Agricultural rooting-promoting microbial agent synergist and preparation method thereof

Abstract

The invention belongs to the technical field of agricultural microorganisms, and particularly relates to an agricultural rooting-promoting microbial agent synergist and a preparation method thereof. The synergist is prepared from the following raw materials in parts by weight: 10 to 20 parts of modified calcium bentonite, 5 to 10 parts of glucose, 10 to 20 parts of rice hull ash, 0.5 to 1 part of morroniside, 2 to 8 parts of calcium superphosphate, 1 to 3 parts of glycine and 0.1 to 0.5 part of multivitamins. A preparation process of the modified calcium bentonite comprises the following steps: adding calcium bentonite into water containing a surfactant, and stirring to obtain pretreated bentonite; adding the pretreated bentonite and cationic cyclodextrin into water, oscillating, and then adding sodium periodate to obtain a product; and adding the product into water, adding chitosan, and stirring to obtain the product. All the components in the synergist are matched for use, so that nutrient substances and places are provided for proliferation and metabolism of microorganisms, and the rooting process of plants is further promoted.

Description

Technical Field

[0001] The invention belongs to the technical field of agricultural microorganisms, and in particular relates to an agricultural root growth-promoting microbial agent synergist and a preparation method thereof. Background Art

[0002] Microbial agents are widely used in agricultural production due to their advantages of increasing agricultural product yields, improving agricultural product quality, reducing fertilizer use, reducing costs, improving soil, and protecting the ecological environment. During seed germination or transplanting, using microbial agents to mix seeds or dip roots can directly or indirectly improve soil, restore soil fertility, prevent soil-borne diseases, maintain the balance of rhizosphere microbial flora, and degrade toxic substances, thereby promoting plant growth.

[0003] At present, there are many kinds of microbial agents in the market and they are used in agricultural production. By summarizing and analyzing a large amount of data, it is found that microbial agents are affected by various factors, such as climate, soil physical and chemical properties, etc., which leads to the inhibition of microbial activity and reproduction ability, and thus cannot play a good role, and also inhibits its promoting effect on plants. Based on this, scholars have proposed to add synergists to microbial agents to enhance the activity of microbial agents in the environment. Existing synergists have problems such as poor promotion effect. Therefore, the present invention provides a new agricultural root-promoting microbial agent synergist to enhance the activity of microorganisms in microbial agents, thereby promoting agricultural development. Summary of the invention

[0004] The first object of the present invention is to provide an agricultural root-promoting microbial agent synergist, which can promote the rooting process of plants.

[0005] The second object of the present invention is to provide a method for preparing an agricultural root growth promoting microbial agent synergist with simple steps.

[0006] In order to achieve the above object, the technical solution adopted by the present invention is:

[0007] An agricultural root growth-promoting microbial agent synergist, comprising the following raw materials in parts by weight: 10-20 parts of modified calcium-based bentonite, 5-10 parts of glucose, 10-20 parts of rice husk ash, 0.5-1 part of morroniside, 2-8 parts of superphosphate, 1-3 parts of glycine, and 0.1-0.5 parts of complex vitamins;

[0008] The preparation process of the modified calcium-based bentonite is as follows:

[0009] (1) adding calcium-based bentonite to water containing a surfactant for stirring and reacting, and obtaining pretreated bentonite after purification;

[0010] (2) adding the bentonite and cationic cyclodextrin pretreated in step (1) to water, shaking, then adding sodium periodate, stirring, and purifying to obtain a product;

[0011] (3) adding the product of step (2) into water, and then adding chitosan, stirring and purifying, thereby obtaining modified calcium-based bentonite.

[0012] Furthermore, in step (1), the mass ratio of calcium bentonite to surfactant is 1:(1.5-3); the surfactant is sodium lauryl sulfonate; and the stirring reaction time is 3-5h.

[0013] Furthermore, in step (2), the mass ratio of the pretreated bentonite, cationic cyclodextrin and sodium periodate in step (1) is 1:(25-35):(1-2); the oscillation time is 5-7h; and the stirring time is 1-2h.

[0014] Furthermore, in step (3), the mass ratio of the product of step (2) to chitosan is 1:(2-4); and the stirring time is 1-2h.

[0015] Furthermore, the preparation process of the cationic cyclodextrin in step (2) is: adding β-cyclodextrin to a sodium hydroxide solution, then adding 3-chloro-2-hydroxypropyltrimethylammonium chloride to carry out a heating reaction, adjusting the pH of the reaction solution to neutral after the reaction is completed, and obtaining the cationic cyclodextrin after purification.

[0016] Furthermore, the mass ratio of the β-cyclodextrin to 3-chloro-2-hydroxypropyltrimethylammonium chloride is 1:(0.5-1).

[0017] Furthermore, the temperature of the heating reaction is 75-85° C., the heating reaction time is 8-10 hours, and the mass concentration of the sodium hydroxide solution is 25-35%.

[0018] Furthermore, the complex vitamin includes vitamin A, vitamin B, vitamin C, and vitamin E; the mass ratio of the vitamin A, vitamin B, vitamin C, and vitamin E is 1:(1-2):(1-2):(2-3).

[0019] The preparation method of the above-mentioned agricultural root growth-promoting microbial agent synergist comprises the following steps:

[0020] According to the weight ratio, all the raw materials can be mixed evenly.

[0021] The beneficial technical effects of the present invention are:

[0022] Modified calcium-based bentonite, morroniglycoside and other ingredients are added to the synergist of the present invention. The modified calcium-based bentonite is pretreated by bentonite and cationic cyclodextrin to form a complex through electrostatic adsorption, and then the hydroxyl groups in β-cyclodextrin are oxidized and cross-linked with the amino groups in chitosan through Schiff base reaction. The rich surface structure of the modified bentonite finally formed can be used as a carrier for microbial attachment, providing a stable microenvironment for the growth of microorganisms, and cooperating with other ingredients in the synergist to provide nutrients for the proliferation and metabolism of microorganisms, promote the activity of microorganisms, and then regulate the number of soil microorganisms, improve the soil, and promote plant rooting. Morroniglycoside can affect the activity of microbial agents by promoting the growth of microorganisms. It can be seen that the combination of the various ingredients in the synergist can provide nutrients and places for the proliferation and metabolism of microorganisms, thereby promoting the rooting process of plants. DETAILED DESCRIPTION

[0023] The following is a further detailed description of the present invention in conjunction with specific preferred embodiments, and it cannot be determined that the specific implementation of the present invention is limited to these descriptions. For those of ordinary skill in the technical field to which the present invention belongs, without departing from the concept of the present invention, several simple deductions or replacements can also be made, all of which should be deemed to belong to the protection scope of the present invention. The specific conditions not indicated in the embodiments are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used, unless otherwise specified, are conventional products obtained through commercial channels.

[0024] Enterobacter ludwigii PC2 (Enterobacter ludwigii) is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number: CGMCC No.21823; Enterobacter cloacae PB7 (Enterobacter cloacae) is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number: CGMCC No.21822.

[0025] Example

[0026] Example 1

[0027] An agricultural root growth promoting microbial agent synergist is composed of the following raw materials in parts by weight: 14 parts of modified calcium-based bentonite, 7 parts of glucose, 12 parts of rice husk ash, 0.8 parts of morroniside, 5 parts of superphosphate, 2 parts of glycine, and 0.4 parts of complex vitamins; the complex vitamins are compounded from vitamin A, vitamin B, vitamin C, and vitamin E in a mass ratio of 1:1.5:1.5:2.5.

[0028] The preparation process of the modified calcium-based bentonite is as follows:

[0029] (1) According to the mass ratio of calcium bentonite to sodium lauryl sulfate of 1:2, calcium bentonite was added to water containing sodium lauryl sulfate, stirred at room temperature for 4 hours, and the pretreated bentonite was obtained after washing and drying.

[0030] (2) According to the mass ratio of β-cyclodextrin and 3-chloro-2-hydroxypropyltrimethylammonium chloride of 1:0.7, β-cyclodextrin is added to a sodium hydroxide solution with a concentration of 30%, and then 3-chloro-2-hydroxypropyltrimethylammonium chloride is added, and the mixture is heated at 80° C. for 9 hours. After the reaction is completed, the pH of the reaction solution is adjusted to neutral, and the reaction solution is filtered, washed, and dried to obtain cationic cyclodextrin.

[0031] According to the mass ratio of the pretreated bentonite in step (1), cationic cyclodextrin and sodium periodate of 1:30:1.5, the pretreated bentonite and cationic cyclodextrin in step (1) are added to water and shaken for 6 hours, and then sodium periodate is added and stirred for 1.5 hours. After washing and drying, the product is obtained.

[0032] (3) According to the mass ratio of the product of step (2) to chitosan of 1:3, the product of step (2) is added to water, and chitosan is then added, and the mixture is stirred for 1.5 hours, and then washed and dried to obtain modified calcium-based bentonite.

[0033] The preparation method of the above-mentioned agricultural root growth-promoting microbial agent synergist is as follows:

[0034] Weigh the raw materials according to the above weight ratio and mix them evenly.

[0035] Example 2

[0036] An agricultural root growth-promoting microbial agent synergist is composed of the following raw materials in parts by weight: 10 parts of modified calcium-based bentonite, 5 parts of glucose, 10 parts of rice husk ash, 0.5 parts of morroniside, 2 parts of superphosphate, 1 part of glycine, and 0.1 parts of complex vitamins; the complex vitamins are compounded from vitamin A, vitamin B, vitamin C, and vitamin E in a mass ratio of 1:1:1:2.

[0037] The preparation process of the modified calcium-based bentonite is as follows:

[0038] (1) According to the mass ratio of calcium bentonite to sodium lauryl sulfate of 1:1.5, calcium bentonite was added to water containing sodium lauryl sulfate, stirred at room temperature for 3 hours, and the pretreated bentonite was obtained after washing and drying.

[0039] (2) According to the mass ratio of β-cyclodextrin and 3-chloro-2-hydroxypropyltrimethylammonium chloride of 1:0.5, β-cyclodextrin is added to a sodium hydroxide solution with a concentration of 25%, and then 3-chloro-2-hydroxypropyltrimethylammonium chloride is added, and the mixture is heated at 75° C. for 10 hours. After the reaction is completed, the pH of the reaction solution is adjusted to neutral, and the reaction solution is filtered, washed, and dried to obtain cationic cyclodextrin.

[0040] According to the mass ratio of the pretreated bentonite in step (1), cationic cyclodextrin and sodium periodate of 1:25:1, the pretreated bentonite and cationic cyclodextrin in step (1) are added to water and shaken for 5 hours, and then sodium periodate is added and stirred for 1 hour. After washing and drying, a product is obtained.

[0041] (3) According to the mass ratio of the product of step (2) to chitosan of 1:2, the product of step (2) is added to water, and then chitosan is added, and after stirring for 1 hour, the mixture is washed and dried to obtain modified calcium-based bentonite.

[0042] The preparation method of the above-mentioned agricultural root growth-promoting microbial agent synergist is as follows:

[0043] Weigh the raw materials according to the above weight ratio and mix them evenly.

[0044] Example 3

[0045] An agricultural root growth promoting microbial agent synergist is composed of the following raw materials in parts by weight: 20 parts of modified calcium-based bentonite, 10 parts of glucose, 20 parts of rice husk ash, 1 part of morroniside, 8 parts of superphosphate, 3 parts of glycine, and 0.5 parts of complex vitamins; the complex vitamins are compounded from vitamin A, vitamin B, vitamin C, and vitamin E in a mass ratio of 1:2:2:3.

[0046] The preparation process of the modified calcium-based bentonite is as follows:

[0047] (1) According to the mass ratio of calcium bentonite to sodium lauryl sulfonate of 1:3, calcium bentonite was added to water containing sodium lauryl sulfonate, stirred at room temperature for 5 hours, and the pretreated bentonite was obtained after washing and drying.

[0048] (2) According to the mass ratio of β-cyclodextrin and 3-chloro-2-hydroxypropyltrimethylammonium chloride of 1:1, β-cyclodextrin is added to a sodium hydroxide solution with a concentration of 35%, and then 3-chloro-2-hydroxypropyltrimethylammonium chloride is added, and the mixture is heated at 85° C. for 8 h. After the reaction is completed, the pH of the reaction solution is adjusted to neutral, and the reaction solution is filtered, washed, and dried to obtain cationic cyclodextrin.

[0049] According to the mass ratio of the pretreated bentonite in step (1), cationic cyclodextrin and sodium periodate of 1:35:2, the pretreated bentonite and cationic cyclodextrin in step (1) are added to water and shaken for 7 hours, and then sodium periodate is added and stirred for 2 hours. After washing and drying, a product is obtained.

[0050] (3) According to the mass ratio of the product of step (2) to chitosan of 1:4, the product of step (2) is added to water, and then chitosan is added, and after stirring for 1-2 hours, the mixture is washed and dried to obtain modified calcium-based bentonite.

[0051] The preparation method of the above-mentioned agricultural root growth-promoting microbial agent synergist is as follows:

[0052] Weigh the raw materials according to the above weight ratio and mix them evenly.

[0053] Comparative Example

[0054] Comparative Example 1

[0055] The difference between Comparative Example 1 and Example 1 is that:

[0056] Calcium-based bentonite was used instead of modified calcium-based bentonite, and the rest was the same as in Example 1.

[0057] Comparative Example 2

[0058] The difference between Comparative Example 2 and Example 1 is that:

[0059] Calcium bentonite, β-cyclodextrin and chitosan are used instead of modified calcium bentonite, and the rest is the same as in Example 1; the amounts of calcium bentonite, β-cyclodextrin and chitosan are consistent with those in Example 1.

[0060] Comparative Example 3

[0061] The difference between Comparative Example 3 and Example 1 is that:

[0062] The remainder is the same as in Example 1 except that morroniside is omitted.

[0063] Test example

[0064] Preparation of microbial inoculants: Enterobacter Ludwigii and Enterobacter cloacae were inoculated into 50 mL LB liquid culture medium, respectively, and cultured in a constant temperature shaking incubator at 30 °C and 200 rpm for 48 h. The two cultured bacterial solutions were diluted to a bacterial concentration of 1 × 10 8 CFU / mL, and then evenly mix the seed solutions of Enterobacter Ludwigii and Enterobacter cloacae at a volume ratio of 1:1 to obtain a microbial agent for later use.

[0065] (1) Microbial activity detection

[0066] 10 g / mL of the synergist obtained in Examples 1-3 and Comparative Examples 1-3 was added to the above-mentioned microbial agent, and the total number of effective viable bacteria therein was determined by the plate count method (refer to the GB 20287-2006 method), and the bacterial survival rate after 60 d and 120 d (the ratio of the total number of effective viable bacteria in a certain period to the initial total number of effective viable bacteria) was calculated, and a blank control group without adding the synergist was set. The results are shown in Table 1.

[0067] (2) Growth performance test

[0068] Experimental setup:

[0069] The blank control group was immersed in the above-mentioned microbial agents only;

[0070] In the test group, 10 g / mL of the synergist obtained in Examples 1-3 and Comparative Examples 1-3 were added to the above microbial agents, and then immersed.

[0071] Select highland barley seeds of uniform size and full grains, then wash with 75% ethanol for 5 minutes, and then rinse with sterile water 2-3 times to obtain sterile seeds. According to the experimental settings, the sterile highland barley seeds are completely immersed in the above-mentioned preparations, and the seeds are taken out after soaking for 2 hours, placed on moistened sterilized filter paper, and cultured in an incubator at 25°C and 85% humidity for 8 days. After the cultivation, the plant height, root length, number of roots, fresh weight and dry weight of each group of seeds are measured. Each group is set with 30 seeds, and the results are shown in Table 2.

[0072] Table 1

[0073]

[0074]

[0075] Table 2

[0076] Grouping Root length (cm) Number of roots (pieces / plant) Plant height (cm) Fresh weight (g / plant) Dry weight (g / plant) Blank control group 9.48 6.21 8.03 0.2219 0.0325 Example 1 Group 10.67 7.86 9.50 0.3007 0.0552 Example 2 Group 10.63 7.72 9.47 0.2951 0.0546 Example 3 Group 10.61 7.64 9.35 0.2874 0.0527 Comparative Example 1 9.77 6.53 8.29 0.2406 0.0350 Comparative Example 2 9.94 6.86 8.52 0.2531 0.0391 Comparative Example 3 9.96 6.90 8.58 0.2618 0.0404

[0077] From Table 1, it can be seen that the number of effective live bacteria in Examples 1-3 did not decrease within 120 days, which shows that the enhancers obtained in Examples 1-3 can improve the activity of microorganisms, which is very beneficial for microbial agents.

[0078] It can be seen from Table 2 that the use of the synergist obtained from Example 1-3 can increase the root length and root number of highland barley seeds, thereby promoting the growth of highland barley. Compared with Example 1-3, Comparative Example 1 uses calcium-based bentonite instead of modified calcium-based bentonite; Comparative Example 2 uses calcium-based bentonite, β-cyclodextrin and chitosan instead of modified calcium-based bentonite; Comparative Example 3 omits morroniside, and the root length and root number of highland barley seeds in Comparative Examples 1-3 are reduced. It can be seen from the above that the modified calcium-based bentonite and morroniside of the present invention can jointly promote the activity of microorganisms, thereby enhancing the function of microbial agents in promoting plant rooting.

[0079] The present invention forms a complex with cationic cyclodextrin after pretreatment of bentonite through electrostatic adsorption, and then oxidizes the hydroxyl groups in β-cyclodextrin and cross-links them with the amino groups in chitosan through Schiff base reaction, and finally forms modified calcium-based bentonite. The rich surface structure of the modified bentonite can be used as a carrier for microbial attachment, providing a stable microenvironment for the growth of microorganisms, and cooperating with other components in the synergist to provide nutrients for the proliferation and metabolism of microorganisms, promote the activity of microbial agents, and then regulate the number of soil microorganisms, improve the soil, and promote plant rooting. Morroniside can affect the activity of microbial agents by promoting the growth of microorganisms. It can be seen that the combination of the components in the synergist can provide nutrients and places for the proliferation and metabolism of microorganisms, thereby promoting the rooting process of plants.

[0080] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit them. The basic principles and main features of the present invention have been described above with specific implementation schemes. On the basis of the present invention, some modifications or replacements may be made thereto, but these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of protection claimed by the present invention.

Claims

1. An agricultural root growth promoting microbial agent synergist, characterized in that: The method comprises the following raw materials in parts by weight: 10-20 parts of modified calcium-based bentonite, 5-10 parts of glucose, 10-20 parts of rice husk ash, 0.5-1 parts of morroniside, 2-8 parts of superphosphate, 1-3 parts of glycine, and 0.1-0.5 parts of multivitamins; The preparation process of the modified calcium-based bentonite is as follows: (1) adding calcium-based bentonite to water containing a surfactant for stirring and reacting, and obtaining pretreated bentonite after purification; (2) adding the bentonite and cationic cyclodextrin pretreated in step (1) to water, shaking, then adding sodium periodate, stirring, and purifying to obtain a product; (3) adding the product of step (2) into water, and then adding chitosan, stirring and purifying, thereby obtaining modified calcium-based bentonite.

2. The agricultural root growth promoting microbial agent synergist as claimed in claim 1, characterized in that: In step (1), the mass ratio of calcium bentonite to surfactant is 1:(1.5-3); the surfactant is sodium lauryl sulfonate; and the stirring reaction time is 3-5h.

3. The agricultural root growth promoting microbial agent synergist as claimed in claim 1, characterized in that: In step (2), the mass ratio of the pretreated bentonite, cationic cyclodextrin and sodium periodate in step (1) is 1:(25-35):(1-2); the oscillation time is 5-7h; and the stirring time is 1-2h.

4. The agricultural root growth promoting microbial agent synergist as claimed in claim 1, characterized in that: In step (3), the mass ratio of the product of step (2) to chitosan is 1:(2-4); and the stirring time is 1-2h.

5. The agricultural root growth promoting microbial agent synergist as claimed in claim 1, characterized in that: The preparation process of the cationic cyclodextrin in step (2) is as follows: β-cyclodextrin is added to a sodium hydroxide solution, and then 3-chloro-2-hydroxypropyltrimethylammonium chloride is added to carry out a heating reaction. After the reaction is completed, the pH of the reaction solution is adjusted to neutral, and the cationic cyclodextrin is obtained after purification.

6. The agricultural root growth promoting microbial agent synergist as claimed in claim 5, characterized in that: The mass ratio of the β-cyclodextrin to 3-chloro-2-hydroxypropyltrimethylammonium chloride is 1:(0.5-1).

7. The agricultural root growth promoting microbial agent synergist as claimed in claim 5, characterized in that: The temperature of the heating reaction is 75-85° C., and the time of the heating reaction is 8-10 hours; the mass concentration of the sodium hydroxide solution is 25-35%.

8. The agricultural root growth promoting microbial agent synergist as claimed in claim 1, characterized in that: The complex vitamin comprises vitamin A, vitamin B, vitamin C and vitamin E; the mass ratio of the vitamin A, vitamin B, vitamin C and vitamin E is 1:(1-2):(1-2):(2-3).

9. The method for preparing the agricultural root growth promoting microbial agent synergist according to any one of claims 1 to 8, characterized in that: The following steps are involved: According to the weight ratio, all the raw materials can be mixed evenly.

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