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Strong phosphorus-dissolving bacillus subtilis, carbon-based microbial compound fertilizer thereof and application

A technology of Bacillus subtilis and compound fertilizer, which is applied in strong phosphorus-soluble Bacillus subtilis and its carbon-based microbial compound fertilizer and its application field, can solve the problems of not being able to fully realize the value of fallen leaves, and achieve the promotion of vegetable growth and quality improvement, yield Strong spore ability, effect of increasing soluble sugar and Vc content

Active Publication Date: 2021-04-09
INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, only turning fallen leaves into biochar is not enough to fully realize the value of fallen leaves. "Leaves return to their roots" is the ultimate destination of resource recycling of fallen leaves

Method used

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  • Strong phosphorus-dissolving bacillus subtilis, carbon-based microbial compound fertilizer thereof and application
  • Strong phosphorus-dissolving bacillus subtilis, carbon-based microbial compound fertilizer thereof and application
  • Strong phosphorus-dissolving bacillus subtilis, carbon-based microbial compound fertilizer thereof and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Isolation and Identification of Bacillus subtilis CNBG-PGPR-1

[0030] (1) Sample dilution and coating

[0031] Take 0.5g soil sample (collected from the farmland around Nanjing, Jiangsu) and suspend and shake in 4.5mL sterile saline, which is 10 -1 Gradient (ten-fold dilution), take 0.5mL from the dilution and suspend and shake in 4.5mL sterile normal saline, which is 10 -2 Gradient (100-fold dilution), and so on until diluted to 10 -6 gradient. Draw 100 μL from the dilutions of the 6 dilution gradients, spread them on the Nutrient Agar (NA) solid medium plate, prepare 2 copies in parallel, invert them, and culture them in a constant temperature incubator at 30°C for 24-36 hours, observe from time to time .

[0032] (2) Strain isolation and purification

[0033] Take out all the plates with colonies grown under the same sample, select a dilution plate with obvious single colonies and a moderate number, pick the colonies with different shapes on the plate...

Embodiment 2

[0056] The generation of embodiment 2 Bacillus subtilis spores in NA medium

[0057] (1) Culture medium preparation

[0058] Liquid medium a: NB liquid medium: peptone 10g, beef extract powder 3g, sodium chloride 5g, agar 15g, distilled water 1L.

[0059] Liquid medium b: LB liquid medium: tryptone 10g, yeast extract powder 5g, sodium chloride 10g, agar 15g, distilled water 1L.

[0060] (2) Spore generation and expansion culture

[0061] Pick a single colony from the plate into a prepared centrifuge tube filled with 5mL of liquid medium a or b, culture at 30°C with shaking at 200rpm. Sampling started at 19h, the smear was dried naturally, fixed by flame heating, 5% malachite green staining solution was added dropwise, heated, and steamed 3-4 times within 30 seconds, do not boil, after cooling, rinse with tap water for 30 Seconds, add 0.5% sand yellow counterstaining solution, after 3 seconds, wash with water, air-dry, count under microscope, see results Figure 4 . Bacter...

Embodiment 3

[0063] Inorganic phosphorus dissolving ability of embodiment 3 bacillus subtilis CNBG-PGPR-1

[0064] (1) Preparation of culture medium

[0065] The composition of inorganic phosphorus liquid medium is glucose 10g, tricalcium phosphate 5g, calcium carbonate 0.1g, magnesium sulfate 0.5g, ammonium sulfate 0.5g, calcium sulfate 0.1g, ferric chloride 0.005g, water 1L.

[0066] (2) Inorganic phosphorus solubility

[0067] Take out the bacterial preservation tube of the strain CNBG-PGPR-1 frozen in the -80°C refrigerator, thaw it on ice until it becomes fluid, shake and mix well, pick up an appropriate amount of bacterial liquid with an inoculation loop, and line the three zones on the NA solid plate. Culture at 30°C for 24±2 hours, pick a single colony and purify it for 1-2 times and make identification and confirmation. After confirmation, pick a single colony and activate it in NA liquid medium, centrifuge to remove the NA medium, and weigh the bacteria sludge Suspend in an equ...

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Abstract

The invention discloses strong phosphorus-dissolving bacillus subtilis, a carbon-based microbial compound fertilizer thereof and application. The bacillus subtilis is preserved in the China General Microbiological Culture Collection center on April 13, 2020, and the preservation number is CGMCC No. 19564. The bacillus subtilis has strong spore production capacity and is kept stable, the content of dissolved inorganic phosphorus reaches 179mg / L, and the degradation rate of lecithin reaches 100% at most. Biomass charcoal prepared from fallen leaves contains high nitrogen, phosphorus and potassium nutrients, and the fallen leaf biomass charcoal is combined with the bacillus subtilis with strong phosphorus dissolving capacity to obtain the fallen leaf carbon-based microbial compound fertilizer. Compared with application of a conventional chemical fertilizer, the application of the fallen leaf carbon-based microbial compound fertilizer can still increase the yield of spinach by 60-64%. Compared with non-application and application of the conventional chemical fertilizer, the application of the fallen leaf carbon-based microbial compound fertilizer significantly increases the content of soluble sugar and Vc of the spinach and reduces the content of nitrate in the spinach, thereby promoting the growth and quality improvement of vegetables.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a strong phosphorus-soluble bacillus subtilis and its carbon-based microbial compound fertilizer and its application. Background technique [0002] Phosphorus is one of the most important nutrients for living things. Phosphorus participates in the structure of many important compounds in plants; phosphorus participates in many metabolic processes in plants; phosphorus can enhance the stress resistance of plants. The content of total phosphorus in my country's soil is relatively high, roughly in the range of 0.44-0.85g / kg, but most of the phosphorus in the soil exists as insoluble calcium phosphate salt, and the available phosphorus content that can be directly absorbed and utilized by plants is very low, so The phosphorus in the soil solution cannot meet the phosphorus demand of the crops in the current season. Since the application of phosphate fertilizers in the 1950s ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N11/14C05G3/00C05G3/80C05G5/12C12R1/125
CPCC05B7/00C05G3/00C12N1/20C12N11/14C05G3/80C05G5/12C05C9/00C05D1/02C05D3/00C05D9/00C05F11/08
Inventor 于金平贾明云吕世鹏周冬琴侯炤琪李琦
Owner INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI