CN102539507A - 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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- CN102539507A CN102539507A CN201110427525XA CN201110427525A CN102539507A CN 102539507 A CN102539507 A CN 102539507A CN 201110427525X A CN201110427525X A CN 201110427525XA CN 201110427525 A CN201110427525 A CN 201110427525A CN 102539507 A CN102539507 A CN 102539507A
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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 a kind of method of utilizing the nitrogen nutrition of microscopic behavior ion flow technology for detection paddy rice and this method in the application of paddy rice nitrogen nutrition aspect efficiently utilizing.
Background technology
Nitrogen is the important composition composition of cellular plasm; Be all teleorganic basic nutrition elements; It is the nutrient that plant is primary, have the call; In processes such as plant structure, energetic supersession and quality formation, bringing into play the effect that can not replace, it has crucial influence to nitrogen-containing compound in plant growth and the body.An amount of 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 outstanding problems such as utilization rate of nitrogen fertilizer (NUE) reduction, severe environmental pollution.
Paddy rice is Chinese topmost cereal crops, has critical role aspect agricultural production and the grain security.And nitrogen is to influence rice yield and the of paramount importance nutrient of quality.Paddy rice directly affects its g and D situation to absorption, assimilation and the transhipment of nitrogen, thereby influences yield and quality.Further investigation Nitrogen Absorption dynamic rule, the adjusting and controlling rice growth and development process that helps to adopt an effective measure improves output and improves quality, can practice thrift fertilizer simultaneously, improve the soil nutrient structure.Microscopic behavior ion flow detection technique as harmless, in real time, dynamically, detection method is to differentiate the direct tool of paddy rice nitrogen nutrition 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 the breeding demand efficiently to the paddy rice nitrogen nutrition, 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 nitrogen form that higher plant absorbs, and plant absorb the selectivity of these two kinds of form nitrogen ions and receive soil physico-chemical property factor affecting such as plant kind, kind, growth and development stage and pH value.Find during research black walnut seedling such as MichaelA, apply the NH of proper ratio
4 +/ NO
3 -Nitrogenous fertilizer can promote (the Michael A. that grows of black walnut better; 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 that nitrate reductase activity and nitrogen-containing compound embody in wooden exudate of black walnut and supply relation) therefore keeps the NH that suits 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 with sky educate 131, sky educates 163, the round-grained rice 8 of pacifying is that experiment material research different cultivars rice root is to NH
4 +, NO
3 -The absorbing rule of ion is efficiently utilized the foundation of estimating screening as the different cultivars rice nutrition.
Microscopic behavior ion flow detection technique is the electrophysiological technique of a comprehensive specific ion/Molecular Detection, and core technology is to utilize the ion selectivity micro-electrode under the situation that does not contact sample, to detect concentration (mM level), the flow velocity (10 of the ion of turnover sample under the control automatically at computer
-12Molcm
-2S
-1) and information such as direction of motion.This technical operation is convenient, fast, real-time, harmless, can truly reflect the vital movement rule that other technologies are difficult to reflect.This technology is that life science provides a kind of visual technological means, and promoting that the mankind find out conventional art can't detected information, has solved many problem in science.
In order in time to grasp the plant growth situation, except that carrying out according to experience outward appearance observes, often with the nitrogen nutrition of aerial part as the foundation of judging the crop nitrogen nutrition situation.Based on traditional nitrogen nutrition detection method complicated operation of chemical composition analytic approach, analytical cycle is long, and poor in timeliness can't accomplish that real-time, the live body during the plant growth detects.
Chlorophyll meter (SPAD) method (Li Gang China, Xue Lihong, You Juan, etc. [J] selected in the leaf position of paddy rice nitrogen and chlorophyll SPAD leaf position characteristic distributions 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 measuring crop chlorophyll level and quick diagnosis crop nitrogen level.But the information faces that chlorophyll meter is gathered is less than normal, and workload is bigger, to measuring standard higher requirement is arranged, and the difficult grasp of diagnostic criteria.The SPAD value can receive the influence 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 that information is obtained fast, an advantage such as economy, operability are good, non-destructive; 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 influencing is a lot; And the canopy color parameter there are differences between crop varieties; Should reduce error as far as possible, make spectral technique more accurate, be applied to the nitrogen diagnosis of crop better as the nitrogen diagnosis method.
Methods such as chemical analysis method commonly used, chlorophyll meter (SPAD) method, spectral technique when as stated, in the past analyzing the crop nitrogen nutrition utilization ratio.As: based on traditional nitrogen nutrition detection method complicated operation of chemical composition analytic approach, analytical cycle is long, and poor in timeliness can't accomplish that real-time, the live body during the plant growth detects; And spectral technique is ripe not enough, and the factor of light absorption and reflection of influencing is a lot, and the canopy color parameter there are differences between crop varieties, and error is bigger.Though above-mentioned these methods have been improved blindness and the excessive problem of nitrogenous fertilizer that the experience fertilising brings to a certain extent, and guide fertilising but still can not carry out the nitrogen nutrition morphological diagnosis easy, quick, real-time, harmless, exactly.Therefore, the method for the new detection paddy rice fungoid damping-off of exploitation has very important significance.
Summary of the invention
The present invention be intended to for seedling cultivation of rice, rice breeding, rice cropping provide a kind of fast, the method for harmless detection paddy rice nitrogen nutrition, efficiently utilize the aspect to be applied to the paddy rice nitrogen nutrition.
Particularly, the present invention provides a kind of method of utilizing microscopic behavior ion flow detection technique to detect the paddy rice nitrogen nutrition, adopts the BIO-001B system to detect.
Said method comprising the steps of:
1) pours into ion filling liquid to being full of said eletrode tip to microelectrode, again the electrode front end is sucked corresponding test ion exchanger;
The electrode that 2) will pass through after step 1) is handled is inserted in the Ag/AgCl electrode wires pedestal of chlorination, and puts into correcting fluid and proofread and correct;
3) get paddy rice seedling to be measured, its root is placed on balance in the assay buffer earlier, detect with the electrode pair paddy rice seedling to be measured after proofreading and correct again;
4) testing result is handled and analyzed.
Further, said method comprising the steps of:
1) pours into ion filling liquid to being full of said eletrode tip 1～1.5cm to microelectrode, again the electrode front end is sucked corresponding test ion exchanger;
The electrode that 2) will pass through after step (1) is handled is inserted in the Ag/AgCl electrode wires pedestal of chlorination, and puts into correcting fluid and proofread and correct;
3) get paddy rice seedling to be measured, its root is placed on balance 30-40min in the assay buffer earlier, detect 10～15min with the electrode pair paddy rice seedling to be measured after proofreading and correct again;
4) testing result is handled and analyzed.
Further, said method comprising the steps of:
1) pours into ion filling liquid to being full of said eletrode tip 1cm to microelectrode, again the electrode front end is sucked corresponding test ion exchanger;
The electrode that 2) will pass through after step (1) is handled is inserted in the Ag/AgCl electrode wires pedestal of chlorination, and puts into correcting fluid and proofread and correct;
3) get paddy rice seedling to be measured, its root is placed on balance 30-40min in the assay buffer earlier, detect 15min with the electrode pair paddy rice seedling to be measured after proofreading and correct again;
4) testing result is handled and analyzed.
In said method, detect the paddy rice of using and be in the seedling phase.
Test ion concentration range in the said microscopic behavior ion flow detection technique in the assay buffer is 0.05～0.15mM.
The paddy rice of detect using is in 1 heart stage of 2 leaves, and the measuring position be outside surface 10～45 μ m places apart from the tip of a root meristematic zone of sharp 100～400 μ m of said paddy rice shoot root.
The length of the front end of electrode suction liquid ion exchanger is in the said step 1):
NH
4 +：20～50μm；
NO
3 -：40～60μm。
Filling liquid in the said step 1) is:
NH
4 +：80～140mM?NH
4Cl；
NO
3 -：80～140mM?CaCl
2。
Said 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；
The described assay buffer of said step 3) 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 said method of utilizing the nitrogen nutrition of microscopic behavior ion flow technology for detection paddy rice in the application of paddy rice nitrogen nutrition aspect efficiently utilizing.
Educating 131 rice seedlings with sky is with reference to material: detect NH
4 +The absorption flow velocity absolute value of ion is greater than 18.44pmolcm
-2S
-1Promptly be 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
-1Promptly be evaluated as NO
3 -The efficient plant of ion utilization.
NH
4 +Ionic 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 that together possesses can be judged as nitrogen nutrition and utilize the heavy duty detergent rice varieties.
Through the NH that records
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 behavior ion flow detection technique can record the paddy rice nitrogen nutrition and efficiently utilizes ion flow information; Through the different water rice varieties relatively, obtain different paddy rice nitrogen nutrition and efficiently utilize ion flow to absorb or release rule, utilize this rule to estimate the paddy rice nitrogen nutrition and efficiently utilize; Thereby realize quick, Non-Destructive Testing to paddy rice nitrogen nutrition utilization; Plant material after the detection can also normal growth, has avoided the loss of precious paddy rice seedling, and the testing result contrast obviously; Method is reliable, for seedling cultivation of rice, rice breeding, rice cropping provide a kind of fast, the new method of harmless detection nitrogen nutrition.
In addition, utilize microscopic behavior ion flow detection technique directly to detect the paddy rice nitrogen nutrition and utilize directly related NH
4+And NO
3-Avoided the deficiency of the whole bag of tricks of indirect detection, directly judged the difference of utilizing of paddy rice nitrogen nutrition, differentiated that the different difference of paddy rice nitrogen nutrition utilization ratio is individual from detecting corresponding ionic absorption speed.
Description of drawings
Fig. 1 utilizes the experimentation figure of the method for microscopic behavior ion flow technology for detection paddy rice nitrogen nutrition for the present invention.
Fig. 2 is for the sky in the embodiment of the 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 for the sky in the embodiment of the 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 for the sky in the embodiment of the 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 for the sky in the embodiment of the invention 1 educates 131, sky educates 163, the round-grained rice 8NO3-average ion flow velocity analysis chart (mean flow rate) of pacifying;
Wherein, ordinate is the ion flow velocity among Fig. 2,4, and unit is pmol.cm
-2.s
-1, horizontal ordinate is the time, unit is second (s); Ordinate is the ion flow velocity among Fig. 3,5, and unit is pmol.cm
-2.s
-1
Embodiment
Following examples are used to explain the present invention, but are not used for limiting scope of the present invention.If do not specialize the conventional means that used technological means is well known to those skilled in the art among the embodiment, the raw materials used commercial goods that is.The rice varieties sky is educated 131 (Heilongjiang Province paddy rice research institutes of land-reclaimable academy of sciences), sky is educated 163 (Heilongjiang Province paddy rice research institutes of land-reclaimable academy of sciences), the round-grained rice 8 (academy of agricultural sciences, Heilongjiang Province Suihua research of agricultural science institute) of pacifying.
Embodiment 1
1. experiment material and condition of culture: for the rice varieties sky is educated 131 (Heilongjiang Province paddy rice research institutes of land-reclaimable academy of sciences), sky is educated 163 (Heilongjiang Province paddy rice research institutes of land-reclaimable academy of sciences), the round-grained rice 8 (academy of agricultural sciences, Heilongjiang Province Suihua research of agricultural science institute) of pacifying; Seed is through 10% sodium hypochlorite sterilization 30min; Behind the distilled water flushing; Place on the double dish of the moistening filter paper of heelpiece, sprout in 25 ℃ of illumination boxs, the back of germinateing adds the Hogland nutrient solution and cultivates; Grow to two leaves during one heart stage, test the ion flow of its root system.
2. experimental apparatus and consumptive material:
The microscopic behavior ion flow detect 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 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 -: ion exchanger (LIX) 50 μ m).Electrode is inserted in the Ag/AgCl electrode wires pedestal of chlorination.Contrast electrode is the low seepage property electrode (WPI) of solid.Electrode is proofreaied and correct in corresponding correcting fluid: NH
4 +58mV, NO
3 --58mV.
Test is preceding with material balance 35min in corresponding test fluid.Then at the local vibration detection apart from the surperficial 20 μ m of paddy rice root tip meristematic zone (apart from the tip of a root 400 μ m), each sample detection 15min.
5. data processing
Ion flow is handled based on Fick ' s diffusion law, and calculates through online software MageFlux-3DIon Flux Plotting System (http://www.xuyue.net/mageflux/).By doing statistical study, utilize Excel2003 to analyze mapping greater than 3 resulting data of independent experiment.
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 absorption of N H pacify
4 +The ion amplitude is different, and its ionic absorption mean flow rate is respectively-24.88pmolcm
-2S
-1,-33.58pmolcm
-2S
-1,-39.74pmolcm
-2S
-1
Like Fig. 4 and shown in 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 big, and the ionic absorption mean flow rate is respectively-102.48pmolcm
-2S
-1,-261.81pmolcm
-2S
-1,-253.38pmolcm
-2S
-1Embodiment 2
1. experiment material and condition of culture are with embodiment 1.
2. experimental apparatus and consumptive material
The microscopic behavior ion flow detect 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 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 -: ion exchanger (LIX) 40 μ m).Electrode is inserted in the Ag/AgCl electrode wires pedestal of chlorination.Contrast electrode is the low seepage property electrode (WPI) of solid.Electrode is proofreaied and correct in corresponding correcting fluid: NH
4 +: 56mV, NO
3 -: 56mV.Test is preceding with material balance 30min in corresponding test fluid.Then at the local vibration detection apart from the surperficial 10 μ m of paddy rice root tip meristematic zone (apart from the tip of a root 100 μ m), each sample detection 10min.
5. data processing
Ion flow is handled based on Fick ' s diffusion law, and calculates through online software MageFlux-3DIon Flux Plotting System (http://www.xuyue.net/mageflux/).By doing statistical study, utilize Excel2003 to analyze data greater than 3 resulting data of independent experiment.
6 experimental evaluation results:
Three kinds of paddy rice skies educate 131, sky educates 163, round-grained rice 8 seedling absorption of N H pacify
4 +The ion amplitude is different, and its ionic absorption mean flow rate is respectively-20.00pmolcm
-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 big, and the ionic absorption mean flow rate is respectively-100.03pmolcm
-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 flow detect 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 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 -: ion exchanger (LIX) 60 μ m).Electrode is inserted in the Ag/AgCl electrode wires pedestal of chlorination.Contrast electrode is the low seepage property electrode (WPI) of solid.Electrode is proofreaied and correct in corresponding correcting fluid: NH
4 +: 60mV, NO
3 -: 60mV.
Test is preceding with material balance 40min in corresponding test fluid.Then at the local vibration detection apart from the surperficial 20 μ m of paddy rice root tip meristematic zone (apart from the tip of a root 400 μ m), each sample detection 40min.
5. data processing
Ion flow is handled based on Fick ' s diffusion law, and calculates through online software MageFlux-3DIon Flux Plotting System (http://www.xuyue.net/mageflux/).By doing statistical study, utilize Excel2003 to analyze data greater than 3 resulting data of independent experiment.
6. experimental evaluation result:
Three kinds of paddy rice skies educate 131, sky educates 163, round-grained rice 8 seedling absorption of N H pacify
4 +The ion amplitude is different, and its ionic absorption mean flow rate is respectively-18.44pmolcm
-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 big, and the ionic absorption mean flow rate is respectively-96.41pmolcm
-2S
-1,-251.76pmolcm
-2S
-1,-241.35pmolcm
-2S
-1
Therefore, to educate 131 rice seedlings with sky be with reference to material in the present invention: detect NH
4 +The absorption flow velocity absolute value of ion is greater than 18.44pmolcm
-2S
-1Promptly be 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
-1Promptly be evaluated as NO
3 -The efficient plant of ion utilization; And NH
4 +Ionic 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 that together possesses can be judged as nitrogen nutrition and utilize the heavy duty detergent rice varieties.Sky in the present embodiment is educated 163 and is nitrogen nutrition and efficiently utilizes the type rice varieties with peaceful round-grained rice 8.
Can find out that by the foregoing description three kinds of paddy rice skies educate 131, sky educates 163, round-grained rice 8 absorption of N O pacify
3 -The ability of ion and NH
4 +The ability of ion all is not quite similar, and explains that the paddy rice of different cultivars is different to the utilization ratio of nitrogenous fertilizer.Utilize the rice varieties sky of reference to educate 131 ion flow testing result, judge that other kinds are to NH as standard
4 +, NO
3 -The difference of the receptivity of ion is differentiated the nitrogen nutrition utilization ratio of different water rice varieties.This method can be used as the efficient evaluation means that utilizes of paddy rice nitrogen nutrition, rationally accurately applies a kind of technological means fast and effectively of different shape nitrogenous fertilizer, also for paddy rice nitrogen nutrition high-efficient breeding harmless, screening technique fast is provided simultaneously.
Though, the present invention has been done detailed description in the preceding text with general explanation and specific embodiments, on basis of the present invention, can to some modifications of do or improvement, 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 (10)
1. a method of utilizing microscopic behavior ion flow detection technique to detect the paddy rice nitrogen nutrition adopts the BIO-001B system to detect, and it is characterized in that it may further comprise the steps:
1) pours into ion filling liquid to being full of said eletrode tip to microelectrode, again the electrode front end is sucked corresponding test ion exchanger;
The electrode that 2) will pass through after step 1) is handled is inserted in the Ag/AgCl electrode wires pedestal of chlorination, and puts into correcting fluid and proofread and correct;
3) get paddy rice seedling to be measured, its root is placed on balance in the assay buffer earlier, detect with the electrode pair paddy rice seedling to be measured after proofreading and correct again;
4) testing result is handled and analyzed.
2. according to the method for utilizing the nitrogen nutrition of microscopic behavior ion flow technology for detection paddy rice described in the claim 1; It is characterized in that; Said step 1) sucks corresponding test ion exchanger with the electrode front end again for to pour into ion filling liquid to being full of said eletrode tip 1～1.5cm to microelectrode; Said step 3) is placed on balance 30-40min in the assay buffer for getting paddy rice seedling to be measured earlier with its root, detects 10～15min with the electrode pair paddy rice seedling to be measured after proofreading and correct again.
3. according to the method for utilizing the nitrogen nutrition of microscopic behavior ion flow technology for detection paddy rice described in the claim 1; It is characterized in that; Said step 1) sucks corresponding test ion exchanger with the electrode front end again for to pour into ion filling liquid to being full of said eletrode tip 1cm to microelectrode; Said step 3) is placed on balance 30-40min in the assay buffer for getting paddy rice seedling to be measured earlier with its root, detects 15min with the electrode pair paddy rice seedling to be measured after proofreading and correct again.
4. according to each described method among the claim 1-3, it is characterized in that, detect the paddy rice of using and be in the seedling phase.
5. according to each described method among the claim 1-3, it is characterized in that the test ion concentration range in the said microscopic behavior ion flow detection technique in the assay buffer is 0.05～0.15mM.
6. according to each described method among the claim 1-3, it is characterized in that the paddy rice of detect using is in 1 heart stage of 2 leaves, the measuring position be outside surface 10～45 μ m places apart from the tip of a root meristematic zone of sharp 100～400 μ m of said paddy rice shoot root.
7. according to each described method among the claim 1-3, it is characterized in that the length of the front end of electrode suction liquid ion exchanger is in the said step 1):
NH
4 +：20～50μm；
NO
3 -：40～60μm。
8. according to each described method among the claim 1-3, it is characterized in that the filling liquid in the said step 1) is:
NH
4 +：100mM?NH
4Cl；
NO
3 -：10mMKNO
3；
Said 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。
9. according to each described method among the claim 1-3, it is characterized in that the described assay buffer of said step 3) 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。
10. the method for utilizing the nitrogen nutrition of microscopic behavior ion flow technology for detection paddy rice described in the claim 1-9 is in the application of paddy rice nitrogen nutrition aspect efficiently utilizing.
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