220mg/LZnSO
47H
2o, 0.090mg/LH
2moO
4solution adjust ph 5.5-6.5, obtains nitrogen stress nutrient solution;
(3) with ultrapure water (18.2M Ω) configuration: 820.7mg/LCa (NO
3)
2, 502.6mg/LKNO
3, 616.2mg/LMgSO
47H
2o, 7.45mg/LNa
2– EDTA, 5.57mg/LFeSO
47H
2o, 149.1mg/LKCl, 2.860mg/LH
3bO
3, 1.015mg/LMnSO
4, 0.079mg/LCuSO
45H
2o, 0.220mg/LZnSO
47H
2o, 0.090mg/LH
2moO
4solution adjust ph 5.5-6.5, obtains scarce phosphorus nutrition liquid;
(4) with ultrapure water (18.2M Ω) configuration: 820.7mg/LCa (NO
3)
2, 616.2mg/LMgSO
47H
2o, 240mg/LNaH
2pO
4, 7.45mg/LNa
2– EDTA, 5.57mg/LFeSO
47H
2o, 424.5mg/LNaNO
3, 2.860mg/LH
3bO
3, 1.015mg/LMnSO
4, 0.079mg/LCuSO
45H
2o, 0.220mg/LZnSO
47H
2o, 0.090mg/LH
2moO
4solution adjust ph 5.5-6.5, obtains potassium deficiency nutrient solution;
(5) with ultrapure water (18.2M Ω) configuration: 820.7mg/LCa (NO
3)
2, 502.6mg/LKNO
3, 616.2mg/LMgSO
47H
2o, 272.2mg/LKH
2pO
4, 2.860mg/LH
3bO
3, 1.015mg/LMnSO
4, 0.079mg/LCuSO
45H
2o, 0.220mg/LZnSO
47H
2o, 0.090mg/LH
2moO
4solution adjust ph 5.5-6.5, obtains iron deficiency nutrient solution.
Step f chlorophyll fluorescence information acquisition step is:
(1) a dark end cycle on plant, before welcoming next periodicity of illumination, ensures the abundant dark adatpation of plant leaf, measures when chlorophyll Photosystem I I (PSII) opens completely;
Penetrate 10-15 μ s with the illumination of wavelength 465-475nm (peak value is 470nm), intensity of illumination 0.03-0.1 μm olm-2s-1, gather the fluorescence information that 680nm place chlorophyll is launched, recording this fluorescent value is Fo simultaneously;
Penetrate 10-15 μ s with the illumination of wavelength 465-485nm (peak value is 470nm), intensity of illumination 4000-5000 μm olm-2s-1, gather the fluorescence information that 680nm place chlorophyll is launched, recording this fluorescent value is Fm simultaneously;
Penetrate 10-30min with the illumination of wavelength 465-485nm (peak value is 470nm), intensity of illumination 200-600 μm olm-2s-1, fully start photosynthesis of plant system.Gather the fluorescence information that 680nm place chlorophyll is launched afterwards, recording this fluorescent value is Fs;
5s is irradiated with the far red light of wavelength 725-755nm (peak value is 730nm), intensity of illumination 200-600 μm olm-2s-1;
Penetrate 10-15 μ s with the illumination of wavelength 465-475nm (peak value is 470nm), intensity of illumination 0.03-0.1 μm olm-2s-1, gather the fluorescence information that 680nm place chlorophyll is launched, recording this fluorescent value is Fo ' simultaneously;
Penetrate 10-15 μ s with the illumination of wavelength 465-485nm (peak value is 470nm), intensity of illumination 4000-5000 μm olm-2s-1, gather the fluorescence information that 680nm place chlorophyll is launched, recording this fluorescent value is Fm ' simultaneously.
The chlorophyll fluorescence diagnosis computing formula of step g plant nutritional deficiency is:
(1) chlorophyll fluorescence Diagnostic parameters Fv/Fm, Fv '/Fm ', Φ PSII; Fv/Fm represents the height of Photochemical Efficiency, and Fv '/Fm ' represents effective Photochemical Efficiency, and Φ PSII represents leaf photosynthesis electron transport rate speed;
(2) chlorophyll fluorescence Diagnostic parameters computing formula: Fv/Fm=(Fm-Fo)/Fm, Fv '/Fm '=(Fm '-Fo ')/Fm ', Φ PSII=(Fm '-Fs)/Fm ';
Fo represents the minimum fluorescence quantum yield of dark adatpation, Fm represents dark adatpation maximum fluorescence output, Fs represents steady-state fluorescence output under light, Fo ' represents the minimum fluorescence quantum yield of photopia, Fm ' represents photopia maximum fluorescence output.
Embodiment 1
The nutrient diagnosis of corn nutritional deficiency
1. the configuration of nutrient solution prescription:
(1) with ultrapure water (18.2M Ω) configuration: 7.45mg/LNa
2– EDTA, 5.57mg/LFeSO
47H
2o, 820.7mg/LCa (NO
3)
2, 502.6mg/LKNO
3, 616.2mg/LMgSO
47H
2o, 272.2mg/LKH
2pO
4, 2.860mg/LH
3bO
3, 1.015mg/LMnSO
4, 0.079mg/LCuSO
45H
2o, 0.220mg/LZnSO
47H
2o, 0.090mg/LH
2moO
4solution adjust ph 5.5-6.5, obtains complete nutrition liquid.
(2) with ultrapure water (18.2M Ω) configuration: 616.2mg/LMgSO
47H
2o, 272.2mg/LKH
2pO
4, 7.45mg/LNa
2– EDTA, 5.57mg/LFeSO
47H
2o, 555mg/LCaCl
2, 372.8mg/LKCl, 2.860mg/LH
3bO
3, 1.015mg/LMnSO
4, 0.079mg/LCuSO
45H
2o, 0.220mg/LZnSO
47H
2o, 0.090mg/LH
2moO
4solution adjust ph 5.5-6.5, obtains nitrogen stress nutrient solution.
(3) with ultrapure water (18.2M Ω) configuration: 820.7mg/LCa (NO
3)
2, 502.6mg/LKNO
3, 616.2mg/LMgSO
47H
2o, 7.45mg/LNa
2– EDTA, 5.57mg/LFeSO
47H
2o, 149.1mg/LKCl, 2.860mg/LH
3bO
3, 1.015mg/LMnSO
4, 0.079mg/LCuSO
45H
2o, 0.220mg/LZnSO
47H
2o, 0.090mg/LH
2moO
4solution adjust ph 5.5-6.5, obtains scarce phosphorus nutrition liquid.
(4) with ultrapure water (18.2M Ω) configuration: 820.7mg/LCa (NO
3)
2, 616.2mg/LMgSO
47H
2o, 240mg/LNaH
2pO
4, 7.45mg/LNa
2– EDTA, 5.57mg/LFeSO
47H
2o, 424.5mg/LNaNO
3, 2.860mg/LH
3bO
3, 1.015mg/LMnSO
4, 0.079mg/LCuSO
45H
2o, 0.220mg/LZnSO
47H
2o, 0.090mg/LH
2moO
4solution adjust ph 5.5-6.5, obtains potassium deficiency nutrient solution.
(5) with ultrapure water (18.2M Ω) configuration: 820.7mg/LCa (NO
3)
2, 502.6mg/LKNO
3, 616.2mg/LMgSO
47H
2o, 272.2mg/LKH
2pO
4, 2.860mg/LH
3bO
3, 1.015mg/LMnSO
4, 0.079mg/LCuSO
45H
2o, 0.220mg/LZnSO
47H
2o, 0.090mg/LH
2moO
4solution adjust ph 5.5-6.5, obtains iron deficiency nutrient solution.
2. will test corn seed distilled water immersion 2.5h, and then use 5% liquor natrii hypochloritis, sterilization 15min, outwells thimerosal, cleans seed 6 times with distilled water;
3. the corn seed after sterilization is placed in the double dish being covered with 2 metafiltration paper, is placed in vernalization under 30 DEG C of conditions;
4. after corn seed germination, consistent for germinating energy corn seed is placed on the meshed flat board of band, flat board is allowed to swim in continued growth in ultrapure water, wherein mesh diameter is less than corn seed particle diameter, prevent corn seed from falling in water, intensity of illumination is set by illumination box: 4000lux, temperature 28 DEG C, light application time 14h, nocturnal temperature 20 DEG C, cultivate 1 week, after Maize Stem minister becomes, the root system of consistent for growing way corn is taken out carefully from mesh, corn stem is put on the spot with sponge, then it is fixed on in the hole of KT plate central authorities, this KT plate is fixed on and cultivates on cup.Be cultivation plant cup of the present invention.The volume cultivating cup is 400ml.
5. in step 1, often kind of nutrient solution arranges 12 and cultivates plant cup, and namely often kind of nutrient solution arranges 12 repetitions.Then milpa is placed in illumination box and arranges intensity of illumination: 4000lux, temperature 28 DEG C, light application time 14h, nocturnal temperature 20 DEG C, be finished pancebrin 0-4 week and cultivate, grow up to after seedling until plant, each nutrient solution prescription of 4-6 week step a is cultivated; .
6. nutritional deficiency and normal growth chlorophyll from maize leaf fluorescence information gather
(1) a dark end cycle on milpa, before welcoming next periodicity of illumination, ensures the abundant dark adatpation of plant leaf, measures when chlorophyll Photosystem I I (PSII) opens completely;
(2) with wavelength 470nm, intensity of illumination 0.05 μm of olm
-2s
-1illumination penetrate 10 μ s, gather the fluorescence information of 680nm place chlorophyll transmitting, recording this fluorescent value is chlorophyll from maize leaf fluorescence parameter Fo simultaneously,
(3) with wavelength 470nm, intensity of illumination 4000 μm of olm
-2s
-1illumination penetrate 10 μ s, gather the fluorescence information of 680nm place chlorophyll transmitting, recording this fluorescent value is chlorophyll from maize leaf fluorescence parameter Fm simultaneously,
(4) with wavelength 470nm, intensity of illumination 250 μm of olm
-2s
-1illumination penetrate 30min, fully start photosynthesis of plant system.Gather the fluorescence information that 680nm place chlorophyll is launched afterwards, recording this fluorescent value is chlorophyll from maize leaf fluorescence parameter Fs,
(5) with wavelength 730nm, intensity of illumination 200 μm of olm
-2s
-1far red light irradiate 5s,
(6) with wavelength 470nm, intensity of illumination 0.05 μm of olm
-2s
-1illumination penetrate 10 μ s, gather the fluorescence information of 680nm place chlorophyll transmitting, recording this fluorescent value is chlorophyll from maize leaf fluorescence parameter Fo ' simultaneously,
(7) with wavelength 470nm, intensity of illumination 4000 μm of olm
-2s
-1illumination penetrate 10 μ s, gather the fluorescence information of 680nm place chlorophyll transmitting, recording this fluorescent value is chlorophyll from maize leaf fluorescence parameter Fm ' simultaneously
7. chlorophyll fluorescence Diagnostic parameters, the computing formula of plant nutritional deficiency
(1) chlorophyll fluorescence Diagnostic parameters Fv/Fm, Fv '/Fm ', Φ PSII
(2) chlorophyll fluorescence Diagnostic parameters computing formula: Fv/Fm=(Fm-Fo)/Fm, Fv '/Fm '=(Fm '-Fo ')/Fm ', Φ PSII=(Fm '-Fs)/Fm '
8. indispensable element N, P, K, Fe wanes and causes chlorophyll from maize leaf fluorescence parameter Fv/Fm, Fv '/Fm ', Φ PSII diagnostic value, as shown in Figure 1
9. the three dimensions diagnosis of nutritional deficiency
(1)
By Fv/Fm, Fv '/Fm ', Φ PSII value input formula a=(ymax-ymin) * (x-xmin)/(xmax-xmin)+ymin,
It is normalized to respectively interval [-1,1].
Y
maxbe 1, y
minfor-1, x
maxbe respectively Fv/Fm, Fv '/Fm ', the maximal value of Φ PSII, x
minbe respectively Fv/Fm, Fv '/Fm ', the minimum value of Φ PSII, x be respectively Fv/Fm, Fv '/Fm ', Φ PSII actual value.
(2) using Fv/Fm, Fv '/Fm ', Φ PSII normalized value as coordinate system x, y, z axle, mapping.Obtain-N ,-P, the volume coordinate of-K ,-Fe and CK is respectively (1.0000,1.0000,1.0000), (0.3547,0.5885,0.4319), (0.3382,0.0338,-0.048), (-0.0226 ,-0.7784 ,-1.0000), (-1.0000,-1.0000 ,-0.7709).
(3) according to 1% fiducial interval, the point in volume coordinate point change 0.01 distance belongs to this nutritional deficiency type.
See that Fig. 2 corn indispensable element N, P, K, Fe wane diagnosis space figure.
Embodiment 2
The nutrient diagnosis of soybean nutritional deficiency
1. the configuration of nutrient solution prescription:
(1) with ultrapure water (18.2M Ω) configuration: 7.45mg/LNa
2– EDTA, 5.57mg/LFeSO
47H
2o, 820.7mg/LCa (NO
3)
2, 502.6mg/LKNO
3, 616.2mg/LMgSO
47H
2o, 272.2mg/LKH
2pO
4, 2.860mg/LH
3bO
3, 1.015mg/LMnSO
4, 0.079mg/LCuSO
45H
2o, 0.220mg/LZnSO
47H
2o, 0.090mg/LH
2moO
4solution adjust ph 5.5-6.5, obtains complete nutrition liquid.
(2) with ultrapure water (18.2M Ω) configuration: 616.2mg/LMgSO
47H
2o, 272.2mg/LKH
2pO
4, 7.45mg/LNa
2– EDTA, 5.57mg/LFeSO
47H
2o, 555mg/LCaCl
2, 372.8mg/LKCl, 2.860mg/LH
3bO
3, 1.015mg/LMnSO
4, 0.079mg/LCuSO
45H
2o, 0.220mg/LZnSO
47H
2o, 0.090mg/LH
2moO
4solution adjust ph 5.5-6.5, obtains nitrogen stress nutrient solution.
(3) with ultrapure water (18.2M Ω) configuration: 820.7mg/LCa (NO
3)
2, 502.6mg/LKNO
3, 616.2mg/LMgSO
47H
2o, 7.45mg/LNa
2– EDTA, 5.57mg/LFeSO
47H
2o, 149.1mg/LKCl, 2.860mg/LH
3bO
3, 1.015mg/LMnSO
4, 0.079mg/LCuSO
45H
2o, 0.220mg/LZnSO
47H
2o, 0.090mg/LH
2moO
4solution adjust ph 5.5-6.5, obtains scarce phosphorus nutrition liquid.
(4) with ultrapure water (18.2M Ω) configuration: 820.7mg/LCa (NO
3)
2, 616.2mg/LMgSO
47H
2o, 240mg/LNaH
2pO
4, 7.45mg/LNa
2– EDTA, 5.57mg/LFeSO
47H
2o, 424.5mg/LNaNO
3, 2.860mg/LH
3bO
3, 1.015mg/LMnSO
4, 0.079mg/LCuSO
45H
2o, 0.220mg/LZnSO
47H
2o, 0.090mg/LH
2moO
4solution adjust ph 5.5-6.5, obtains potassium deficiency nutrient solution.
(5) with ultrapure water (18.2M Ω) configuration: 820.7mg/LCa (NO
3)
2, 502.6mg/LKNO
3, 616.2mg/LMgSO
47H
2o, 272.2mg/LKH
2pO
4, 2.860mg/LH
3bO
3, 1.015mg/LMnSO
4, 0.079mg/LCuSO
45H
2o, 0.220mg/LZnSO
47H
2o, 0.090mg/LH
2moO
4solution adjust ph 5.5-6.5, obtains iron deficiency nutrient solution.
2. will test soya seeds distilled water immersion 2h, and then use 8% hydrogen peroxide solution, sterilization 20min, outwells thimerosal, cleans seed 8 times with distilled water;
3. the soya seeds after sterilization is placed in the double dish being covered with 3 metafiltration paper, is placed in vernalization under 25 DEG C of conditions;
4. after soya seeds germinates, consistent for germinating energy soya seeds is placed on the meshed flat board of band, its flat board is allowed to swim in continued growth in ultrapure water, wherein mesh diameter is less than soya seeds particle diameter, prevent soya seeds from falling in water, intensity of illumination is set by illumination box: 2000lux, temperature 25 DEG C, light application time 12h, nocturnal temperature 18 DEG C, cultivate 1 week, after soybean stem minister becomes, the root system of consistent for growing way soybean seedling is taken out carefully from mesh, soybean stem is put on the spot with sponge, then it is fixed on in the hole of KT plate central authorities, this KT plate is fixed on and cultivates on cup.Be cultivation plant cup of the present invention.The volume cultivating cup is 500ml.
5. in step 1, often kind of nutrient solution arranges 25 and cultivates plant cup, and namely often kind of nutrient solution arranges 25 repetitions.Then soybean plant strain is placed in illumination box and arranges intensity of illumination: 3000lux, temperature 5 DEG C, light application time 12h, nocturnal temperature 18 DEG C, be finished pancebrin 0-3 week and cultivate, grow up to after seedling until plant, each nutrient solution prescription of 3-5 week step a is cultivated; .
6. nutritional deficiency and normal growth bean plant leaf chlorophyll fluorescence information gather
(1) a dark end cycle on soybean plant strain, before welcoming next periodicity of illumination, ensures the abundant dark adatpation of soybean plant strain blade, measures when chlorophyll Photosystem I I (PSII) opens completely;
(2) with wavelength 470nm, intensity of illumination 0.05 μm of olm
-2s
-1illumination penetrate 10 μ s, gather the fluorescence information of 680nm place chlorophyll transmitting, recording this fluorescent value is soybean leaves chlorophyll fluorescence parameters Fo simultaneously,
(3) with wavelength 470nm, intensity of illumination 4000 μm of olm
-2s
-1illumination penetrate 10 μ s, gather the fluorescence information of 680nm place chlorophyll transmitting, recording this fluorescent value is soybean leaves chlorophyll fluorescence parameters Fm simultaneously,
(4) with wavelength 470nm, intensity of illumination 250 μm of olm
-2s
-1illumination penetrate 30min, fully start photosynthesis of plant system.Gather the fluorescence information that 680nm place chlorophyll is launched afterwards, recording this fluorescent value is soybean leaves chlorophyll fluorescence parameters Fs,
(5) with wavelength 730nm, intensity of illumination 200 μm of olm
-2s
-1far red light irradiate 5s,
(6) with wavelength 470nm, intensity of illumination 0.05 μm of olm
-2s
-1illumination penetrate 10 μ s, gather the fluorescence information of 680nm place chlorophyll transmitting, recording this fluorescent value is soybean leaves chlorophyll fluorescence parameters Fo ' simultaneously,
(7) with wavelength 470nm, intensity of illumination 4000 μm of olm
-2s
-1illumination penetrate 10 μ s, gather the fluorescence information of 680nm place chlorophyll transmitting, recording this fluorescent value is soybean leaves chlorophyll fluorescence parameters Fm ' simultaneously
7. chlorophyll fluorescence Diagnostic parameters, the computing formula of plant nutritional deficiency
(1) chlorophyll fluorescence Diagnostic parameters Fv/Fm, Fv '/Fm ', Φ PSII;
(2) chlorophyll fluorescence Diagnostic parameters computing formula: Fv/Fm=(Fm-Fo)/Fm, Fv '/Fm '=(Fm '-Fo ')/Fm ', Φ PSII=(Fm '-Fs)/Fm '.
8. indispensable element N, P, K, Fe wanes and causes chlorophyll from maize leaf fluorescence parameter Fv/Fm, Fv '/Fm ', Φ PSII diagnostic value, see Fig. 3.
9. the three dimensions diagnosis of nutritional deficiency
(1)
By Fv/Fm, Fv '/Fm ', Φ PSII value input formula a=(ymax-ymin) * (x-xmin)/(xmax-xmin)+ymin, it is normalized to respectively interval [-1,1].
Y
maxbe 1, y
minfor-1, x
maxbe respectively Fv/Fm, Fv '/Fm ', the maximal value of Φ PSII, x
minbe respectively Fv/Fm, Fv '/Fm ', the minimum value of Φ PSII, x be respectively Fv/Fm, Fv '/Fm ', Φ PSII actual value.
(2) using Fv/Fm, Fv '/Fm ', Φ PSII normalized value as coordinate system x, y, z axle, mapping.Obtain-N ,-P, the volume coordinate of-K ,-Fe and CK is respectively (1.0000,1.0000,1.0000), (0.1133,0.3290,0.3143), (-0.5234,-0.0584,0.0114), (-0.3513 ,-0.1541 ,-0.2883), (-1.0000,-1.0000 ,-1.0000).
(3) according to 1% fiducial interval, the point in volume coordinate point change 0.01 distance belongs to this nutritional deficiency type, as Fig. 4.
To those skilled in the art, obviously the invention is not restricted to the details of above-mentioned one exemplary embodiment, and when not deviating from spirit of the present invention or essential characteristic, the present invention can be realized in other specific forms.Therefore, no matter from which point, all should embodiment be regarded as exemplary, and be nonrestrictive, scope of the present invention is limited by claims instead of above-mentioned explanation, and all changes be therefore intended in the implication of the equivalency by dropping on claim and scope are included in the present invention.
In addition, be to be understood that, although this instructions is described according to embodiment, but not each embodiment only comprises an independently technical scheme, this narrating mode of instructions is only for clarity sake, those skilled in the art should by instructions integrally, and the technical scheme in each embodiment also through appropriately combined, can form other embodiments that it will be appreciated by those skilled in the art that.
Claims (4)
1. the chlorophyll fluorescence diagnostic method that wanes of a Plants essential nutrient element, is characterized in that, comprise the steps:
The configuration of the some nutrient solution prescriptions of a;
B, by test plants seed distilled water immersion 2-3h, then with 5-10% liquor natrii hypochloritis or 3-5% hydrogen peroxide solution sterilization 10-20min, outwells thimerosal, cleans seed 5-8 time with distilled water;
Seed after sterilization is placed in the double dish being covered with layer 2-3 filter paper by c, is placed in vernalization under 25-30 DEG C of condition;
After d germination, consistent for germinating energy seed is placed on the meshed flat board of band, its flat board is allowed to swim in continued growth in ultrapure water, wherein mesh diameter is less than seed particle size, prevent seed from falling in water, intensity of illumination is set by illumination box: 0-4000lux, temperature 4-30 DEG C, continuous parameters is adjustable, arrange plant optimum cultivation temperature to cultivate 1 week, after axis minister becomes, the root system of consistent for growing way plant is taken out carefully from mesh, plant stem is put on the spot with sponge, then it is fixed on in the hole of KT plate central authorities, this KT plate is fixed on and cultivates on cup, be cultivation plant cup of the present invention, the volume cultivating cup is 400-600ml,
In step e a, often kind of nutrient solution arranges 10-30 and cultivates plant cup, namely often kind of nutrient solution arranges 10-30 repetition, then plant is placed in illumination box and intensity of illumination is set: 0-4000lux, temperature 4-30 DEG C, continuous parameters is adjustable, arranges plant optimum cultivation temperature and cultivates, and is finished pancebrin 0-4 week and cultivates, grow up to after seedling until plant, each nutrient solution prescription of 4-8 week step a is cultivated;
F nutritional deficiency and the information acquisition of normal growth plant leaf blade chlorophyll fluorescence;
The chlorophyll fluorescence diagnosis of g plant nutritional deficiency calculates.
2. the chlorophyll fluorescence diagnostic method that wanes of a Plants essential nutrient element according to claim 1, is characterized in that, being configured to of some nutrient solutions in step a:
(1) with ultrapure water (18.2M Ω) configuration: 7.45mg/LNa
2– EDTA, 5.57mg/LFeSO
47H
2o, 820.7mg/LCa (NO
3)
2, 502.6mg/LKNO
3, 616.2mg/LMgSO
47H
2o, 272.2mg/LKH
2pO
4, 2.860mg/LH
3bO
3, 1.015mg/LMnSO
4, 0.079mg/LCuSO
45H
2o, 0.220mg/LZnSO
47H
2o, 0.090mg/LH
2moO
4solution adjust ph 5.5-6.5, obtains complete nutrition liquid;
(2) with ultrapure water (18.2M Ω) configuration: 616.2mg/LMgSO
47H
2o, 272.2mg/LKH
2pO
4, 7.45mg/LNa
2– EDTA, 5.57mg/LFeSO
47H
2o, 555mg/LCaCl
2, 372.8mg/LKCl, 2.860mg/LH
3bO
3, 1.015mg/LMnSO
4, 0.079mg/LCuSO
45H
2o, 0.220mg/LZnSO
47H
2o, 0.090mg/LH
2moO
4solution adjust ph 5.5-6.5, obtains nitrogen stress nutrient solution;
(3) with ultrapure water (18.2M Ω) configuration: 820.7mg/LCa (NO
3)
2, 502.6mg/LKNO
3, 616.2mg/LMgSO
47H
2o, 7.45mg/LNa
2– EDTA, 5.57mg/LFeSO
47H
2o, 149.1mg/LKCl, 2.860mg/LH
3bO
3, 1.015mg/LMnSO
4, 0.079mg/LCuSO
45H
2o, 0.220mg/LZnSO
47H
2o, 0.090mg/LH
2moO
4solution adjust ph 5.5-6.5, obtains scarce phosphorus nutrition liquid;
(4) with ultrapure water (18.2M Ω) configuration: 820.7mg/LCa (NO
3)
2, 616.2mg/LMgSO
47H
2o, 240mg/LNaH
2pO
4, 7.45mg/LNa
2– EDTA, 5.57mg/LFeSO
47H
2o, 424.5mg/LNaNO
3, 2.860mg/LH
3bO
3, 1.015mg/LMnSO
4, 0.079mg/LCuSO
45H
2o, 0.220mg/LZnSO
47H
2o, 0.090mg/LH
2moO
4solution adjust ph 5.5-6.5, obtains potassium deficiency nutrient solution;
(5) with ultrapure water (18.2M Ω) configuration: 820.7mg/LCa (NO
3)
2, 502.6mg/LKNO
3, 616.2mg/LMgSO
47H
2o, 272.2mg/LKH
2pO
4, 2.860mg/LH
3bO
3, 1.015mg/LMnSO
4, 0.079mg/LCuSO
45H
2o, 0.220mg/LZnSO
47H
2o, 0.090mg/LH
2moO
4solution adjust ph 5.5-6.5, obtains iron deficiency nutrient solution.
3. the chlorophyll fluorescence diagnostic method that wanes of a Plants essential nutrient element according to claim 1, it is characterized in that, step f chlorophyll fluorescence information acquisition step is:
(1) a dark end cycle on plant, before welcoming next periodicity of illumination, ensures the abundant dark adatpation of plant leaf, measures when chlorophyll Photosystem I I (PSII) opens completely;
Penetrate 10-15 μ s with the illumination of wavelength 465-475nm (peak value is 470nm), intensity of illumination 0.03-0.1 μm olm-2s-1, gather the fluorescence information that 680nm place chlorophyll is launched, recording this fluorescent value is Fo simultaneously;
Penetrate 10-15 μ s with the illumination of wavelength 465-485nm (peak value is 470nm), intensity of illumination 4000-5000 μm olm-2s-1, gather the fluorescence information that 680nm place chlorophyll is launched, recording this fluorescent value is Fm simultaneously;
Penetrate 10-30min with the illumination of wavelength 465-485nm (peak value is 470nm), intensity of illumination 200-600 μm olm-2s-1, fully start photosynthesis of plant system.Gather the fluorescence information that 680nm place chlorophyll is launched afterwards, recording this fluorescent value is Fs;
5s is irradiated with the far red light of wavelength 725-755nm (peak value is 730nm), intensity of illumination 200-600 μm olm-2s-1;
Penetrate 10-15 μ s with the illumination of wavelength 465-475nm (peak value is 470nm), intensity of illumination 0.03-0.1 μm olm-2s-1, gather the fluorescence information that 680nm place chlorophyll is launched, recording this fluorescent value is Fo ' simultaneously;
Penetrate 10-15 μ s with the illumination of wavelength 465-485nm (peak value is 470nm), intensity of illumination 4000-5000 μm olm-2s-1, gather the fluorescence information that 680nm place chlorophyll is launched, recording this fluorescent value is Fm ' simultaneously.
4. the chlorophyll fluorescence diagnostic method that wanes of a Plants essential nutrient element according to claim 1, is characterized in that, the chlorophyll fluorescence diagnosis computing formula of step g plant nutritional deficiency is:
(1) chlorophyll fluorescence Diagnostic parameters Fv/Fm, Fv '/Fm ', Φ PSII; Fv/Fm represents the height of Photochemical Efficiency, and Fv '/Fm ' represents effective Photochemical Efficiency, and Φ PSII represents leaf photosynthesis electron transport rate speed;
(2) chlorophyll fluorescence Diagnostic parameters computing formula: Fv/Fm=(Fm-Fo)/Fm, Fv '/Fm '=(Fm '-Fo ')/Fm ', Φ PSII=(Fm '-Fs)/Fm ';
Fo represents the minimum fluorescence quantum yield of dark adatpation, Fm represents dark adatpation maximum fluorescence output, Fs represents steady-state fluorescence output under light, Fo ' represents the minimum fluorescence quantum yield of photopia, Fm ' represents photopia maximum fluorescence output.
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