Preparation and application of a salt-tolerant Bacillus subtilis for disease prevention and growth and its inoculum

A technology of Bacillus subtilis and microbial inoculants, applied in the field of agricultural microorganisms, can solve the problems that plants cannot normally absorb water, fertilizer, gas, heat, destroy soil structure, aeration, reduce soil microbial diversity, etc., and reduce chemical fertilizers. Application rate, beneficial to industrial production, high salt tolerance effect

Active Publication Date: 2021-04-30
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Soil salinization reduces soil microbial diversity, destroys soil structure and aeration, makes plants unable to absorb water, fertilizer, gas, heat and other conditions normally, and reduces agricultural and forestry productivity

Method used

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  • Preparation and application of a salt-tolerant Bacillus subtilis for disease prevention and growth and its inoculum
  • Preparation and application of a salt-tolerant Bacillus subtilis for disease prevention and growth and its inoculum
  • Preparation and application of a salt-tolerant Bacillus subtilis for disease prevention and growth and its inoculum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Isolation of HG-15 strain

[0058] In this study, soil samples were obtained from farmland, lakeside, saline-alkali land, and oil fields in the Yellow River Delta region. The soil samples were divided into 4 groups, and 10 g of soil in each group was placed in a 250 mL triangular flask filled with 50 mL of sterile water. Fully disperse the glass beads, fully shake in a shaker at 32°C and 120r / min for 30min, and dilute 10% respectively according to the dilution coating method. -1 、10 -2 、10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 Gradients, each gradient was repeated three times, respectively spread to PDA medium, inorganic phosphate solution medium, potassium solution medium, Congo red medium, and skim milk medium, and cultured at a constant temperature of 32°C for 2-5 days. Pick a single colony, streak it into a pure culture, and save it.

[0059] The PDA medium components are: 200.0g potato, 5.0g beef extract, 20.0g glucose, 15.0g agar, (NH 4 ) 2 SO 4 1.0g, MgSO ...

Embodiment 2

[0066] Identification of the HG-15 strain

[0067] (1) Cell morphology and colony characteristics

[0068] The colony of HG-15 strain on the PDA plate was yellowish, the surface was rough and opaque, and the edges were irregular. With the prolongation of culture time, the surface of the colony formed tiny wrinkles, and the shape of the bacteria observed under the microscope was rod-shaped. The HG-15 strain is a Gram-positive bacterium, and the spores are elliptical to columnar, located in the center of the bacterium, and the bacterium does not expand after spore formation.

[0069] (2) Physiological and biochemical characteristics

[0070] The physiological and biochemical characteristics of the HG-15 strain are shown in Table 1. The aerobic performance of the strain is facultative anaerobic, Gram reaction, glucose oxidative fermentation test, contact enzyme, starch hydrolysis test, V-P test, salt tolerance test, phenylalanine amino acid deaminase test and citrate utilizatio...

Embodiment 3

[0084] HG-15 strain rep-PCR gene fingerprint.

[0085] 1. Genomic DNA extraction of HG-15 strain

[0086] Using TIANGEN Bacterial Genomic DNA Extraction Kit DP302

[0087] (1) Take 1 mL of bacterial culture solution, centrifuge at 10,000 rpm for 1 min, and absorb the supernatant as much as possible;

[0088] (2) Add 110 μL TE buffer and 70 μL lysozyme to the cell pellet, and treat at 37°C for more than 30 minutes;

[0089] (3) Add 20 μL of Proteinase K solution to the tube and mix well;

[0090] (4) Add 200 μL of buffer solution GB and shake for 15 sec, place at 70°C for 10 min, the solution should become clear, and briefly centrifuge for 10-15 sec to remove water droplets on the inner wall of the tube cap;

[0091] (5) Add 220 μL of absolute ethanol, shake and mix well for 15 seconds, at this time flocculent precipitation may appear, briefly centrifuge for 10-15 seconds to remove water droplets on the inner wall of the tube cap;

[0092] (6) Add the solution obtained in t...

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PUM

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Abstract

The invention relates to the preparation and application of a salt-resistant, disease-preventing, growth-promoting Bacillus subtilis and its bacterial agent; the HG‑15 strain has high salt tolerance and can stably colonize in saline-alkali soil; it can be used in liquid water-soluble fertilizers. , survive in solid water-soluble fertilizers and maintain a high survival rate, and can be mixed with organic fertilizers, compound fertilizers, water-soluble fertilizers, controlled-release fertilizers and other fertilizers; It has good control effect on root rot diseases of crops and other soil-borne diseases; it can significantly increase the content of available phosphorus and available potassium in the soil, and reduce the application of phosphorus and potassium fertilizers; Simple, low cost, and conducive to industrial production and transportation.

Description

technical field [0001] The invention provides the preparation and application of a strain of salt-tolerant, disease-preventing, growth-promoting Bacillus subtilis and its bacterial agent, belonging to the technical field of agricultural microorganisms. Background technique [0002] Soil salinization has seriously affected agricultural and forestry production, and is one of the four major ecological and environmental problems facing the world today. Salinized soil is an important land resource. The world's salinized soil area reaches 1.0×10 9 hm 2 , accounting for about 30% of the total land area. The proportion of salinized soil in China is much higher than the world average, about 1.7×10 7 hm 2 , Nearly 1 / 3 of the irrigation areas have soil salinization problems. Soil salinization reduces soil microbial diversity, destroys soil structure and aeration, makes plants unable to absorb water, fertilizer, air, heat and other conditions normally, and reduces agricultural and f...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/22A01P3/00C05F11/08C12R1/125
CPCA01N63/10C05F11/08C12N1/20C12N1/205C12R2001/125
Inventor 刘训理纪超王晓辉隋君康李倩王长栋张嘉淼
Owner SHANDONG AGRICULTURAL UNIVERSITY
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