A plant growth regulator and its application in promoting plant growth, increasing plant yield and enhancing plant stress resistance

Abstract

The application discloses a plant growth regulator and application thereof in promoting plant growth, improving plant yield and enhancing plant stress resistance. An active ingredient of the plant growth regulator disclosed by the application is gamma-aminobutyric acid. It is found through experiments that the plant growth regulator has the advantages of promoting plant growth, improving plant yield, improving drought resistance of plants, improving pest stress resistance of plants and improving low nitrogen stress resistance of plants, and the gamma-aminobutyric acid as a natural amino acid derivative has been certified by the US EPA and the Ministry of Health of China, and is safe for humans and mammals. Therefore, the plant growth regulator has high agricultural application and popularization value.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biotechnology, and particularly relates to a plant growth regulator and application thereof in promoting plant growth, improving plant yield and enhancing plant stress resistance, in particular to a plant growth regulator containing gamma-aminobutyric acid and application thereof in promoting plant growth, improving plant yield and enhancing plant stress resistance. BACKGROUND

[0002] Crops will be subjected to various adverse environmental pressures and stresses in the field growth environment, including severe weather conditions and harmful biological stresses, and the use of plant growth regulators is an efficient, economical and environmentally friendly technical measure to improve crop yield and enhance stress resistance. As an important branch of pesticides, plant growth regulators have been widely used in crop field management. At present, there are various types of plant growth regulators used in the field, but many of them still have the disadvantages of single function, large side effects, low efficiency and difficulty in degradation. Therefore, the development of new green and efficient plant growth regulators is the current research focus. However, the development of new plant growth regulators is a complex work that requires manpower and material resources,

[0003] Gamma-aminobutyric acid is a four-carbon non-protein amino acid, which is a chemical substance widely existing in organisms, and belongs to a natural plant signal substance, physiological metabolism regulating substance and nutrient substance. SUMMARY

[0004] The application provides a plant growth regulator.

[0005] The active ingredient of the plant growth regulator provided by the application is gamma-aminobutyric acid.

[0006] Further, the concentration of gamma-aminobutyric acid in the plant growth regulator can be 50-100 mM, and specifically can be 50 mM or 100 mM.

[0007] Further, the plant growth regulator can further contain a gamma-aminobutyric acid receptor agonist, and the gamma-aminobutyric acid receptor agonist can be specifically Muscimol or Baclofen.

[0008] Further, the plant growth regulator can further contain other components, such as synergists, solubilizers, sticking agents and the like.

[0009] In a specific embodiment of the application, the plant growth regulator is composed of gamma-aminobutyric acid and water.

[0010] The plant growth regulator has all or part of the following functions:

[0011] P1, promoting plant growth;

[0012] P2, improving plant yield;

[0013] P3, improving plant drought resistance;

[0014] P4, improving plant pest stress resistance;

[0015] P5, improving plant low nitrogen stress resistance.

[0016] Another object of the present application is to provide a novel use of the above plant growth regulator.

[0017] The present application provides the use of the above plant growth regulator in any one of P1-P5:

[0018] P1, promoting plant growth;

[0019] P2, improving plant yield;

[0020] P3, improving plant drought resistance;

[0021] P4, improving plant pest stress resistance;

[0022] P5, improving plant low nitrogen stress resistance.

[0023] The present application also provides a method for using the above plant growth regulator.

[0024] The method for using the above plant growth regulator provided by the present application comprises the step of preparing a solution of the plant growth regulator and spraying it.

[0025] Further, the spraying is foliar spraying.

[0026] Still further, when the plant is alfalfa, the plant growth regulator is foliar sprayed at the 3-leaf stage or the 6-leaf stage.

[0027] When the plant is tobacco, the plant growth regulator is foliar sprayed at the 4-true-leaf stage.

[0028] The use of the above method in any one of P1-P5 also falls within the protection scope of the present application:

[0029] P1, promoting plant growth;

[0030] P2, improving plant yield;

[0031] P3, improving plant drought resistance;

[0032] P4, improving plant pest stress resistance;

[0033] P5, improving plant low nitrogen stress resistance.

[0034] In any of the above-mentioned applications or methods, the promoting plant growth specifically manifests as increasing plant height and / or increasing plant leaf area index.

[0035] The improving drought resistance of the plant specifically manifests as increasing plant height and / or increasing plant leaf area index and / or increasing plant aboveground biomass and / or increasing plant underground biomass under drought stress.

[0036] The improving insect stress resistance of the plant can be improving aphid stress resistance (improving aphid feeding stress resistance) or improving thrips stress resistance (improving thrips feeding stress resistance). The improving aphid stress resistance or the improving thrips stress resistance specifically manifests as increasing plant leaf area index under aphid or thrips stress.

[0037] The improving low nitrogen stress resistance of the plant specifically manifests as increasing plant growth rate under nitrogen deficiency stress.

[0038] In any of the above-mentioned applications or methods, the plant can be a monocotyledon or a dicotyledon, and the dicotyledon can be alfalfa (such as Zhongmu No. 4) or tobacco (such as K326), and the monocotyledon can be corn (such as Denghai 605).

[0039] In any of the above-mentioned plant growth regulators or applications or methods, the structural formula of the gamma-aminobutyric acid is shown as Formula I.

[0040]

[0041] The structural formula of the Muscimol is shown as Formula IV.

[0042]

[0043] The structural formula of the Baclofen is shown as Formula V.

[0044]

[0045] The present application provides a plant growth regulator, and the active ingredient of the plant growth regulator is gamma-aminobutyric acid. Through experiments, it is found that the plant growth regulator has the functions of promoting plant growth, improving plant yield, improving drought resistance of the plant, improving insect stress resistance of the plant, and improving low nitrogen stress resistance of the plant. Therefore, the plant growth regulator of the present application has high agricultural application and promotion value. BRIEF DESCRIPTION OF DRAWINGS

[0046] Figure 1 Effect of gamma-GABA on plant height and leaf area index of alfalfa

[0047] Figure 2 Effects of γ-GABA on alfalfa plant height, leaf area index, aboveground biomass and underground biomass under drought stress.

[0048] Figure 3 Effects of γ-GABA on alfalfa leaf area index under pest stress.

[0049] Figure 4 Effects of γ-GABA on tobacco leaf area index and yield.

[0050] Figure 5 Effects of γ-GABA on corn growth rate and yield under nitrogen deficiency stress. DETAILED DESCRIPTION

[0051] The application will be further described in conjunction with the specific embodiments, and the examples given are only to illustrate the application, and are not intended to limit the scope of the application. The examples provided below can serve as a guide for further improvement by those skilled in the art, and do not in any way constitute a limitation on the application.

[0052] The experimental methods in the following examples are all conventional methods, and are carried out according to the techniques or conditions described in the literature in the art or according to the product instructions, unless otherwise specified. The materials, reagents, etc. used in the following examples can be obtained commercially, unless otherwise specified.

[0053] The γ-GABA in the following examples is a product of aladdin, with the product number A104200-2.5kg, and the structural formula is shown as formula I.

[0054]

[0055] The Bicuculline in the following examples is a product of MedChemExpress, with the product number HY-N0219Bicuculline, and the structural formula is shown as formula II.

[0056]

[0057] The Benzodiazepines in the following examples is a product of Bio-rad, with the product number 1100-5004, and the structural formula is shown as formula III.

[0058]

[0059] The Muscimol in the following examples is a product of TRC, with the product number M815000, and the structural formula is shown as formula IV.

[0060]

[0061] Baclofen in the following examples is a product of Macklin / McCline, item no. A830520-25g, and has the structure shown in Formula V.

[0062]

[0063] Example 1, Use of γ-GABA in Promoting the Growth of Alfalfa

[0064] I. Test Site and Materials

[0065] Medicago sativa L. (Zhongmou No. 4) was selected as the test variety for field tests in alfalfa fields in Yinchuan, Ningxia, China.

[0066] II. Test Method

[0067] Alfalfa seedlings at the 3-leaf stage with uniform growth were selected and planted in all test plots with a row spacing of 75 cm and a plant spacing of 30 cm. The area of each test plot was set to 30 m 2 . All test plots were randomly grouped, and 3 repeated test plots were set for each alfalfa treatment group. All test plots were managed according to conventional cultivation methods, and the cultivation conditions were kept consistent.

[0068] The following treatment groups were set according to whether γ-GABA was sprayed or not:

[0069] Control group (Control): Water was sprayed on the alfalfa seedlings as a foliar spray treatment, and spraying was performed every 5 days.

[0070] GABA 50 treatment group: γ-Aminobutyric acid (γ-GABA) was dissolved in water to prepare a γ-GABA solution with a concentration of 50 mM, and the alfalfa seedlings were sprayed with the γ-GABA solution with a concentration of 50 mM as a foliar spray treatment, and spraying was performed every 5 days.

[0071] GABA 100 treatment group: γ-GABA was dissolved in water to prepare a γ-GABA solution with a concentration of 100 mM, and the alfalfa seedlings were sprayed with the γ-GABA solution with a concentration of 100 mM as a foliar spray treatment, and spraying was performed every 5 days.

[0072] The height and leaf area index of alfalfa were measured for 30 days. The leaf area index of alfalfa was measured by LAI-2200C plant canopy analyzer. The measurement was carried out at 19:00 in the afternoon on sunny days. The specific measurement method is as follows: select 270° view cover cap, measurement point is located in the middle row of each plot, measurement point position is approximately S-shaped. When measuring, read 1 A value on the canopy and 5 B values under the canopy, try to reduce the time difference between reading on and under the canopy, repeat three times, and calculate the average LAI value, which is the leaf area index of the treatment.

[0073] The measured data of alfalfa height and leaf area index were statistically analyzed. All test data were analyzed by SPSS 25.0. The analysis method was ANOVA for difference analysis, and P<0.05 indicated significant difference.

[0074] III. Test results

[0075] The results showed that exogenous γ-GABA spraying had a significant promoting effect on alfalfa growth, which was embodied in that γ-GABA produced a significant difference in alfalfa plant height with the control group after 10 days of application Figure 1 A); γ-GABA produced no significant difference in alfalfa leaf area index with γ-GABA 50mM and γ-GABA 100mM concentrations after 10 days of application; but after 15 days of application, γ-GABA increased the leaf area index of alfalfa, which was significantly different from the control group Figure 1 B). It was shown that exogenous γ-GABA spraying significantly improved the growth rate of alfalfa plants, and the leaf area index was not significantly different between γ-GABA 50mM and γ-GABA 100mM concentrations, but was significantly higher than the control group. It was shown that γ-GABA could significantly promote the growth of alfalfa.

[0076] Example 2, Application of γ-GABA in improving drought resistance of alfalfa

[0077] I. Test site and materials

[0078] Alfalfa (Zhongmou No. 4) was selected as the test variety for cultivation test in Yinchuan City, Ningxia, China.

[0079] II. Test method

[0080] Alfalfa seedlings with uniform size at the 3-leaf stage were selected for potting test in the test plot, and the area of each test plot was set to 30m 2 . All test plots were randomly grouped, and 4 repeated test plots were set for each alfalfa treatment group. All test plots were managed according to the conventional cultivation management method, and the cultivation conditions were kept consistent.

[0081] According to whether drought treatment is carried out and whether γ-GABA is sprayed, the following various treatment groups are set up:

[0082] Irrigation (Neg. Control) treatment group: The soil moisture content is ensured by weighing method, the soil moisture content is ensured to be 75% ± 5% of the maximum field water holding capacity, and no plant growth regulator is applied.

[0083] Drought (Pos. Control) treatment group: The soil moisture content is ensured by weighing method, the soil moisture content is kept at 35% ± 5% of the maximum field water holding capacity, and no plant growth regulator is applied.

[0084] GABA 50 treatment group: The soil moisture content is ensured by weighing method, the soil moisture content is kept at 35% ± 5% of the maximum field water holding capacity, and the alfalfa seedlings are sprayed with a γ-GABA solution (solvent is water) with a concentration of 50 mM every 5 days.

[0085] GABA 100 treatment group: The soil moisture content is ensured by weighing method, the soil moisture content is kept at 35% ± 5% of the maximum field water holding capacity, and the alfalfa seedlings are sprayed with a γ-GABA solution (solvent is water) with a concentration of 100 mM every 5 days.

[0086] The plant height, leaf area index, aboveground biomass and Underground biomass.

[0087] The data related to plant height, leaf area index, aboveground biomass and underground biomass are statistically analyzed. All test data are analyzed by SPSS 25.0. The analysis method is ANOVA for difference analysis, and P < 0.05 indicates significant difference.

[0088] III. Test results

[0089] The results show that exogenous γ-GABA can significantly improve the ability of alfalfa to cope with drought stress, which is specifically manifested as follows: under drought stress, the plant height, leaf area index, aboveground biomass and underground biomass of alfalfa are significantly reduced, but the exogenous γ-GABA spraying significantly improves the plant height, leaf area index, aboveground biomass and underground biomass of alfalfa. Figure 2 ). It is proved that γ-GABA can significantly improve the drought tolerance of alfalfa.

[0090] Example 3, application of γ-GABA in improving the ability of alfalfa to resist insect stress

[0091] I. Test site and materials

[0092] In China, alfalfa (Zhongmou No. 4) is selected as the test variety for potting test in Yinchuan City, Ningxia.

[0093] II. Experimental Methods

[0094] Select 3-leaf stage alfalfa seedlings with uniform growth size for pot experiment in test plots, and set the area of each test plot to 30 m 2 All test plots are randomly grouped, and 4 repeated test plots are set for each alfalfa treatment group. All test plots are managed according to the conventional cultivation management method, and the cultivation conditions are kept consistent. The alfalfa seedlings are cultured to 6-leaf stage for insect introduction experiment.

[0095] According to whether the insect introduction treatment is performed and whether γ-GABA is sprayed, the following various treatment groups are set:

[0096] Negative control (Control): 100 aphids or thrips are introduced on the leaves of each pot of alfalfa, and then the net is covered, and no plant growth regulator is applied.

[0097] GABA treatment group (aphid): 100 aphids are introduced on the leaves of each pot of alfalfa, and then the net is covered, and the alfalfa seedlings are treated with γ-GABA solution with a concentration of 50 mM (solvent is water) by foliar spraying, and the spraying is performed every 5 days.

[0098] GABA treatment group (thrips): 100 thrips are introduced on the leaves of each pot of alfalfa, and then the net is covered, and the alfalfa seedlings are treated with γ-GABA solution with a concentration of 50 mM (solvent is water) by foliar spraying, and the spraying is performed every 5 days.

[0099] The alfalfa in all test plots is continuously observed for 30 days, and the leaf area index is measured.

[0100] The measured leaf area index data is statistically analyzed. All test data is analyzed using SPSS 25.0. The analysis method uses ANOVA for difference analysis, and P<0.05 indicates significant difference.

[0101] III. Test Results

[0102] The results show that under aphid stress, exogenous γ-GABA spraying can significantly increase the leaf area index of alfalfa, which increases by 46.8% (a) compared with the control; similarly, under thrips stress, exogenous γ-GABA spraying can significantly increase the leaf area index of alfalfa, which increases by 49.8% (b) compared with the control. Figure 3 Figure 3 Therefore, exogenous γ-GABA can significantly enhance the resistance of alfalfa to the feeding stress of pests.

[0103] Example 4, Application of γ-GABA in promoting tobacco growth and increasing tobacco yield

[0104] I. Experimental site and materials ​​

[0105] Tobacco (K326) was selected as the test variety for field experiments in tobacco fields in Yimen County, Yunnan Province, China.

[0106] II. Test method

[0107] Four tobacco seedlings with uniform growth size at the four-leaf stage were selected and evenly planted in all test plots in the tobacco field, with a planting density of 1000 plants per mu. The area of each test plot was set to 30 m 2 All test plots were randomly grouped, and three repeated test plots were set for each tobacco treatment group. All test plots were managed according to the conventional cultivation management method, and the cultivation conditions were kept consistent.

[0108] According to the type of plant growth regulator sprayed, the following treatment groups were set up:

[0109] Control group (Control): Water was used for foliar spraying treatment of tobacco seedlings, and spraying was performed every 5 days.

[0110] GABA 50 treatment group: γ-GABA was dissolved in water to prepare a γ-GABA solution with a concentration of 50 mM, and the tobacco seedlings were treated with foliar spraying with the γ-GABA solution with a concentration of 50 mM, and spraying was performed every 5 days.

[0111] GABA + Bicuculline treatment group: γ-GABA was dissolved in water to prepare a γ-GABA solution with a concentration of 50 mM, and Bicuculline was dissolved in water to prepare a Bicuculline solution with a concentration of 50 mM, and the tobacco seedlings were treated with foliar spraying with the γ-GABA solution with a concentration of 50 mM and the Bicuculline solution with a concentration of 50 mM at the same time, and spraying was performed every 5 days.

[0112] GABA + Benzodiazepines treatment group: γ-GABA was dissolved in water to prepare a γ-GABA solution with a concentration of 50 mM, and Benzodiazepines was dissolved in water to prepare a Benzodiazepines solution with a concentration of 50 mM, and the tobacco seedlings were treated with foliar spraying with the γ-GABA solution with a concentration of 50 mM and the Benzodiazepines solution with a concentration of 50 mM at the same time, and spraying was performed every 5 days.

[0113] GABA+Muscimol treatment group: γ-GABA was dissolved in water to prepare a γ-GABA solution with a concentration of 50 mM, Muscimol was dissolved in water to prepare a Muscimol solution with a concentration of 50 mM, and tobacco seedlings were simultaneously sprayed with the γ-GABA solution with a concentration of 50 mM and the Muscimol solution with a concentration of 50 mM, once every 5 days.

[0114] GABA+Baclofen treatment group: γ-GABA was dissolved in water to prepare a γ-GABA solution with a concentration of 50 mM, Baclofen was dissolved in water to prepare a Baclofen solution with a concentration of 50 mM, and tobacco seedlings were simultaneously sprayed with the γ-GABA solution with a concentration of 50 mM and the Baclofen solution with a concentration of 50 mM, once every 5 days.

[0115] Tobacco in all test plots was observed continuously for 50 days, and leaf area index and yield were determined.

[0116] The data related to the determined leaf area index and yield were statistically analyzed. All test data were analyzed using SPSS 25.0. The analysis method was ANOVA for difference analysis, and P < 0.05 indicated that there was a significant difference Different.

[0117] III. Test Results

[0118] The results showed that exogenous GABA could significantly increase the leaf area index and yield of tobacco; the simultaneous application of GABA and receptor inhibitors (Bicuculline or Benzodiazepines) significantly reduced the leaf area index and yield of tobacco; the simultaneous application of GABA and agonists (Muscimol or Baclofen) significantly increased the leaf area index and yield of tobacco Figure 4 ). It was shown that γ-GABA could significantly promote the growth of tobacco and increase the yield of tobacco.

[0119] Example 5, Application of γ-Aminobutyric Acid in Improving the Low Nitrogen Stress Resistance of Corn

[0120] I. Test Site and Materials

[0121] Corn (Denghai 605) was selected as the test variety in a corn field in Kaifeng City, Henan Province, China for field testing.

[0122] II. Test Method

[0123] Uniformly planted 3-leaf stage corn seedlings with the same size in all test plots in the corn field, with a planting density of 4000 plants per mu. The area of each test plot was set to 30 m2 All test plots were randomly grouped, and 3 repeated test plots were set for each corn treatment group. All test plots were managed according to the conventional cultivation management method, and no additional nitrogen fertilizer was applied, i.e. all test plots were subjected to nitrogen deficiency treatment, and other cultivation conditions were kept consistent.

[0124] The following various treatment groups were set according to whether γ-GABA was sprayed or not:

[0125] Control group (Neg. control): water was used to spray corn seedlings, and spraying was performed once every 5 days.

[0126] GABA 50 treatment group: γ-GABA was dissolved in water to prepare a γ-GABA solution with a concentration of 50 mM, and the corn seedlings were sprayed with the γ-GABA solution with a concentration of 50 mM, and spraying was performed once every 5 days.

[0127] All test plots of corn were continuously observed for 50 days, and the growth rate and yield of corn were measured. The increase in height was measured using the fixed plant measurement method, and then the average was calculated. At the beginning of the experiment, 5 corn plants with similar growth heights were randomly selected in the test plot, their positions were marked, and their straight heights were measured from the ground to the top of the plant. The same plants were measured every 5 days. The growth rate was calculated based on the measured plant height and the number of growth days.

[0128] The data related to the measured growth rate and yield of corn were statistically analyzed. All test data were analyzed using SPSS 25.0. The analysis method was ANOVA for difference analysis, and P < 0.05 indicated a significant difference.

[0129] III. Test Results

[0130] The results showed that under the condition of nitrogen deficiency, compared with the control group, the additional application of γ-GABA could significantly promote the growth rate of corn ( Figure 5 a); and after exogenous supplementation of γ-GABA, the yield of corn had no significant difference with the control ( Figure 5 b). Therefore, γ-GABA can significantly improve the ability of corn to resist low nitrogen stress.

[0131] The application has been described in detail. For those skilled in the art, the application can be implemented in a wider range under the same parameters, concentrations and conditions without departing from the spirit and scope of the application and without unnecessary experiments. Although the application gives a special example, it should be understood that the application can be further improved. In summary, according to the principle of the application, the application intends to include any change, use or improvement of the application, including changes made by conventional techniques known in the art, which deviates from the range disclosed in the application. Some basic features can be applied within the scope of the following attached claims.

Claims

1. Use of a plant growth regulator in improving the ability of a plant to resist insect stress, wherein the active ingredient of the plant growth regulator is γ-aminobutyric acid; the ability to resist insect stress is the ability to resist aphid stress or the ability to resist thrips stress; the plant is alfalfa; the concentration of γ-aminobutyric acid in the plant growth regulator is 50-100 mM.

2. Use according to claim 1, characterized in that: the plant growth regulator further comprises a γ-aminobutyric acid receptor agonist.

3. Use according to claim 2, characterized in that: the γ-aminobutyric acid receptor agonist is Muscimol or Baclofen.

4. Use according to claim 1 or 2, characterized in that: the plant growth regulator further comprises a synergist or a solubilizer or a spreading agent.

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