Method for improving content of colloidal phosphorus in soil and application of kidney-shaped worm in preparation of biological fertilizer for improving content of colloidal phosphorus in soil

Abstract

The invention discloses a method for increasing the content of colloidal phosphorus in soil and application of kidney-shaped worms to preparation of a biological fertilizer for increasing the content of colloidal phosphorus in soil. The method comprises the step of inoculating the biological fertilizer containing the kidney-shaped worms into the soil. By the adoption of the technical scheme, the effect of improving colloidal phosphorus in the soil is remarkable, operation is easy, complex and expensive instruments and equipment are not needed, the cost and technical difficulty are greatly reduced, the gain effect is remarkable, the effect can be obtained within a short time, and an innovative solution is provided for phosphorus activation in the black soil area.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of agricultural bio-fertilizer, in particular to a method for increasing the content of colloidal phosphorus in soil and application of Nephromma sp. to preparing bio-fertilizer for increasing the content of colloidal phosphorus in soil. BACKGROUND

[0002] Soil colloidal phosphorus, i.e. soil phosphorus combined with inorganic mineral colloids such as iron and aluminum oxides, clay minerals, organic colloids such as organic macromolecules and microorganisms, and organic-inorganic composite colloids to form a fine particulate state phosphorus (1nm-1000nm), which includes molybdenum blue reaction colloidal phosphorus and non-molybdenum blue reaction colloidal phosphorus, and the molybdenum blue reaction colloidal phosphorus can be absorbed and utilized by plants. Colloidal phosphorus is of great significance to the process of soil phosphorus cycle, especially in fertile soils such as black soil. First, colloids can diffuse to the surrounding phase through Brownian motion, while colloidal phosphorus moves by its colloidal carrier, is easily absorbed by plant roots, supplies phosphorus for plant growth, and enhances the biological availability of phosphorus. On the other hand, colloidal phosphorus has strong migration ability and can enter deep soil by soil pores or with water migration. This characteristic greatly affects the spatial distribution and ecological effects of phosphorus, and has important significance that cannot be ignored for maintaining soil fertility and ecological balance. At present, the main technologies for regulating soil colloidal phosphorus include conservation tillage, application of organic fertilizer, etc. The above technologies change the phosphorus occurrence form and migration and transformation in soil by reducing tillage disturbance and increasing soil organic matter input, but the above technologies generally have problems such as high economic cost, slow results, etc. SUMMARY

[0003] In view of the above technical problems, the present application discloses a method for increasing the content of colloidal phosphorus in soil and application of Nephromma sp. to preparing bio-fertilizer for increasing the content of colloidal phosphorus in soil, which is also a colloidal phosphorus efficient enrichment method based on protozoan stimulation method, and solves the bottleneck problems of current soil colloidal phosphorus enrichment technology, such as strong professional dependence, high operation cost and insignificant efficiency improvement.

[0004] To this end, the technical scheme adopted by the present application is as follows:

[0005] A method for increasing the content of colloidal phosphorus in soil, inoculating bio-fertilizer containing Nephromma sp. into soil.

[0006] By adopting the technical scheme, the content of colloidal phosphorus in soil can be increased, and the problems of complex operation, high cost, low efficiency and long period in the prior art can be effectively solved.

[0007] As a further improvement of the present application, the Nephromma sp. is Harbin Nephromma sp. screened from black soil, and then obtained by purification and large-scale culture.

[0008] As a further improvement of the present application, after screening, the kidney-shaped worms are cultured with wheat grain leaching solution to obtain living culture solution, and then purified.

[0009] As a further improvement of the present application, the purification comprises culturing the living kidney-shaped worms by monoclonal method.

[0010] As a further improvement of the present application, the large-scale culture is carried out in a cell culture plate, with wheat grain leaching solution as food, and in a constant temperature incubator at 25℃±2℃.

[0011] As a further improvement of the present application, the kidney-shaped worms separated from black soil are placed in a constant temperature incubator at 25℃±2℃, fed with wheat grain leaching solution, and subjected to preliminary culture for 1-7 days; then, under the same temperature condition, the large-scale culture is continued for 1-7 days, thereby completing the large-scale culture.

[0012] As a further improvement of the present application, 90-110 kidney-shaped worms are inoculated per gram of soil. Further, 100 kidney-shaped worms are inoculated per gram of soil. In practical application, the soil quality is calculated according to the following formula:

[0013] Soil quality (g) = soil bulk density (g / cm 3 ) x soil area (cm 2 ) x soil depth (cm).

[0014] The present application discloses application of kidney-shaped worms in preparation of biological fertilizer for increasing content of colloidal phosphorus in soil.

[0015] As a further improvement of the present application, the kidney-shaped worms are Harbin kidney-shaped worms screened from black soil, and then obtained by purification and large-scale culture.

[0016] As a further improvement of the present application, the kidney-shaped worms separated from black soil are placed in a constant temperature incubator at 25℃±2℃, fed with wheat grain leaching solution, and subjected to preliminary culture for 1-7 days; then, under the same temperature condition, the large-scale culture is continued for 1-7 days, thereby completing the large-scale culture.

[0017] Compared with the prior art, the present application has the following beneficial effects:

[0018] By using the technical solution of the present application, the effect of increasing content of colloidal phosphorus in soil is remarkable, and the operation is simple, without the need of complex and expensive instruments and equipment, thereby greatly reducing the cost and technical difficulty, and remarkably increasing the effect, so that the effect can be obtained in a short time, and an innovative solution is provided for activation of phosphorus in black soil area and control of phosphorus pollution in agriculture. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1is a live body graph of the kidney-shaped worm screened in the embodiment of the present application.

[0020] Figure 2 is an ammoniacal silver staining graph of the kidney-shaped worm screened in the embodiment of the present application.

[0021] Figure 3 is an evolutionary tree graph of the kidney-shaped worm in the embodiment of the present application.

[0022] Figure 4 is a graph of the influence of the kidney-shaped worm on the soil colloidal phosphorus content in the embodiment of the present application.

[0023] Figure 5 is a graph of the influence of the kidney-shaped worm on the soil molybdenum blue reaction phosphorus content in the embodiment of the present application.

[0024] Figure 6 is a graph of the influence of the kidney-shaped worm on the proportion of soil molybdenum blue reaction phosphorus content to colloidal phosphorus content in the embodiment of the present application. DETAILED DESCRIPTION

[0025] The preferred embodiment of the present application is further described in detail below.

[0026] The soil sample used in the following examples is black soil.

[0027] A method for increasing the content of colloidal phosphorus in soil inoculates a biological fertilizer containing Harbin kidney-shaped worms into the soil. The Harbin kidney-shaped worms are screened from black soil and then obtained by purification and large-scale culture. Specifically, it comprises:

[0028] Step S1, preparing soil leaching solution

[0029] Take 30g of black soil sample, put it in a beaker, add 100mL of distilled water, boil for 10min to 15min, stand for precipitation, cool, and filter the suspension to obtain the soil leaching solution.

[0030] Step S2, live body culture and screening by "non-submergence culture method".

[0031] Take another 5g of black soil sample, wrap it with gauze and put it into a culture dish, add soil leaching solution, and submerge the sample at 2 / 3 of the leaching solution. The selected protozoa in the black soil are cultured in a 25℃ constant temperature incubator for 7 days to obtain the live body culture solution.

[0032] Step S3, monoclonal culture of protozoa

[0033] Take one of the selected protozoa with a glass capillary tube, put it in a sterile cell culture plate, and add sterilized distilled water at 2 / 3 of the well plate. The protozoa are placed in a 25℃ constant temperature incubator and fed with wheat leaching solution.

[0034] Step S4, species identification

[0035] The screened protozoa were identified by morphology and molecular biology. The live image of the screened Nephromma is shown in Figure 1 The Nephromma was stained with ammoniacal silver and the image is shown in Figure 2 The phylogenetic tree of the Nephromma is shown in Figure 3

[0036] Step S5, protozoa expansion

[0037] The Nephromma after monoclonal culture was cultured in a sterile cell culture plate and placed in a 25℃ constant temperature incubator for culture.

[0038] Step S6, black soil inoculation

[0039] The expanded Nephromma was added to the black soil, with 100 individuals per gram of soil.

[0040] The control group was not added with protozoa but only with distilled water. After 30 days of culture, the colloidal phosphorus content in the black soil was compared and the results are shown in Figure 4 The results of the molybdenum basket reaction colloidal phosphorus content comparison are shown in Figure 5 The results of the soil molybdenum blue reaction phosphorus content comparison are shown in Figure 6 It can be seen that the technical solution of the embodiment can quickly increase the colloidal phosphorus content in the black soil, especially the molybdenum basket reaction colloidal phosphorus content, which increases to 90.1%.

[0041] The above is a further detailed description of the present application in combination with specific preferred embodiments, and the specific implementation of the present application should not be limited to these descriptions. For ordinary skilled persons in the technical field to which the present application belongs, without departing from the concept of the present application, a number of simple deductions or substitutions can be made, which should be considered as falling within the protection scope of the present application.​

Claims

1. A method for increasing the colloidal phosphorus content in soil, characterized in that: Inoculate the soil with bio-fertilizer containing kidney-shaped insects.

2. The method for increasing colloidal phosphorus content in soil according to claim 1, characterized in that: The kidney-shaped parasites were selected from Harbin kidney-shaped parasites in black soil using a "non-submerged culture method," and then purified and expanded through culture.

3. The method for increasing colloidal phosphorus content in soil according to claim 2, characterized in that: After screening, the kidney-shaped parasites were cultured in wheat grain extract to obtain a live culture medium, which was then purified.

4. The method for increasing colloidal phosphorus content in soil according to claim 2, characterized in that: The purification process includes culturing the live culture medium using a monoclonal method.

5. The method for increasing colloidal phosphorus content in soil according to claim 4, characterized in that: The expanded culture was obtained by culturing kidney-shaped parasites in wheat grain extract in cell culture plates and then culturing them in large quantities in a constant temperature incubator at 25℃±2℃.

6. The method for increasing colloidal phosphorus content in soil according to claim 1, characterized in that: The kidney-shaped insects isolated from the black soil were placed in a constant temperature incubator at 25℃±2℃ and fed with wheat grain extract for a preliminary culture period of 1-7 days. Subsequently, under the same temperature conditions, the culture was expanded for another 1-7 days to complete the expansion process.

7. The method for increasing colloidal phosphorus content in soil according to any one of claims 1 to 6, characterized in that: Inoculate 90-110 kidney-shaped worms per gram of soil.

8. The application of kidney-shaped insects in the preparation of bio-fertilizers that increase colloidal phosphorus content in soil, characterized by: The bio-fertilizer is used to increase the colloidal phosphorus content in the soil.

9. The application of the kidney-shaped insect according to claim 8 in the preparation of bio-fertilizer that increases colloidal phosphorus content in soil, characterized in that: The kidney-shaped parasites mentioned were selected from Harbin kidney-shaped parasites from black soil, and then obtained through purification and large-scale culture.

10. The application of the protozoan according to claim 8 in the preparation of bio-fertilizers that increase colloidal phosphorus content in soil, characterized in that: The kidney-shaped insects isolated from the black soil were placed in a constant temperature incubator at 25℃±2℃ and cultured with wheat grain extract for 1-7 days for initial culture. Subsequently, under the same temperature conditions, the culture was expanded for another 1-7 days to complete the expansion process.

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