CN102435590B - Method for confirming actinic light intensity in chlorophyll fluorescence induction curve measurement - Google Patents
Method for confirming actinic light intensity in chlorophyll fluorescence induction curve measurement Download PDFInfo
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Abstract
The invention provides a method for confirming actinic light intensity in chlorophyll fluorescence induction curve measurement, which comprises the following steps of: obtaining plant leaf blades, shading environment light, setting multiple light intensity gradients, irradiating for a preset time with each light intensity gradient and measuring practical photosynthetic efficiency and relative electron transmission rate; drawing a response curve of the relative electron transmission rate, which is changed along the light intensity and fitting the response curve to obtain semi-saturated light intensity; taking the semi-saturated light intensity as the actinic light intensity for measuring the chlorophyll fluorescence induction curve; calculating a difference value between a maximal fluorescence value after light adaptation and a practical fluorescence value and a difference value between a maximal fluorescence value after dark adaptation and a basic fluorescence value after dark adaptation, and calculating the ratio of the two difference values; judging if the ratio is in a preset range, if so, confirming the current light intensity as the actinic light intensity, and if the ratio is less/more than the lower limit/the upper limit of the preset range, reducing/increasing the light intensity, re-measuring the chlorophyll fluorescence induction curve and calculating the ratio. The method can measure typical chlorophyll fluorescence induction curve.
Description
Technical field
The present invention relates to the plant physiology technical field, specifically, the present invention relates to definite method that a kind of chlorophyll fluorescence is induced actinic light intensity in the curved measurement.
Background technology
Photosynthesis is most important chemical reaction in the plant physiology metabolic process, and it utilizes the luminous energy splitting water to emit oxygen, assimilates simultaneously the carbon dioxide synthesis of glucose, is the basis that all life activity can be carried out on the earth.Photosynthetic measurement and research are the focuses in the fields such as plant physiology, plant ecology, agronomy, forestry, gardening, Ecological Physiology in Algae always.
Photosynthetic measurement mainly comprises gas exchange, several technology such as modulation chlorophyll fluorescence, Differential absorbance and photosynthetic oxygen evolution.Wherein modulate the live body probe that the chlorophyll fluorescence technology is called as photosynthesis research, the photosynthetic primary reaction processes such as it can not only reflect that luminous energy absorbs, excitation energy transmission and photochemical reaction, and with the foundation of electronics transmission, proton gradient and ATP is synthetic and CO
2The process such as fixing is relevant.Most photosynthesis change in process all can reflect by the modulation chlorophyll fluorescence, and its mensuration does not need smudge cells, does not injure biosome, therefore change to reflect that by the research chlorophyll fluorescence photosynthetic variation is a kind of easy, quick, reliable method, (Papageorgiou GC, Govindjee.Chlorophyll a Fluorescence:a Signature of Photosynthesis.Dordrecht:Springer have been obtained using very widely in international scientific research circle; 2004.).
It is one of standard method of modulation chlorophyll fluorescence measurement that chlorophyll fluorescence is induced curve.Germany scientist Kautsky in 1931 observes for the first time chlorophyll fluorescence and induces phenomenon, and proposes fluorescence induction process and CO
2Fixing closely related (Lichtenthaler HK.The Kautsky effect:60years of chlorophyll fluorescence induction kinetics.Photosynthetica, 1992,27:45-55.).After this in half a century, scientist induces curve to carry out extensive and deep research to chlorophyll fluorescence, and it is theoretical progressively to have formed the photosynthesis fluorescence induction, be widely used in photosynthesis research, but the instrument that adopts can only be measured under indoor condition of covering surround lighting fully, has greatly limited the use of chlorophyll fluorescence technology.
Nineteen eighty-three, Germany scientist Schreiber has designed pulse-amplitude-modulation chlorophyll fluorescence instrument (Pulse-Amplitude-Modulation Chlorophyll Fluorometer, be called for short PAM) (Schreiber U, Bilger W, Schliwa U.Continuous recording of photochemical and non-photochemical chlorophyll fluorescence quenching with a new type of modulation fluorometer.Photosynthesis Research, 1986,10:51-62.), and commercially produced by internationally famous photosynthesis instrument manufacturer Germany WALZ company.The invention of PAM has greatly promoted popularizing of chlorophyll fluorescence technology, expand to rapidly the field (Lin Shiqing such as plant ecology, agronomy, forestry, Ecological Physiology in Algae, wetland ecology, genetic breeding from traditional plant physiology, Xu Chunhui, Zhang Qide, Xu Li, Mao Dazhang, Kuang Tingyun. the application of chlorophyll fluorescence kinetics in physiological responses of plants to anti-environment, ecology and agricultural modernization. BULLETIN OF BOTANY Vol., 1992,9 (1): 1-16; Chen Yizhu, Li Xiaoping, Xia Li, Guo Junyan. the chlorophyll fluorescence technology is coerced application in the research at plant environment. tropical and subtropical plant journal, 1995,3 (4): 79-86; Guo Yupeng, Zheng Xia, Wang Xinyu, Cao Ziyi. the application of chlorophyll fluorescence technology in the photosynthetic mutant of screening. Acta Prataculture, 2009,18 (6): 226-234; Zhang Jingping, Huang Xiaoping. the application of chlorophyll fluorescence technology in the sea grass ecological study. marine environment science, 2009,28 (6): 772-778).In China, the PAM series modulation chlorophyll fluorescence instrument that present R﹠D institutions at different levels are using is above 500.
It is the most classical method during the modulation chlorophyll fluorescence is measured that chlorophyll fluorescence is induced curve, plant leaf blade at first will be through after a while dark adatpation so that all electronic gates on the photosynthetic electron transport chain all be in open state when measuring, open subsequently a very weak modulation of intensity and measure light (Measuring Light, ML) be used for exciting the basic fluorescence Fo that produces after the dark adatpation, then open the maximum fluorescence Fm of a duration after the rapid closeall electronic gate of saturation pulse (Saturation Pulse, SP) of 0.4-1s obtains dark adatpation.Postpone approximately after the fluorescent value such as 40s is reduced near the Fo, the actinic light (Actinic Light) of opening a constant intensity carries out photosynthetic induction, and lasting irradiation a few minutes.When actinic light is opened, open a maximum fluorescence Fm ' after the saturation pulse measurement photopia every 30s.Then close actinic light, open electronics that far-red light (Far-red light, FR) promotes to be accumulated in the electronic gate place as early as possible toward the Photosystem I transmission, measure the minimum fluorescence Fo ' after the photopia.
Induce the measurement of curve by above-mentioned chlorophyll fluorescence, can draw following parameter (Han Boping, Han Zhiguo pay Xiang. the algae photosynthesis mechanism and model .2003. .P.57-78 of Science Press; Schreiber U.Pulse-Amplitude-Modulation (PAM) fluorometry and saturation pulse method:an overview.In:Chlorophyll Fluorescence:a Signature of Photosynthesis.Edited by Papageorgiou GC, Govindjee, Springer; 2004:279-319; Kramer DM, Johnson G, Kiirats O, Edwards GE.New fluorescence parameters for the determination of QA redox state and excitation energy fluxes.Photosynthesis Research, 2004,79:209-218):
Basic fluorescence Fo after the dark adatpation;
Maximum fluorescence Fm after the dark adatpation;
Real-time fluorescence F;
Maximum fluorescence Fm ' after the photopia;
Minimum fluorescence Fo ' after the photopia;
Maximum photosynthetic efficiency Fv/Fm=(Fm-Fo)/Fm;
Actual photosynthesis efficient Y (II)=(Fm '-F)/Fm ';
Relative electronics transfer rate rETR=PAR*Y (II) * 0.84*0.5;
Photochemical quenching qP=(Fm '-F)/Fv '=1-(F-Fo ')/(Fm '-Fo ') or qL=(Fm '-F)/(Fm '-Fo ') Fo '/F=qPFo '/F;
Non-Photochemical quenching qN=(Fv-Fv ')/Fv=1-(Fm '-Fo ')/(Fm-Fo) or NPQ=(Fm-Fm ')/Fm '=Fm/Fm '-1;
The quantum yield Y (NPQ) that the modulability energy dissipates=1-Y (II)-1/ (NPQ+1+qL (Fm/Fo-1));
The quantum yield Y (NO)=1/ (NPQ+1+qL (Fm/Fo-1)) that non-modulability energy dissipates etc.
These fluorescence parameters all have typical biological significance, have a very wide range of applications.For example when the Fv/Fm of plant＞0.8, illustrate that plant is in the physiological status of health; Otherwise then be forced.
In addition, a series of intensity gradient from low to high and difference irradiation processing 10-30s are set, then open saturation pulse and measure actual photosynthesis efficient Y (II) and Relative electron transport rate rETR, can draw the response curve that rETR changes with light intensity, this is referred to as fast light curve (White AJ, Critchley C.Rapid light curves:A new fluorescence method to assess the state of the photosynthetic apparatus.Photosynthesis Research, 1999,59:63-72; Ralph PJ, Gademann R.Rapid light curves:A powerful tool to assess photosynthetic activity.Aquatic Botany, 2005,82:222-237.).Can utilize equation P=Pm (1-e to the fast light curve
-α PAR/Pm) e
-β PAR/Pm(Plat T, Gallegos CL, Harrison WG.Photoinhibition of photosynthesis in natural assemblages of marine phytoplankton.Journal of Marine Research, 1980,38:687-701.) carry out nonlinear fitting, wherein P represents photosynthetic rate, be Relative electron transport rate rETR, maximum potential Relative electron transport rate rETRmax during inhibition that the Pm representative is unglazed, α represents initial slope, β is that light suppresses parameter, and PAR represents namely intensity of illumination of photosynthetically active radiation intensity.Then can calculate semi-saturation light intensity Ik=Pm/ α.
When measuring chlorophyll fluorescence and inducing curve, require to measure light must be enough a little less than, be not enough to cause photosynthesis, common intensity is less than 1 μ mol m
-2s
-1Require saturation pulse intensity can all turn off all Photosynthetic Electron doors in moment enough by force, higher plant is generally greater than 8000 μ mol m
-2s
-1, algae is generally greater than 4000 μ mol m
-2s
-1Far-red light intensity generally can meet the demands by the default setting of instrument.Only having the selection of actinic light intensity, is puzzlement scientific research personnel's a difficult problem.Generally only provide an actinic light intensity in the bibliographical information, but this actinic light intensity of specifically how determining, whether this actinic light intensity is suitable, then do not point out.If the selection of actinic light intensity is improper, then can directly have influence on the parameters such as Y (II), rETR, qP, qL, qN, NPQ, Y (NPQ), Y (NO) whether representative to the scientific experimentation of design.For example field (beam intensity ratio is higher) corn is subject to behind the drought stress Y (II) and can showing and reduce, if induce curve but adopted lower actinic light intensity to measure chlorophyll fluorescence, the Y that then draws (II) decline degree can cause test findings not have representativeness well below the real standard in field.
Summary of the invention
Technical matters to be solved by this invention provides definite method that a kind of chlorophyll fluorescence is induced actinic light intensity in the curved measurement, induces curve for the correct measurement chlorophyll fluorescence suitable method is provided.
For solving the problems of the technologies described above, the invention provides definite method that a kind of chlorophyll fluorescence is induced actinic light intensity in the curved measurement, comprise step:
A. obtain the plant leaf blade that needs test, in the situation that shelter from surround lighting, a plurality of intensity gradient from low to high are set, and each opens respectively a saturation pulse after the described intensity gradient irradiation schedule time, measures actual photosynthesis efficient and Relative electron transport rate under the described light intensity;
B. draw the response curve that described Relative electron transport rate changes with light intensity, and described response curve is carried out nonlinear fitting, obtain the semi-saturation light intensity;
C. with described semi-saturation light intensity or near the light intensity of described semi-saturation light intensity as actinic light intensity, measure according to a conventional method chlorophyll fluorescence and induce curve;
D. after described chlorophyll fluorescence induces curved measurement to finish, calculate maximum fluorescence value and the difference between the real-time fluorescence value and the maximum fluorescence value after the dark adatpation and the difference between the basic fluorescent value after the dark adatpation after the photopia, and calculate the ratio of above-mentioned two differences;
E. judge whether described ratio is in the preset range, if so, determine that then current light intensity is described actinic light intensity; If described ratio returns step C after, then reducing described light intensity less than the lower limit of described preset range; If greater than the upper limit of described preset range, then raising, described ratio returns step C after the described light intensity; So circulation is until described ratio is in the described preset range.
Preferably, described plant leaf blade is that live body healthy leaves or its petiole are immersed in the stripped healthy leaves in the water.
Preferably, described intensity gradient from low to high is 8～20.
Preferably, the schedule time of each described intensity gradient irradiation is 10～30 seconds.
Preferably, measuring described chlorophyll fluorescence induces the requirement of curve to comprise:
At least the value of the maximum fluorescence after latter two photopia is basic identical, to prove that this light intensity is to the photosynthetic stable state that reached of inducing.
Preferably, the preset range of described ratio is 0.33～0.66.
More preferably, described ratio is 0.5.
Preferably, reduce or the amplitude of the described light intensity that raises is each 1 intensity gradient.
Compared with prior art, the present invention has the following advantages:
(1) the actinic light intensity of utilizing method of the present invention to determine can guarantee that control sample (healthy plant leaf blade) can measure very typical chlorophyll fluorescence and induce curve, guarantees the appropriate design of whole scientific experimentation and obtains representational data;
(2) method of the present invention is implemented simple, fast and easy, workable;
(3) method of the present invention has wide range of applications, and the chlorophyll fluorescence that is applicable to all higher plants and algae is induced curved measurement.
Description of drawings
Above-mentioned and other feature, character and advantage of the present invention will become more obvious by the description below in conjunction with drawings and Examples, wherein:
Fig. 1 is the method flow diagram that definite chlorophyll fluorescence is induced curved measurement actinic light intensity that is used for of one embodiment of the invention;
Fig. 2 is that the chlorophyll fluorescence of the epipremnum aureum of one embodiment of the invention is induced curve;
Fig. 3 is that the chlorophyll fluorescence of creeping oxalis under different actinic light intensity of one embodiment of the invention induced curve;
Fig. 4 is that the chlorophyll fluorescence of camphor tree under different actinic light intensity of one embodiment of the invention induced curve;
Fig. 5 is that the chlorophyll fluorescence of Alternanthera philoxeroides under different actinic light intensity of one embodiment of the invention induced curve.
Embodiment
The invention will be further described below in conjunction with specific embodiments and the drawings; set forth in the following description more details so that fully understand the present invention; but the present invention obviously can implement with multiple this description ground alternate manner that is different from; those skilled in the art can be in the situation that do similar popularization, deduction without prejudice to intension of the present invention according to practical situations, therefore should be with content constraints protection scope of the present invention of this specific embodiment.
Fig. 1 is the method flow diagram that definite chlorophyll fluorescence is induced curved measurement actinic light intensity that is used for of one embodiment of the invention.As shown in Figure 1, this chlorophyll fluorescence induces that definite method of actinic light intensity can comprise step in the curved measurement:
Execution in step S101 obtains the plant leaf blade that needs test, and this plant leaf blade can be immersed in stripped healthy leaves in the water for live body healthy leaves or petiole.In the situation that shelter from surround lighting, setting example is such as 8～20 intensity gradient from low to high, each intensity gradient irradiation is opened respectively a saturation pulse after 10～30 second schedule time, measures actual photosynthesis efficient Y (II) and Relative electron transport rate rETR under the light intensity.
Execution in step S102 draws the response curve that Relative electron transport rate rETR changes with light intensity, and response curve is carried out nonlinear fitting, obtains semi-saturation light intensity Ik.
Execution in step S103, with semi-saturation light intensity Ik or near the light intensity of semi-saturation light intensity Ik as actinic light intensity, measure according to a conventional method chlorophyll fluorescence and induce curve.Wherein, measuring chlorophyll fluorescence induces the requirement of curve to comprise: the maximum fluorescence value Fm ' after latter two photopia is basic identical at least, and this shows that this light intensity is to the photosynthetic stable state that reached of inducing.
Execution in step S104, after chlorophyll fluorescence induces curved measurement to finish, calculate maximum fluorescence value Fm ' after the photopia and the difference DELTA F between the real-time fluorescence value F (Δ F=Fm '-F) and the difference Fv (Fv=Fm-Fo) between the basic fluorescent value Fo after the maximum fluorescence value Fm after the dark adatpation and the dark adatpation, and calculate the ratios delta F/Fv of above-mentioned two differences.
Execution in step S105 judges whether ratios delta F/Fv is in the preset range, and this preset range can be set as 0.33～0.66, take 0.5 as best.If so, illustrate that then this actinic light intensity is suitable, execution in step S106 determines that current light intensity is actinic light intensity; If ratios delta F/Fv is less than the lower limit of preset range, then execution in step S107 returns execution in step S103 after the reduction light intensity (actinic light intensity); If ratios delta F/Fv is greater than the upper limit of preset range, then execution in step S108 returns step S103 after the rising light intensity (actinic light intensity).Wherein, the amplitude of reduction or rising light intensity can be each 1 intensity gradient.Certainly, if numerical difference between is apart from larger, then those skilled in the art also can reduce or a plurality of intensity gradient that raise according to actual conditions at every turn.
So circulation until ratios delta F/Fv is in the preset range, thereby is determined actinic light intensity.
Ultimate principle of the present invention is: Fm ' peak has reflected that with respect to the height at Fm peak the luminous energy that absorbs is used for carrying out the ratio height of photochemistry energy conversion.Fm ' peak value is higher, illustrates that the luminous energy that absorbs is higher for the ratio of carrying out the conversion of photochemistry energy, and vice versa.Δ F/Fv can be more directly perceived, quantitative description Fm ' with respect to the height of Fm.When Δ F/Fv greater than 0.66 the time, the height that Fm ' peak is described is in close proximity to Fm, photosynthetic organs can be chemical energy with the most light energy conversions that absorb very easily, illustrate that the actinic light undercapacity of this moment is to transfer all photosynthetic organs performance maximum capacities, the otherness of different disposal in the scientific experimentation (such as in various degree drought stress, heat stress etc.) is difficult to embody, the significance difference opposite sex may not observed out or not have to the significance difference opposite sex of the fluorescence parameter that in other words, should observe because actinic light intensity arranged low.When Δ F/Fv less than 0.33 the time, the height that Fm ' peak is described is very low, photosynthetic organs can only be chemical energy with the light energy conversion of a very little part, show that photosynthetic organs may be subject to light and suppress injury, the control sample that should be in normal condition has been subjected to the light inhibition, can cause the design of scientific experimentation unreasonable, data do not have representativeness.
Fig. 2 is that the chlorophyll fluorescence of the epipremnum aureum of one embodiment of the invention is induced curve.Get healthy live body epipremnum aureum (Scindapsus aureun) (Fv/Fm＞0.8), clamp blade with binary channels PAM-100 measuring system DUAL-PAM-100 (Walz, Germany), cover whole blade with black cloth.10 intensity gradient (9,89,127,211,342,522,696,861,1064 and 1323 μ mol m from low to high are set in instrument software DualPAM v1.9
-2s
-1), and the irradiation time that each intensity gradient is set be 20s, then measure according to a conventional method the fast light curve.Measure the equation P=Pm (1-e that carries with software after finishing
-α PAR/Pm) e
-β PAR/PmCarry out nonlinear fitting, draw semi-saturation light intensity Ik=142 μ mol m
-2s
-1Select in the light intensity tabulation and the immediate light intensity 127 μ mol m of Ik
-2s
-1As actinic light intensity, then to measure according to a conventional method chlorophyll fluorescence and induce curve, the chlorophyll fluorescence that draws is as shown in Figure 2 induced curve.Two last among Fig. 2 Fm ' values are basic identical, show to measure when finishing photosynthetic inducing reached stable state.The Δ F/Fv=0.62 that calculates according to Fig. 2, within the 0.33-0.66 scope, therefore 127 μ mol m
-2s
-1More satisfactory actinic light intensity for epipremnum aureum.
Fig. 3 is that the chlorophyll fluorescence of creeping oxalis under different actinic light intensity of one embodiment of the invention induced curve.Get healthy window box oxalis (Oxalis corymbosa DC.) blade (Fv/Fm＞0.8), with binary channels PAM-100 measuring system DUAL-PAM-100 (Walz, Germany) clamp blade, petiole is immersed in the water, covers whole blade with black cloth.10 intensity gradient (49,89,127,211,342,522,696,861,1064 and 1323 μ mol m from low to high are set in instrument software DualPAM v1.9
-2s
-1), and the irradiation time that each intensity gradient is set be 20s, then measure according to a conventional method the fast light curve.Measure the equation P=Pm (1-e that carries with software after finishing
-α PAR/Pm) e
-β PAR/PmCarry out nonlinear fitting, draw semi-saturation light intensity Ik=263 μ mol m
-2s
-1Select respectively in the light intensity tabulation light intensity 127 μ mol m far below Ik
-2s
-1, near the light intensity 211 μ mol m of Ik
-2s
-1, be higher than the light intensity 342 μ mol m of Ik
-2s
-1With the light intensity 522 μ mol m far above Ik
-2s
-1Carry out chlorophyll fluorescence as actinic light intensity and induce curved measurement, the result who draws is respectively shown in Fig. 3 A-Fig. 3 D.When actinic light intensity is 127 μ mol m
-2s
-1The time, inducing the Δ F/Fv=0.79 (Fig. 3 A) of curve, Δ F/Fv illustrates that greater than 0.66 actinic light intensity is excessively low; When actinic light intensity is 211 μ mol m
-2s
-1The time, induce the Δ F/Fv=0.46 (Fig. 3 B) of curve, within the 0.33-0.66 scope, be more satisfactory actinic light intensity; When actinic light intensity is 342 μ mol m
-2s
-1The time, induce the Δ F/Fv=0.31 (Fig. 3 C) of curve, close to 0.33, actinic light intensity is slightly higher; When actinic light intensity is 522 μ mol m
-2s
-1The time, inducing the Δ F/Fv=0.10 (Fig. 3 D) of curve, Δ F/Fv illustrates that much smaller than 0.33 actinic light intensity is too high, has caused light to suppress.By above-mentioned comparison, can find out obviously that different actinic light intensity can cause chlorophyll fluorescence to induce the remarkable difference of curve, only having when satisfying the light intensity of Δ F/Fv between 0.33-0.66 the time (is 211 μ mol m in the present embodiment
-2s
-1) just be fit to as actinic light intensity.
Embodiment 3
Fig. 4 is that the chlorophyll fluorescence of camphor tree under different actinic light intensity of one embodiment of the invention induced curve.Get healthy camphor tree (Cinnamomum camphora (L.) Presl.) blade (Fv/Fm＞0.8), with portable modulation chlorophyll fluorescence instrument MINI-PAM (Walz, Germany) leaf folder 2030-B clamps blade, and petiole is immersed in the water, covers whole blade with black cloth.8 intensity gradient (116,207,312,427,620,816,1143 and 1504 μ mol m from low to high are set in MINI-PAM software WinControl v3.18
-2s
-1), and the irradiation time that each intensity gradient is set be 20s, then measure according to a conventional method the fast light curve.Measure the equation P=Pm (1-e that carries with software after finishing
-α PAR/Pm) e
-β PAR/PmCarry out nonlinear fitting, draw semi-saturation light intensity Ik=286 μ mol m
-2s
-1With in the light intensity tabulation close to the light intensity 312 μ mol m of Ik
-2s
-1Go out chlorophyll fluorescence induction curve (Fig. 4 A) as the actinic light ionization meter, wherein Δ F/Fv=0.27 is too high less than 0.33 explanation light intensity.Select low one grade light intensity 207 μ mol m
-2s
-1Go out chlorophyll fluorescence induction curve (Fig. 4 B) as the actinic light ionization meter, wherein Δ F/Fv=0.44 between 0.33-0.66, illustrates 207 μ mol m
-2s
-1The more satisfactory actinic light intensity of camphor leaf.
Embodiment 4
Fig. 5 is that the chlorophyll fluorescence of Alternanthera philoxeroides under different actinic light intensity of one embodiment of the invention induced curve.Get healthy Alternanthera philoxeroides (Alternanthera philoxeroides (Mart.) Griseb.) blade (Fv/Fm＞0.8), with portable modulation chlorophyll fluorescence instrument MINI-PAM (Walz, Germany) leaf folder 2030-B clamps blade, petiole is immersed in the water, covers whole blade with black cloth.8 intensity gradient (160,230,345,481,724,991,1510 and 2166 μ mol m from low to high are set in MINI-PAM software WinControlv3.18
-2s
-1), and the irradiation time that each intensity gradient is set be 20s, then measure according to a conventional method the fast light curve.Measure the equation P=Pm (1-e that carries with software after finishing
-α PAR/Pm) e
-β PAR/PmCarry out nonlinear fitting, draw semi-saturation light intensity Ik=447 μ mol m
-2s
-1With in the light intensity tabulation close to the light intensity 481 μ mol m of Ik
-2s
-1With 345 μ mol m
-2s
-1Go out chlorophyll fluorescence induction curve (Fig. 4) as the actinic light ionization meter respectively.When actinic light intensity is 481 μ mol m
-2s
-1The time, Δ F/Fv=0.41 (Fig. 5 A); When actinic light intensity is 345 μ mol m
-2s
-1The time, Δ F/Fv=0.54 (Fig. 5 B).No matter adopt 481 μ mol m
-2s
-1Or 345 μ mol m
-2s
-1As actinic light intensity, the Δ F/Fv that draws is between 0.33-0.66, illustrate that they are more satisfactory actinic light intensity, this has illustrated that from the side Alternanthera philoxeroides can both carry out desirable photosynthesis in larger range of light intensity, may be one of mechanism of its biotic intrusion.
Compared with prior art, the present invention has the following advantages:
(1) the actinic light intensity of utilizing method of the present invention to determine can guarantee that control sample (healthy plant leaf blade) can measure very typical chlorophyll fluorescence and induce curve, guarantees the appropriate design of whole scientific experimentation and obtains representational data;
(2) method of the present invention is implemented simple, fast and easy, workable;
(3) method of the present invention has wide range of applications, and the chlorophyll fluorescence that is applicable to all higher plants and algae is induced curved measurement.
Although the present invention with preferred embodiment openly as above, it is not to limit the present invention, and any those skilled in the art can make possible change and modification without departing from the spirit and scope of the present invention.Therefore, every content that does not break away from technical solution of the present invention, all falls within the protection domain that claim of the present invention defines any modification, equivalent variations and modification that above embodiment does according to technical spirit of the present invention.
Claims (7)
1. a chlorophyll fluorescence is induced definite method of actinic light intensity in the curved measurement, comprises step:
A. obtain the plant leaf blade that needs test, in the situation that shelter from surround lighting, a plurality of intensity gradient from low to high are set, and each opens respectively a saturation pulse after the described intensity gradient irradiation schedule time, measures actual photosynthesis efficient and Relative electron transport rate under the described light intensity;
B. draw the response curve that described Relative electron transport rate changes with light intensity, and described response curve is carried out nonlinear fitting, obtain the semi-saturation light intensity;
C. with described semi-saturation light intensity or near the light intensity of described semi-saturation light intensity as actinic light intensity, measure according to a conventional method chlorophyll fluorescence and induce curve;
D. after described chlorophyll fluorescence induces curved measurement to finish, calculate maximum fluorescence value and the difference between the real-time fluorescence value and the maximum fluorescence value after the dark adatpation and the difference between the basic fluorescent value after the dark adatpation after the photopia, and calculate the ratio of above-mentioned two differences;
E. judge whether described ratio is in the preset range, and described preset range is 0.33～0.66, if so, determine that then current light intensity is described actinic light intensity; If described ratio returns step C after, then reducing described light intensity less than the lower limit of described preset range; If greater than the upper limit of described preset range, then raising, described ratio returns step C after the described light intensity; So circulation is until described ratio is in the described preset range.
2. definite method of actinic light intensity according to claim 1 is characterized in that, described plant leaf blade is that live body healthy leaves or petiole are immersed in the stripped healthy leaves in the water.
3. definite method of actinic light intensity according to claim 1 is characterized in that, described intensity gradient from low to high is 8～20.
4. definite method of actinic light intensity according to claim 3 is characterized in that, the schedule time of each described intensity gradient irradiation is 10～30 seconds.
5. definite method of actinic light intensity according to claim 1 is characterized in that, measures described chlorophyll fluorescence and induces the requirement of curve to comprise:
At least the value of the maximum fluorescence after latter two photopia is basic identical, to prove that this light intensity is to the photosynthetic stable state that reached of inducing.
6. definite method of actinic light intensity according to claim 1 is characterized in that, described ratio is 0.5.
7. definite method of actinic light intensity according to claim 1 is characterized in that, reduces or the amplitude of the described light intensity that raises is each 1 intensity gradient.
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