CN111574299B

CN111574299B - Bacterial fertilizer for repairing plant leaf antioxidant system and preparation method thereof

Info

Publication number: CN111574299B
Application number: CN202010468592.5A
Authority: CN
Inventors: 苏清海
Original assignee: Pulege Xiamen Biotechnology Co ltd
Current assignee: Pulege Xiamen Biotechnology Co ltd
Priority date: 2020-05-28
Filing date: 2020-05-28
Publication date: 2022-07-26
Anticipated expiration: 2040-05-28
Also published as: CN111574299A

Abstract

The invention provides a bacterial fertilizer for repairing an antioxidant system of plant leaves and a preparation method thereof, and relates to the technical field of agriculture. The preparation method of the bacterial fertilizer comprises the following steps: obtaining compound strain fermentation liquor; inoculating the composite strain fermentation liquor into a silybum marianum meal culture medium, and performing anaerobic fermentation to obtain a fermentation product; filtering the fermentation product to obtain silybum marianum meal fermentation liquor; adding bacillus and trace elements into the silybum marianum liquid to obtain the bacterial fertilizer. The compound strain comprises lactobacillus and yeast. According to the invention, silybum marianum meal is subjected to solid state fermentation by lactic acid bacteria and yeast, macromolecular antioxidant substances in silybum marianum are degraded, and the content of soluble substances is increased. When the bacterial manure is applied to plant leaves, plant-derived antioxidant substances can be added, the antioxidant system of the plant leaves can be balanced, the plant growth can be promoted, and the disease resistance can be enhanced.

Description

Bacterial fertilizer for repairing plant leaf antioxidant system and preparation method thereof

Technical Field

The invention relates to the technical field of agriculture, and in particular relates to a bacterial fertilizer for repairing a plant leaf antioxidant system and a preparation method thereof.

Background

A set of antioxidant systems responsible for scavenging the production of reactive oxygen species exists in plants. Under normal plant growth conditions, the antioxidant system is responsible for bringing the production and scavenging of reactive oxygen species into a state of dynamic equilibrium. However, when the plant is in a stress condition, such as drought, low temperature, heavy metals, damage and the like, the capacity of an antioxidant system in the plant body for removing active oxygen is reduced, and the dynamic balance of the active oxygen is broken. Stress such as drought, low temperature, heavy metals, injury, etc., responds to stress on the leaves of the plant, adversely affecting the growth of the plant. In order to improve the stress resistance of plants and promote the growth of plants, chemical fertilizers are widely used. However, excessive use of fertilizers is liable to cause soil destruction and environmental pollution.

At present, in order to reduce the use of fertilizers and pesticides, make up the deficiency of nutrient absorption of root systems and prevent and cure diseases, foliar fertilizers which directly act on plant leaf surfaces are generally selected. However, most of the foliar fertilizers on the market at present are added with pesticides, hormones and the like to promote crop growth and resist diseases and insect pests, so that safety problems are easily caused. And there is no product for the repair of the antioxidant system of plant leaves.

Disclosure of Invention

The invention aims to provide a preparation method of bacterial manure for repairing an antioxidant system of plant leaves, which adopts a waste resource-silybum marianum meal as a culture medium and utilizes compound strains for fermentation to obtain the bacterial manure suitable for plant leaf surfaces.

The invention also aims to provide a bacterial fertilizer for repairing the plant leaf antioxidant system, which contains microbial flora and a fermentation product of silybum marianum meal and has good antioxidant effect on the plant leaves.

The technical problem to be solved by the invention is realized by adopting the following technical scheme.

The invention provides a preparation method of bacterial fertilizer for repairing a plant leaf antioxidant system, which comprises the following steps:

s1, obtaining a composite strain fermentation liquid, wherein the composite strain comprises lactic acid bacteria and saccharomycetes;

s2, inoculating the composite strain fermentation liquor to a silybum marianum meal culture medium, and performing anaerobic fermentation to obtain a fermentation product;

s3, filtering the fermentation product to obtain silybum marianum meal fermentation liquor;

s4, adding bacillus and trace elements into the silybum marianum liquid to obtain the bacterial manure.

The invention provides a bacterial fertilizer for repairing an antioxidant system of plant leaves, which is prepared according to the preparation method.

The bacterial fertilizer for repairing the plant leaf antioxidant system and the preparation method thereof have the beneficial effects that:

bacterial fertilizers, also known as microbial fertilizers, contain not only nutrients required for crop growth, but also a large number of microbial flora. The method takes the waste resource of the silybum marianum meal as the fermentation raw material, and utilizes the composite strain fermentation liquor of the lactic acid bacteria and the microzyme for fermentation, so that the preparation method is simple and efficient, and the cost is low. The obtained fermentation broth can increase plant-derived antioxidant substances, help balance antioxidant system of plant leaves, promote plant growth, enhance disease resistance, and reduce pesticide consumption. In addition, bacillus and trace elements are added into the fermentation liquor, and the trace elements are used for providing sufficient nutrient components for plant growth. The bacillus can propagate and fix the value on the plant leaves, and effectively block the infection of plant pathogenic microorganisms.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and those skilled in the art can also obtain other related drawings based on the drawings without inventive efforts.

FIG. 1 is a flow chart of the preparation of bacterial manure of example 1 of the present invention;

FIG. 2 is a graph showing the results of the germination test of seeds in test example 2 of the present invention;

FIG. 3 is a photograph of tea leaves before bacterial manure was applied in test example 3 of the present invention;

FIG. 4 is a photograph of tea leaves three days after the bacterial manure was applied in test example 3 of the present invention;

FIG. 5 is a photograph of a horse hoof tomato to which no bacterial manure was applied in test example 4 of the present invention;

FIG. 6 is a photograph of a horse hoof tomato to which bacterial manure was applied in test example 4 of the present invention.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are conventional products which are not indicated by manufacturers and are commercially available.

The bacterial manure for repairing the plant leaf antioxidant system and the preparation method thereof according to the embodiment of the present invention are specifically described below.

The embodiment provides a preparation method of bacterial manure for repairing an antioxidant system of plant leaves, which comprises the following steps:

s1, obtaining the composite strain fermentation liquor, wherein the composite strain comprises lactobacillus and saccharomycetes.

In this embodiment, the composite strain fermentation broth is prepared by the following steps: inoculating the composite strain into a strain culture medium, and culturing at 35-40 ℃ for 24-80 h to obtain the strain. Further preferably, the strain culture medium comprises 1-4% of molasses, 0.5-2% of sodium chloride, 0.5-2% of maltose, 1-5% of peptone and the balance of water by mass fraction. The composite strain fermentation liquor with high activity is obtained by fermenting and culturing the strain.

Further, in the present embodiment, the inoculation amount of the lactic acid bacteria and the yeast is 1-4: 1 to 4. Preferably, the inoculation amount of the lactic acid bacteria and the yeast is 2: 1. By regulating and controlling the difference of the inoculation amount and adjusting the flora composition of the composite strain fermentation liquor, the degradation efficiency of the silybum marianum meal is higher and the active ingredients are more under the condition that lactic acid bacteria account for more dominance.

Further, the lactic acid bacteria are selected from one or more of enterococcus faecalis, lactobacillus acidophilus, lactobacillus plantarum, streptococcus lactis and enterococcus faecium. The yeast is selected from one or two of Saccharomyces cerevisiae and Candida utilis.

Specifically, in the embodiment of the present invention, the Lactobacillus species are ACCC10180 Enterococcus faecalis (Enterococcus faecalis), ACCC10637 Lactobacillus acidophilus (Lactobacillus acidophilus), ACCC11118 Lactobacillus plantarum (Lactobacillus plantarum), ACCC10653 Streptococcus lactis (Streptococcus lactis), and ACCC06037 Enterococcus faecium (Enterococcus faecalis). The yeast strain is ACCC20065 Saccharomyces cerevisiae (Saccharomyces cerevisiae) and ACCC20059 Candida utilis (Candida utilis). All strains were purchased from the China agricultural culture Collection of microorganisms.

Lactic acid bacteria such as enterococcus faecalis and yeasts such as Saccharomyces cerevisiae can produce a large amount of vitamins, organic acids, protease, growth promoting factors, etc., and can ferment the silybum marianum meal, stimulate the secretion of active oxygen in the silybum marianum meal, and metabolize to generate antioxidant components, amino acids, organic acids, coenzyme factors, etc. The silybum marianum meal is fermented by compounding the specific strains. The lactobacillus and the yeast have mutual promotion effect in the growth process, and have good symbiotic basis. The co-fermentation can improve the fermentation speed and accelerate the accumulation of metabolites. Lactic acid produced in the growth process of the lactic acid bacteria can provide a good acid growth environment for yeast, and the yeast also has a promotion effect on the growth of the lactic acid bacteria.

S2, inoculating the composite strain fermentation liquid obtained in the step S1 into a silybum marianum dreg culture medium, and performing anaerobic fermentation to obtain a fermentation product.

Further, in this step, the silybum marianum pulp culture medium includes silybum marianum pulp, molasses, magnesium sulfate, and water. Preferably, the molasses, the magnesium sulfate and the water are respectively 3-6%, 0.1-2% and 35-40% of the mass fraction of the silybum marianum meal. The invention takes the silybum marianum meal as a main fermentation raw material, adds a proper amount of molasses, magnesium sulfate and water, and uses the compound strain of lactobacillus and saccharomycetes for fermentation, thus being capable of degrading the silybum marianum meal, obtaining a large amount of antioxidant substances, improving the content of soluble substances and improving the antioxidant effect of bacterial manure on plants.

Furthermore, in the step, the inoculation amount of the composite strain fermentation liquor in the silybum marianum meal culture medium is 1-5 wt%. The inoculation amount of the composite strain fermentation liquid is regulated and controlled so as to well control the fermentation degree and ensure the quality of the fermentation product.

Further, in the step, the anaerobic fermentation parameters are as follows: and (3) carrying out closed fermentation at the fermentation temperature of 25-40 ℃ for 60-80 h.

S3, filtering the fermentation product obtained in the step S2 to obtain the silybum marianum dreg fermentation liquid.

The silybum marianum meal is the residue of herbal silybum marianum after silymarin extraction, is used as a waste resource, has high yield and is easy to cause the pollution to the surrounding environment. At present, the silybum marianum meal is mainly applied as a feed additive, and the silybum marianum meal is generally fermented to be high-protein feed. The invention utilizes the composite strain to ferment the silybum marianum meal, so that the antioxidant ingredients and other active ingredients in the silybum marianum meal are degraded and dissolved out. Meanwhile, in the process of fermenting the silybum marianum meal by the composite strains, active substances and sterilizing substances can be generated, the immunity and disease resistance of plant lamps are enhanced, and the occurrence of diseases is indirectly reduced. The fermentation residues are removed by squeezing and filtering, the fermentation liquor is reserved and used as bacterial manure, and the fermentation residues left after squeezing can be further used as soil fertilizer or animal feed.

S4, adding bacillus and trace elements into the silybum marianum liquid obtained in the step S3 to obtain bacterial manure.

Further, in the step, the addition amount of the bacillus is 50-100 ⁹ cfu/ml. The bacillus is one or more selected from bacillus subtilis, bacillus megaterium and bacillus amyloliquefaciens. The bacillus can form nutrition competition and space site competition with pathogenic bacteria, and through the multiplication and colonization of the bacillus, the plant is protected from being infected by the pathogenic bacteria, and the diffusion of the pathogenic bacteria in the plant body is prevented.

Furthermore, in the step, the addition amount of the trace elements is 2-4% of the mass fraction of the silybum marianum meal fermentation liquor, and the trace elements are selected from one or more of Fe, Mn, Cu, Zu, Mo and B. And trace elements are added to further strengthen the nutrition supplement to the plants and promote the benign growth of the plants.

It is understood that the addition of trace elements, for example, by adding ammonium molybdate to provide Mo, by adding boric acid to provide B, by adding chelated iron to provide Fe, etc. In the present embodiment, the amount of the trace element added is 2 to 4% based on the amount of the trace element added.

The embodiment of the invention also provides bacterial fertilizer for repairing the plant leaf antioxidant system, and the bacterial fertilizer is prepared by the preparation method.

The features and properties of the present invention are described in further detail below with reference to examples.

Example 1

The bacterial fertilizer for repairing the antioxidant system of the plant leaves is prepared according to the following steps:

(1) seed culture: 2 wt% of molasses, 0.5 wt% of sodium chloride, 1 wt% of maltose and 3 wt% of peptone are added into sterilized water to prepare a strain culture medium. Then inoculating according to the inoculation amount of the lactobacillus and the yeast being 2:1, and culturing the inoculated lactobacillus and the yeast at 37 ℃ for 48h to obtain the compound strain fermentation liquor.

(2) Fermentation: adding 1 wt% of molasses, 0.5 wt% of magnesium sulfate and 40 wt% of water into silybum marianum meal, sterilizing at 90 deg.C for 30min, and using as fermentation medium. And uniformly mixing the composite strain fermentation liquor with 5 wt% of inoculum size, uniformly mixing with a fermentation medium, filling into a fermentation barrel, sealing, and carrying out anaerobic fermentation. The fermentation temperature is controlled at 30 ℃; and (5) fermenting for 72 hours to finish the fermentation.

(3) Squeezing fermented product, filtering to obtain filtrate, and mixing with the filtrate at a ratio of 50% ⁹ Adding bacillus megaterium powder according to the cfu/ml content, and adding trace elements according to the 2 wt% content, wherein the trace elements comprise Fe, Mn, Cu, Zu, Mo and B. And (4) uniformly mixing to obtain the bacterial manure.

Example 2

(1) seed culture: 2 wt% of molasses, 0.5 wt% of sodium chloride, 1 wt% of maltose and 3 wt% of peptone are added into sterilized water to prepare a strain culture medium. Then inoculating according to the inoculation amount of 1:1 of the lactobacillus and the microzyme, and culturing the inoculated lactobacillus and the microzyme at 37 ℃ for 48 hours to obtain the composite strain fermentation liquor.

(2) And (3) fermentation: adding 1 wt% of molasses, 0.5 wt% of magnesium sulfate and 40 wt% of water into silybum marianum meal, sterilizing at 90 deg.C for 30min, and using as fermentation medium. And uniformly mixing the composite strain fermentation liquor with 5 wt% of inoculum size, uniformly mixing with a fermentation medium, filling into a fermentation barrel, sealing, and carrying out anaerobic fermentation. Controlling the fermentation temperature at 30 ℃; and (5) fermenting for 72 hours to finish the fermentation.

(3) Squeezing fermented product after fermentation, filtering to obtain filtrate, and processing at 60 deg.C ⁹ Adding bacillus megaterium powder according to the cfu/ml content, and adding trace elements according to the 2 wt% content, wherein the trace elements comprise Fe, Mn, Cu, Zu, Mo and B. And (4) uniformly mixing to obtain the bacterial manure.

Comparative example 1

The page fertilizer provided by the comparative example is obtained through the following steps:

(1) seed culture: 2 wt% of molasses, 0.5 wt% of sodium chloride, 1 wt% of maltose and 3 wt% of peptone are added into sterilized water to prepare a strain culture medium. Then inoculating according to the inoculation amount of the lactobacillus and the microzyme of 2:1, and culturing the inoculated lactobacillus and the microzyme at 37 ℃ for 48h to obtain the composite strain fermentation liquor.

(2) And (3) fermentation: 1 wt% of molasses, 0.5 wt% of magnesium sulfate and 40 wt% of water were added to a mixture of soybean meal and straw powder, and sterilized at 90 ℃ for 30min to serve as a fermentation medium. And uniformly mixing the composite strain fermentation liquor with 5 wt% of inoculum size, uniformly mixing with a fermentation medium, filling into a fermentation barrel, sealing, and carrying out anaerobic fermentation. Controlling the fermentation temperature at 30 ℃; and (5) fermenting for 72 hours to finish the fermentation.

(3) Squeezing fermented product, filtering to obtain filtrate, and mixing with the filtrate at a ratio of 50 ⁹ Adding bacillus megaterium powder according to the cfu/ml content, and adding trace elements according to the 2 wt% content, wherein the trace elements comprise Fe, Mn, Cu, Zu, Mo and B. And (4) uniformly mixing to obtain the foliar fertilizer.

Comparative example 2

(2) 50 portions of the strain in the composite strain fermentation liquor ⁹ Adding bacillus megaterium powder according to the cfu/ml content, and adding trace elements according to the 2 wt% content, wherein the trace elements comprise Fe, Mn, Cu, Zu, Mo and B. And (4) uniformly mixing to obtain the foliar fertilizer.

Test example 1

In a Fujian rice planting base, the products in example 1, example 1 and comparative example 1 are respectively used, foliar spraying is carried out 8 days after rice transplanting to serve as a test group, the dilution is 300 times, and the test group is sprayed with 1L of the product per 3 mu. The rice seeds, planting means, fertilization method and management method of all the test fields are consistent. Meanwhile, a test field without foliar fertilizer is taken as a control group.

TABLE 1

Group of	Control group	Example 1	Example 2	Comparative example 1	Comparative example 2
						Average tiller number (number/point)	11	19	19	14	14
Increase of yield	/	30.1％	27.4％	6.0％	5.3％
						Disease prevention and control rate	/	93.4％	91.8％	61.8％	67.7％

Disease control rate (1-test group disease rate/control group disease rate) × 100%

Test example 2

In the germination test of seeds (Shanghai green) performed in a greenhouse, the products of example 1, example 1 and comparative example 1 were sprayed on the surfaces of the seeds, and the spraying amount of each test group was kept uniform as a test group. Non-sprayed seeds were used as a control group. All other experimental conditions of the seeds remained the same.

Observing for 7 days, determining the germination rate, and determining the root system activity by adopting a TTC method, wherein the test results are as follows:

TABLE 2

Group of	Control group	Example 1	Example 2	Comparative example 1	Comparative example 2
						Germination rate	73.0％	92.7％	93.5％	83.2％	84.7％
Root system activity (mg/g. h)	0.12	0.31	0.26	0.12	0.13
						Chlorophyll increase rate	/	22.2％	20.9％	3.6％	2.2％

As shown in FIG. 2, which is a graph of the germination results of the seeds of example 1, it can be seen that the germination rate of the seeds is high and the chlorophyll content is high after the bacterial manure of the present invention is used, so that the germination quality of the seeds is greatly improved.

Test example 3

The bacterial manure prepared in example 1 was applied to a tea leaf plantation site by spraying on leaf surfaces of tea leaves, and fig. 3 and 4 are photographs comparing tea leaves before and three days after the bacterial manure application. Test results show that after the bacterial manure disclosed by the embodiment of the invention is used, the growth of leaves of tea is obvious, the leaves are oily green and bright, the pesticide dosage is reduced by 70%, the yield is improved by 13.5%, and the economic benefit is increased by 12%.

Test example 4

The bacterial manure prepared in example 1 was applied to a corm-shaped tomato plantation site by spraying on the leaf surface of the tomato, fig. 5 is a photograph of a corm-shaped tomato without the bacterial manure, and fig. 6 is a photograph of a corm-shaped tomato with the bacterial manure prepared in example 1. The horseshoe tomatoes without bacterial manure showed few fruits, the leaves of the plants were slightly yellow, the number of surface dehiscent fruits per plant was about 2, the average number of fruits per plant was 7, and the average fruit diameter was 3.4 cm. After the bacterial manure of the embodiment is used, leaves are oily green, no dry tips exist, no diseases are seen, no cracks are seen on the surfaces of the tomatoes, the number of the results is obviously increased, the average number of the results per plant is 15, and the average diameter of the fruits is 7.2 cm.

The embodiments described above are some, not all embodiments of the invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be obtained by a person skilled in the art without inventive step based on the embodiments of the present invention, are within the scope of protection of the present invention.

Claims

1. A preparation method of bacterial manure for repairing an antioxidant system of plant leaves is characterized by comprising the following steps:

s1, obtaining a compound strain fermentation liquid, wherein the compound strain comprises lactic acid bacteria and saccharomycetes;

s4, adding bacillus and trace elements into the silybum marianum liquid to obtain the bacterial manure;

in step S2, the anaerobic fermentation parameters are: carrying out closed fermentation at the temperature of 25-40 ℃ for 60-80 h;

wherein, in step S1, the lactic acid bacteria are selected from one or more of enterococcus faecalis, lactobacillus acidophilus, lactobacillus plantarum, streptococcus lactis and enterococcus faecium; the yeast is selected from one or two of saccharomyces cerevisiae and candida utilis;

in the step S2, the inoculation amount of the composite strain fermentation liquor in the silybum marianum meal culture medium is 1-5%;

wherein in the step S4, the addition amount of the bacillus is 50-100 ⁹ cfu/ml is selected from one or more of bacillus subtilis, bacillus megaterium and bacillus amyloliquefaciens;

in the step S4, the adding amount of the trace elements is 2-4% of the mass fraction of the silybum marianum meal fermentation liquor, and the trace elements are selected from one or more of Fe, Mn, Cu, Zu, Mo and B;

in step S2, the silybum marianum cake culture medium includes silybum marianum cake, molasses, magnesium sulfate, and water;

in the silybum marianum meal culture medium, the molasses, the magnesium sulfate and the water respectively account for 3-6% of the silybum marianum meal, 0.1-2% of the silybum marianum meal and 35-40% of the silybum marianum meal by mass;

in step S1, the composite strain fermentation broth is prepared by the following steps: inoculating the composite strain into a strain culture medium, and culturing at 35-40 ℃ for 24-80 h, wherein the strain culture medium comprises 1-4% of molasses, 0.5-2% of sodium chloride, 0.5-2% of maltose, 1-5% of peptone and the balance of water according to mass fraction;

wherein, the inoculation amount of the lactobacillus and the yeast is 2: 1.

2. a bacterial fertilizer for repairing the antioxidant system of plant leaves, which is prepared by the preparation method of claim 1.