CN114938814A - Use of surfactant and composition thereof in promoting plant root growth - Google Patents
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
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- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
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- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
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Abstract
The invention relates to a surfactant and application of a composition thereof in promoting plant root growth, belonging to the technical field of biological stimulin. The surfactant comprises surfactant with fatty acyl length of C12-C16. The invention provides a surfactant or a composition containing the surfactant for preparing liquid post-treatment soil or plant body. The invention provides a new application of surfactant and a composition thereof in promoting the growth of plant roots. Experiments prove that the surfactant and the composition thereof have remarkable effects of improving the fresh weight and the yield of root systems of crops such as tomatoes, wheat, melons and the like, and can be applied to agricultural production.
Description
Technical Field
The invention belongs to the technical field of biological stimulin, and particularly relates to a surfactant and application of a composition thereof in promoting plant root growth.
Background
The surfactin is a cyclic lipopeptide type biosurfactant, has good surface/interface activity, and is one of the most studied biosurfactants with the strongest surface activity. The surfactant was first found in Bacillus subtilis culture and was subsequently identified in various Bacillus fermentation broths, such as Bacillus amyloliquefaciens and Bacillus belgii. The structure is that a heptapeptide is connected with a beta-hydroxyl of a fatty acid chain carbon atom through an inner ester bond. The surfactant widely found in fermentation broth is mostly C 13 、C 14 、C 15 、C 16 Lipopeptides of four different length aliphatic chains generally have the hydrophobic amino acid residues in the peptide loop at positions 2, 3, 4, 6 and 7, and the glutamic and aspartic acid residues at positions 1 and 5, respectively. Furthermore, the researchers found that the number of carbon atoms of beta-hydroxy fatty acid in the surfactant structure was 12 to 17, and L-Leu at C terminal was substituted by L-Ile or L-Val. The research finds that the surfactant has biological activities of resisting bacteria, viruses and mycoplasma, and preventing and treating homoptera or coleoptera insects. In addition, the surfactant can induce plants to generate immune response and improve the resistance of grapes to downy mildew. However, up to now, no studies have been made to evaluate the effect of root application of high purity surfactant (95% or more) on crop root growth and yield of agricultural products.
The good root system is the basis for the plants to absorb nutrition, maintain growth and ensure the yield of agricultural products. Therefore, the search for active substances with a pronounced root-promoting ability is one of the important research and development directions. Plant growth regulators such as indolebutyric acid, diethyl aminoethyl hexanoate, triacontanol and the like (according to pesticide management in China) have a certain root growth promoting effect, but the exertion of the physiological activities of the substances is closely related to the dosage, the use time and the weather condition of the substances, and obvious phytotoxicity is usually caused by improper use. Biological stimulants such as humic acid, alginic acid, fish protein and the like have a certain rooting function, but generally comprise dozens of or even hundreds of components, and the activity of the product is greatly influenced by raw materials and processes. There is no document reporting the use of surfactin to promote root growth.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a surfactant and application of a composition thereof in promoting the growth of plant roots, so as to solve the problems. The surfactin family lipopeptide has a definite structure, is easy to biodegrade, can inhibit part of root diseases, and is suitable for being developed into novel rooting-promoting and yield-increasing biostimulant.
The technical scheme of the invention is as follows:
use of a surfactant comprising a fatty acyl group having a length C for promoting plant root growth 12 -C 16 The surfactant of (1).
Said surfactant comprises a fatty acyl group of length C 12 -C 16 The surfactant of (1) is named as C 12 -surfactant, C 13 -surfactant, C 14 -surfactant, C 15 -surfactant, C 16 -a surfactant.
As a specific embodiment, the plant includes tomato, pepper, cucumber, melon, wheat, rice, corn, apple, banana, citrus, peach, pear, grape.
As a specific embodiment, the application method of the surfactant is to prepare the surfactant into liquid to treat soil or plant bodies.
In a specific embodiment, the mass concentration of the surfactant in the liquid is 1.0-900 mg/L.
The use of a composition containing the surfactant in promoting the growth of plant roots, wherein the composition comprises the surfactant and chitosan oligosaccharide.
As a specific embodiment, the chitosan oligosaccharide is a polymer with deacetylation degree of 75-90% and polymerization degree of 6-20 (purity is more than or equal to 85%), and is obtained by degrading chitosan through an enzymatic method, a chemical method or a physical method.
As a specific embodiment, the plant includes tomato, pepper, cucumber, melon, wheat, rice, corn, apple, banana, citrus, peach, pear, grape.
As a specific embodiment, the composition is applied by formulating the composition as a liquid post-treatment soil or plant material.
In a specific embodiment, the mass concentration of the surfactant in the liquid is 1.0-900 mg/L.
In a specific embodiment, in the surfactant-containing composition, the mass ratio of the surfactant to the chitosan oligosaccharide is 3.5-25.0: 1.0.
The beneficial effects of the invention are as follows:
the invention provides a new application of surfactant and a composition thereof in promoting the growth of plant roots. Experiments prove that the surfactant and the composition thereof have remarkable effects of improving the fresh weight and the yield of root systems of crops such as tomatoes, wheat, melons and the like, and can be applied to agricultural production.
Drawings
In order to more clearly illustrate the embodiments or technical solutions in the prior art of the present invention, the drawings used in the description of the embodiments or prior art will be briefly described below, and it is obvious for those skilled in the art that other drawings can be obtained based on these drawings without creative efforts.
FIG. 1 is a comparison of the surfactant prepared in example 1 of the present invention and a surfactant standard in a UPLC spectrum; wherein the upper panel is a surfactant standard and the lower panel is the surfactant prepared in example 1.
FIG. 2 is C prepared according to example 1 of the present invention 12 MS spectrum of surfactant.
FIG. 3 is C prepared according to example 1 of the present invention 13 MS spectrum of surfactant.
FIG. 4 is C prepared according to example 1 of the present invention 14 MS spectrum of surfactant.
FIG. 5 is C prepared according to example 1 of the present invention 15 MS spectrum of surfactant.
FIG. 6 is C prepared according to example 1 of the present invention 16 MS spectrum of surfactant.
FIG. 7 is a graph comparing root growth of tomatoes in example 2 of the present invention.
FIG. 8 is a comparison of melon seedlings grown in example 3 of the present invention.
FIG. 9 is a graph showing a comparison of growth of triticale in example 4 of the present invention.
Detailed Description
In order to make those skilled in the art better understand the technical solution of the present invention, the technical solution in the embodiment of the present invention will be clearly and completely described below with reference to the drawings in the embodiment of the present invention, and it is obvious that the described embodiment is only a part of the embodiment of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
Preparation of surfactin
(1) Bacillus subtilis ATCC21332 was subjected to scale-up culture to obtain a fermentation broth.
(2) And centrifuging the fermentation liquor to remove thalli to obtain a supernatant.
(3) Concentrating the supernatant by 3 times with a rotary evaporator, adjusting pH of the feed liquid to 6.5 with 4mol/L sulfuric acid, adding equal volume of 95% (V/V) ethanol into the concentrated solution, shaking to mix the feed liquid sufficiently, allowing the protein in the concentrated solution to denature into floccule with the ethanol, centrifuging to remove the flocculent protein, and obtaining the clear liquid.
(4) Removing ethanol from the clear liquid by a rotary evaporator, and then adjusting the pH value of the feed liquid to 2.0 by using 4mol/L sulfuric acid to precipitate surfactant. And centrifuging, collecting the precipitate, performing acid washing on the precipitate for 2 times by adopting a sulfuric acid aqueous solution with the pH value of 2 because impurities in part of fermentation liquor still remain in the product, centrifuging, collecting the precipitate, and performing freeze drying to obtain a surfactant sample.
(5) The surfactant samples were qualitatively analyzed by UPLC-MS, while an external surfactant standard (purchased from Sigma) was used as a control, and the results of comparison are shown in fig. 1. In FIG. 1, the upper diagram shows the surface activityThe lower panel shows the surfactant prepared in example 1. As can be seen by comparison, the standard contains four peaks, each of which is a fatty acyl group with a length of C 12 ～C 15 The surfactant of (1). The peak position of the surfactant prepared in example 1 was consistent with that of the standard; the peak at RT ═ 12.6min is the fatty acyl length C 16 The surfactant of (1). And determining the surfactant and the homologue thereof contained in the detection sample by a full-scan mode.
The MS spectra of the surfactant sample are shown in FIGS. 2 to 6.
The mass spectrometric detection results of the purified samples are given in table 1 below:
TABLE 1 Mass Spectrometry results of samples
| Number of Compounds | m/z[M-H]- | |
| 1 | 992.6310 | C 12 - |
| 2 | 1006.6455 | C 13 -surfactin |
| 3 | 1020.6612 | C 14 - |
| 4 | 1034.6779 | C 15 - |
| 5 | 1052.6875 | C 16 -surfactin |
Example 2
Application of surfactant in promoting growth and increasing yield of tomato root system
The test was carried out in 2020 in glass greenhouse of Jingbo agrichemical technologies GmbH, and the test material was cherry tomato. Mixing turfy soil (purchased in flower market) and sand according to the proportion of 1: 1, uniformly mixing and loading the mixture into a seedling raising tray of 54cm multiplied by 27cm multiplied by 6cm, uniformly sowing seeds into the seedling raising tray, covering a soil layer with the thickness of about 0.5cm, watering from the bottom, and performing seed germination and seedling raising under the condition of sufficient sunlight at the temperature of 24-28 ℃. Selecting seedlings with consistent growth vigor after the tomato plants grow to 6-8 leaves, transplanting the seedlings into flowerpots (the diameter of each flowerpot is 30cm, the height of each flowerpot is 15cm, and each pot is filled with 9.0kg of field soil), keeping the growth conditions of each pot of seedlings consistent, applying base fertilizer after seedling delay, wherein the base fertilizer is compound fertilizer (N-P) 2 O 5 -K 2 O15-15-15), according to the dosage of 500kg/hm 2 And (4) application.
The experiment was set up with 3 treatments, 30 seedlings each, 10 seedlings in one group, for a total of three replicates. (1) Treatment group a was a placebo control, applied with a water soluble fertilizer. 20-20-20 (N-P) 2 O 5 -K 2 O) diluting the water-soluble fertilizer by 300 times, and applying 100mL of the water-soluble fertilizer diluent to the root of each tomato seedling. (2) Treatment group B was applied with mineral potassium fulvate (purchased on the market, humic acid content 50%). Adding 3000mL of 20-20-20 (N-P) 2 O 5 -K 2 O) adding 10g of mineral source potassium fulvate into 300 times of diluent of the water-soluble fertilizer, stirring to completely dissolve the mineral source potassium fulvate to obtain a humic acid-containing water-soluble fertilizer solution, and applying 100mL of the solution to the root of each tomato seedling. (3) Treatment group C was administered the surfactant prepared in example 1. Adding 3000mL of 20-20-20 (N-P) 2 O 5 -K 2 O) 300-time dilution of water-soluble fertilizer0.30g of surfactant is added and stirred to be completely dissolved to obtain a water-soluble fertilizer solution with the surfactant content of 100mg/L, and 100mL of the solution is applied to the roots of each tomato seedling.
After 20 days of the application of the water-soluble fertilizer, 6 plants were randomly selected from each treatment group, soil at the roots was carefully washed off, and the fresh weight of the roots and the fresh weight of the ground were measured. Another 3 plants were taken for each treatment, the roots were washed in an ice-water bath and 0.5g of fresh root tip samples were taken, and the activity of the roots in the different treatment groups was determined by TTC staining. The specific results of the root growth ratio shown in FIG. 7 are shown in Table 2 below:
TABLE 2 results of the measurements
| Treatment group | Fresh weight of root (g) | Root system activity (mg. g) -1 ·h -1 ) | Fresh weight on ground (g) |
| A | 128.6 | 7.3 | 197.9 |
| B | 158.1 | 9.8 | 237.4 |
| C | 240.7 | 12.5 | 302.3 |
As can be seen from the data in table 2 and fig. 7, the surfactant can strongly stimulate the growth of the tomato root system, the fresh root weight is respectively increased by 87.2% and 52.2% compared with the control group (treatment a) and the humic acid treatment group (treatment B), and the root system keeps high activity. The good root system enhances the absorption capacity of the plant to nutrition, thereby promoting the aboveground biomass accumulation; the fresh weight of the ground of the surfactant treated group is respectively increased by 52.8 percent and 27.3 percent compared with the ground of the control group and the humic acid treated group.
The remaining 21 tomatoes in each treatment group continue to grow, and the water and fertilizer application amount and application time are kept completely consistent. The treatment group B and the treatment group C are respectively applied with water-soluble fertilizers containing humic acid and surfactant (the dosage of humic acid and surfactant is the same as that of the water-soluble fertilizers) once in the young fruit period, and the treatment group A is only applied with the same amount of water-soluble fertilizers. And tomato yield and quality were investigated during the fruiting period (four fruits before production). The results are given in table 3 below:
TABLE 3 tomato yield and quality index results for different treatment groups
a The higher the solid acid ratio, the sweeter the fruit. b The higher the vitamin C content, the better the fruit quality.
The data in table 3 show that the yield of tomatoes in the surfactant-treated group is increased by 22.9% and 11.5% respectively compared with A, B in the treated group. The quality of the tomatoes in the surface active agent treatment group is obviously improved compared with the tomatoes in the other two groups, and the taste is obviously improved. Tomato yield and quality data are positively correlated with root growth, which is highly consistent with the long-term observation in agricultural production, i.e., good root systems are the basis for healthy growth and high yield of crops.
Example 3
Application of surfactant and composition thereof in promoting rooting of melons
The test was carried out in 2021 in glass greenhouse of Jingbo agrichemical technologies, Inc., and the test material was melon (Boyang No. 9). Uniformly mixing turfy soil and sand according to a ratio of 1: 1, loading the mixture into a seedling tray with the thickness of 54cm multiplied by 27cm multiplied by 6cm, uniformly sowing seeds into the seedling tray, covering a soil layer with the thickness of about 0.5cm, watering from the bottom, and carrying out seed germination and seedling raising under the condition of sufficient sunlight at the temperature of 24-28 ℃. After the melon plants grow to 2-4 leaves, seedlings with consistent growth vigor are selected and transplanted into flowerpots (the diameter of each flowerpot is 15cm, the height of each flowerpot is 11cm, and each pot is filled with 1.0kg of field soil). The pH value of the soil is 7.36, and the contents of organic matters, alkaline hydrolysis nitrogen, quick-acting phosphorus and quick-acting potassium are respectively 25.14g/kg, 135.71mg/kg, 32.25mg/kg and 121.29 mg/kg.
The experiment was set up for a total of 7 treatments, 30 seedlings each, 10 seedlings in one group, for a total of three replicates. And 5 days after transplanting, a Jingbo 'plant-times health' water-soluble fertilizer and different biostimulants are applied to roots. Wherein D is control group, and only water soluble fertilizer diluent is applied, and the water soluble fertilizer diluent has a content of 20-20-20 (N-P) 2 O 5 -K 2 O) comprises monoammonium phosphate, urea and potassium sulfate as main components. E-J is that different biostimulants are respectively added into the water-soluble fertilizer diluent, and the experimental design is shown in a table 4:
TABLE 4 Experimental design
After 15 days from the application of the water-soluble fertilizer, 6 plants were randomly selected from each treatment group, carefully washed off the soil at the roots, and the fresh weight of the roots and the fresh weight on the ground were measured according to the design of table 4. Another 3 plants were taken for each treatment, the roots were washed in an ice-water bath and 0.5g of fresh root tip samples were taken, and the activity of the roots in the different treatment groups was determined by TTC staining. The growth rate of melons in different treatment groups is shown in fig. 8, and the results are shown in the following table 5:
TABLE 5 comparison of root systems and growth indices of melons in different treatment groups
| Treatment group | Average fresh root weight (g) | Root system activity (mg. g) -1 ·h -1 ) | Fresh weight on ground (g) |
| D | 2.85 | 5.73 | 5.67 |
| E | 3.76 | 7.52 | 6.61 |
| F | 4.65 | 9.32 | 8.62 |
| G | 3.94 | 8.11 | 7.51 |
| H | 5.46 | 10.87 | 11.82 |
| I | 3.47 | 6.33 | 5.72 |
| J | 3.75 | 6.96 | 6.38 |
The data in table 5 show that: (1) compared with a control group, the surface active element, the chitosan oligosaccharide and the humic acid can obviously promote the growth of root systems and overground parts of melon seedlings and enhance the activity of the root systems, and the surface active element is best in performance. (2) The combination ratio of the surfactant to the chitosan oligosaccharide is 10:1, and the obvious synergistic effect is achieved; when the proportion of the chitosan oligosaccharide in the composition is higher, the chitosan oligosaccharide and the composition have certain antagonistic effect. (3) The surfactant and humic acid are combined (50mg/L +1617mg/L) to generate obvious antagonistic effect.
Example 4
Use of surfactin for increasing wheat yield
The test is carried out in Pongjia experimental base of Jingbo agro-chemical science and technology Limited in 2020, and the test time is 10 months to 2021 months and 6 months in 2020. Fertilizing and soil preparation before sowing are carried out according to conventional operation. The pH value of the soil is 7.24, and the contents of organic matters, alkaline hydrolysis nitrogen, quick-acting phosphorus and quick-acting potassium are respectively 16.33g/kg, 145.12mg/kg, 37.67mg/kg and 131.57 mg/kg. The wheat variety is 22 Jimai, the seeding rate is 10 kg/mu, and the seeding area is 10 mu.
The test set up two treatment groups (5 acres per treatment group): performing conventional management on a control group (treatment group K); and (3) spraying 50mg/L of surfactant to the experimental group (treatment group L) in the main leaf period and the booting period of the wheat respectively on the basis of conventional management. The fresh weight of wheat roots (green turning period), the tiller number and the second internode length are tracked and measured, and the yield of the wheat of the two treatment groups is measured according to DB 32/T2856-. The results of the different treatment groups for ear growth are shown in fig. 9 and are shown in table 6 below:
TABLE 6 comparison of wheat growth index and yield in different treatment groups
As can be seen from the data in fig. 9 and table 6, spraying surfactant can promote tillering of wheat and shorten the second internode length (which helps to reduce lodging). The theoretical yield of wheat in the experimental group is increased by 15.2 percent compared with that in the control group.
Although the present invention has been described in detail by referring to the drawings in connection with the preferred embodiments, the present invention is not limited thereto. Various equivalent modifications or substitutions can be made on the embodiments of the present invention by those skilled in the art without departing from the spirit and scope of the present invention, and these modifications or substitutions are within the scope of the present invention/any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (10)
1. The application of the surfactant in promoting the growth of plant roots is characterized in that: said surfactant comprises a fatty acyl group of length C 12 -C 16 The surfactant of (1).
2. Use according to claim 1, wherein the plant comprises tomato, pepper, cucumber, melon, wheat, rice, corn, apple, banana, citrus, peach, pear, grape.
3. The use according to claim 1, wherein the surfactant is applied by formulating the surfactant as a liquid for post-treatment of soil or plant material.
4. The use according to claim 3, wherein the surfactant concentration in the liquid is 1.0 to 900mg/L by mass.
5. Use of a composition comprising the surfactant of claim 1 for promoting root growth in plants, wherein: the composition comprises the surfactant and chitosan oligosaccharide.
6. Use according to claim 5, characterized in that: the chitosan oligosaccharide is a polymer with deacetylation degree of 75-90% and polymerization degree of 6-20.
7. Use according to claim 5, wherein the plant comprises tomato, pepper, cucumber, melon, wheat, rice, corn, apple, banana, citrus, peach, pear, grape.
8. Use according to claim 5, wherein the composition is applied by formulating the composition as a liquid for post-treatment of the soil or plant material.
9. Use according to claim 8, characterized in that: the mass concentration of the surfactant in the liquid is 1.0-900 mg/L.
10. Use according to claim 5, characterized in that: in the composition containing the surfactant, the mass ratio of the surfactant to the chitosan oligosaccharide is 3.5-25.0: 1.0.
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