CN114557259A - Vegetable seedling culture medium containing high-activity growth-promoting bacteria and preparation method and application thereof - Google Patents
Vegetable seedling culture medium containing high-activity growth-promoting bacteria and preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to the technical field of seedling raising substrates, in particular to a vegetable seedling raising substrate containing high-activity growth-promoting bacteria and a preparation method and application thereof. The preparation method provided by the invention adopts a step-by-step expansion mode to mix the bacterium powder of the growth-promoting bacteria and the seedling substrate, and specifically, the bacterium powder is added into a carrier, and then the carrier containing the growth-promoting bacteria is added into the seedling substrate. The invention also provides a method for determining the type of the carrier, the particle size of the carrier, the moisture content of the matrix, the mixing speed and the mixing time aiming at different bacterial powders. In the vegetable seedling substrate obtained by the preparation method provided by the invention, the variation coefficient of growth-promoting bacteria is below 5%, and the vegetable seedling substrate obtained by the preparation method provided by the invention can be used for cultivating vegetable seedlings with high growth state consistency, and is beneficial to the unified management of vegetable seedling.
Description
Technical Field
The invention relates to the technical field of seedling raising substrates, in particular to a vegetable functional seedling raising substrate containing growth-promoting bacteria, and a preparation method and application thereof.
Background
The substrate is used for replacing soil in the seedling culture process of the vegetables, so that proper conditions of fertilizer, water, gas and the like are provided for the growth of seedlings, and the conventional seedling culture substrate is produced by mixing turfy soil, perlite, vermiculite and other materials serving as raw materials according to a certain proportion. In order to ensure the quality of vegetable seedlings in actual production, chemical fertilizers and pesticides are inevitably used in the seedling raising process, and residues of the chemical fertilizers and the pesticides are a great hidden danger of food safety.
The plant growth-promoting rhizobacteria are microorganisms which are beneficial to the growth of plants, can promote the growth of the plants, relieve plant diseases and increase the capability of the plants to resist biotic stress and abiotic stress. Therefore, the plant growth-promoting rhizobacteria are added into the seedling culture substrate to prepare the vegetable seedling culture substrate containing the growth-promoting bacteria, so that the vegetable seedling culture substrate has the functions of promoting growth, preventing diseases and increasing resistance, and the use of chemical fertilizers and pesticides can be effectively reduced while the seedling culture quality is ensured.
In the process of adding the plant growth-promoting rhizobacteria into the seedling substrate, the adding mode and the adding amount are main influence factors on the functionality of the plant growth-promoting rhizobacteria. The adding mode of the plant growth-promoting rhizobacteria mainly influences the uniformity of the growth-promoting bacteria in the matrix, and the uneven distribution of the growth-promoting bacteria can cause the growth condition of seedlings to be different in the seedling raising process, thereby causing seedling raising accidents. In addition, because the bacteria content of the plant rhizosphere growth promoting microbial inoculum is high, the addition amount in the practical application process is relatively small, the difficulty of mixing is increased by directly adding the plant rhizosphere growth promoting microbial inoculum, the stirring time needs to be increased, and the production efficiency is reduced in enterprise production.
Disclosure of Invention
The invention aims to provide a vegetable seedling substrate with uniformly distributed growth-promoting bacteria. In the seedling culture substrate containing growth-promoting bacteria in the prior art, the seedling growth condition in the seedling culture process is different due to the uneven distribution of the growth-promoting bacteria, so that seedling culture accidents are caused.
In a first aspect, the invention provides a preparation method of a vegetable seedling substrate containing growth-promoting bacteria. By adopting the method provided by the invention, both single growth-promoting bacteria and mixed growth-promoting bacteria can be uniformly distributed in the seedling culture substrate.
In the prior art, in order to make the growth-promoting bacteria uniformly distributed, the stirring time is generally increased. However, when the stirring time is increased, the growth-promoting bacteria are uniformly distributed, the production efficiency is also reduced, the seedling raising cost is increased, and the volume weight, the porosity and the like of the matrix are reduced due to the overlong stirring and mixing time, so that the water and fertilizer holding capacity of the matrix is reduced, the plants are easy to fall down, and the growth of seedlings is not facilitated.
In the prior art, in the using process of the fungus powder, the adsorbent is used for helping the fungus powder to be more uniform in the matrix, but the adsorbent has certain defects, firstly, the adsorbent is additionally used to increase the cost in the production process, secondly, the adsorbent is additionally added substance, the additional addition has influence on the physicochemical property of the seedling matrix and is not beneficial to actual seedling culture, for example, calcium bicarbonate powder is used for fungus powder adsorption, but after the fungus powder is used, the influence on the pH value of the seedling matrix is large, so that the matrix is alkaline and is not suitable for seedling culture of most vegetables.
The invention discovers that the uniformity of the distribution of the growth-promoting bacteria can be more effectively improved by using a certain component in the seedling culture substrate as a carrier, mixing the component with the bacteria powder and adopting a step-by-step amplification method.
Specifically, the preparation method of the seedling substrate containing growth-promoting bacteria comprises the steps of mixing the growth-promoting bacteria and a carrier for the first time, and mixing the carrier containing the growth-promoting bacteria and the seedling substrate for the second time; the carrier is one of the components in the seedling substrate.
In the preparation method provided by the invention, the weight ratio of the carrier to the seedling substrate is 1: (15-25). In the prior art, before the bacterial powder is added to the seedling substrate, the bacterial powder and a small amount of seedling substrate are mixed and then added. However, experiments show that the problem of uneven distribution of growth-promoting bacteria cannot be solved by the aid of the adding mode. Furthermore, the invention discovers that the variation coefficient of the growth-promoting bacteria is the lowest after the carrier is mixed with the bacterial powder and then the carrier containing the bacterial powder is mixed with the seedling culture substrate; the carrier used in the invention is only one component in the breeding substrate.
Based on the idea that the growth-promoting bacteria need to be mixed with the carrier firstly, the invention further defines indexes which have correlation with the variation coefficient of the growth-promoting bacteria, such as the types of the carrier, the particle size of the carrier, the water content of the seedling substrate, and the rotating speed and the time during mixing.
In the preparation method provided by the invention, before the first mixing, the type of the carrier and the particle size of the carrier are determined.
In the preparation method provided by the invention, before the second mixing, the moisture content of the seedling substrate and the rotation speed of the second mixing are determined.
In the preparation method provided by the invention, the water content of the seedling substrate is 30-35% during the second mixing; in the seedling raising substrate, the volume ratio of turfy soil: vermiculite: the perlite is (2-3): (1-2): 1-2).
In the preparation method provided by the invention, the rotating speed is 30-40r/min and the mixing time is 5-8min during the first mixing or the second mixing.
More specifically, in the preparation method provided by the invention, during the first mixing or the second mixing, the mixing mode is clockwise rotation for 20-30s, and anticlockwise rotation for 20-30 s; clockwise and anticlockwise alternately, and circulating for 4-5 times.
In a second aspect, the invention provides a seedling substrate containing growth-promoting bacteria, which is prepared by the preparation method, wherein the variation coefficient of the growth-promoting bacteria in the seedling substrate is less than 5%.
In the seedling raising substrate containing growth-promoting bacteria, carriers are turfy soil, perlite and vermiculite; the grain diameter of the turfy soil is less than 1 mm; in the seedling raising matrix containing the growth-promoting bacteria, the breeding matrix is turfy soil, vermiculite and perlite, wherein the turfy soil: vermiculite: the volume ratio of the perlite is (2-3) to (1-2).
According to the understanding of the technical personnel in the field, the invention also claims the application of the seedling substrate containing the growth-promoting bacteria in improving the industrial seedling raising efficiency.
In a third aspect, the invention provides a method for detecting the variation coefficient of growth-promoting bacteria in a seedling substrate, which comprises the steps of sampling the seedling substrate containing the growth-promoting bacteria, wherein the sampling number is more than or equal to 5, and adding the sampled sample into physiological saline to obtain a bacterial liquid; in an ultra-clean workbench, carrying out gradient dilution on the bacterial liquid, coating the diluted bacterial liquid on an LB flat plate, and culturing to obtain bacterial colonies; and taking the average value of the colonies of the samples as the number of the growth-promoting bacteria, and calculating the variation coefficient of the growth-promoting bacteria in the same sample.
The invention has the beneficial effects that:
by adopting the method provided by the invention, the uniformity of the growth-promoting bacteria in the seedling substrate is obviously improved, and particularly, the variation coefficient of the growth-promoting bacteria in the prepared seedling substrate is less than 5%. Meets the requirement of industrial seedling culture.
Drawings
FIG. 1 is a schematic diagram of sampling in example 1 of the present invention.
FIG. 2 is a line graph showing the growth-promoting bacteria variation coefficients of different vector types in example 1 of the present invention.
FIG. 3 is a line graph showing the variation coefficient of growth-promoting bacteria when different turfy soil particle sizes are used as carriers in example 1 of the present invention.
FIG. 4 is a line graph showing the variation coefficient of growth-promoting bacteria when the water content of the substrate is different in example 1 of the present invention.
FIG. 5 is a line graph showing the growth-promoting bacteria variation coefficient when the second mixing rotation speed is different in example 1 of the present invention.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.
By adopting the method provided by the invention, the most suitable carrier type and carrier particle size can be found for any bacterium powder, so that the invention provides a general method for preparing the seedling culture substrate containing growth-promoting bacteria.
Example 1
The bacterial powder used in this example is a mixed bacterial powder containing various bacteria, and is in the form of powder, specifically, the bacterial powder is produced by the laboratory and contains 1 × 10 bacteria11CFU/g; the seedling culture medium is a mixed medium of turfy soil, vermiculite and perlite in a ratio of 3:1:1(v/v/v), wherein the turfy soil is domestic turfy soil, vermiculite and pearlPerlite was purchased from Beijing flower market (particle size 3-6 mm).
This example provides a process for exploring a method for preparing a vegetable seedling substrate containing high-activity growth-promoting bacteria, as follows:
1. type of support
Comparison group (i): namely turfy soil, vermiculite and perlite mixed matrix with the ratio of 3:1:1(v/v/v) as an inoculums group
Processing group two: directly adding the bacterial powder into a seedling culture substrate, and mixing for the second time to be used as a carrier-free group;
processing group III: mixing the bacterial powder with turfy soil, namely mixing the bacterial powder with a turfy soil carrier, adding the turfy soil carrier containing growth-promoting bacteria into the mixed seedling culture substrate, and mixing for the second time to obtain a turfy soil group;
treatment group iv: mixing the bacterial powder with perlite, namely mixing the bacterial powder with a perlite carrier, adding the perlite carrier containing growth-promoting bacteria into the mixed seedling substrate, and mixing for the second time to obtain a perlite group;
a treatment group is: mixing the bacterial powder and vermiculite, namely mixing the bacterial powder and a vermiculite carrier, adding the vermiculite carrier containing growth promoting bacteria into the mixed seedling culture substrate, and mixing for the second time to obtain a vermiculite group;
treatment groups are: the bacterial powder is firstly mixed with the seedling substrate, namely the bacterial powder is firstly mixed with the seedling substrate, then the substrate containing the growth-promoting bacteria is added into the mixed seedling substrate, and the mixture is mixed for the second time to be used as the substrate group.
And during the second mixing, adding the carrier containing the growth-promoting bacteria into the seedling substrate, mixing at a fixed rotating speed, and rotating for 30s clockwise and 30s anticlockwise. The clockwise and anticlockwise are alternately carried out, the circulation is carried out for 8 times, and the total time is 8 minutes.
In the second mixing process, 1min, 2min, 3min, 4min, 5min, 6min, 7min and 8min are taken as time points each time, the mixing is stopped, the seedling substrate is uniformly loaded into a 72-hole plug tray, five-point sampling is carried out on the loaded seedling substrate, samples obtained at the five sampling points are respectively marked as a sample 1, a sample 2, a sample 3, a sample 4 and a sample 5, 25ml of sample is taken at each point, and 40 samples are taken in each treatment. The sampling schematic is shown in fig. 1. And (4) calculating the number of the growth-promoting bacteria in each sample, and calculating the coefficient of variation.
Specifically, the calculation steps of the coefficient of variation are as follows:
adding 25ml of seedling substrate sample into sterilized physiological saline containing 225ml (a bottle is pre-filled with a proper amount of sterile glass beads), shaking for 20min at 200r/min in a 15 ℃ shaking box, performing gradient dilution in an ultra-clean workbench, sucking 100 mul of diluent with proper gradient, dropwise adding the diluent into an LB flat plate, uniformly coating the surface of the LB flat plate with a sterile coating rod, culturing for 24h at 37 ℃, and counting the number of colonies on the flat plate to serve as the number of growth-promoting bacteria contained in the sample. Each gradient was repeated 3 times, and the mean value (M) was obtained, and the standard deviation (S) and Coefficient of Variation (CV) of the growth-promoting bacteria contained in the sample at each time point were calculated.
The coefficient of variation of different carrier types obtained by the method is shown in figure 2, and the result in figure 2 shows that the coefficient of variation of the growth-promoting bacteria is the minimum when the turfy soil is used as a carrier, so that the turfy soil is determined to be used as the carrier for preparing the growth-promoting bacteria vegetable seedling substrate.
2. Particle size of the support
Mix fungus powder and particle diameter for being less than 1 mm's turfy soil, be greater than 1mm and be less than 2 mm's turfy soil, be greater than 2mm and be less than 5 mm's turfy soil, be less than 5 mm's turfy soil respectively, by mass ratio, the proportion of fungus powder and turfy soil is 1:25, mixing;
then mixing the carrier containing the bacterial powder with a seedling culture substrate; and (3) mixing, sampling, and calculating the growth-promoting bacterium count and the growth-promoting bacterium variation coefficient in the sample in the same step 1. When different turfy soil grain diameters are used as carriers, the variation coefficient of the growth-promoting bacteria is shown in figure 3. The results in fig. 3 show that peatmoss of less than 1mm is suitable as a carrier because: (1) the turfy soil with the grain diameter smaller than 1mm has the lowest final coefficient of variation; (2) the variation coefficient of the turfy soil with the grain diameter less than 1mm is reduced most rapidly in the mixing process.
3. Moisture content of the substrate
Mixing the bacterial powder with turfy soil with the particle size less than 1mm, mixing the carrier containing the bacterial powder with seedling culture medium with the water content of 15%, 20%, 25%, 30% and 35%, respectively, and mixing, sampling, counting and calculating the method.
The variation coefficient of the growth-promoting bacteria when the moisture content of the substrate was different is shown in FIG. 4. The results in fig. 4 show that the moisture content of the seedling substrate when mixed is the best at 35%, the final coefficient of variation is the lowest and the coefficient of variation decreases the fastest.
4. Rotation speed at the time of second mixing
Mixing the bacterial powder with turfy soil with the particle size less than 1mm, adding the carrier containing the bacterial powder into a seedling culture substrate with the water content of 35%, mixing at the rotating speeds of 20r/min, 25r/min, 30r/min, 35r/min, 40r/min and 45r/min respectively, and carrying out mixing, sampling, counting and calculating by the same method as the above.
The variation coefficient of the growth-promoting bacteria when the rotation speed is different in the second mixing is shown in fig. 5. The results in fig. 5 show that the effect is best when the rotation speed of the second mixing is 40r/min, the final coefficient of variation is the lowest, and at the 5 th min of mixing, the coefficient of variation is 4.90%, and the mixing time is the shortest, so that the production requirement is met.
In addition, in practical experiments, it is found that if the mixing time is too long, such as 8min, or the rotating speed is too high, such as 45r/min, even if the variation coefficient can meet the requirement of less than 5%, the prepared seedling raising substrate has poor properties, such as volume weight, porosity and the like, and the vegetable seedling raising substrate is not beneficial to the growth of seedlings.
Example 2
The bacterial powder and the seedling substrate used in this example were the same as those in example 1.
In this example, the results obtained in example 1 were investigated to prepare a growth-promoting fungus-containing vegetable seedling substrate. Specifically, the carrier used in this example is peatmoss smaller than 1 mm; the moisture content of the substrate used in this example was 35%; the mixing speed of the present embodiment is 40 r/min.
The embodiment provides a preparation method of a vegetable seedling substrate containing high-activity growth-promoting bacteria, which comprises the following steps:
(1) the turfy soil with the particle size of less than 1mm is adopted as a carrier, and is mixed with growth-promoting bacterium powder for the first time, and the bacterium powder comprises the following components in percentage by mass: the mixing ratio of the carriers is 1: 25; adding the bacterial powder into the carrier, mixing at a fixed rotation speed, rotating clockwise for 30s, rotating anticlockwise for 30s, and alternately rotating clockwise and anticlockwise for 8 times, wherein the total time is 8 minutes. Obtaining the carrier containing the growth-promoting bacteria.
(2) Mixing a substrate with the moisture content of 35% with the carrier containing the growth-promoting bacteria obtained in the step (1) in a mixing ratio of 1: 25; the rotation speed during mixing is 40r/min, clockwise rotation is 30s, and anticlockwise rotation is 30 s. And (4) alternately performing clockwise rotation and anticlockwise rotation, circulating for 5 times, and obtaining the vegetable seedling substrate containing the growth-promoting bacteria in 5 minutes.
By using the same variation coefficient calculation method as that in example 1, the variation coefficient of the growth-promoting bacteria in the vegetable seedling substrate containing the growth-promoting bacteria prepared in this example is calculated to be 4.77%. Meets the production requirements of the functional seedling raising matrix and meets the requirements of industrial seedling raising.
Thus, this example demonstrates that the type of carrier, the particle size of the carrier, the moisture content of the seedling substrate, the rotation speed at the time of mixing, and the mixing time during the mixing can be determined by the method of example 1. Finally, the requirements of the production regulation of the composite functional seedling substrate that the variation coefficient of the growth-promoting bacteria in the vegetable seedling substrate containing the growth-promoting bacteria is below 5 percent after the mixing in a short time can be achieved.
Example 3
The growth-promoting bacterium powder in the embodiment is lucokwei, and the bacterium content is 2 multiplied by 1010CFU/g, available from Zhongnong Lvkang (Beijing) Biotechnology, Inc.; in the embodiment, the method provided in embodiment 1 is adopted, the most suitable carrier obtained is turfy soil, the particle size is less than 1mm, the water content of the matrix is 35%, the mixing speed in the embodiment is 35r/min, and the mixing time is 6 min.
In this embodiment, a vegetable seedling substrate containing growth-promoting bacteria is obtained by the same preparation method as in embodiment 2, and the variation coefficient of the vegetable seedling substrate containing growth-promoting bacteria obtained in this embodiment is 4.89%, which meets the industrial seedling requirement.
Example 4
The growth-promoting bacterium powder in the embodiment is bacillus subtilis, and the bacterium content is 4 multiplied by 1010CFU/g, available from Shandong LvLong Biotech, Inc.; in the embodiment, the method provided in embodiment 1 is adopted, the most suitable carrier obtained is turfy soil, the particle size is less than 1mm, the water content of the matrix is 35%, the mixing speed in the embodiment is 30r/min, and the mixing time is 8 min.
In the embodiment, the same preparation method as that in the embodiment 2 is adopted to obtain the vegetable seedling substrate containing the growth-promoting bacteria, and the variation coefficient of the vegetable seedling substrate containing the growth-promoting bacteria obtained in the embodiment is 4.93%, so that the vegetable seedling substrate meets the industrial seedling requirement.
Comparative example 1
The comparative example researches the existence of the carrier in the preparation of the vegetable seedling substrate containing the growth promoting bacteria, and the calculation method of the variation coefficient of the growth promoting bacteria adopted by the example is the same as that of the example 1, and the comparative example discovers that the variation coefficient of the growth promoting bacteria cannot be remarkably reduced by prolonging the mixing time under the condition of not using the carrier in the mixing process and is far higher than the variation coefficient of the carrier, so that the condition of mixing without the carrier is not considered when the vegetable seedling substrate containing the growth promoting bacteria is prepared.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (10)
1. A preparation method of a seedling substrate containing growth-promoting bacteria is characterized by comprising the steps of mixing the growth-promoting bacteria and a carrier for the first time, and mixing the carrier containing the growth-promoting bacteria and the seedling substrate for the second time; the carrier is one of the components in the seedling substrate.
2. The preparation method according to claim 1, wherein the weight ratio of the carrier to the seedling substrate is 1: (15-25).
3. The production method according to claim 2, wherein the type of the carrier used and the particle size of the carrier are determined before the first mixing.
4. The preparation method according to claim 3, wherein the moisture content of the seedling substrate and the rotation speed of the second mixing are determined before the second mixing.
5. The preparation method according to claim 4, wherein the moisture content of the seedling substrate used in the second mixing is 30-35%; in the seedling raising substrate, turfy soil: vermiculite: the volume ratio of the perlite is (2-3) to (1-2).
6. The method according to any one of claims 1 to 5, wherein the rotation speed is 30 to 40r/min and the mixing time is 5 to 8min at the time of the first mixing or the second mixing.
7. A seedling substrate containing growth-promoting bacteria, which is prepared by the preparation method of any one of claims 1 to 6, and the variation coefficient of the growth-promoting bacteria in the seedling substrate is less than 5%.
8. The growth-promoting bacterium-containing seedling substrate according to claim 7, wherein in the seedling substrate, the carrier is turfy soil, perlite or vermiculite; the grain diameter of the turfy soil is less than 1 mm.
9. Use of a growth-promoting bacterium-containing seedling substrate according to any one of claims 7 to 8 for improving industrial seedling raising efficiency.
10. A method for detecting the variation coefficient of growth-promoting bacteria in a seedling substrate is characterized in that the seedling substrate containing the growth-promoting bacteria is sampled, the number of the samples is more than or equal to 5, and the samples obtained by sampling are added into physiological saline to obtain a bacterial liquid; in an ultra-clean workbench, carrying out gradient dilution on the bacterial liquid, coating the diluted bacterial liquid on an LB flat plate, and culturing to obtain bacterial colonies; and taking the average value of the colonies of the samples as the number of the growth-promoting bacteria, and calculating the variation coefficient of the growth-promoting bacteria in the same sample.
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