CN114303836A - Method for determining rice stress resistance regulation and control optimization strategy - Google Patents
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Abstract
The invention discloses a method for determining a rice stress-resistant regulation and control optimization strategy, which applies different nitrogen source application schemes to rice seedlings; constructing a matrix I for cultivating rice stress resistance regulation and control by different nitrogen sources in rice tissues, and processing the matrix I to obtain a matrix II-a matrix V; comparing the values in the matrix I respectively to obtain the contribution of the inorganic nitrogen and the organic nitrogen which are applied in combination to the free Pro; comparing the values in the matrix II, the matrix III and the matrix V to obtain the contribution of the inorganic nitrogen or the organic nitrogen which is independently applied to the free Pro; according to the method, a strategy model for regulating and controlling chromium stress of rice is obtained by screening different nitrogen sources by taking free Pro as a biomarker, and stress resistance regulation and control optimization strategies of the rice at different growth stages are determined. The method for determining the rice stress-resistance regulation and control optimal strategy simplifies the various experimental processes of botany by constructing the strategy model, and provides a scientific and effective strategy determination method for the field of rice stress-resistance regulation and control.
Description
Technical Field
The invention belongs to the technical field of botany, and relates to a determination method of a rice stress-resistance regulation and control optimization strategy.
Background
Chromium (Cr) is an important industrial raw material and is widely used in industries such as petroleum refining, leather tanning, textile manufacturing, electroplating, dye processing, and the like. According to statistics, the annual average consumption of Cr in the world is about 1250 ten million tons at present, and the accumulated Cr in the early stage is about 11054 hundred million tons, so that the Cr pollution in the environment is more and more serious. The national soil pollutant survey bulletin issued by the ministry of environmental protection in 2014, 4, 17 shows that the total exceeding rate of national soil pollution is 16.1%, wherein the exceeding rate of Cr point is 1.1%. Plants are producers in the food chain and also the most important component in the ecosystem, and regulation of plant growth and development by exogenous green regulators under pollution stress is a hot issue of great concern.
Nitrogen is a major element essential for plant growth, among which nitrate Nitrogen (NO)3 -) And ammonium Nitrogen (NH)4 +) Is the most easily absorbed inorganic nitrogen by plants. Proline (Pro) is an amino acid with multiple functions in plants, which not only relieves biotic and abiotic stress, but also plays an important role in maintaining amino acid balance in plants. Thus, Pro is also often used as a biomarker in response to the stress intensity of plants. Glutamic acid (Glu) and arginine (Arg) are precursors in 2 synthetic Pro pathways in plants, and NO3 -And NH4 +Assimilation in plants is also closely related to the amount of Glu and Arg stored.
The invention uses free Pro as biomarker to screen different nitrogen sources (inorganic nitrogen source: NO) under chromium stress3 -And NH4 +(ii) a Organic nitrogen source: glu and Arg) to culture rice, and provides selectivity for rice adapting to chromium pollution.
Disclosure of Invention
In order to achieve the aim, the invention provides a method for determining a rice stress-resistant regulation and control optimization strategy, which is characterized in that free proline is used as a biomarker to screen different nitrogen source combinations under chromium stress to culture the rice stress-resistant regulation and control optimization strategy, so that the toxicity of pollutants to plants is relieved, and the problems in the prior art are solved.
The technical scheme adopted by the invention is that the method for determining the rice stress-resistant regulation and control optimization strategy comprises the following steps:
step 1: applying different nitrogen source application schemes to the rice seedlings under the stress of various concentrations of Cr (III); the nitrogen source administration regimen comprises: any one or a combination of any two of organic nitrogen-free treatment, arginine treatment, glutamic acid treatment, inorganic nitrogen-free treatment, nitrate nitrogen treatment and ammonium nitrogen treatment; determining the content of free Pro in rice seedling tissues in each nitrogen source application scheme;
step 2: constructing a matrix I for cultivating rice stress resistance regulation and control by using any two combinations of organic nitrogen-free treatment, arginine treatment, glutamic acid treatment, inorganic nitrogen-free treatment, nitrate nitrogen treatment and ammonium nitrogen treatment, wherein the matrix I is formed by the following matrixes:
the rows and columns of the matrix are each denoted by riAnd cjRepresents, i, j ═ 1,2, 3; a specific element of the matrix is represented by aijRepresents, i, j ═ 1,2, 3;&represents the combined administration of two nitrogen sources; in matrix I, a11Indicates the free Pro content in the absence of organic nitrogen treatment and in the absence of inorganic nitrogen treatment; a is12Represents the free Pro content in arginine pretreatment and in the absence of inorganic nitrogen treatment; a is13Represents the free Pro content in the glutamic acid pretreatment and in the absence of inorganic nitrogen treatment; a is21Indicates the free Pro content in the case of treatment without organic nitrogen and treatment with nitrate nitrogen; a is22Represents the free Pro content when arginine pretreatment and nitrate nitrogen addition treatment are carried out; a is23Represents the free Pro content when the glutamic acid is pretreated and the nitrate nitrogen is added; a is31Indicates the free Pro content in the case of treatment without organic nitrogen and treatment with ammonium nitrogen; a is32Represents the free Pro content in arginine pretreatment and ammonium nitrogen addition treatment; a is33Represents the free Pro content when glutamic acid is pretreated and ammonium nitrogen is added;
and step 3: r is performed on the matrix I2-r1，r3-r1，r3-r2Performing three times of primary row transformation to obtain a matrix II; c is performed on the matrix I2-c1，c3-c1，c3-c2Performing three times of primary column transformation to obtain a matrix III; will be provided withSubtracting the matrix II from the matrix III to obtain a matrix IV; from a in the matrix IV11、a12、a21、a22Forming a matrix V;
and 4, step 4: comparing matrix I with a respectively22、a23、a32、a33The magnitude of the numerical value, resulting in the magnitude of the contribution of the combined administration of inorganic nitrogen and organic nitrogen to free Pro;
comparing matrix II third row elements a separately3jThird column element a of matrix IIIi3And matrix V in a11、a12、 a21、a22The amount of free Pro compared to the amount of contribution of inorganic nitrogen or organic nitrogen administered alone;
and 5: according to the method of the step 4, obtaining the contribution size of the nitrogen source to the free Pro by the single and combined application in the rice seedling tissues under different stress concentrations; according to the contribution of the nitrogen sources to free Pro by independent and combined application in rice seedling tissues under various stress concentrations, a strategy model for screening different nitrogen sources to culture rice to regulate chromium stress by taking the free Pro as a biomarker is obtained, and stress resistance regulation optimization strategies of the rice at different growth stages are determined according to the contribution model.
Further, in step 3, the matrix II is specifically as follows:
in matrix II, r1Represents the contribution of nitrate nitrogen treatment to free Pro; r is2Represents the contribution of ammonium nitrogen treatment to free Pro; r is3Represents the contribution of the difference in free Pro from the ammonium nitrogen treatment and the nitrate nitrogen treatment.
Further, in step 3, the matrix III is as follows:
in matrix III, c1Represents the contribution of arginine treatment to free Pro; c. C2Represents the contribution of glutamate treatment to free Pro; c. C3Represents the contribution representing the difference in free Pro from glutamate treatment and arginine treatment.
Further, in step 3, the matrix IV is as follows:
in matrix IV, a11Represents the contribution of arginine treatment to the difference in free Pro from nitrate nitrogen treatment; a is12Represents the contribution of glutamic acid treatment and nitrate nitrogen treatment to the difference of free Pro; a is13Represents the contribution of the difference of the glutamic acid treatment, the arginine treatment and the nitrate nitrogen treatment to the free Pro; a is21Represents the contribution of arginine treatment to the difference in free Pro from ammonium nitrogen treatment; a is22Represents the contribution of glutamic acid treatment and ammonium nitrogen treatment to the difference of free Pro; a is23Represents the contribution of the difference of free Pro by glutamic acid treatment, arginine treatment and ammonium nitrogen treatment; a is31Represents the contribution of arginine treatment, nitrate nitrogen treatment and ammonium nitrogen treatment to the difference of free Pro; a is32Represents the contribution of glutamic acid treatment, nitrate nitrogen treatment and ammonium nitrogen treatment to the difference of free Pro; a is33The contribution of the difference between glutamic acid treatment and arginine treatment, nitrate nitrogen treatment, and ammonium nitrogen treatment to the free Pro was shown.
Further, in step 3, the matrix V is as follows:
further, in step 4, the third row elements a of the matrix II are compared respectively3jThird column element a of matrix IIIi3And matrix V in a11、a12、a21、a22The amount of contribution of inorganic nitrogen or organic nitrogen to free Pro compared to the amount of contribution of inorganic nitrogen or organic nitrogen administered alone is obtained by:
comparing the application of ammonium nitrogen alone with the application of a single element according to the sign of the value of each element in the third row of matrix IIThe amount of contribution of nitrate nitrogen alone to free Pro; comparing the amount of contribution of glutamic acid alone to arginine alone to free Pro according to the positive and negative values of the elements in the third column of matrix III; according to a of the matrix V11Positive and negative values, comparing the contribution of arginine alone to free Pro versus nitrate nitrogen alone; according to a of the matrix V12The magnitude of the contribution to free Pro of glutamic acid alone versus nitrate nitrogen alone; according to a of the matrix V21Positive and negative values, comparing the amount of contribution of arginine alone to free Pro versus ammonium nitrogen alone; according to a of the matrix V22The magnitude of the contribution to free Pro compared to glutamic acid alone versus ammonium nitrogen alone;
the magnitude of the contribution of inorganic or organic nitrogen alone to free Pro is obtained from the magnitude of the contribution of ammonium nitrogen alone and nitrate nitrogen alone to free Pro, the magnitude of the contribution of glutamic acid alone and arginine alone to free Pro, the magnitude of the contribution of arginine alone and nitrate nitrogen alone to free Pro, the magnitude of the contribution of glutamic acid alone and nitrate nitrogen alone to free Pro, the magnitude of the contribution of arginine alone and ammonium nitrogen alone to free Pro, and the magnitude of the contribution of glutamic acid alone and ammonium nitrogen alone to free Pro.
The invention has the beneficial effects that: according to the embodiment of the invention, free proline is used as a biomarker, a single or combined regulator is simply and efficiently selected, the toxicity of pollutants to plants is greatly relieved, a matrix I for stress resistance regulation and control of different nitrogen sources cultivated rice in rice tissues can be constructed by only one test of nine treatments, the matrix I is subjected to matrix transformation and treatment such as primary row transformation, primary column transformation, matrix subtraction and the like, the contribution of each nitrogen source in each nitrogen source application scheme to stress resistance regulation and control of rice can be analyzed according to the size and the positive and negative of numerical values in the transformed matrix, a strategy model for screening different nitrogen sources cultivated rice to regulate and control chromium stress by using free Pro as the biomarker is further obtained, and stress resistance regulation and control optimization strategies of rice at different growth stages are determined according to the contribution model. According to the method for determining the rice stress-resistance regulation and control optimal strategy, disclosed by the embodiment of the invention, through the construction of the strategy model, the test process of a great variety of botany is simplified, the rice stress-resistance regulation and control efficiency is effectively improved, and a scientific and effective strategy determination method is provided for the field of rice stress-resistance regulation and control.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
1. Experimental protocol
1.1 preparation of Rice seedlings
The rice seeds are soaked in water for 12 hours and then put into a disposable cup for sand culture, and the planting density is about 30-40 seeds per cup. The cultivation is carried out in an artificial climate box with the temperature of 25 ℃, the humidity of 65 percent and the illumination intensity of 20000 lux. Pour modified ISO8692 nutrient solution (table 1). After the rice grows for 16 days, cleaning sandy soil at the roots of the rice, and selecting rice seedlings with the same individual size for later use.
Table 1 modified ISO8692 nutrient solution formula
| Serial number | Reagent | Concentration of | Serial number | Reagent | Concentration of |
| 1 | KNO3 | 2823.9μmol/L | 8 | H3BO3 | 2992.1nmol/L |
| 2 | MgCl2·6H2O | 59.0μmol/L | 9 | MnCl2·4H2O | 2097.0nmol/L |
| 3 | CaCl2·2H2O | 122.4μmol/L | 10 | Na2MoO4·2H2O | 28.9nmol/L |
| 4 | MgSO4·7H2O | 60.9μmol/L | 11 | CuSO4·2H2O | 0.1nmol/L |
| 5 | KH2PO4 | 246.0μmol/L | 12 | ZnSO4 | 22.0nmol/L |
| 6 | NaHCO3 | 1785.5μmol/L | 13 | CoCl2·6H2O | 6.3nmol/L |
| 7 | Fe-EDTA | 10.0μmol/L |
1.2 design of the experiment
CrCl was selected for this test3As major contaminants, the screened rice seedlings were cultured in the following treatment solutions:
(1) cr (III) -N treatment [ (-N)o)&(-NI)]: the rice seedlings were placed in 50mL nitrogen-free nutrient solution (nitrogen starvation, nitrogen-free nutrient solution means removal of KNO)3Modified ISO8692 nutrient solution) of (1), the concentration of cr (iii) in the nitrogen-free nutrient solution: 0. 12.0, 24.0 and 40.0mg Cr/L.
(2) Cr (III) -N + Arg treatment [ (+ N)Arg)&(-NI)]: rice seedlings were placed in 3mM Arg solution for 12h and then placed in 50mL Erlenmeyer flasks without nitrogen at the concentration of Cr (III): 0. 12.0, 24.0 and 40.0mg Cr/L.
(3) Cr (III) -N + Glu treatment [ (+ N)Glu)&(-NI)]: rice seedlings were placed in 10mM Glu solution for 12h and then placed in 50mL conical flasks without nitrogen nutrient at the concentration of Cr (III): 0. 12.0, 24.0 and 40.0mg Cr/L.
(4)Cr(III)+NO3 -Treatment of
Placing the rice seedlings in 50mL of KNO-containing seedlings3(39.5mg N/L) nutrient solution in conical flask containing KNO3Concentration of cr (iii) in nutrient solution: 0. 12.0, 24.0 and 40.0mg Cr/L.
(5)Cr(III)+NO3 -+ Arg treatment
The rice seedlings are placed in 3mM Arg solution for 12h and then placed in 50mL KNO-containing solution3(39.5mg N/L) nutrient solution in conical flask containing KNO3Concentration of cr (iii) in nutrient solution: 0. 12.0, 24.0 and 40.0mg Cr/L.
(6)Cr(III)+NO3 -+ Glu treatment
The rice seedlings are placed in 10mM Glu solution for 12h and then placed in 50mL KNO-containing solution3(39.5mg N/L) nutrient solution in conical flask containing KNO3Concentration of cr (iii) in nutrient solution: 0. 12.0, 24.0 and 40.0mg Cr/L.
(7)Cr(III)+NH4 +Treatment of
Placing the rice seedlings in 50mL of solution containing NH4NH in a Cl (39.5mg N/L) nutrient solution conical flask4Concentration of cr (iii) in Cl nutrient solution: 0. 12.0, 24.0 and 40.0mg Cr/L.
(8)Cr(III)+NH4 ++ Arg treatment
The rice seedlings are placed in 3mM Arg solution for 12h and then placed in 50mL NH-containing solution4NH in a Cl (39.5mg N/L) nutrient solution conical flask4Concentration of cr (iii) in Cl nutrient solution: 0. 12.0, 24.0 and 40.0mg Cr/L.
(9)Cr(III)+NH4 ++ Glu treatment
The rice seedlings are placed in 10mM Glu solution for 12h and then placed in 50mL NH-containing solution4NH in a Cl (39.5mg N/L) nutrient solution conical flask4Concentration of cr (iii) in Cl nutrient solution: 0. 12.0, 24.0 and 40.0mg Cr/L. The conical flask is wrapped by tinfoil paper for shading treatment, so that the water loss is reduced to the maximum extent and the growth of algae is inhibited. There were 4 biological replicates per treatment and the exposure time was 3 days.
1.3 content of free proline in different tissues of Rice
After the stress of each treatment group is finished for 3 days, accurately weighing a certain amount of plant tissue samples (the weighed weight of root tissues (fresh weight) is 0.2 g, the weighed fresh weight of leaf tissues (fresh weight) is 0.2 g), placing the plant tissue samples into a precooled mortar, firstly adding 2.5mL of sulfosalicylic acid aqueous solution (the mass percent of sulfosalicylic acid is 3%), grinding and homogenizing the plant tissue samples, then transferring the plant tissue samples into a 10mL centrifuge tube, cleaning the mortar with 2.5mL of sulfosalicylic acid aqueous solution (the mass percent of sulfosalicylic acid is 3%), pouring cleaning liquid into the centrifuge tube, and centrifuging the plant tissue samples (11000rpm) for 15min at a low temperature (4 ℃); taking 2mL of the supernatant into a 10mL glass tube, sequentially adding 2mL of glacial acetic acid and 2mL of acidic ninhydrin solution (1.25g of ninhydrin is dissolved in 30mL of glacial acetic acid and 20mL of phosphoric acid, heating at 70 ℃ for dissolving, and storing in a refrigerator for later use), carrying out boiling water bath for 1h, then carrying out ice bath for 5min, adding 4mL of toluene, fully shaking, standing for 30min, taking the supernatant for color comparison at 520nm, taking toluene as a blank control, and calculating to obtain the content of free proline in the plant sample, wherein the numerical unit of the content of the free proline is (mu g/g FW, and the FW refers to fresh weight).
2. Results of the experiment
The results of the changes in the content of free Pro in rice cultivated under Cr (III) stress by different nitrogen sources are shown in Table 2.
Cr (III) -N treatment: the content of free Pro in both the root and the leaf is increased;
cr (III) -N + Arg treatment: the content of free Pro in both the root and the leaf is increased;
cr (III) -N + Glu treatment: the content of free Pro in both the root and the leaf is increased;
Cr(III)+NO3 -and (3) treatment: the content of free Pro in the blade is increased, and the content of free Pro in the root is not obviously changed;
Cr(III)+NO3 -+ Arg treatment: the free Pro content in the roots and leaves is reduced;
Cr(III)+NO3 -+ Glu treatment: the content of free Pro in the blade is reduced, and the content of free Pro in the root is increased;
Cr(III)+NH4 +and (3) treatment: the content of free Pro in both the root and the leaf is increased;
Cr(III)+NH4 ++ Arg treatment: the content of free Pro in the blade is reduced, and the change of the root is not obvious;
Cr(III)+NH4 ++ Glu treatment: the content of free Pro in the leaves is reduced, and the change of the roots is not obvious.
From the above experimental results, it has not been possible to quickly judge what kind of nitrogen source (nitrogen source alone: NO)3 -、NH4 +Arg, Glu; or a combination of nitrogen sources: NO3 -+NH4 +、NO3 -+Arg、NO3 -+Glu、NH4 ++Arg、 NH4 ++ Glu, Arg + Glu, are the optimal nitrogen sources for Cr (III) stress to control the content of free Pro in rice seedlings. Therefore, it is necessary to construct a preferable strategy model for screening different nitrogen sources under Cr (III) stress to cultivate stress resistance regulation of rice.
TABLE 2 variation of free Pro content in Rice cultivated under Cr (III) stress with different Nitrogen sources
3. Optimal strategy model for culturing rice and regulating chromium stress by screening different nitrogen sources with free proline as biomarker
3.1 modeling Process of stress-resistant regulation and control optimization strategy for culturing Rice with different Nitrogen sources under chromium stress
The content of free Pro in the plant body is closely related to the supply type of exogenous nitrogen. Such as: in NO3 -Or NH4 +The content of free Pro in cultured rice seedlings is obviously different, and the supply of exogenous Glu and Arg also has obvious influence on the content of free Pro in the rice seedlings. The model is established based on the influence of independent or combined cultivation of different nitrogen sources on the change of the content of free Pro in rice seedlings, the change of the content of free Pro in the rice seedlings by independent or combined cultivation of different nitrogen sources under different Cr (III) stress concentrations is simulated, and the independent or combined cultivation of rice by using the free Pro as a biomarker is researched3 -And NH4 +(ii) a Organic nitrogen source: glu and Arg) preferred strategies for modulating Cr (III) stress.
Definition of different concentrations of cr (iii) under stress, different nitrogen sources were applied in rice seedlings, alone or in combination: treatment without organic Nitrogen (-N)O) Arginine treatment (+ N)Arg) Glutamic acid treatment (+ N)Glu) Inorganic nitrogen treatment (-N)I) Nitrate nitrogen treatment
Ammonium nitrogen treatment
Then the matrix is obtained:
the rows and columns of the matrix are each denoted by riAnd cjDenotes (i, j ═ 1,2,3), and the specific elements of the matrix are represented by a, respectivelyijRepresents (i, j ═ 1,2,3), in a matrix "&"denotes the combined administration of two nitrogen sources, thenIn this case:
a11indicates the free Pro content in the absence of organic nitrogen treatment and in the absence of inorganic nitrogen treatment;
a12represents the free Pro content in arginine pretreatment and in the absence of inorganic nitrogen treatment;
a13represents the free Pro content in the glutamic acid pretreatment and in the absence of inorganic nitrogen treatment;
a21indicates the free Pro content without organic nitrogen treatment and with the addition of nitrate nitrogen;
a22represents the free Pro content when arginine is pretreated and nitrate nitrogen is added;
a23represents the free Pro content when glutamic acid is pretreated and nitrate nitrogen is added;
a31indicates the free Pro content in the case of no organic nitrogen treatment and addition of ammonium nitrogen;
a32represents the free Pro content when arginine was pretreated and ammonium nitrogen was added;
a33indicates the free Pro content when glutamic acid was pretreated and ammonium nitrogen was added.
If need be compared, the matrix (r) is respectively subjected to three primary row transformations: r is2-r1，r3-r1，r3-r2Obtaining three matrixes, namely a matrix II, a matrix III and a matrix IV:
r in combined matrix 21R in matrix c1R in matrix r2Obtaining a matrix fifth:
at this time, matrix c
r1Represents the contribution of nitrate nitrogen treatment to free Pro;
r2represents the contribution of ammonium nitrogen treatment to free Pro;
r3represents the difference between ammonium nitrogen treatment and nitrate nitrogen treatment in the amount of free Pro.
And then, performing three times of primary column transformation on the matrix (i): c. C2-c1，c3-c1，c3-c2Three matrices are obtained, and the matrices are combined by the same method to obtain:
at this time, matrix is
c1Represents the contribution of arginine treatment to free Pro;
c2represents the contribution of glutamate treatment to free Pro;
c3represents the contribution of the difference in free Pro from the glutamic acid treatment and the arginine treatment.
Matrix c is subtracted from matrix c to obtain matrix c:
wherein-Noand-NIIs set to 0.
Extraction of the fractions in which analysis is required: a is11、a12、a21、a22Forming a new matrix (b):
at this time in matrix (b)
a11Represents the contribution of arginine treatment to the difference in free Pro from nitrate nitrogen treatment;
a12represents the contribution of glutamic acid treatment and nitrate nitrogen treatment to the difference of free Pro;
a21represents the contribution of arginine treatment to the difference in free Pro from ammonium nitrogen treatment;
a22represents the contribution of glutamate treatment to the difference in free Pro as compared to ammonium nitrogen treatment.
If the contribution of inorganic nitrogen or organic nitrogen to free Pro needs to be compared with that of inorganic nitrogen or organic nitrogen alone, only the third row element a needs to be compared with the matrix3jMatrix of the third column element ai3And matrix r where11、a12、a21、a22The numerical value of (c).
3.2 modeling results
The results of measuring the content of free Pro in the leaves and roots of rice seedlings are shown in the following tables 3 and 4, respectively:
TABLE 3 content of free Pro (μ g/g FW) in young leaves of rice plants stressed by Cr (III) at different concentrations
TABLE 4 free Pro content (μ g/g FW) in rice seedling roots stressed by Cr (III) at different concentrations
Taking 3 Control data in the leaves of table 3 as an example, a matrix is established and compared, and the matrix is:
from the matrix r', it can be seen that, in the absence of cr (iii) stress, the contribution of the combined application of inorganic nitrogen and organic nitrogen to free Pro is in the order:
and (3) performing three primary row transformations on the matrix (r') respectively: r is2-r1，r3-r1，r3-r2R in the combined matrix 21R in matrix c1R in matrix r2Obtaining a matrix of:
from the matrix, the third row, the contribution of inorganic nitrogen alone to free Pro when no cr (iii) stress was added:
and (3) performing three primary column transformations on the matrix (r') respectively: c. C2-c1，c3-c1， c3-c2Three matrices are obtained, and the matrices are combined by the same method to obtain a matrix of:
from the matrix, the third column, the contribution of organic nitrogen alone to free Pro when no cr (iii) stress was added: n is a radical ofArg>NGlu(ii) a Subtracting matrix from matrix to obtain matrix to extract needed parts: a is11、a12、a21、a22Forming a new matrix (v):
from the matrix, it can be seen that the contribution of inorganic nitrogen or organic nitrogen alone to free Pro without cr (iii) stress addition:
taken together, the contribution of inorganic or organic nitrogen alone to free Pro in leaves when no stress was added was:
following this approach, the contribution of inorganic nitrogen and organic nitrogen to free Pro at other stress concentrations, either in combination or separately, was obtained as shown in Table 5 below.
TABLE 5 contribution of inorganic and organic Nitrogen to Rice seedling Pro under stress of different concentrations of Cr (III) in combination or separately
According to the contribution of the nitrogen sources to free Pro by independent and combined application in rice seedling tissues under various stress concentrations, a strategy model for screening different nitrogen sources to culture rice to regulate chromium stress by taking the free Pro as a biomarker is obtained, and stress resistance regulation optimization strategies of the rice at different growth stages are determined according to the contribution model.
As can be seen from Table 5, in both rice roots and leaves, and in different concentrations of Cr (III) stress,
+NArgthe N source combination has the largest contribution to free Pro and can be used as the preferable combination N source in future experiments and practical application; while the nitrogen source test group was administered alone,
the contribution to free Pro in Cr (III) -stressed rice seedling leaves is the greatest, NArgThe method has the greatest contribution to Cr (III) stress of free Pro in rice seedling roots, so that different nitrogen source applying strategies can be adopted in different growth stages of rice to meet the requirement of Cr (III) stress.
All the embodiments in the present specification are described in a related manner, and the same and similar parts among the embodiments may be referred to each other, and each embodiment focuses on the differences from the other embodiments.
The above description is only for the preferred embodiment of the present invention, and is not intended to limit the scope of the present invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention shall fall within the protection scope of the present invention.
Claims (7)
1. The method for determining the rice stress-resistance regulation and control optimization strategy is characterized by comprising the following steps of:
step 1: applying different nitrogen source application schemes to the rice seedlings under the stress of various concentrations of Cr (III); determining the content of free Pro in rice seedling tissues in each nitrogen source application scheme;
step 2: constructing a matrix I for cultivating rice stress resistance regulation and control by using any two combinations of organic nitrogen-free treatment, arginine treatment, glutamic acid treatment, inorganic nitrogen-free treatment, nitrate nitrogen treatment and ammonium nitrogen treatment, wherein the matrix I comprises the following components:
in matrix I, the rows and columns of the matrix are each denoted by riAnd cjRepresents, i, j ═ 1,2, 3; a specific element of the matrix is represented by aijRepresents, i, j ═ 1,2, 3;&represents the combined administration of two nitrogen sources; in matrix I, a11Indicates the free Pro content in the absence of organic nitrogen treatment and in the absence of inorganic nitrogen treatment; a is12Represents the free Pro content in arginine pretreatment and in the absence of inorganic nitrogen treatment; a is13Represents the free Pro content in the glutamic acid pretreatment and in the absence of inorganic nitrogen treatment; a is21Indicates the free Pro content in the case of treatment without organic nitrogen and treatment with nitrate nitrogen; a is22Represents the free Pro content when arginine pretreatment and nitrate nitrogen addition treatment are carried out; a is23Represents the free Pro content when the glutamic acid is pretreated and the nitrate nitrogen is added; a is31Indicates the free Pro content in the case of treatment without organic nitrogen and treatment with ammonium nitrogen;a32represents the free Pro content in arginine pretreatment and ammonium nitrogen addition treatment; a is33Represents the free Pro content when glutamic acid is pretreated and ammonium nitrogen is added;
and step 3: r is performed on the matrix I2-r1，r3-r1，r3-r2Performing three times of primary row transformation to obtain a matrix II; c is performed on the matrix I2-c1，c3-c1，c3-c2Performing three times of primary column transformation to obtain a matrix III; subtracting the matrix II from the matrix III to obtain a matrix IV; from a in the matrix IV11、a12、a21、a22Forming a matrix V;
and 4, step 4: comparing matrix I with a respectively22、a23、a32、a33The magnitude of the numerical value, resulting in the magnitude of the contribution of the combined administration of inorganic nitrogen and organic nitrogen to free Pro;
comparing matrix II third row elements a separately3jThird column element a of matrix IIIi3And matrix V in a11、a12、a21、a22The amount of free Pro compared to the amount of contribution of inorganic nitrogen or organic nitrogen administered alone;
and 5: according to the method of the step 4, obtaining the contribution size of the nitrogen source to the free Pro by the single and combined application in the rice seedling tissues under different stress concentrations; according to the contribution of the nitrogen sources to free Pro by independent and combined application in rice seedling tissues under various stress concentrations, a strategy model for screening different nitrogen sources to culture rice to regulate chromium stress by taking the free Pro as a biomarker is obtained, and stress resistance regulation optimization strategies of the rice at different growth stages are determined according to the contribution model.
2. The method for determining rice stress-tolerance-controlling preference strategy as claimed in claim 1, wherein the nitrogen source application scheme comprises: any one or a combination of any two of organic nitrogen-free treatment, arginine treatment, glutamic acid treatment, inorganic nitrogen-free treatment, nitrate nitrogen treatment, and ammonium nitrogen treatment.
3. The method for determining the rice stress-resistance control optimization strategy according to claim 1, wherein in the step 3, the matrix II is specifically as follows:
in matrix II, r1Represents the contribution of nitrate nitrogen treatment to free Pro; r is2Represents the contribution of ammonium nitrogen treatment to free Pro; r is3Represents the contribution of the difference in free Pro from the ammonium nitrogen treatment and the nitrate nitrogen treatment.
4. The method for determining the rice stress-resistance control optimization strategy according to claim 1, wherein in the step 3, the matrix III is as follows:
in matrix III, c1Represents the contribution of arginine treatment to free Pro; c. C2Represents the contribution of glutamate treatment to free Pro; c. C3Represents the contribution representing the difference in free Pro from glutamate treatment and arginine treatment.
5. The method for determining the rice stress-resistance control optimization strategy according to claim 1, wherein in the step 3, the matrix IV is as follows:
in matrix IV, a11Represents the contribution of arginine treatment to the difference in free Pro from nitrate nitrogen treatment; a is12Represents the contribution of glutamic acid treatment and nitrate nitrogen treatment to the difference of free Pro; a is13Represents the contribution of the difference of the glutamic acid treatment, the arginine treatment and the nitrate nitrogen treatment to the free Pro; a is21Represents the contribution of arginine treatment to the difference in free Pro from ammonium nitrogen treatment; a is22Represents the contribution of glutamic acid treatment and ammonium nitrogen treatment to the difference of free Pro; a is23Represents the contribution of the difference of free Pro by glutamic acid treatment, arginine treatment and ammonium nitrogen treatment; a is31Represents the contribution of arginine treatment, nitrate nitrogen treatment and ammonium nitrogen treatment to the difference of free Pro; a is32Represents the contribution of glutamic acid treatment, nitrate nitrogen treatment and ammonium nitrogen treatment to the difference of free Pro; a is33The contribution of the difference between glutamic acid treatment and arginine treatment, nitrate nitrogen treatment, and ammonium nitrogen treatment to the free Pro was shown.
6. The method for determining the rice stress-resistance control optimization strategy according to claim 1, wherein in the step 3, the matrix V is as follows:
7. the method for determining the rice stress-resistance control optimization strategy as claimed in claim 1, wherein in step 4, the third row element a of the respective comparison matrix II3jThird column element a of matrix IIIi3And matrix V in a11、a12、a21、a22The amount of contribution of inorganic nitrogen or organic nitrogen to free Pro compared to the amount of contribution of inorganic nitrogen or organic nitrogen administered alone is obtained by:
comparing the contribution of ammonium nitrogen alone with nitrate nitrogen alone to free Pro according to the sign of the value of each element in the third row of matrix II; comparing the amount of contribution of glutamic acid alone to arginine alone to free Pro according to the positive and negative values of the elements in the third column of matrix III; according to a of the matrix V11Positive and negative values, comparing the contribution of arginine alone to free Pro versus nitrate nitrogen alone; according to a of the matrix V12The magnitude of the contribution to free Pro of glutamic acid alone versus nitrate nitrogen alone; according to momentA of the matrix V21Positive and negative values, comparing the amount of contribution of arginine alone to free Pro versus ammonium nitrogen alone; according to a of the matrix V22The magnitude of the contribution to free Pro compared to glutamic acid alone versus ammonium nitrogen alone;
the magnitude of the contribution of inorganic or organic nitrogen alone to free Pro is obtained from the magnitude of the contribution of ammonium nitrogen alone and nitrate nitrogen alone to free Pro, the magnitude of the contribution of glutamic acid alone and arginine alone to free Pro, the magnitude of the contribution of arginine alone and nitrate nitrogen alone to free Pro, the magnitude of the contribution of glutamic acid alone and nitrate nitrogen alone to free Pro, the magnitude of the contribution of arginine alone and ammonium nitrogen alone to free Pro, and the magnitude of the contribution of glutamic acid alone and ammonium nitrogen alone to free Pro.
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