CN102539507B - Method for evaluating nitrogen nutrition of paddy by micro dynamic ion flux technique and application thereof - Google Patents
Method for evaluating nitrogen nutrition of paddy by micro dynamic ion flux technique and application thereof Download PDFInfo
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Abstract
The invention provides a method for evaluating nitrogen nutrition of paddy by micro dynamic ion flux technique and an application thereof. The method comprises the following steps of: (1) injecting an ion filling liquid into a microelectrode until the tip of the microelectrode is full of the liquid, and then sucking a corresponding testing ion exchanger to the front end of the electrode; (2) inserting the electrode which is treated in the step (1) in a chlorinated Ag/AgCl electrode wire base and correcting through a correcting liquid; (3) putting the root of a paddy seedling to be tested in a test buffer solution until balanced, and then detecting the to-be-tested paddy seedling with the corrected electrode; and (4) processing and analyzing the detection results. The method is mainly applied to paddy, achieves the purpose of noninvasively and rapidly detecting nitrogen-absorbing capability of paddy with short detection time and high detection accuracy, and provides the significant difference in net ion flux of absorbing for detecting nitrogen in different forms in different varieties of paddy seedlings. The method is simple and reliable and provides a new approach to noninvasive and rapid screening of high nitrogen nutrition paddy cultivars.
Description
Technical field
The invention belongs to agricultural technology field, specifically, relate to the application aspect efficiently utilizing at Nitrogen Nutrition of Paddy Rice Plant of a kind of method of utilizing microscopic behavior ion current technology for detection Nitrogen Nutrition of Paddy Rice Plant and the method.
Background technology
Nitrogen is the important composition composition of cellular plasm, all teleorganic basic nutrition elements, it is the nutrient that plant is primary, have the call, bringing into play the effect that can not replace in the processes such as plant structure, energetic supersession and quality formation, it has very important impact to nitrogen-containing compound in plant growth and body.Appropriate nitrogen can promote growing of vegetable nutritorium and reproductive organs, increases greenery area, strengthens the accumulation of photosynthesis and nutriment.And excessively applied nitrogen causes the outstanding problems such as utilization rate of nitrogen fertilizer (NUE) reduction, severe environmental pollution.
Paddy rice is Chinese topmost cereal crops, has very important status aspect agricultural production and grain security.And nitrogen is to affect the of paramount importance nutrient of yield and quality of rice.Paddy rice directly affects its g and D situation to absorption, assimilation and the transhipment of nitrogen, thereby affects yield and quality.Further investigation Nitrogen Absorption dynamic rule, the adjusting and controlling rice growth and development process that contributes to adopt an effective measure, improve output and improve quality, can save fertilizer simultaneously, improve the soil nutrient structure.Microscopic dynamic ion flow detecting technology as harmless, in real time, dynamically, detection method is to differentiate the direct tool of Nitrogen Nutrition of Paddy Rice Plant utilization factor fast, utilizes this technology for detection different cultivars paddy rice to different shape nitrogen NH
4 +, NO
3 -receptivity, be intended to estimate efficiently the breeding demand for Nitrogen Nutrition of Paddy Rice Plant, for the nitrogenous fertilizer high-yield and high-efficiency provides theoretical foundation and technical support.
NH
4 +and NO
3 -be two kinds of main forms of nitrogens that higher plant absorbs, and plant absorb on the selectivity of these two kinds of form nitrogen ions soil physico-chemical property factors such as being subject to plant kind, kind, growth and development stage and pH value and affect.Find during the research black walnut seedling such as MichaelA, apply the NH of proper ratio
4 +/ NO
3 -nitrogenous fertilizer can promote better (the Michael A. that grows of black walnut, Nicodemus K.Nitrate reductase activity and nitrogencompounds in xylem exudate of Juglans nigra seedlings:relation tonitrogen source and supply[J] .Trees, 2008,22:685-695.).(nitrogenous source and recharge connection that in wooden exudate of black walnut, nitrate reductase activity and nitrogen-containing compound embody) therefore, keeps suitable NH at the different times of growth and development of plants
4+/ NO
3-ratio has important effect, so utilize root system of plant to NH
4 +, NO
3 -the receptivity of ion instructs the rational application of fertilizer, realizes that efficient utilization of nitrogen becomes possibility, the present invention take that sky educates 131, sky educates 163, the round-grained rice 8 of pacifying as experiment material research different cultivars rice root to NH
4 +, NO
3 -the absorbing rule of ion, efficiently utilize the foundation of estimating screening as the different cultivars rice nutrition.
Microscopic dynamic ion flow detecting technology is the electrophysiological technique of a comprehensive specific ion/Molecular Detection, and core technology is in the situation that computer utilizes concentration (mM level), the flow velocity (10 of ion/molecular selectivity micro-electrode in the ion/molecular that does not contact sample detection turnover sample under automatically controlling
-12molcm
-2s
-1) and the information such as direction of motion.This technical operation is convenient, fast, real-time, harmless, can truly reflect that other technologies are difficult to the vital movement rule of reflection.This technology provides a kind of visual technological means for life science, promotes that the mankind find out the information that conventional art can't detect, and has solved many problem in science.
In order to grasp in time the plant growth situation, except carrying out the outward appearance observation according to experience, often using the nitrogen nutrition of aerial part as the foundation of judgement crop nitrogen nutrition situation.Traditional nitrogen nutrition detection method complicated operation based on the chemical composition analytic approach, analytical cycle is long, and poor in timeliness can't accomplish that real-time, the live body during plant growth detects.
Chlorophyll meter (SPAD) method (Li Gang China, Xue Lihong, You Juan, etc. [J] selected in the leaf position of Nitrogen in Rice and chlorophyll SPAD Leaf distribution characteristics and nitrogen diagnosis. Scientia Agricultura Sinica, 2007,40 (6): 1127-1134.; Arregui L M, Lasa B, Lafarga A, Iranneta I, Baroja E, Quemada M.Evaluation of chlorophyll meters astools for N fertilization in winter wheat under humid Mediterraneanconditions is (under the condition of moist Mediterranean, chlorophyll content is as the appraisal tool of nitrogen application) .Eur J Agron, 2006,24:140-148.) can Fast Measurement crop chlorophyll level and quick diagnosis crop nitrogen level.But the information faces that chlorophyll meter gathers is less than normal, and workload is larger, to measuring standard, higher requirement is arranged, and the difficult grasp of diagnostic criteria.The SPAD value can be subject to the impact of time, place, kind, soil property, crop growing season and growing environment.
Spectral technique (Tan Changwei, Zhuan Hengyang, Zhou Qingbo. the spectroscopic diagnostics research [J] of rice leaf nutrition. Yangzhou University's journal: agricultural and life science version, 2009,30 (2): 50-53.; Zhang Lizhou etc. the application [J] of Digital image technology in the summer corn Nitrogen Nutrition Diagnosis. Chinese Ecological Agriculture journal, 2010,18 (6): 1340-1344.; Li Hongjun etc. the Applied Digital image carries out the research [J] of image analysis method in the wheat nitrogen nutrient diagnosis. the Chinese Ecological Agriculture journal, 2011,19 (1): 155-159.) have acquisition of information fast, the advantage such as good, the non-destructive of economy, operability, also can develop into remote Diagnosis Technology, latest developments are comparatively quick.Yet spectral technique is ripe not enough, and the factor of light absorption and reflection of affecting is a lot, and between crop varieties, the canopy color parameter there are differences, should reduce error as far as possible, make spectral technique more accurate as the nitrogen diagnosis method, be applied to better the nitrogen diagnosis of crop.
The methods such as chemical analysis method commonly used, chlorophyll meter (SPAD) method, spectral technique while as mentioned above, in the past analyzing the crop nitrogen nutrition utilization ratio.As: the traditional nitrogen nutrition detection method complicated operation based on the chemical composition analytic approach, analytical cycle is long, and poor in timeliness can't accomplish that real-time, the live body during plant growth detects; And spectral technique is ripe not enough, and the factor of light absorption and reflection of affecting is a lot, and between crop varieties, the canopy color parameter there are differences, and error is larger.Although above-mentioned these methods have been improved blindness and the excessive problem of nitrogenous fertilizer that empiric fertilization brings to a certain extent, but still can not carry out the nitrogen nutrition morphological diagnosis easy, quick, real-time, harmless, exactly, and guide fertilising.Therefore, the method for developing new detection paddy rice fungoid damping-off has very important significance.
Summary of the invention
The present invention is intended to provide for seedling cultivation of rice, rice breeding, rice cropping a kind of method of quick, harmless detection Nitrogen Nutrition of Paddy Rice Plant, to be applied to Nitrogen Nutrition of Paddy Rice Plant, efficiently utilizes aspect.
Particularly, the invention provides a kind of method of utilizing microscopic dynamic ion flow detecting technology to detect Nitrogen Nutrition of Paddy Rice Plant, adopt the BIO-001B system to detect.
Said method comprising the steps of:
1) pour into ion filling liquid to being full of described eletrode tip to microelectrode, then the electrode front end is sucked to corresponding test ion exchanger;
2) will be through step 1) electrode after processing is inserted in the Ag/AgCl electrode wires pedestal of chlorination, and puts into correcting fluid and proofread and correct;
3) get rice seedlings to be measured, its root first is placed on to balance in assay buffer, then detected with the electrode pair rice seedlings to be measured after proofreading and correct;
4) testing result is processed and analyzed.
Further, said method comprising the steps of:
1) pour into ion filling liquid to being full of described eletrode tip 1～1.5cm to microelectrode, then the electrode front end is sucked to corresponding test ion exchanger;
2) electrode after step (1) is processed is inserted in to the Ag/AgCl electrode wires pedestal of chlorination, and puts into correcting fluid and proofread and correct;
3) get rice seedlings to be measured, its root first is placed on to balance 30-40min in assay buffer, then detected 10～15min with the electrode pair rice seedlings to be measured after proofreading and correct;
4) testing result is processed and analyzed.
Further, said method comprising the steps of:
1) pour into ion filling liquid to being full of described eletrode tip 1cm to microelectrode, then the electrode front end is sucked to corresponding test ion exchanger;
2) electrode after step (1) is processed is inserted in to the Ag/AgCl electrode wires pedestal of chlorination, and puts into correcting fluid and proofread and correct;
3) get rice seedlings to be measured, its root first is placed on to balance 30-40min in assay buffer, then detected 15min with the electrode pair rice seedlings to be measured after proofreading and correct;
4) testing result is processed and analyzed.
In described method, detect the paddy rice of use in Seedling Stage.
Test ion concentration range in described microscopic dynamic ion flow detecting technology in assay buffer is 0.05～0.15mM.
The paddy rice of detect using is in 21 heart stages of leaf, outside surface 10～45 μ m places of the Meristernatic zone that measuring position be the described rice seedlings tip of a root 100～400 μ m of distance.
Described step 1) in, the length of the front end of electrode suction liquid ion exchanger is:
NH
4 +：20～50μm；
NO
3 -：40～60μm。
Described step 1) the filling liquid in is:
NH
4 +：80～140mM?NH
4Cl；
NO
3 -：80～140mM?CaCl
2。
Described step 2) correcting fluid in is:
NH
4 +: 0.05～0.15mM NH
4cl and 0.5～1.5mM NH
4cl;
NO
3 -: 0.05～0.15mM KNO
3with 0.5～1.5mM KNO
3;
The Nernst equation of proofreading and correct rear electrode calculates ideal value and is:
NH
4 +：56～60mV；
NO
3 -：56～60mV；
Described step 3) described assay buffer is:
NH
4 +：0.8～1.2mM?KCl、0.05～0.15mM?CaCl
2、0.05～0.15mMNH
4Cl。
NO
3 -：0.05～0.15mM?KNO
3、0.8～1.2mM?KCl、0.05～0.15mMCaCl
2。
The present invention also provides the application aspect efficiently utilizing at Nitrogen Nutrition of Paddy Rice Plant of the described method of utilizing microscopic behavior ion current technology for detection Nitrogen Nutrition of Paddy Rice Plant.
The sky of take is educated 131 rice seedlings as with reference to material: detect NH
4 +the absorption flow velocity absolute value of ion is greater than 18.44pmolcm
-2s
-1be evaluated as NH
4 +the efficient plant of ion utilization; Detect NO
3 -the absorption flow velocity absolute value of ion is greater than 96.41pmolcm
-2s
-1be evaluated as NO
3 -the efficient plant of ion utilization.
NH
4 +ions Absorption flow velocity absolute value is greater than 18.44pmolcm
-2s
-1and NO
3 -the absorption flow velocity absolute value of ion is greater than 96.41pmolcm
-2s
-1the material together possessed can be judged as nitrogen nutrition and utilize the heavy duty detergent rice varieties.
By the NH recorded
4+ion and NO
3-the absorption flow velocity of ion, can instruct the rational application of fertilizer, realizes that efficient utilization of nitrogen becomes possibility.
The present invention utilizes microscopic dynamic ion flow detecting technology can record Nitrogen Nutrition of Paddy Rice Plant and efficiently utilizes ion current information, by different rice varieties relatively, obtaining different Nitrogen Nutrition of Paddy Rice Plants efficiently utilizes ion current to absorb or release rule, utilizing this rule to estimate Nitrogen Nutrition of Paddy Rice Plant efficiently utilizes, it is quick that thereby realization utilizes Nitrogen Nutrition of Paddy Rice Plant, Non-Destructive Testing, plant material after detection can also normal growth, avoided the loss of precious rice seedlings, the testing result contrast obviously, method is reliable, for seedling cultivation of rice, rice breeding, it is a kind of quick that rice cropping provides, the new method of harmless detection nitrogen nutrition.
In addition, utilize microscopic dynamic ion flow detecting technology direct-detection Nitrogen Nutrition of Paddy Rice Plant to utilize directly related NH
4+and NO
3-avoid the deficiency of the whole bag of tricks of indirect detection, directly, from detecting the difference of utilizing of corresponding Ions Absorption speed judgement Nitrogen Nutrition of Paddy Rice Plant, differentiated the different difference individuality of Nitrogen Nutrition of Paddy Rice Plant utilization ratio.
The accompanying drawing explanation
Fig. 1 is the experimentation figure that the present invention utilizes the method for microscopic behavior ion current technology for detection Nitrogen Nutrition of Paddy Rice Plant.
Fig. 2 is that the sky in the embodiment of the present invention 1 educates 131, sky educates 163, the dynamic absorption analysis figure of round-grained rice 8NH4+ ion (real-time flow rate) pacifies;
Fig. 3 is that the sky in the embodiment of the present invention 1 educates 131, sky educates 163, the round-grained rice 8NH4+ average ion flow velocity analysis chart (mean flow rate) of pacifying;
Fig. 4 is that the sky in the embodiment of the present invention 1 educates 131, sky educates 163, the dynamic absorption analysis figure of round-grained rice 8NO3-ion (real-time flow rate) pacifies;
Fig. 5 is that the sky in the embodiment of the present invention 1 educates 131, sky educates 163, the round-grained rice 8NO3-average ion flow velocity analysis chart (mean flow rate) of pacifying;
Wherein, in Fig. 2,4, ordinate is the ion flow velocity, and unit is pmol.cm
-2.s
-1, horizontal ordinate is the time, unit is second (s); In Fig. 3,5, ordinate is the ion flow velocity, and unit is pmol.cm
-2.s
-1.
Embodiment
Following examples are used for the present invention is described, but are not used for limiting the scope of the invention.If do not specialize, the conventional means that in embodiment, technological means used is well known to those skilled in the art, the raw materials used commercial goods that is.The rice varieties sky is educated 131 (Heilongjiang Province rice research institutes of land-reclaimable academy of sciences), sky is educated 163 (Heilongjiang Province rice research institutes of land-reclaimable academy of sciences), the round-grained rice 8 (Heilongjiang Academy of Agricultural Sciences's Suihua Institute of agricultural sciences) of pacifying.
Embodiment 1
1. experiment material and condition of culture: for the rice varieties sky is educated 131 (Heilongjiang Province rice research institutes of land-reclaimable academy of sciences), sky is educated 163 (Heilongjiang Province rice research institutes of land-reclaimable academy of sciences), the round-grained rice 8 (Heilongjiang Academy of Agricultural Sciences's Suihua Institute of agricultural sciences) of pacifying, seed is through 10% sodium hypochlorite sterilization 30min, after distilled water flushing, be placed on the double dish of the moistening filter paper of heelpiece, in 25 ℃ of illumination boxs, sprout, after germination, add the Hogland nutrient solution to cultivate, grow to two leaves during one heart stage, test the ion current of its root system.
2. experimental apparatus and consumptive material:
The microscopic behavior ion current detects with non-damage micrometering system (BIO-001B, YoungerUSA Sci.& Tech.Corp., USA), system software imFlux, the glass microelectrode tip diameter is 3 μ m.
3. experiment reagent and solution:
(1) liquid ion exchanger comprises:
NH
4 +：Ammonium?ionophore?I-cocktail?A；Sigma-Aldrich，Louis，MO?63103，USA.
NO
3 -：Xuyue(Beijing)Sci.&Tech.Co.，Ltd.Beijing，100080，P.R.China.
(2) assay buffer composition:
NH
4 +ion: 0.1mM NH
4cl, 1mM KCl, 0.1mM CaCl
2
NO
3 -ion: 0.1mM KNO
3, 1mM KCl, 0.1mM CaCl
2
(3) electrode filling liquid:
NH
4 +：100mM?NH
4Cl
NO
3 -：10mM?KNO
3
(4) correcting fluid:
NH
4 +ion: 0.05mM and 0.5mM NH
4cl
NO
3 -ion: 0.05mM and 0.5mM KNO
3
4. experiment content and method:
Experimentation is as shown in Figure of description 1.
Filling liquid to the eletrode tip that pours into the corresponding test ion of 1cm from electrode end is full of, and front end sucks suitable length (NH
4 +: 30 μ m; NO
3 -: 50 μ m) ion exchanger (LIX).Electrode is inserted in to the Ag/AgCl electrode wires pedestal of chlorination.Contrast electrode is the low seepage electrode (WPI) of solid.Electrode is proofreaied and correct in corresponding correcting fluid: NH
4 +58mV, NO
3 --58mV.
Test is front by material balance 35min in corresponding test fluid.Then in the local vibration detection of the surperficial 20 μ m of distance paddy rice root tip meristematic zone (apart from the tip of a root 400 μ m), each sample detection 15min.
5. data are processed
Ion current is processed based on Fick ' s diffusion law, and calculates by online software MageFlux-3DIon Flux Plotting System (http://www.xuyue.net/mageflux/).Do statistical study by being greater than 3 resulting data of independent experiment, utilize Excel2003 to analyze mapping.
6 experimental evaluation results:
As shown in Figures 2 and 3, three kinds of paddy rice skies educate 131, sky educates 163, round-grained rice 8 seedling of pacifying absorb NH
4 +ion amplitude difference, be respectively-24.88pmolcm of its Ions Absorption mean flow rate
-2s
-1,-33.58pmolcm
-2s
-1,-39.74pmolcm
-2s
-1.
As shown in Figure 4 and Figure 5, three kinds of paddy rice skies educate 131, sky educates 163, round-grained rice 8 seedling of pacifying are to NO
3 -absorptive amplitude is very large, be respectively-102.48pmolcm of Ions Absorption mean flow rate
-2s
-1,-261.81pmolcm
-2s
-1,-253.38pmolcm
-2s
-1.Embodiment 2
1. experiment material and condition of culture are with embodiment 1.
2. experimental apparatus and consumptive material
The microscopic behavior ion current detects with non-damage micrometering system (BIO-001B, YoungerUSA Sci.& Tech.Corp., USA), system software imFlux, the glass microelectrode tip diameter is 2 μ m.
3. experiment reagent and solution:
(1) liquid ion exchanger comprises:
NH
4 +：Ammonium?ionophore?I-cocktail?A；Sigma-Aldrich，Louis，MO?63103，USA.
NO
3 -：Xuyue(Beijing)Sci.&Tech.Co.，Ltd.Beijing，100080，P.R.China.
(2) assay buffer composition:
NH
4 +ion: 0.05mM NH
4cl, 0.8mM KCl, 0.05mM CaCl
2
NO
3 -ion: 0.05mM KNO
3, 0.8mM KCl, 0.05mM CaCl
2
(3) electrode filling liquid:
NH
4 +：100mM?NH
4Cl
NO
3 -：10mM?KNO
3
(4) correcting fluid:
NH
4 +ion: 0.15mM and 1.5mMNH
4cl
NO
3 -ion: 0.15mM and 1.5mM KNO
3
4. experiment content and method:
Experimentation is as shown in Figure of description 1.
Filling liquid to the eletrode tip that pours into the corresponding test ion of 1.5cm from electrode end is full of, and front end sucks suitable length (NH
4 +: 20 μ m; NO
3 -: 40 μ m) ion exchanger (LIX).Electrode is inserted in to the Ag/AgCl electrode wires pedestal of chlorination.Contrast electrode is the low seepage electrode (WPI) of solid.Electrode is proofreaied and correct in corresponding correcting fluid: NH
4 +: 56mV, NO
3 -: 56mV.Test is front by material balance 30min in corresponding test fluid.Then in the local vibration detection of the surperficial 10 μ m of distance paddy rice root tip meristematic zone (apart from the tip of a root 100 μ m), each sample detection 10min.
5. data are processed
Ion current is processed based on Fick ' s diffusion law, and calculates by online software MageFlux-3DIon Flux Plotting System (http://www.xuyue.net/mageflux/).Do statistical study by being greater than 3 resulting data of independent experiment, utilize Excel2003 to analyze data.
6 experimental evaluation results:
Three kinds of paddy rice skies educate 131, sky educates 163, round-grained rice 8 seedling of pacifying absorb NH
4 +ion amplitude difference, be respectively-20.00pmolcm of its Ions Absorption mean flow rate
-2s
-1,-31.01pmolcm
-2s
-1,-35.22pmolcm
-2s
-1.
Three kinds of paddy rice skies educate 131, sky educates 163, round-grained rice 8 seedling of pacifying are to NO
3 -absorptive amplitude is very large, be respectively-100.03pmolcm of Ions Absorption mean flow rate
-2s
-1,-260.55pmolcm
-2s
-1,-251.74pmolcm
-2s
-1.
Embodiment 3
1. experiment material and condition of culture are with embodiment 1.
2. experimental apparatus and consumptive material:
The microscopic behavior ion current detects with non-damage micrometering system (BIO-001B, YoungerUSA Sci.& Tech.Corp., USA), system software imFlux, the glass microelectrode tip diameter is 4 μ m.
3. experiment reagent and solution:
(1) liquid ion exchanger comprises:
NH
4 +：Ammonium?ionophore?I-cocktail?A；Sigma-Aldrich，Louis，MO?63103，USA
NO
3 -：Xuyue(Beijing)Sci.&Tech.Co.，Ltd.Beijing，100080，P.R.China.
(2) assay buffer composition:
NH
4 +ion: 0.15mM NH
4cl, 1.2mM KCl, 0.15mM CaCl
2
NO
3 -ion: 0.15mM KNO
3, 1.2mM KCl, 0.15mM CaCl
2
(3) electrode filling liquid:
NH
4 +：100mM?NH
4Cl
NO
3 -：10mM?KNO
3
(4) correcting fluid:
NH
4 +ion: 0.15mM and 1.5mMNH
4cl
NO
3 -ion: 0.15mM and 1.5mM KNO
3
4. experiment content and method:
Experimentation is as shown in Figure of description 1.
Filling liquid to the eletrode tip that pours into the corresponding test ion of 1cm from electrode end is full of, and front end sucks suitable length (NH
4 +: 50 μ m; NO
3 -: 60 μ m) ion exchanger (LIX).Electrode is inserted in to the Ag/AgCl electrode wires pedestal of chlorination.Contrast electrode is the low seepage electrode (WPI) of solid.Electrode is proofreaied and correct in corresponding correcting fluid: NH
4 +: 60mV, NO
3 -: 60mV.
Test is front by material balance 40min in corresponding test fluid.Then in the local vibration detection of the surperficial 20 μ m of distance paddy rice root tip meristematic zone (apart from the tip of a root 400 μ m), each sample detection 40min.
5. data are processed
Ion current is processed based on Fick ' s diffusion law, and calculates by online software MageFlux-3DIon Flux Plotting System (http://www.xuyue.net/mageflux/).Do statistical study by being greater than 3 resulting data of independent experiment, utilize Excel2003 to analyze data.
6. experimental evaluation result:
Three kinds of paddy rice skies educate 131, sky educates 163, round-grained rice 8 seedling of pacifying absorb NH
4 +ion amplitude difference, be respectively-18.44pmolcm of its Ions Absorption mean flow rate
-2s
-1,-28.15pmolcm
- 2s
-1,-36.65pmolcm
-2s
-1.
Three kinds of paddy rice skies educate 131, sky educates 163, round-grained rice 8 seedling of pacifying are to NO
3 -absorptive amplitude is very large, be respectively-96.41pmolcm of Ions Absorption mean flow rate
-2s
-1,-251.76pmolcm
-2s
-1,-241.35pmolcm
-2s
-1.
Therefore, the present invention be take sky and is educated 131 rice seedlings as with reference to material: detect NH
4 +the absorption flow velocity absolute value of ion is greater than 18.44pmolcm
-2s
-1be evaluated as NH
4 +the efficient plant of ion utilization; Detect NO
3 -the absorption flow velocity absolute value of ion is greater than 96.41pmolcm
-2s
-1be evaluated as NO
3 -the efficient plant of ion utilization; And NH
4 +ions Absorption flow velocity absolute value is greater than 18.44pmolcm
-2s
-1and NO
3 -the absorption flow velocity absolute value of ion is greater than 96.41pmolcm
-2s
-1the material together possessed can be judged as nitrogen nutrition and utilize the heavy duty detergent rice varieties.Sky in the present embodiment educate 163 and peaceful round-grained rice 8 be nitrogen nutrition and efficiently utilize the type rice varieties.
As can be seen from the above-described embodiment, three kinds of paddy rice skies educate 131, sky educates 163, the round-grained rice 8 of pacifying absorbs NO
3 -the ability of ion and NH
4 +the ability of ion all is not quite similar, and the utilization ratio difference of the paddy rice of different cultivars to nitrogenous fertilizer is described.Utilize the rice varieties sky of reference to educate 131 ion current testing result as standard, judge that other kinds are to NH
4 +, NO
3 -the difference of the receptivity of ion is differentiated the nitrogen nutrition utilization ratio of different rice varieties.This method can be used as the efficient evaluation means utilized of Nitrogen Nutrition of Paddy Rice Plant, rationally accurately applies a kind of technological means fast and effectively of different shape nitrogenous fertilizer, also for the Nitrogen Nutrition of Paddy Rice Plant high-efficient breeding, provides harmless, screening technique fast simultaneously.
Although above the present invention is described in detail with a general description of the specific embodiments, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements, all belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (7)
1. a method of utilizing microscopic dynamic ion flow detecting technology to detect Nitrogen Nutrition of Paddy Rice Plant, adopt the BIO-001B system to detect, and it is characterized in that, it comprises the following steps:
1) pour into ion filling liquid to being full of described eletrode tip to microelectrode, then the electrode front end is sucked to corresponding test ion exchanger;
2) electrode after step 1) is processed is inserted in to the Ag/AgCl electrode wires pedestal of chlorination, and puts into correcting fluid and proofread and correct;
3) get rice seedlings to be measured, its root first is placed on to balance in assay buffer, then detected with the electrode pair rice seedlings to be measured after proofreading and correct;
4) testing result is processed and analyzed;
Wherein, described step 1) is for to pour into ion filling liquid to being full of described eletrode tip 1～1.5cm to microelectrode, then the electrode front end is sucked to corresponding test ion exchanger;
Described step 3), for getting rice seedlings to be measured, first is placed on balance 30-40min in assay buffer by its root, then is detected 10～15min with the electrode pair rice seedlings to be measured after proofreading and correct;
Described assay buffer is:
NH
4 +：0.8～1.2mM?KCl、0.05～0.15mM?CaCl
2、0.05～0.15mMNH
4Cl；
NO
3 －：0.05～0.15mM?KNO
3、0.8～1.2mM?KCl、0.05～0.15mMCaCl
2。
2. according to the method for utilizing microscopic behavior ion current technology for detection Nitrogen Nutrition of Paddy Rice Plant described in claim 1, it is characterized in that, described step 1) is for to pour into ion filling liquid to being full of described eletrode tip 1cm to microelectrode, then the electrode front end is sucked to corresponding test ion exchanger; Described step 3), for getting rice seedlings to be measured, first is placed on balance 30-40min in assay buffer by its root, then is detected 15min with the electrode pair rice seedlings to be measured after proofreading and correct.
3. method according to claim 1 and 2, is characterized in that, detects the paddy rice of use in Seedling Stage.
4. method according to claim 1 and 2, is characterized in that, the paddy rice of detect using is in 21 heart stages of leaf, outside surface 10～45 μ m places of the Meristernatic zone that measuring position be the described rice seedlings tip of a root 100～400 μ m of distance.
5. method according to claim 1 and 2, is characterized in that, in described step 1), the length of the front end of electrode suction liquid ion exchanger is:
NH
4 +：20～50μm；
NO
3 －：40～60μm。
6. method according to claim 1 and 2, is characterized in that, the filling liquid in described step 1) is:
NH
4 +：100mM?NH
4Cl；
NO
3 －：10mMKNO
3；
Described step 2) correcting fluid in is:
NH
4 +: 0.05～0.15mM NH
4cl and 0.5～1.5mM NH
4cl;
NO
3 -: 0.05～0.15mM KNO
3with 0.5～1.5mM KNO
3;
The Nernst equation of proofreading and correct rear electrode calculates ideal value and is:
NH
4 +：56～60mV;
NO
3 －：56～60mV。
7. the application aspect efficiently utilizing at Nitrogen Nutrition of Paddy Rice Plant of the method for utilizing microscopic behavior ion current technology for detection Nitrogen Nutrition of Paddy Rice Plant described in claim 1-6.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN2045494U (en) * | 1988-12-10 | 1989-10-04 | 陆文林 | Multifunction soil analysis apparatus |
| CN1186237A (en) * | 1996-12-26 | 1998-07-01 | 中国科学院长春应用化学研究所 | Preparation of metal ion sensor |
| CN101813662A (en) * | 2009-02-19 | 2010-08-25 | 清华大学 | Method for detecting ammonia-nitrogen content in water, and special device thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2045494U (en) * | 1988-12-10 | 1989-10-04 | 陆文林 | Multifunction soil analysis apparatus |
| CN1186237A (en) * | 1996-12-26 | 1998-07-01 | 中国科学院长春应用化学研究所 | Preparation of metal ion sensor |
| CN101813662A (en) * | 2009-02-19 | 2010-08-25 | 清华大学 | Method for detecting ammonia-nitrogen content in water, and special device thereof |
Non-Patent Citations (2)
| Title |
|---|
| 几种黑杨无性系苗期氮素吸收的特点;田东梅;《中国优秀硕士学位论文全文数据库 农业科技辑》;20111015(第10期);3-6,13,14 * |
| 田东梅.几种黑杨无性系苗期氮素吸收的特点.《中国优秀硕士学位论文全文数据库 农业科技辑》.2011,(第10期),3-6,13,14. |
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