CN110872518B

CN110872518B - Acid soil conditioner and preparation method and application thereof

Info

Publication number: CN110872518B
Application number: CN201911142509.9A
Authority: CN
Inventors: 马金霞; 谭寿湖; 张琦敏; 林振业; 梁春吉
Original assignee: Guangxi Jinsui Ecological Technology Group Co ltd
Current assignee: Guangxi Jinsui Ecological Technology Group Co ltd
Priority date: 2019-11-20
Filing date: 2019-11-20
Publication date: 2021-10-08
Anticipated expiration: 2039-11-20
Also published as: CN110872518A

Abstract

The invention discloses an acid soil conditioner, wherein the total bacterial quantity of alcaligenes faecalis in 1 ml of the acid soil conditioner is more than or equal to 50 hundred million. The invention also discloses a preparation method and application of the acid soil conditioner. The soil conditioner of the invention takes the live alcaligenes faecalis as functional bacteria, and not only can effectively improve the pH value of acid soil, reduce the EC value of the soil, inhibit the growth and reproduction of pathogenic bacteria and improve the structure of microbial community in the soil by the life activity of the bacteria, but also can release fixed nutrient elements, thus fundamentally improving the soil and not damaging the ecological environment.

Description

Acid soil conditioner and preparation method and application thereof

Technical Field

The invention belongs to the technical field of soil improvement, and particularly relates to an acid soil conditioner and a preparation method and application thereof.

Background

The acid soil is a general term for soil with a pH value of less than 6.5, comprises brick red soil, red soil, yellow soil, dry red soil and other soils, and is mainly distributed in tropical and subtropical regions in China. The method is mainly characterized in that the soil acidification is caused by large and concentrated precipitation, strong leaching effect and large loss of alkaline salts such as calcium, magnesium, potassium and the like. Lime application, burnt manure application, large fertilizer application and the like in traditional agriculture are also important reasons for soil acidification. The harm of soil acidification mainly comprises the following points: firstly, after soil acidification, the quantity of beneficial microorganisms in soil is reduced, the growth and the activity of the beneficial microorganisms are inhibited, and even the microbial population in the soil is changed to influence the growth of crops; secondly, the soil acidification can cause the fixation of nutrient elements (P), thus aggravating the soil hardening and influencing the extension of root systems; thirdly, soil acidification can promote release, activation and dissolution of certain toxic elements, thereby influencing the yield and quality of crops.

At present, the main methods for improving acid soil are as follows: the quicklime is used for neutralizing acidity, planting green manure, increasing organic matters, applying organic fertilizers and farmyard manure, increasing irrigation times, applying alkaline fertilizers and the like. However, the methods have great defects, such as the use of quicklime for neutralizing acidity can destroy the microbial population structure of soil, the application of organic fertilizer needs great amount to exert the effect, the application of alkaline fertilizer is limited, and the improvement effect is not obvious.

Alcaligenes faecalis is a gram-negative bacterium widely existing in soil and water, can inhibit the growth of microorganisms such as staphylococcus aureus, pseudomonas aeruginosa, proteus vulgaris and the like, is harmless to human bodies, can produce some unnatural amino acids by various types of the bacteria, and is widely applied to the current pharmaceutical industry. In addition, the bacteria can also be applied to wastewater treatment to decompose organic matters in wastewater and realize water purification. However, no precedent exists for preparing the acid soil conditioner by using the alcaligenes faecalis.

Disclosure of Invention

The object of the present invention is to solve at least the above drawbacks and to provide advantages which will be explained later.

To achieve these objects and other advantages of the present invention, there is provided an acid soil conditioner, by which the pH of acid soil is raised.

Another object of the present invention is to provide an acid soil conditioner, which can reduce the EC value of soil.

It is still another object of the present invention to provide an acid soil conditioner, which can suppress the growth of pathogenic bacteria in soil.

In other words, the invention discloses an acid soil conditioner, wherein the total bacterial quantity of the alcaligenes faecalis in 1 ml of the acid soil conditioner is more than or equal to 50 hundred million.

The soil conditioner of the invention takes the live alcaligenes faecalis as functional bacteria, and through the life activities of the alcaligenes faecalis strains, the pH value of acid soil can be effectively improved, so that the growth and the propagation of pathogenic bacteria are inhibited, the structure of microbial communities in the soil is improved, the soil can release fixed nutrient elements, the EC value of the soil is reduced, the soil is fundamentally improved, and the ecological environment is not damaged.

The invention also discloses a preparation method of the acid soil conditioner, which comprises the following steps:

step one, obtaining an alcaligenes faecalis stock, after thawing, streaking and separating on an NA culture medium plate, and culturing for 18-36 hours at 32-37 ℃ to obtain a single colony of the alcaligenes faecalis; step two, selecting a single colony of the alcaligenes faecalis, inoculating the single colony in an NB liquid culture medium, and performing shake culture at the temperature of 32-37 ℃ and the rpm of 180-250 for 16-24 hours to obtain an activated seed solution; step three, taking the activated seed solution according to the volume ratio of 0.5-4% and transferring the activated seed solution into a liquid fermentation culture medium, and carrying out shake cultivation for 16-24 hours at the temperature of 32-37 ℃ and the rotation speed of 180-250 rpm to obtain a large amount of seed solution; transferring a large amount of the seed liquid obtained in the step three to a liquid fermentation culture medium according to the volume ratio of 0.5-4%, performing high-density fermentation culture in a fermentation tank, and stopping fermentation when the total bacterial quantity is more than or equal to 50 hundred million/ml to obtain a high-density liquid microbial agent; step five, directly taking the obtained high-density liquid microbial agent as the acid soil conditioner; or the acid soil conditioner is obtained after being matched with a carrier.

The acid soil conditioner of the invention takes alcaligenes faecalis as a strain stock, and the alcaligenes faecalis is activated and then is subjected to step-by-step enlarged culture to obtain high-density alcaligenes faecalis bacterial liquid, namely the acid soil conditioner. The preparation process is simple, the raw material consumption is low, the cost is low, and the preparation method is very suitable for large-scale production.

Preferably, the NA medium is obtained by: respectively taking 3-5 parts of beef extract, 8-10 parts of peptone, 3-5 parts of sodium chloride, 16-20 parts of agar and 1000 parts of water, mixing, and adjusting the pH value to 6.5-7.5 to obtain the NA culture medium.

Preferably, the NB liquid medium is obtained by: respectively taking 3-5 parts of beef extract, 8-10 parts of peptone, 3-5 parts of sodium chloride and 1000 parts of water, mixing, and adjusting the pH value to 6.5-7.5 to obtain the NB liquid culture medium.

Preferably, the liquid fermentation medium is obtained by: respectively taking 10-30 parts of glucose, 10-30 parts of corn steep liquor and 1000 parts of water, mixing and adjusting the pH value to 6.5-7.5 to obtain the liquid fermentation medium.

Preferably, peat soil is used as a carrier, and after the high-density liquid microbial agent is adsorbed, the peat soil is dried until the water content is less than or equal to 35 percent, so that the acid soil conditioner is obtained.

Preferably, the acid soil conditioner is obtained by mixing an organic material serving as a carrier with a high-density liquid microbial agent.

Preferably, the acid soil conditioner is obtained by mixing a liquid or solid nutrient and a high-density liquid microbial agent.

The invention takes peat soil or organic materials or liquid and solid nutrient substances as carriers to adsorb the alcaligenes faecalis and maintain the basic metabolism of the alcaligenes faecalis, and after the acid soil conditioner is applied to a field, the alcaligenes faecalis can better utilize the nutrient substances in the alcaligenes faecalis to continue to maintain life activities and further adapt to the external environment.

The invention also discloses an application of the alcaligenes faecalis as an acid soil conditioner.

The invention has the advantages that:

1. the acid soil conditioner of the invention takes the live alcaligenes faecalis as functional bacteria, and not only can effectively improve the pH value of acid soil through the life activities of the bacteria, but also can inhibit the growth and the propagation of pathogenic bacteria, improve the structure of microbial communities in the soil, enable the soil to release fixed nutrient elements, reduce the EC value of the soil, fundamentally improve the soil and cannot damage the ecological environment.

2. The acid soil conditioner of the invention takes alcaligenes faecalis as a strain stock, and the alcaligenes faecalis is activated and then is subjected to step-by-step enlarged culture to obtain high-density alcaligenes faecalis bacterial liquid, namely the acid soil conditioner. The preparation process is simple, the raw material consumption is low, the cost is low, and the preparation method is very suitable for large-scale production.

3. The invention takes peat soil or organic materials or liquid and solid nutrient substances as carriers to adsorb the alcaligenes faecalis and maintain the basic metabolism of the alcaligenes faecalis, and after the acid soil conditioner is applied to a field, the alcaligenes faecalis can better utilize the nutrient substances in the alcaligenes faecalis to continue to maintain life activities and further adapt to the external environment.

4. The acid soil conditioner of the invention takes the viable bacteria of the alcaligenes faecalis as raw materials, the raw materials have wide sources and low cost, the content of the viable bacteria is high after the step-by-step enlarged culture, and the acid soil conditioner can be produced on a large scale.

Detailed Description

The present invention is described in further detail below to enable those skilled in the art to practice the invention with reference to the description.

Example 1

An acid soil conditioner, wherein the total bacterial quantity of alcaligenes faecalis in 1 ml of the acid soil conditioner is more than or equal to 50 hundred million.

A preparation method of an acid soil conditioner comprises the following steps:

step one, obtaining an alcaligenes faecalis stock, after thawing, streaking and separating on an NA culture medium plate, and culturing for 24 hours at 35 ℃ to obtain a single colony of the alcaligenes faecalis.

And step two, selecting a single colony of the alcaligenes faecalis, inoculating the single colony in an NB liquid culture medium, and performing shake culture at 35 ℃ and 200rpm for 20 hours to obtain an activated seed solution.

Step three, taking the activated seed solution according to the volume ratio of 2 percent, transferring the activated seed solution into a liquid fermentation culture medium, and carrying out shake cultivation for 20 hours at 35 ℃ and 200rpm to obtain a large amount of seed solution.

Step four, transferring a large amount of the seed liquid obtained in the step three to a liquid fermentation culture medium according to the volume ratio of 2%, performing high-density fermentation culture in a fermentation tank, and stopping fermentation when the total bacterial load is more than or equal to 50 hundred million/ml to obtain the acid soil conditioner.

Example 2

A preparation method of an acid soil conditioner comprises the following steps:

step one, obtaining an alcaligenes faecalis stock, after thawing, streaking and separating on an NA culture medium plate, and culturing for 24 hours at 37 ℃ to obtain a single colony of the alcaligenes faecalis.

And step two, selecting a single colony of the alcaligenes faecalis, inoculating the single colony in an NB liquid culture medium, and performing shake culture at 37 ℃ and 200rpm for 18 hours to obtain an activated seed solution.

Step three, taking the activated seed solution according to the volume ratio of 1 percent, transferring the activated seed solution into a liquid fermentation culture medium, and carrying out shake cultivation for 18 hours at 37 ℃ and 200rpm to obtain a large amount of seed solution.

Step four, transferring a large amount of the seed liquid obtained in the step three to a liquid fermentation culture medium according to the volume ratio of 0.5%, performing high-density fermentation culture in a fermentation tank, and stopping fermentation when the total bacterial load is more than or equal to 50 hundred million/ml to obtain the acid soil conditioner.

Example 3

A preparation method of an acid soil conditioner comprises the following steps:

And step two, selecting a single colony of the alcaligenes faecalis, inoculating the single colony in an NB liquid culture medium, and carrying out shake culture at 32 ℃ and 200rpm for 36 hours to obtain an activated seed solution.

Step three, taking the activated seed solution according to the volume ratio of 4 percent, transferring the activated seed solution into a liquid fermentation culture medium, and carrying out shake cultivation for 24 hours at the temperature of 32 ℃ and the speed of 200rpm to obtain a large amount of seed solution.

Step four, transferring a large amount of the seed liquid obtained in the step three to a liquid fermentation culture medium according to the volume ratio of 4%, performing high-density fermentation culture in a fermentation tank, and stopping fermentation when the total bacterial load is more than or equal to 50 hundred million/ml to obtain the acid soil conditioner.

Example 4

A preparation method of an acid soil conditioner comprises the following steps:

Step four, transferring the mass of seed liquid obtained in the step three to a liquid fermentation culture medium according to the volume ratio of 2%, and performing high-density fermentation culture in a fermentation tank until the total bacterial load is more than or equal to 50 hundred million/ml, so as to obtain the high-density liquid microbial agent.

And step five, adsorbing the high-density liquid microbial agent by using peat soil as a carrier, and drying until the water content is less than or equal to 35% to obtain the acid soil conditioner.

Example 5

A preparation method of an acid soil conditioner comprises the following steps:

And step five, mixing the high-density liquid microbial agent with an organic material as a carrier, and drying until the water content is less than or equal to 35% to obtain the acid soil conditioner.

Example 6

A preparation method of an acid soil conditioner comprises the following steps:

the method comprises the following steps: obtaining an alcaligenes faecalis stock, after thawing, streaking and separating on an NA culture medium plate, and culturing for 24 hours at 35 ℃ to obtain a single colony of the alcaligenes faecalis.

And step five, mixing the high-density liquid microbial agent with a liquid nutrient substance as a carrier to obtain the acid soil conditioner.

Example 7

A preparation method of an acid soil conditioner comprises the following steps:

And step five, mixing the high-density liquid microbial agent with solid nutrient substances as carriers to obtain the acid soil conditioner.

Example 8

The indoor soil improvement effect test:

step one, selecting acid soil with the pH value of 4.5 as test soil, and screening to remove large particles to obtain planting soil.

And step two, adopting a microwave sterilization method to sterilize the acid soil conditioner prepared in the example 1 to be used as a substrate. And performing aseptic inspection on the sterilized acidic soil conditioner to obtain aseptic growth.

Step three, adding water to the substrate obtained in the step two to dilute by 50 times to obtain a first diluent; the acid soil conditioner prepared in example 1 was diluted 50 times with water to obtain a second diluted solution.

And step four, respectively taking 3 parts of 1 kg of planting soil for grouping treatment. Wherein, equal amount of clear water is added into the first planting soil as a first group; adding an equal amount of the first diluent into the second planting soil to serve as a second group; and adding an equivalent amount of second diluent into the third planting soil to serve as a third group.

And step five, placing the three groups of planting soil subjected to different treatments for 25 days in the same environment.

And sixthly, repeating the test for 3 times, detecting indexes of pH, organic matters, alkaline hydrolysis nitrogen, available phosphorus, quick-acting potassium, bacteria, fungi, actinomycetes and EC of soil of each group, calculating and recording an average value, wherein the result is shown in a table 1:

table 1: index for improving indoor soil

As can be seen from the data in Table 1, the acid soil conditioner prepared by the invention has good conditioning effect on acid soil, wherein the most obvious index is soil pH, and the pH value of the soil can be improved from 4.5 to 6.7; and secondly, the EC value of the soil can be obviously reduced, and the organic matter content in the soil before and after the test is obviously reduced. N, P, K indexes in the soil are all improved, which shows that the acid soil conditioner prepared by the invention can activate the nutrition in the soil. The change trend of soil microorganism indexes conforms to the theory, namely the acid soil is beneficial to the growth and the propagation of fungi in the soil and is unfavorable for bacteria and actinomycetes, and the pH value of the soil is increased by applying the soil conditioner, so that the population structures of the soil bacteria, the fungi and the actinomycetes are influenced.

Example 9

Soil bacteriostasis effect test:

step one, selecting acid soil with the pH value of 5.5 as test soil, and screening to remove large particles to obtain the test soil.

And step two, culturing pathogenic fusarium by using a potato and potato culture medium (PDA) to obtain a pathogenic bacterium liquid containing a large number of spores.

And step three, sterilizing the acid soil conditioner prepared in the example 1 by adopting a microwave sterilization method to serve as a substrate. And performing aseptic inspection on the sterilized acidic soil conditioner to obtain aseptic growth.

Step four, adding water to dilute the substrate obtained in the step three by 50 times to obtain a first diluent; the acid soil conditioner prepared in example 1 was diluted 50 times with water to obtain a second diluted solution.

Step five, adding the fusarium liquid into the standby soil, uniformly stirring, and then evenly dividing the soil into A, B, C trisections for treatment; wherein group A is added with a second diluent such as soil; adding a first diluent with the same amount of soil into the group B; and adding clean water with the same amount as soil into the group C.

And step six, adopting a dilution coating method to periodically track and detect the bacterial load of pathogenic fusarium in the soil. 3 replicates were made and averaged, the results are shown in Table 2:

table 2: table of change in the amount of soil pathogenic Fusarium (cfu/g)

Item

Day 0

10 days

20 days

30 days

40 days

50 days

Group A

5.5E+5

1.1E+5

3.3E+4

7.5E+3

3.6E+3

1.0E+3

Group B

5.1E+5

3.8E+5

8.1E+5

9.7E+5

2.5E+6

2.4+E6

Group C

5.7E+5

6.6E+5

7.2E+5

8.6E+5

2.0E+6

1.8+E6

As can be seen from the data in Table 2, the content of pathogenic fusarium in the soil of the test group A shows a significant decline trend; the content of pathogenic fusarium in the soil of the test group B is slightly reduced in the first 10 days and then shows an ascending trend; the test group C is a blank group, the content of pathogenic fusarium in the soil is in an increasing trend, and the natural growth condition of pathogenic bacteria in the soil is reflected. The acidic soil conditioner prepared by the invention can effectively inhibit the growth of pathogenic fusarium in the acidic soil.

Example 10

The field soil improvement effect test:

step one, selecting a plot which is flat in terrain, is planted with a variety of Wei-cheap B6 bananas and is subjected to bud drawing as a test area at a certain company headquarter base, and randomly dividing the plot into 3 groups.

Step three, performing conventional fertilization on the first group of bananas; adding the substrate obtained in the second step on the basis of conventional fertilization to the second group of bananas in an amount of 4 liters/mu; the third group of bananas is added with the acid soil conditioner prepared in example 1 in an amount of 4 liters/acre on the basis of conventional fertilization. The application method of the matrix and the acid soil conditioner comprises the following steps: calculated by 4 liters per mu, the fertilizer is diluted by 100 times and applied by irrigating roots.

And step four, after the test is started, all groups are uniformly managed according to a conventional banana planting method, and only at the beginning of the test, the groups are respectively treated according to the test design until the test is finished for 90 days.

Step five, repeating the test for 3 times, detecting indexes of pH, organic matters, alkaline hydrolysis nitrogen, available phosphorus, quick-acting potassium, bacteria, fungi, actinomycetes and EC of soil of each group, calculating and recording an average value, and obtaining a result shown in a table 3:

table 3: index for improving field soil

As can be seen from the data in Table 3, the acid soil conditioner prepared by the invention has good conditioning effect on acid soil, wherein the most significant index is soil pH. The pH value of the soil can be increased from 5.8 to 6.9; and secondly, the EC value of the soil can be obviously reduced, and the organic matter content in the soil before and after the test is obviously reduced. The N, K index in the soil has a downward trend, while the P index has an upward trend, which shows that the pH of the soil is increased to release the P element fixed by the soil, but the P content of the soil is increased because the banana has less demand for P. The change trend of soil microorganism indexes conforms to the theory, namely the acid soil is beneficial to the growth and the propagation of fungi in the soil and is unfavorable for bacteria and actinomycetes, and the pH value of the soil is increased by applying the soil conditioner, so that the population structures of the soil bacteria, the fungi and the actinomycetes are influenced.

Example 11

Vegetable yield increase effect test:

step one, selecting a vegetable field, and dividing the vegetable field into 4 test areas with the specification of 3.2m multiplied by 10 m.

And step two, adopting a microwave sterilization method to sterilize the acid soil conditioner prepared in the embodiment 3 to be used as a substrate. And performing aseptic inspection on the sterilized acidic soil conditioner to obtain aseptic growth.

Step three, after the vegetable field is ridged, scattering organic fertilizer as base fertilizer according to the amount of 600 kg/mu in the first test area; the second test area sprays the acid soil conditioner prepared in the embodiment 3 in an amount of 5 liters/mu on the basis of the first test area; spraying the substrate obtained in the second step on the basis of the first test area in a third test area in an amount of 5 liters/mu; the fourth test area was not fertilized.

Step four, sowing equal amount of leaf tip rape pod heart in four test areas at the same time; after 15 days of sowing, dressing urea once in an amount of 25 kg/mu except the fourth test area; the field management of each test area is the same until the heart of the leaf mustard green is harvested.

And step five, removing old leaves and rotten leaves after the core of the leaf tip rape is collected respectively, and weighing. Wherein, the four test areas are harvested on the same day.

Wherein, the vegetable yield increase effect test sets up 3 times of repeated tests, and the result is as shown in Table 4:

table 4: vegetable yield increasing effect (kilogram)

Item	First test zone	Second test area	Third test zone	Fourth test area
					Repeat test 1	79.2	88.5	81.5	59.8
Repeat test 2	82.1	86.9	79.9	63.0
					Repeat test 3	77.9	89.5	82.5	62.3
Average yield	79.7	88.3	81.3	62.3

As can be seen from the data in Table 4, the acid soil conditioner prepared by the invention has extremely obvious effect on yield increase of the leaf tip rape heart.

While embodiments of the invention have been disclosed above, it is not intended to be limited to the uses set forth in the specification and examples. It can be applied to all kinds of fields suitable for the present invention. Additional modifications will readily occur to those skilled in the art. The invention is therefore not to be limited to the specific details described herein, without departing from the general concept as defined by the appended claims and their equivalents.

Claims

1. The preparation method of the acid soil conditioner is characterized by comprising the following steps:

step one, obtaining an alcaligenes faecalis stock, after thawing, streaking and separating on an NA culture medium plate, and culturing for 18-36 hours at 32-37 ℃ to obtain a single colony of the alcaligenes faecalis;

step two, selecting a single colony of the alcaligenes faecalis, inoculating the single colony in an NB liquid culture medium, and performing shake culture at the temperature of 32-37 ℃ and the rpm of 180-250 for 16-24 hours to obtain an activated seed solution;

step three, taking the activated seed solution according to the volume ratio of 0.5-4% and transferring the activated seed solution into a liquid fermentation culture medium, and carrying out shake cultivation for 16-24 hours at the temperature of 32-37 ℃ and the rotation speed of 180-250 rpm to obtain a large amount of seed solution;

transferring a large amount of the seed liquid obtained in the step three to a liquid fermentation culture medium according to the volume ratio of 0.5-4%, performing high-density fermentation culture in a fermentation tank, and stopping fermentation when the total bacterial quantity is more than or equal to 50 hundred million/ml to obtain a high-density liquid microbial agent;

step five, directly taking the obtained high-density liquid microbial agent as the acid soil conditioner; or the acid soil conditioner is obtained after being matched with the carrier;

wherein the total bacterial amount of the alcaligenes faecalis in 1 ml of the acid soil conditioner is more than or equal to 50 hundred million.

2. The method for preparing an acid soil conditioner according to claim 1, wherein the NA medium is obtained by:

respectively taking 3-5 parts of beef extract, 8-10 parts of peptone, 3-5 parts of sodium chloride, 16-20 parts of agar and 1000 parts of water, mixing, and adjusting the pH value to 6.5-7.5 to obtain the NA culture medium.

3. The method for preparing an acid soil conditioner according to claim 1, wherein the NB liquid medium is obtained by:

respectively taking 3-5 parts of beef extract, 8-10 parts of peptone, 3-5 parts of sodium chloride and 1000 parts of water, mixing, and adjusting the pH value to 6.5-7.5 to obtain the NB liquid culture medium.

4. The method for preparing an acid soil conditioner according to claim 1, wherein the liquid fermentation medium is obtained by:

respectively taking 10-30 parts of glucose, 10-30 parts of corn steep liquor and 1000 parts of water, mixing and adjusting the pH value to 6.5-7.5 to obtain the liquid fermentation medium.

5. The method for preparing an acid soil conditioner as claimed in claim 1, wherein the acid soil conditioner is obtained by using peat soil as a carrier, adsorbing the high-density liquid microbial agent, and drying until the water content is less than or equal to 35%.

6. The method for producing an acid soil improvement agent according to claim 1, wherein the acid soil improvement agent is obtained by mixing an organic material as a carrier with a high-density liquid microbial agent.

7. The method for producing an acid soil improvement agent according to claim 1, wherein the acid soil improvement agent is obtained by mixing a liquid or solid nutrient with a high-density liquid microbial agent.