Composition based on methyl carboline acid and application of composition in stress resistance and growth promotion of plants

Abstract

The invention belongs to the technical field of biology, and particularly discloses application of several compositions based on methyl carboline acid in stress resistance and growth promotion of plants. The invention solves the problems of single function, poor effect, large dosage, high cost and the like of the plant regulator in the market, and the methyl carboline acid, paclobutrazol, alginate oligosaccharide and diethyl aminoethyl hexanoate are compounded for use under a specific concentration, so that the growth of crops can be obviously promoted, and the yield and quality of the crops can be improved. The methyl carboline acid and the gamma-aminobutyric acid are compounded for use under a specific concentration, so that the influence of saline-alkali stress on the growth of crops such as wheat and cucumber can be remarkably relieved; the methyl carboline acid and the 5-aminolevulinic acid are compounded for use under a specific concentration, so that the growth of crops such as tomatoes and peanuts at a low temperature can be promoted. The stress-resistant and growth-promoting composition provided by the invention can be used for remarkably improving the self resistance of plants under a proper concentration, effectively promoting the growth and development of the plants and improving the yield and quality of crops.

Description

Technical Field

[0001] The invention belongs to the technical field of plant growth regulators, and in particular relates to a composition based on methylcarbolinic acid and its application in plant stress resistance and growth promotion. Background Art

[0002] During the growth process of plants, they are susceptible to abiotic stresses such as low temperature and salinity, which affect plant growth and cause crop yield reduction, and have a significant impact on the distribution and productivity of agricultural crops. This impact further poses a challenge to food safety and the sustainable development of agriculture. With the development of modern agriculture, people's demand for agricultural product output and quality has increased, and the long-term irrational use of traditional pesticides and fertilizers has caused harm to the ecological environment and food security. Safe, green and efficient plant cultivation products have become an inevitable trend in agricultural development.

[0003] Plant growth regulators include plant hormones, natural compounds and synthetic compounds that have physiological effects on plant growth and development. There are many types of plant growth regulators. At present, there are hundreds of registered plant growth regulators in my country. However, the plant growth regulator products on the market are of varying quality, with problems such as single function, poor effect, large dosage and high cost. Methylcarboline acid is a newly discovered plant growth regulator, and there is still little research on it. There is even less research on its combination with other plant growth regulators in the field of plant stress resistance and growth promotion.

[0004] Chinese patent document CN117204433A (application number: 201510059660.1) discloses the application and preparation method of the compound 1-methyl-1,234-tetrahydro-β-carboline 3-carboxylic acid. The patent document discloses the preparation and application of the compound methylcarboline acid obtained by separation and purification from the polysaccharide mushroom YXI. The patent document discloses that the compound has the effects of drought resistance, antiviral resistance and promoting plant growth and yield when combined with fertilizers, but does not conduct in-depth research on the stress resistance and growth-promoting effects of methylcarboline acid combined with other plant regulators. Summary of the invention

[0005] In order to solve the defects of the prior art, the present invention uses a plant growth regulator, methylcarboline acid, in combination with conventional plant growth regulators for stress resistance and growth promotion, which can improve the plant's own resistance, effectively promote plant growth, and have a significant synergistic effect. The technical solution provided by the present invention is as follows:

[0006] As a first aspect of the present invention, there is provided a composition based on methylcarbolinic acid, comprising methylcarbolinic acid and other plant growth regulators, wherein the other plant growth regulators are selected from one or more of brown algae oligosaccharides, paclobutrazol, aminoethyl aminobutyric acid, γ-aminobutyric acid (GABA), and 5-aminolevulinic acid. The effective concentration of methylcarbolinic acid is 1 to 200 ng / mL.

[0007] In another aspect, the present invention also provides the use of a composition based on methylcarbolinic acid in plant stress resistance and growth promotion.

[0008] Preferably, the methylcarboline acid is obtained by separation and purification from the Pholiota adiposa strain (Pholiota adiposa) YX1, and the preservation number of the Pholiota adiposa strain (Pholiota adiposa) YX1 is CGMCC No. 21077. The prior patent "Application and preparation method of compound 1-methyl-1,2,3,4-tetrahydro-β-carboline-3-carboxylic acid" (application number: CN202310972464.8) promotes crop growth, increases crop yield, and improves crop resistance to drought stress.

[0009] The present invention verifies the application of methylcarboline acid (1-methyl-1,2,3,4-tetrahydro-β-carboline-3-carboxylic acid) compounded with different plant regulators in plant stress resistance, growth promotion and synergy enhancement.

[0010] In one embodiment of the present invention, a composition comprising methylcarbolinic acid, brown algae oligosaccharides, paclobutrazol, and aminoethyl hexanoate is provided, wherein the effective concentration of the methylcarbolinic acid is 1 to 200 ng / mL, preferably 10 ng / mL, the effective concentration of paclobutrazol (15%) is 0.5 to 10 mg / mL, preferably 2.50 mg / mL, the effective concentration of brown algae oligosaccharides is 0.25 to 5 mg / mL, preferably 0.5 mg / mL, and the effective concentration of aminoethyl hexanoate DA-6 is 5 to 30 mg / L, preferably 10 mg / L.

[0011] Furthermore, the present invention provides the effect of a composition comprising methylcarbolinic acid, brown alginic oligosaccharide, paclobutrazol and diethylaminoethyl hexanoate in promoting plant growth, wherein the plants include Chinese cabbage, lettuce and the like.

[0012] As a highly active seaweed extract obtained by degrading macromolecular alginate, brown algae oligosaccharides can act as an excitant to induce tobacco suspended cells to produce excessive reactive oxygen in a short period of time, thereby initiating the signal transduction program of intracellular reactive oxygen, stimulating the increase of antioxidant enzyme activity in the plant, and maintaining intracellular free radicals at a low level, thereby reducing plant cell membrane damage, reducing electrolyte extravasation, and improving plant stress resistance. Adding 0.2% to 1.0% brown algae oligosaccharides during corn planting can significantly improve the fertilizer utilization rate and corn yield of corn. Exogenous brown algae oligosaccharides can promote crop growth and development, thereby improving yield and quality.

[0013] Paclobutrazol is a plant growth regulator that can delay the growth of basal leaves, promote tillering, promote carbohydrate output, and increase the content of chlorophyll, protein and nucleic acid in plants by regulating the levels of endogenous hormones and carbon and nitrogen nutrients in plants. It has the effects of delaying plant growth, inhibiting stem elongation, shortening internodes, promoting plant tillering, increasing plant stress resistance, and improving yield.

[0014] Aminoethyl hexanoate (DA-6) is a broad-spectrum plant growth regulator. After being absorbed by the plants, it can increase the content of chlorophyll, protein, and nucleic acid in the plants; improve photosynthetic efficiency, improve the carbon and nitrogen metabolism of the plants, enhance the plants' absorption of water and fertilizer, and regulate the water balance in the plants. It has the effects of significantly increasing the yield, improving resistance, and improving quality of many crops.

[0015] In one embodiment of the present invention, a composition comprising methylcarbolinic acid and γ-aminobutyric acid is provided, wherein the effective concentration of the methylcarbolinic acid is 1 to 200 ng / mL, preferably 10 ng / mL, and the effective concentration of the γ-aminobutyric acid is 0.1 mg / mL to 5.0 mg / mL, preferably 0.5 mg / mL.

[0016] Furthermore, the present invention provides the use of a composition comprising methylcarbolinic acid and γ-aminobutyric acid in the resistance of plants to salt-alkali stress, and the plants include cucumber and wheat.

[0017] γ-Aminobutyric acid (GABA) is a four-carbon non-protein amino acid widely found in animals, plants and microorganisms. A large number of studies have shown that GABA can act as a signal substance or energy substance, participate in various physiological and biochemical reactions of plants, regulate plant growth and development and response to adverse stress, and exogenous GABA can also be absorbed by plants as an alternative to organic nitrogen fertilizer, promote photosynthesis and affect energy metabolism, promote plant nutritional growth and reproductive growth, improve plant resistance to different adversities, and promote the absorption of trace elements by plants.

[0018] In one embodiment of the present invention, a composition comprising methylcarboline acid and 5-aminolevulinic acid is provided, wherein the effective concentration of the methylcarboline acid is 1 to 200 ng / mL, preferably 10 ng / mL, and the effective concentration of the 5-aminolevulinic acid is 0.05 to 10 mg / mL, preferably 0.20 mg / mL.

[0019] Furthermore, the present invention provides the use of a composition based on methylcarboline acid containing 5-aminolevulinic acid in the resistance of plants to low temperature stress, wherein the plants include tomatoes and peanuts.

[0020] 5-Aminolevulinic acid (ALA) was first discovered in the culture medium of Chlorella vulgaris. It is a natural amino acid widely present in living cells of bacteria, fungi, animals and plants. ALA can enhance the tolerance of plants to low temperatures through various mechanisms. Exogenous ALA can enhance the activity of antioxidant enzyme systems and the efficiency of photosynthesis, increase the content of osmotic substances and polyamines, activate the signal transduction of NO and γ-aminobutyric acid, protect organelles, enhance chlorophyll synthesis and lutein cycle, and regulate plant secondary metabolism and mineral element absorption, thereby improving plant low temperature tolerance. Exogenous ALA can also enhance the low temperature tolerance of plant seedlings by regulating glutathione metabolism, β-alanine metabolism, phenylalanine metabolism and starch content.

[0021] After adopting this technical solution, 1-methyl-1,2,3,4-tetrahydro-β-carboline-3-carboxylic acid (methylcarboline acid) has good stress resistance and growth-promoting effects on plants.

[0022] Mechanism of methylcarbolinic acid to resist low temperature: It increases the activity of oxidases such as superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) in plants under low temperature environment, reduces the content of hydrogen peroxide and malondialdehyde (MDA), alleviates cell membrane damage, stabilizes cell membrane structure, improves the antioxidant capacity of plant membrane, reduces cell osmotic regulation, and reduces the degree of cell membrane damage. Mechanism of methylcarbolinic acid to resist salt and alkali: It increases the content of free proline and soluble sugar in plants, improves its osmotic regulation ability, reduces MDA content, increases the activity of antioxidant enzymes to regulate the content of abscisic acid in plants, and thus improves the salt and alkali tolerance of plants.

[0023] Methylcarbolinic acid, brown algae oligosaccharides, γ-aminobutyric acid, 5-aminolevulinic acid, paclobutrazol and ammonium chloride are all plant growth regulators with the effects of promoting growth, resisting stress and controlling growth. Their combination can not only make up for the shortcomings, but also have their own unique functions. The combination can enhance the effect, expand the function, reduce the dosage of a single dose and save costs. They show synergistic effects under plant growth and adverse stress.

[0024] The beneficial effects of the present invention are as follows:

[0025] 1. The present invention uses multiple tests to determine the optimal ratio of three plant conditioning agents: methylcarboline acid, paclobutrazol (15%), brown algae oligosaccharide and aminoethyl hexanoate DA-6, and verifies their significant role in promoting plant growth, increasing yield and improving quality;

[0026] 2. The combined application of methylcarboline acid with γ-aminobutyric acid and 5-aminolevulinic acid was found to improve the resistance of plants to low temperature and salinity;

[0027] 3. The absolute dosage is small. Experiments show that 0.1g of methylcarbolinic acid mixed with paclobutrazol (15%), brown algae oligosaccharides, aminoethyl ester DA-6, γ-aminobutyric acid and 5-aminolevulinic acid can be sprayed on 1 mu of crops, and it has obvious growth-promoting and stress-resistant effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] The accompanying drawings in the specification, which constitute a part of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0029] Figure 1 The figure shows the growth-promoting phenotypes of lettuce after the combination of methylcarbolinic acid and three compositions in Example 3 (the first row on the left is the water control, the second on the left is the single-dose treatment of methylcarbolinic acid, and the third on the left is the single-dose treatment of paclobutrazol; the second row on the left is the single-dose treatment of brown algae oligosaccharide, the second on the left is the single-dose treatment of aminoethyl hexanoate, and the third on the left is the combination treatment of methylcarbolinic acid with brown algae oligosaccharide, paclobutrazol, and aminoethyl hexanoate);

[0030] Figure 2 The figure shows the effect of the combined use of methylcarbolinic acid and γ-aminobutyric acid in Example 4 on the germination characteristics of wheat seeds under salt stress conditions (from left to right in the figure: clear water control under normal conditions, clear water control under salt stress conditions, single-dose treatment of γ-aminobutyric acid under salt conditions, single-dose treatment of methylcarbolinic acid under salt conditions, and combined treatment of γ-aminobutyric acid and methylcarbolinic acid under salt conditions);

[0031] Figure 3 The phenotype of cucumber seedlings under saline-alkali stress conditions after the combination of methylcarboline acid and γ-aminobutyric acid in Example 4 is shown (the left 1 in the figure is a clear water control under normal conditions, the left 2 is a clear water control under saline-alkali conditions, the left 3 is a single-dose treatment of γ-aminobutyric acid under saline-alkali conditions, the left 4 is a single-dose treatment of methylcarboline acid, and the left 5 is a combination treatment of γ-aminobutyric acid and methylcarboline acid);

[0032] Figure 4 The figure shows the effect of the combination of methylcarboline acid and 5-aminolevulinic acid on the phenotype of tomatoes under low temperature stress in Example 5 (left 1 is a water control at room temperature, left 2 is a water control at low temperature, left 3 is a single-dose treatment of 5-aminolevulinic acid at low temperature, left 4 is a single-dose treatment of methylcarboline acid at low temperature, and left 5 is a combination treatment of 5-aminolevulinic acid and methylcarboline acid at low temperature);

[0033] Figure 5The figure shows the phenotype of peanut plants under low temperature stress conditions after the combination of methylcarboline acid and 5-aminolevulinic acid in Example 5 (left 1 is a water control, left 2 is a single-dose 5-aminolevulinic acid treatment, left 3 is a single-dose methylcarboline acid treatment, and left 4 is a combination of 5-aminolevulinic acid and methylcarboline acid treatment);

[0034] Figure 6 The figure shows the contents of MDA, proline, soluble sugar and SOD in cucumber leaves under saline-alkali stress conditions after the combination of methylcarboline acid and γ-aminobutyric acid in Example 4;

[0035] Figure 7 The figure shows the contents of MDA, proline, soluble sugar and SOD in peanut seedlings under low temperature stress after the combination of methylcarboline acid and 5-aminolevulinic acid in Example 5. DETAILED DESCRIPTION

[0036] It should be noted that the following detailed descriptions are exemplary and are intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which the present invention belongs.

[0037] The fatty agaricus YX1 was first disclosed in patent CN113637594B "A fatty agaricus YX1, a cultivation method and its application". It was collected from the wild forest of Mount Tai and obtained by separation, purification and culture. The fatty agaricus (Pholiotaadiposa) YX1 was deposited in the General Microbiology Center of China Microorganism Culture Collection Committee (CGMCC for short, address: No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences) on December 1, 2020, with the deposit number CGMCC NO.21077.

[0038] Example 1 Preparation of Methylcarboline Acid

[0039] The polypore agaricus YX1 is fermented and cultured at 24°C and 180 r / min to obtain a fermentation broth, and the fermentation broth is centrifuged to obtain fermentation mycelium; the mycelium is washed, dried, weighed, and crushed, the crushed mycelium is mixed with ethanol and ultrasonically treated to obtain an extract, which is then freeze-dried and dissolved in water to prepare a solution, which is adsorbed using a macroporous resin, and the resin adsorbate is eluted with ethanol, and then the adsorbate is separated using 80% acetonitrile, and the supernatant is collected. The supernatant is separated by HPLC to obtain the compound 1-methyl-1,2.3.4-tetrahydro-B-carboline-3-carboxylic acid.

[0040] Example 2 Experiment on effective concentration of methylcarboline acid

[0041] (1) Experimental design:

[0042] This experiment was an indoor potted plant experiment. Arabidopsis, wheat, corn, rice, tomato, pepper, soybean, and peanut were selected as test materials. Ten treatments were set up, namely, water control (CK), methylcarboline acid 1ng / mL, 5ng / mL, 10ng / mL, 20ng / mL, 50ng / mL, 100ng / mL, 150ng / mL, 200ng / mL, and 250ng / mL. Take an appropriate amount of sterilized nutrient soil and put it into the flowerpot. The flowerpot has a diameter of 15cm and a height of 20cm. Sow 5 seeds or tubers of the above plants in each flowerpot, evenly cover a layer of nutrient soil, and manage each treatment uniformly. After the plants emerge for one week, each flowerpot is watered with 50mL of methylcarboline acid of different concentrations, and the control treatment is watered with an equal amount of water, and watered once every two weeks. Measure the fresh weight of the plants one month after emergence.

[0043] (2) Test results:

[0044] The results are shown in Table 1. The concentration of methylcarboline acid in the range of 1-200 ng / mL showed different degrees of promotion on the growth of different crops. When the concentration of methylcarboline acid reached 250 ng / mL, it showed an inhibitory effect on the growth of a few plants, and had no obvious growth-promoting effect on most plants. In the range of 1-200 ng / mL, the fresh weight of most plants was optimal when the concentration of methylcarboline acid was 10 ng / mL. At this concentration, the fresh weight of soybeans, peanuts, wheat, corn, and peppers increased by 13.81%, 24.54%, 30.99%, 23.75%, and 34.72% respectively compared with CK.

[0045] Table 1 Effects of different concentrations of methylcarboline on the fresh weight (g) of different plants

[0046]

[0047] Example 3 Growth-promoting effect of a composition of methylcarboline acid and various plant regulators on plants

[0048] 1. Screening of the concentration of compound of methylcarboline acid, brown algal oligosaccharide, paclobutrazol and aminoethyl hexadecene

[0049] (1) Experimental design:

[0050] The pakchoi seedlings were planted and the film was removed after two weeks. Lettuce seedlings of the same size were selected and transplanted into small pink pots. Every 6 pots were placed in a tray. After 5 days of seedling acclimatization, the treatment solution was sprayed to make the treatment solution evenly adhere to the surface of the lettuce leaves. Methylcarboline acid, brown algae oligosaccharide, paclobutrazol, and aminoethyl hexadecene were set at three different concentrations, and the pakchoi yield was measured after harvest, in order to obtain the best concentration combination of the four planting adjustments for promoting the growth and yield of pakchoi.

[0051] (2) Test results

[0052] Table 2 Effects of different concentrations of compound treatments on the growth of Chinese cabbage

[0053]

[0054] As shown in Table 2, compared with CK, the four agents applied alone or in combination at different concentrations promoted the growth of pakchoi to varying degrees. Among them, the combination of 10 ng / mL methylcarbolinic acid, 2.50 mg / mL 15% paclobutrazol, 0.5 mg / mL brown algae oligosaccharide, and 10 mg / L aminoethyl hexadecene had the most significant promoting effect on pakchoi. The fresh weight of pakchoi reached 11.31 g, which was 48.23%, 12.99%, 30.15%, 26.09%, and 18.18% higher than that of CK treatment and single application of methylcarbolinic acid, 15% paclobutrazol, brown algae oligosaccharide, and aminoethyl hexadecene, respectively.

[0055] 2. Compound agents promote the growth and development of lettuce

[0056] (1) Experimental design

[0057] The optimal compound concentrations of the four planting agents in the previous experiment were selected for subsequent experimental verification. The experiment set up 6 treatments, namely, water control (CK), 10ng / mL methylcarbolinic acid (methylcarbolinic acid), 2.5mg / mL 15% paclobutrazol (paclobutrazol), 0.5mg / mL brown algae oligosaccharide (brown algae oligosaccharide), 10mg / L aminoethyl hexanoate (aminoethyl hexanoate), methylcarbolinic acid + brown algae oligosaccharide + paclobutrazol + aminoethyl hexanoate (combination). Lettuce was selected as the test crop, and the growth indicators and quality indicators such as the number of lettuce leaves, the maximum leaf area, the leaf SPAD value, and the fresh weight were measured and counted after harvest.

[0058] (2) Test results

[0059] As shown in Table 3 and Figure 1 As shown in the results, compared with CK, the single application of the four agents promoted the growth of lettuce to varying degrees and improved the quality of lettuce. Among them, methylcarbophosphate and brown algae oligosaccharide had a better effect on the growth of lettuce, paclobutrazol and methylcarboline acid had the best effect on the yield of lettuce, and aminoethyl ...

[0060] Table 3 Effects of different treatments on lettuce growth

[0061]

[0062] Note: Different lowercase letters indicate significant differences at the 0.05 level.

[0063] Example 4: Methylcarboline acid combined with γ-aminobutyric acid to improve crop resistance to salt-alkali stress

[0064] 1. Promote the germination characteristics of wheat seeds under salt stress

[0065] (1) NaCI salt stress concentration screening

[0066] Six NaCI concentration gradients were set, namely 0, 50, 100, 150, 200, and 250 g / L. Wheat seeds (Jimai 22) of uniform size and fullness were selected, disinfected and dried, and soaked for 20 hours. Two layers of sterile filter paper were placed in the culture dish, and 10 mL of salt solution of different concentrations was added to each dish. The filter paper was replaced and 10 mL of solution was added again every day; each treatment was repeated 3 times, and 30 soaked seeds were added in each repetition. The culture temperature condition was 23 ° C (16 / 8h), and the culture was protected from light. From the day of seed germination, the total number of seed germinations and the number of newly germinated seeds were counted regularly every day. After culturing to the 7th day, various indicators were measured, and the test concentration of the NaCI solution was finally determined to be 200 mmol / L.

[0067] (2) Experimental design

[0068] There were five treatment groups, including normal treatment (CK0), salt treatment group (CK1), 0.5mg / mL γ-aminobutyric acid (GABA), 10ng / mL methylcarboline acid (methylcarboline acid), and 0.5mg / mL γ-aminobutyric acid + 10ng / mL methylcarboline acid (GABA+methyl). CK0 and CK1 were treated with water, and the other three groups were treated with agents. After 20h, the seeds in the CK0 group were treated with 10ml water, and the seeds in the CK1 group and the other three agent groups were treated with an equal volume of 200mmol / L NaCI solution, 30 seeds per dish, and repeated three times. The total number of germinated seeds and the number of newly germinated seeds were counted regularly every day. After 7d, 10 seedlings were taken from each dish to measure the root length and bud length, and the indicators were calculated according to the following formula:

[0069] Germination rate = number of seeds germinated within 7 days / total number of seeds

[0070] Germination potential = number of seeds germinated within 3 days / total number of seeds

[0071] Germination index = ∑ daily germination number / corresponding germination days

[0072] Seedling activity index = germination index x 7 d sprout length

[0073] Salt tolerance germination index = germination index of treatment / germination index of control

[0074] (3) Test results

[0075] The germination of wheat seeds in different treatment groups is shown in Table 4. Figure 2 It can be seen that the germination and growth of wheat seeds were significantly inhibited under salt stress, and all indicators of CK1 were significantly reduced. Compared with CK1, the three groups of drug treatments alleviated the impact of salt stress on wheat to a certain extent. Among them, all indicators of wheat treated with methylcarboline acid combined with γ-aminobutyric acid were significantly improved. The germination potential, germination rate, radicle length, embryo length, germination index, vitality index, drought resistance germination index and drought resistance activity index of wheat were increased by 31.10%, 8.74%, 35.42%, 37.76%, 7.05%, 47.46%, 6.59% and 47.92% respectively compared with CK1. From the germination indicators of wheat seeds under salt stress, the germination indicators of methylcarboline acid combined with GABA were significantly improved compared with those of single application of GABA and methylcarboline acid, indicating that the effect of methylcarboline acid combined with γ-aminobutyric acid on improving the salt tolerance of plants is significantly higher than that of single application of GABA and methylcarboline acid.

[0076] Table 4 Germination indexes of wheat seeds under different treatments under salt stress

[0077]

[0078]

[0079] 2. Promote the growth and development of cucumber seedlings under saline-alkali stress

[0080] (1) Experimental design

[0081] Cucumbers were sown and raised normally, and when they grew to the stage of four leaves and one heart, cucumber seedlings with uniform growth were selected for transplanting. The experiment set up 5 treatment groups, namely normal treatment (CK0), saline-alkali treatment group (CK1), saline-alkali + 10ng / mL methylcarboline acid (methylcarboline acid), saline-alkali + 0.5mg / mL γ-aminobutyric acid (GABA) saline-alkali + 10ng / mL methylcarboline acid + 0.5mg / mL γ-aminobutyric acid (GABA+methyl), and each treatment was repeated 5 times. The normal treatment group was treated with clean water, and the saline-alkali treatment group was added with 150mM saline-alkali solution (NaCI:Na2SO4:NaHCO3:NaCO3=9:1:9:1), and the saline-alkali test group was flushed with the corresponding treatment solution. Regularly observe and record the yellowing growth, and determine the growth indicators. The cucumber leaves were retained to determine the malondialdehyde content, proline content, catalase activity and abscisic acid content.

[0082] (2) Test results

[0083] The test results showed that the cucumber seedlings in the normal treatment group (CK0) grew normally, while the cucumbers in the salt stress treatment groups grew slowly, and the leaves wilted and yellowed. After 28 days of cultivation, the cucumber seedlings were harvested and the indexes were measured. Figure 3 It can be seen that compared with CK1, single application of methylcarboline acid and γ-aminobutyric acid can promote the growth of cucumber seedlings under saline-alkali stress, and all growth indicators of cucumber are generally improved, among which the maximum leaf area and aboveground fresh weight are significantly increased. The effect of methylcarboline acid combined with γ-aminobutyric acid on the growth of cucumber seedlings under salt stress is better than that of single application, among which the maximum leaf area, aboveground fresh weight and number of flower buds of cucumber are significantly improved, which are 48.97%, 58.20% and 38.89% higher than CK, 28.61%, 42.88% and 25.00% higher than γ-aminobutyric acid, and 14.74%, 9.50% and 51.52% higher than methylcarboline acid. It shows that methylcarboline acid combined with γ-aminobutyric acid has a significant synergistic effect on the growth of cucumber seedlings under salt stress.

[0084] Table 5 Growth indexes of cucumber seedlings under different treatments under saline-alkali stress

[0085]

[0086]

[0087] like Figure 6 As shown in Figure 2, compared with CK0, under saline-alkali stress conditions, the MDA content in cucumber leaves increased, and the MDA content in cucumber seedlings treated with CK1 was the highest, reaching 0.1056 umol·L -1 The MDA content in cucumber seedlings treated with methylcarbolinic acid combined with γ-aminobutyric acid was significantly lower than that in CK1, decreasing by 59.94%; the MDA content was reduced by 10.98% and 16.08% compared with that treated with methylcarbolinic acid and γ-aminobutyric acid alone, respectively; the proline, CAT activity and abscisic acid content in cucumber seedlings treated with methylcarbolinic acid combined with γ-aminobutyric acid were significantly increased compared with CK1, increasing by 64.56%, 40.46% and 624.32% respectively compared with CK1, indicating that the combination of methylcarbolinic acid and γ-aminobutyric acid can improve the osmotic regulation ability of cucumber, reduce cell oxidative damage, regulate the abscisic acid content, and thus improve the salt-alkali resistance of cucumber.

[0088] Example 5 Effect of the combination of methylcarboline acid and 5-aminolevulinic acid on crop growth under low temperature stress

[0089] 1. Promote the growth of tomatoes under low temperature stress

[0090] (1) Experimental design

[0091] After sterilizing the substrate, mix it thoroughly, put it into the seedling tray, flatten the surface, and plant tomato seeds at a sowing depth of about 0.5cm. Cultivate it in a culture room at a temperature of 23℃, a humidity of 50%-60%, and a light of 6000-7000lx. Select disease-free and insect-free seedlings with uniform growth to transplant when they grow to the four-leaf and one-heart stage. After 3 days of seedling acclimatization, spray the seeds for testing. There are five treatments: normal temperature treatment (CK0), low temperature treatment (CK1), low temperature + 10ng / mL methylcarboline acid (methylcarboline acid), low temperature + 5-aminolevulinic acid-0.2mg / mL (ALA), and low temperature + 10ng / mL methylcarboline acid + 0.2mg / mL 5-aminolevulinic acid (ALA+methyl). Spray the seeds once every two days, and perform low temperature treatment 12h after the second spraying. Place them in two intelligent incubators respectively, with normal temperature: 23℃ (16 / 8); low temperature treatment: 4℃ (16 / 8). There were 4 replicates for each treatment, and various growth indicators of tomatoes were measured after harvest.

[0092] (2) Test results

[0093] From Table 6 and Figure 4 It can be seen that under low temperature stress, the growth of tomatoes is inhibited, the plants grow short, and the leaves wilt and turn yellow. Compared with the single application of methylcarboline acid and 5-aminolevulinic acid, the combination of methylcarboline acid and 5-aminolevulinic acid has different degrees of improvement in various tomato indicators. The combination of methylcarboline acid and 5-aminolevulinic acid has a significant synergistic effect, and the plant height, root length and SPAD value of tomatoes are increased by 28.54%, 38.55% and 75.93% respectively compared with CK1.

[0094] Table 6 Growth indexes of tomatoes under different treatments under low temperature stress

[0095]

[0096] 2. Promote the growth of peanuts under low temperature stress

[0097] (1) Experimental design

[0098] The nutrient matrix was placed in the planting basket of the Pengbo model, and 4 soaked peanut seeds of each treatment were sown in each planting basket, and each treatment was repeated 4 times. A certain amount of water was added to the culture bottle, and the distance between the water level and the bottom of the planting basket was kept at about 2 cm. It was placed in a 10 / 4℃ incubator for low-temperature treatment for 7 days, and then transferred to room temperature for restoration of culture. The emergence of peanuts was recorded, and the emergence rate of peanuts and growth indicators such as plant height, root length, and fresh weight were measured. Peanut leaf samples were retained to measure the malondialdehyde content, proline content, soluble sugar content, superoxide dismutase activity, and catalase activity of peanuts.

[0099] (2) Data statistics and analysis

[0100] Table 7 Growth of peanut seedlings under different treatments under low temperature stress

[0101]

[0102] From Table 7 and Figure 5 As shown in the results, both single application and compound treatment of 5-aminolevulinic acid and methylcarboline acid can alleviate the effect of low temperature on the growth of peanut seedlings, and the compound effect of the two is significantly better than single application. After 5-aminolevulinic acid and methylcarboline acid were combined, the plant height, root length, stem thickness, fresh weight of aboveground part and fresh weight of underground part increased by 54.20%, 89.92%, 9.69%, 48.39% and 37.84% respectively compared with CK. The plant height of peanuts treated with 5-aminolevulinic acid and methylcarboline acid increased by 24.20% and 11.38% respectively compared with single application of the two, the root length of peanuts increased by 58.32% and 27.82% respectively compared with single application of the two, and the stem thickness of peanuts increased by 4.62% and 2.52% respectively compared with single application of the two. It can be seen that compared with single application, 5-aminolevulinic acid and methylcarboline acid significantly increased the plant height, root length and stem thickness of peanut seedlings under low temperature, and promoted the growth of peanut seedlings under low temperature.

[0103] like Figure 7 As shown in the results, under low temperature stress, the MDA content in peanut seedlings treated with 5-aminolevulinic acid combined with methylcarbolic acid was significantly reduced, 44.70% less than that of CK; the proline and soluble sugar contents in peanut seedlings treated with 5-aminolevulinic acid combined with methylcarbolic acid were significantly increased, 96.90% and 84.85% higher than that of CK, respectively; the SOD and CAT activities of the combined treatment increased by 68.21% and 62.56% compared with CK, respectively; the SOD and CAT activities of 5-aminolevulinic acid alone increased by 51.40% and 27.68% compared with CK, respectively; the SOD and CAT activities of methylcarbolic acid alone increased by 59.26% and 34.24% compared with CK, respectively. This indicates that compared with the single application of the two, 5-aminolevulinic acid combined with methylcarbolic acid can improve the cold resistance of peanuts by increasing the activity of peanut antioxidant enzymes, reducing cell oxidative damage, and maintaining cell osmotic pressure balance, thereby promoting the growth and development of peanut seedlings under low temperature stress.

[0104] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A composition based on methylcarbolinic acid, comprising methylcarbolinic acid and another plant growth regulator, characterized in that: The another plant growth regulator is selected from at least one of brown algae oligosaccharide, paclobutrazol, diethylaminoethyl hexanoate, γ-aminobutyric acid, and 5-aminolevulinic acid.

2. The composition based on methylcarbolinic acid according to claim 1, characterized in that The effective concentration of methylcarbolinic acid is 1 to 200 ng / mL.

3. The composition based on methylcarbolinic acid according to claim 1, characterized in that The methylcarboline acid is obtained by separation and purification of the Pholiota adiposa strain YX1, and the preservation number of the Pholiota adiposa strain YX1 is CGMCC No.21077.

4. The composition based on methylcarboline acid according to claim 1, characterized in that The invention comprises methylcarbolinic acid, brown algae oligosaccharide, paclobutrazol and aminoethyl aminoethyl hexanoate, wherein the effective concentration of the methylcarbolinic acid is 1-200 ng / mL, the effective concentration of paclobutrazol 15% is 0.5-10 mg / mL, the effective concentration of brown algae oligosaccharide is 0.25-5 mg / mL, and the effective concentration of aminoethyl aminoethyl hexanoate DA-6 is 5-30 mg / L.

5. Effect of the methylcarbolinic acid-based composition of claim 4 in promoting plant growth, wherein the plants include Chinese cabbage and lettuce.

6. The composition based on methylcarboline acid according to claim 1, characterized in that It comprises methylcarbolinic acid and γ-aminobutyric acid, wherein the effective concentration of the methylcarbolinic acid is 1-200 ng / mL, preferably 10 ng / mL, and the effective concentration of the γ-aminobutyric acid is 0.1 mg / mL-5.0 mg / mL, preferably 0.5 mg / mL.

7. Use of the composition based on methylcarbolinic acid according to claim 6 in resisting saline-alkali stress in plants, wherein the plants include cucumber and wheat.

8. The composition based on methylcarbolinic acid according to claim 1, characterized in that It comprises methylcarboline acid and 5-aminolevulinic acid, wherein the effective concentration of the methylcarboline acid is 1-200 ng / mL, preferably 10 ng / mL, and the effective concentration of the 5-aminolevulinic acid is 0.05-10 mg / mL, preferably 0.20 mg / mL.

9. Use of the composition based on methylcarbolinic acid according to claim 8 in the resistance of plants to low temperature stress, wherein the plants include tomatoes and peanuts.

10. The use according to any one of claims 5, 7 and 9, characterized in that: The composition improves the activity of plant oxidase, increases the proline content, reduces the malondialdehyde content, and regulates the abscisic acid content, thereby improving the salt-alkali tolerance of plants; It can also improve the activity of plant oxidases, increase the content of osmotic regulating substances, reduce the content of malondialdehyde, and improve the ability of plants to resist low temperature stress.

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