CN113030220B - Method for evaluating shade resistance and lodging resistance of soybeans by measuring hydrogen ion flow of stem epidermal cells - Google Patents
Method for evaluating shade resistance and lodging resistance of soybeans by measuring hydrogen ion flow of stem epidermal cells Download PDFInfo
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Abstract
The invention is suitable for the technical field of plant shade-tolerant lodging-resistant evaluation, and provides a method for evaluating soybean shade-tolerant lodging resistance by measuring a stalk epidermal cell hydrogen ion flow. The method comprises the following steps: polishing the surface of the soybean stalk to form a polished part in the original growing environment of the soybean; balancing the ion exchange of epidermal cells at the polished part by adopting a balance buffer solution; measuring the hydrogen ion current of the epidermal cells at the polished part; and carrying out shade-resistant lodging-resistant analysis evaluation according to the hydrogen ion current measurement result. The invention measures the hydrogen ion flow condition of the soybean stem epidermis under normal illumination and shading environment through non-damage micro-measurement technology under the original growth environment of plants, thereby judging the relative strength of shading and lodging resistance of different soybean varieties.
Description
Technical Field
The invention belongs to the technical field of plant shade resistance and lodging resistance evaluation, and particularly relates to a method for evaluating soybean shade resistance and lodging resistance by measuring a hydrogen ion flow of stem epidermal cells.
Background
Soybean (Glycine max) is native to china and is an important source of edible vegetable proteins and fats worldwide. In recent years, with the improvement of the living standard of people, the demand of China for soybeans is rapidly increased, the contradiction between supply and demand is increasingly prominent, and the improvement of the yield of soybeans becomes an urgent need for guaranteeing national food safety. Close planting and intercropping are important means for improving the yield per unit of soybean and effectively utilizing the planting space, however, the mutual shading among plants can cause the remarkable change of the ambient light environment, so that the soybean has obvious shading reaction (such as the reduction of leaf angle, the elongation and thinning of stems and petioles, the early blooming, the reduction of branches and the like), and the popularization and the application of two cultivation modes are seriously restricted. In addition, the elongation and the thinning of the stems of soybean plants are the main reasons for causing the lodging of the soybeans, which seriously affect the yield and the quality of the soybeans at the later stage, so that the evaluation of the shade resistance and the lodging resistance of the soybeans is very important.
In the prior art, the evaluation methods of soybean shade tolerance are mainly to measure physiological indexes indoors after field agronomic character survey or field sampling, for example, in related literature 1 (wuxialing, longhaiyu, poplar peak, liuwei, junjiu, yangqi, yanwenyu. Screening of soybean seedling shade tolerance comprehensive evaluation and identification indexes [ J ]. Chinese agricultural science, 2015,48 (13): 2497-2507.), 24 morphological physiological indexes in the field are reported to be investigated, a regression equation is established, and the comprehensive evaluation of soybean seedling shade tolerance can be carried out by using four indexes of dry leaf weight, conductivity, plant height and maximum fluorescence yield in the dark; in related literature 2 (yancaiqiong, hu bao, wu navy, qin wenting, zhang jijun, liuqi, liuwei nationality, yanwenyu, liujiang, black bean germplasm shade tolerance evaluation and response of root system to weak light stress [ J ]. Ecological agriculture report in china, 2017,25 (06): 893-902.), five indexes of transpiration rate, plant height, leaf dry weight, maximum fluorescence intensity and initial fluorescence intensity are reported as evaluation basis of black bean seedling shade tolerance; in related literature 3 (li chun hong, yao xingdong, jubaotao, zhugueh, royal sea english, zhuyijun, and ao, yue mei, xipu, song shuhong. Screening of analysis and identification indexes of soybean shade tolerance of different genotypes [ J ]. Chinese agricultural science, 2014,47 (15): 2927-2939.), related literature 4 (zhao yinyue, jenzhang and ming, shimada, dan alone, king iron army, yunnan intercropping soybean shade tolerance comprehensive evaluation and identification indexes screening [ J ]. Chinese oil crop academic press, 2019,41 (01): 81-91.), all reports that indexes related to soybean yield and yield constituent factors can be used as identification indexes of soybean shade tolerance; in related document 5 (LIU, wei-guo, REN, et al. Effect of shade stress on lignin biosyntheses in soybean stem [ J ]. Journal of Integrated Agriculture culture, 2018.), related document 6 (Deng Yuchuan, liuwei nation, yuan, jin, junlin, dubo, yangyu. Relation between synthetic metabolism of stalk cellulose and lodging resistance in relay intercropping soybean seedling stage [ J ]. Applied ecology newspaper, 2016,27 (02): 469-476.), the shade-resistant and lodging-resistant properties of soybean seedling stage are reported to be related to the content of lignocellulose in stalk.
In summary, currently, the shading resistance of the soybeans is mainly evaluated from the aspects of seedling stage characteristics, yield constituent factors, stalk lignocellulose content and the like, the soybean is required to be planted in the field by the evaluation methods, the research process is complex, the operation is not easy, and the research period is long; the shade-resistant lodging-resistant soybean material cannot be quickly and accurately identified under indoor conditions; the real growth condition of the plant under the shading condition can not be reflected; can not realize the dynamic detection of the living biological materials and cause waste to the treasure experiment materials. Therefore, the development of a new shade tolerance evaluation method is of great significance to a method for screening lodging-resistant crops.
Disclosure of Invention
In order to solve the problems, the invention provides a method for evaluating the shade resistance and lodging resistance of soybeans by measuring the hydrogen ion flow of stem epidermal cells.
The invention is realized in such a way, and a method for evaluating the shade resistance and lodging resistance of soybeans by measuring the hydrogen ion flow of stalk epidermal cells comprises the following steps:
step S1: polishing the surface of the soybean stalk to form a polished part in the original soybean growth environment;
step S2: balancing the ion exchange of epidermal cells at the polished part by adopting a balance buffer solution;
and step S3: measuring the hydrogen ion flow of the epidermal cells at the polished part;
and step S4: and carrying out shade-resistant lodging-resistant analysis evaluation according to the hydrogen ion current measurement result.
Further, in the step S1, the skins of the soybean stalks are polished by sand paper.
Further, the sand paper is 1000 meshes.
Further, in the step S2, an equilibrium buffer is used to balance the ion exchange of the epidermal cells at the polished part, including the following steps:
step S2-1: wetting the wrap with the equilibration buffer;
step S2-2: winding the wrap around the polished portion of the soybean stem;
step S2-3: during the balancing process, the balancing buffer solution is added to the wrappage until the ion exchange of the epidermal cells of the polished part reaches the balance.
Further, the equilibration buffer comprises CaCl 2 MES solution.
Further, in step S2-3, the time intervals for adding the equilibration buffer to the wrap during equilibration are equal.
Further, during equilibration, the equilibration buffer is added to the wrap at intervals of 25min-30 min.
Further, in the step S2-3, the amount of the equilibration buffer added to the wrap at each time is equal during equilibration.
Further, in the step S2-3, the time for the epidermal cell ion exchange of the polished part to reach equilibrium is positively correlated with the polishing degree of the plant stalks.
Further, in the step S3, the hydrogen ion flow of the epidermal cells at the polished part is measured by using a non-damage micrometering technique.
Further, in step S4, the evaluation of shade-tolerant lodging-resistant analysis based on the hydrogen ion current measurement result includes the following steps:
step S4-1: and analyzing the correlation between the flow velocity of the hydrogen ions and the internode length and the plant height of the soybean, wherein the correlation coefficient is marked as P, P is more than 0.7, which shows a high positive correlation, P is more than 0.4 and less than 0.7, which shows a medium positive correlation, and P is more than 0.2 and less than 0.4, which shows a weak positive correlation.
Compared with the prior art, the invention has the technical effects that:
(1) According to the invention, the stalk epidermis can be polished and the ions can be balanced in the original growing environment of the plant, the integrity and the normal physiological state of the plant are maintained, the hydrogen ion change condition of the plant in the environment is measured, and the measurement result is more accurate;
(2) In the invention, the epidermis of the plant stem is polished by sand paper, so that the cuticle and the wax layer are removed, and the microelectrode can detect the hydrogen ion flow in epidermal cells;
(3) In the invention, a non-damage micrometering technology is adopted, the hydrogen ion flow conditions (including flow velocity and flow direction) of the internal physiological state of the plant under the real growth environment can be dynamically measured and obtained in real time on the premise of not damaging the plant stem sample, the sample does not need to be extracted and marked on a substance to be measured, and the real growth state of the plant can be reflected;
(4) The method for screening the shade-tolerant lodging-resistant soybeans provided by the invention is simple and reliable, is easy to operate, can quickly and accurately measure the change of hydrogen ion flow in plant tissues under indoor conditions, thereby judging the relative strength of shade resistance and lodging resistance of different soybean varieties,
drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments of the present invention or in the description of the prior art will be briefly described below, and it is obvious that the drawings described below are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to the drawings without creative efforts. In the quantitative tests in the following examples, more than three times of repeated experiments are set, and the results are averaged. The totality of MES is known as 2- (N-morpholino) ethanesulfonic acid.
FIG. 1 is a flow chart of the method for screening shade-tolerant lodging-resistant soybeans by measuring the hydrogen ion current of epidermal cells of plant stalks provided by the embodiment of the invention;
FIG. 2 is a schematic view of a glass microelectrode provided by an embodiment of the present invention;
FIG. 3 is a schematic view showing the positions of electrodes and stems when a glass microelectrode is used for detection;
FIG. 4 is a graph of the real-time hydrogen ion flow rate analysis of C103 and E9-2 under different light environments according to an embodiment of the present invention;
FIG. 5 is a graph of hydrogen ion average flow velocity analysis of C103 and E9-2 under different light environments according to an embodiment of the present invention;
FIG. 6 is a diagram showing the phenotype of C103 plants grown under different light environments;
FIG. 7 is a diagram showing the phenotype of E9-2 plants grown under different light environments;
FIG. 8 is a statistical representation of plant heights and second internode lengths of C103 and E9-2 grown under different light environments, provided by the examples of the present invention.
Description of the drawings:
1-glass microelectrodes; 2-Ag/AgCl electrode wire base; 3-an electrolyte; 4-liquid ion exchanger;
Detailed Description
Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like reference numerals refer to the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the drawings are illustrative and intended to be illustrative of the invention and are not to be construed as limiting the invention.
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to the accompanying drawings and embodiments.
The auxin can stimulate a proton pump on a cell membrane to be activated, promote the discharge of hydrogen ions in cells, so that the cell wall is acidified and relaxed, and further, the extension of organs is promoted, therefore, the influence of the environment of the plant on the growth and development of the plant can be researched by measuring the extracellular hydrogen ion flow of the plant cytoplasm. The plant adapts to unfavorable light environment by changing the shape, the most remarkable characteristic is that the plant can capture more light by the elongation of the stem or the petiole, but the over-elongated stem makes the plant easy to fall, therefore, the invention takes the soybean stem as a research object, and evaluates the relative strength of the shade resistance and the lodging resistance of the soybean by measuring the hydrogen ion of the epidermis of the soybean stem, in particular, the invention relates to a method for evaluating the shade resistance and the lodging resistance of the soybean by measuring the hydrogen ion flow of the epidermal cell of the stem according to the flow chart shown in figure 1, and the method comprises the following steps:
the experimental materials are soybean varieties C103 and E9-2 with different shading sensitivity degrees, the soybean varieties are provided by ecological research institute of Sichuan university of agriculture, the source of the soybean varieties is not limited, wherein the C103 is a shading sensitive variety, and the length of the shaded stalks is longer; e9-2 is a shade-insensitive variety, and the stalks under shade have shorter elongation. C103 and E9-2 are firstly sprouted for 1.5 days at 25 ℃ and then sowed in Pingshi soil, and the PPFD of the seedlings is 450 umol.m-2.s- 1 Growing under normal light, moving the true leaves to 150 umol.m PPFD after the leaves are unfolded -2 ·s -1 The soybean varieties grow in a shading environment, then the change condition of the hydrogen ion flow of the stalk epidermal cells of the C103 and the E9-2 under the shading condition is tested, and the relative strength of the shading and lodging resistance of different soybean varieties is judged according to the flow rate of the hydrogen ion.
The specific measurement method comprises the following steps:
step S1: polishing the surface of the soybean stalk to form a polished part in the original soybean growth environment;
step S2: balancing the ion exchange of epidermal cells at the polished part by adopting a balance buffer solution;
and step S3: measuring the hydrogen ion flow of epidermal cells at the polished part;
and step S4: and carrying out analysis and evaluation on the shading resistance according to the hydrogen ion flow measurement result.
In the step S1 and the step S2, the soybeans C103 and E9-2 are both carried out in the original growing environment, so that the integrity and the normal physiological state of soybean plants are kept, the measurement result is more accurate, and the growth condition of the soybean plants can be more truly reflected by the measurement result.
Furthermore, because the soybean stalks are mature and hard, rich in lignin and poor in water permeability, and the epidermal cells contain thick cuticle or wax layer, the hydrogen ion flow of the epidermal cells cannot be directly measured, the epidermis of the soybean stalks needs to be polished to expose young and tender epidermal cells, and the young and tender epidermal cells are detected by the probe. In the invention, C103 and E9-2 are sampled after growing for 17 days in an illumination incubator, 6 soybean varieties are taken from each soybean variety and kept in the growing environment before sampling, then the epidermis at the middle part of the first internode of the soybean stalk is gently and uniformly polished by abrasive paper, and the surface cuticle and the wax layer are removed to form a polished part. The mesh number, the grinding time and the grinding times of the abrasive paper are all determined by the keratinization degree of the plant stem tissue, in the embodiment, 1000-mesh abrasive paper is adopted, and the grinding times are 75 times.
Furthermore, after the soybean stalks are polished by sand paper in the above steps, certain damage may be caused to the soybean plants, so that the flow rate and the flow direction of hydrogen ions in the soybean stalks are affected, and therefore, after the polishing treatment of the soybean stalks is completed, the polished parts of the soybean stalks must be immediately balanced by adopting a balance buffer solution, so that the ion exchange of epidermal cells of the soybean stalks is balanced.
The method for balancing the plant stem sample comprises the following steps:
step S2-1: wetting the wrapping object with a balance buffer solution, so that the balance buffer solution in the gauze is in a state of being about to drip but not dripping, and the wrapping object is made of a material with good liquid absorption performance and liquid storage performance, such as gauze, sponge and the like, but is not limited to this, in the embodiment, the wrapping object is the gauze;
step S2-2: the soybean stalks are still kept in the growing environment before sampling, and then the wrappage wetted by the equilibrium buffer solution in the step S2-1 is wound on the polished parts of the soybean stalks, the winding tool and the winding mode of the gauze are not limited, and the candy line is adopted in the embodiment;
step S2-3: and (3) changing the ion concentration of the balance buffer solution in the wrapper along with the balance time, and adding the balance buffer solution into the wrapper at intervals of 25-30 min to ensure that the epidermal cell ion exchange of the polished part is completely balanced, preferably adding 3mL of the balance buffer solution into the wrapper wound around the polished part of the soybean stem by using an injector at intervals of 30min until the epidermal cell ion exchange of the soybean stem reaches balance.
In the step S2-1 to the step S2-3, both the C103 and the E9-2 are carried out in the original growing environment, and quantitative balance buffer solution is continuously added to the wrappage in the balance process, so that the measurement result is more accurate, and the real physiological state of the soybean plant can be reflected more.
Further, the time for the epidermal cell ion exchange at the polished part to reach equilibrium is positively correlated with the polishing degree of the plant stalks, which is not limited in the present invention, and in this embodiment, the time for the epidermal cell ion exchange at the polished part of the soybean stalks to reach equilibrium is 3 hours.
Further, in the above step S2-1 to step S2-3, the equilibration buffer used comprises 0.1mM CaCl with pH =5.8 2 And 0.5mM MES solution.
Further, a non-damage micrometering technology is adopted to measure the hydrogen ion flow of the epidermal cells at the polished part, and the specific method is as follows:
the ion detector used was a non-invasive micrometering system (BI 0-001B, young USA Sci. & Tech. Crop., USA), the system software was imFlux V2.0, and the microelectrode tip used was a glass microelectrode 1 with a tip diameter of 2-4 um.
(1) Fabrication of microelectrodes
As shown in FIG. 2, an electrolyte 3 of 15mM NaCl and 40mM KH having a pH =7 was injected into the glass micro-electrode 1 until the tip of the glass micro-electrode 1 was filled by 0.8cm to 1.8cm 2 PO 4 Keeping the front end of the glass microelectrode 1 under a microscope, and sucking 40-50um of liquid ion exchanger 4 to complete the manufacture of the glass microelectrode 1, wherein the liquid ion exchanger 4The product number is NMT-HC-11.
(2) Installing silver chloride wire
Sleeving the glass microelectrode 1 prepared in the step (1) into a chloridized Ag/AgCl electrode wire base 2, and in order to ensure the measurement accuracy, putting the glass microelectrode 1 into a high-concentration correction liquid and a low-concentration correction liquid for correction, wherein the two correction liquids are different in pH value (namely different in hydrogen ion concentration), wherein the high-concentration correction liquid comprises 0.1mM CaCl with the pH =5.6 2 And 0.5mM MES solution, the low concentration correction fluid comprises 0.1mM CaCl with pH =6.6 2 And 0.5mM MES solution, and after the correction was completed, the Nernst equation of the electrode calculated as the ideal value H + ：55-60mV。
(3) Measurement of hydrogen ion flux
Putting the balanced soybean stalks into a test buffer solution, and then putting the soybean stalks under a microscope of a non-damage micrometering system; observing the epidermis edge under microscope, searching for the region with intact cell structure, selecting well-formed cells as the detection point, wherein the test buffer comprises 0.1mM CaCl with pH =5.8 2 And 0.5mM MES solution.
As shown in FIG. 3, a measurement data was recorded by aligning the tip of the glass microelectrode 1 at the detection points 30um away from the cell and moving the glass microelectrode 1 by 30um for 5s, and the observation time at each detection point was 10min, and then the measurement was performed again at a position 800um away from the previous detection point until all the detection points were measured. The glass microelectrode 1 is used for measuring the hydrogen ion concentration of two points of the soybean stalk at different distances and finally converting the hydrogen ion concentration into the hydrogen ion flow speed.
The invention adopts non-damage micrometering technology, can dynamically measure and obtain the ion flow condition of the plant in real physiological state in real time on the premise of not damaging the stem sample of the plant, and does not need to extract the sample and mark the substance to be measured.
(4) Data processing and shade-fastness analysis evaluation
Processing the measured experimental data by using Fick's diffusion law, calculating by using online software, namely, mage Flux-3Dion Flux marking System (http:// www.xueyue. Net/mageflux), performing statistical analysis, and analyzing and mapping by using Excel2010 to obtain a hydrogen ion real-time flow velocity analysis chart of C103 and E9-2 under different illumination environments, wherein the hydrogen ion real-time flow velocity analysis chart is shown in figure 4; the analysis graphs of the average hydrogen ion flow rates of C103 and E9-2 under different illumination environments are obtained according to the experimental results in FIG. 4, and shown in FIG. 5, wherein LC is C103 soybean under normal illumination, LE is E9-2 soybean under normal illumination, SC is C103 soybean under shading condition, and SE is E9-2 soybean under shading condition in FIGS. 4 and 5.
As can be seen from fig. 5: under normal illumination, the measured hydrogen ion flow rate is negative no matter how sensitive the tested soybean variety is to shading, which indicates that under normal illumination, the C103 and E9-2 stem epidermal cells absorb hydrogen ions from the test buffer solution. Wherein the average flow velocity of hydrogen ions of the shading-sensitive LC stems is-2.5 pmol cm -2 ·s -1 The average flow rate of hydrogen ions of the shade-insensitive LE stems was-15 pmol cm -2 ·s -1 。
As can be seen from fig. 5: under the shading condition, the measured hydrogen ion flow rate is a positive value no matter how sensitive the tested soybean variety is to the shading, which indicates that the C103 and E9-2 stem epidermal cells discharge hydrogen ions to the environment under the shading condition. Wherein the average flow velocity of hydrogen ions of the shading-sensitive SC stalks is 19.91pmol cm -2 ·s -1 The average hydrogen ion flow rate of the SE stems insensitive to shading was 13.08pmol cm -2 ·s -1 。
Further, in step S4, the shading resistance analysis and evaluation is performed based on the hydrogen ion current measurement result, specifically as follows:
fig. 6 is a schematic phenotype diagram of a C103 plant after growing for 20 days under normal illumination and shading conditions, respectively, fig. 7 is a schematic phenotype diagram of an E9-2 plant after growing for 20 days under normal illumination and shading conditions, respectively, plant heights and second internode lengths of C103 and E9-2 plants under normal illumination and shading conditions can be respectively calculated according to fig. 6 and 7, see fig. 8, the abscissa in fig. 8 is soybean under different illumination conditions, and the ordinate is second internode length and plant height of soybean.
As can be seen from FIG. 8, for the C103 plant, the second internode length under normal illumination is 2.61cm, the plant height is 15.01cm, the second internode length under the shading condition is 10.54cm, and the plant height is 32.99cm, compared with the two, the second internode length of the C103 plant under the shading condition is increased by 10.54cm, the increase is 403.8%, the plant height is increased by 17.98cm, and the increase is 119.8%; aiming at the E9-2 plant, the second internode length under normal illumination is 0.83cm, the plant height is 10.10cm, the second internode length under the shading condition is 3.60cm, and the plant height is 15.63cm, compared with the two, the second internode length of the E9-2 plant under the shading condition is increased by 2.77cm, the amplification is 333.7 percent, the plant height is increased by 5.53cm, and the amplification is 54.8 percent; therefore, no matter how sensitive the tested soybean variety is to shading, the stem and the plant height of the soybean plant show obvious increase under the shading condition, and the increase degree of the stem and the plant height of the shading sensitive C103 soybean is far higher than that of the shading insensitive E9-2 soybean, so that the epidermal cells of the soybean stem emit hydrogen ions to the environment under the shading condition, and the increase of the stem and the plant height of the soybean plant can be promoted.
Further, in step S4-1, performing Person correlation analysis on the hydrogen ion flow rate, the soybean internode length and the plant height by using SPSS 20 software according to the experimental data of the stem epidermal hydrogen ion flow rate, the soybean second internode length and the plant height measured under the shading conditions, wherein the correlation coefficient is recorded as P, the significance of P is recorded as M, if the P value is greater than 0, the positive correlation is obtained, and if the P value is less than 0, the negative correlation is obtained, if the P value is greater than 0.7, the positive correlation of the height is obtained, if the P value is greater than 0.4, the P is less than 0.7, the moderate positive correlation is obtained, and if the P value is greater than 0.2, the P is less than 0.4, the weak positive correlation is obtained; if M is between 0.01 and 0.05, the correlation is significant and is represented by x, if M is less than 0.01, the correlation is very significant and is represented by x, specifically, the correlation coefficient P1=0.834 between the hydrogen ion flow rate of the stalk epidermis and the soybean second internode length, the significance of P is represented by x, the correlation coefficient P2=0.850 between the hydrogen ion flow rate of the stalk epidermis and the soybean plant height, and the significance of P is represented by x, thereby indicating that the second internode length, the plant height and the hydrogen ion flow rate height in the stalk epidermis of the soybean plant are positively correlated, i.e., the larger the hydrogen ion flow rate in the stalk epidermis, the longer the soybean second internode length and the plant height are increased, the larger the degree of influence of shading is, the weaker the shading resistance of the soybean plant is, and therefore, the relative strength of shading resistance of the soybean plant can be judged by measuring the hydrogen ion flow rate in the stalk epidermis.
In conclusion, under the shading condition, a large amount of hydrogen ions are discharged from shading-sensitive soybeans to acidify cell walls, so that the elongation of stalks and the increase of plant height are promoted, the stalks are easy to lodging, and the shading-insensitive soybeans can ensure that the pH value of the cell wall environment is appropriate and have strong shading and lodging resistance by reducing the discharge of the hydrogen ions, so that the measurement of the flowing condition of the hydrogen ions of the epidermal cells of the plant stalks under the shading condition can be used as a basis for simply and effectively evaluating the relative strength of the shading and lodging resistance of the soybeans.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (8)
1. A method for evaluating the shade resistance and lodging resistance of soybeans by measuring the hydrogen ion flow of stalk epidermal cells is characterized by comprising the following steps:
step S1: polishing the surface of the soybean stalk to form a polished part in the original growing environment of the soybean;
step S2: balancing the ion exchange of epidermal cells at the polished part by adopting a balance buffer solution;
and step S3: measuring the hydrogen ion current of the epidermal cells at the polished part;
and step S4: carrying out shade tolerance and lodging resistance analysis evaluation according to the hydrogen ion flow measurement result;
the equilibrium buffer solution comprises CaCl2 and MES solution;
in the step S4, the shade-tolerant lodging-resistant analysis and evaluation is performed according to the hydrogen ion current measurement result, and the method includes the following steps:
step S4-1: and analyzing the correlation between the flow rate of the hydrogen ions and the internode length and the plant height of the soybeans, recording the correlation coefficient as P, wherein P is more than 0.7, indicating high positive correlation, P is more than 0.4 and less than 0.7, indicating moderate positive correlation, and P is more than 0.2 and less than 0.4, indicating weak positive correlation.
2. The method for evaluating the shade resistance and lodging resistance of soybeans according to the measurement of the hydrogen ion current of the stalk epidermal cells of claim 1, wherein in the step S1, the epidermis of the stalks of the soybeans is sanded by sand paper.
3. The method for evaluating the shade-tolerant lodging-resistant property of soybean according to the measurement of the hydrogen ion current of the stalk epidermal cells of claim 2, wherein the sandpaper is 1000 mesh.
4. The method for evaluating soybean shade-tolerant lodging-resistant property by measuring the hydrogen ion current of the stalk epidermal cells as claimed in claim 1, wherein in the step S2, the epidermal cell ion exchange of the polished part is balanced by using an equilibrium buffer, and the method comprises the following steps:
step S2-1: wetting the wrap with the equilibration buffer;
step S2-2: winding the wrapper around the polished part of the soybean stalk;
step S2-3: during the balancing process, the balancing buffer solution is added to the wrappage until the epidermal cell ion exchange of the polished part reaches the balance.
5. The method for evaluating soybean shade-tolerance and lodging-resistance by measuring the hydrogen ion current of the stalk epidermal cells as claimed in claim 4, wherein the equilibration buffer is added to the wrapper every 25min-30min during equilibration.
6. The method for evaluating soybean shade tolerance and lodging resistance by measuring the flow of hydrogen ions of stalk epidermal cells as claimed in claim 4, wherein in the step S2-3, the amount of the equilibration buffer added to the wrap is equal each time during equilibration.
7. The method for evaluating the shade-tolerant lodging-resistant performance of soybean according to the measurement of the flow of hydrogen ions of the epidermal cells of the stalks in the step S2-3, wherein the time for the ion exchange of the epidermal cells at the ground part to reach the equilibrium is positively correlated with the grinding degree of the plant stalks.
8. The method for evaluating soybean shade resistance and lodging resistance by measuring the hydrogen ion current of the stalk epidermal cells according to claim 1, wherein in the step S3, the hydrogen ion current of the epidermal cells at the polished part is measured by adopting a non-damage micrometering technology.
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