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Preparation and application of a salt-tolerant antagonist m4-1 and its bacterial agent

A technology of salt tolerance and bacterial agent, applied in the field of agricultural microorganisms, can solve the problems of low vegetation survival rate and agricultural production efficiency, poor physical and chemical properties of saline-alkali soil, affecting agricultural and forest production, etc. The effect of industrialized production and short production cycle

Active Publication Date: 2021-05-14
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The area of ​​saline-alkali land in my country is huge, the physical and chemical properties of saline-alkali soil are poor, the ecological environment is harsh, and the number and types of soil microorganisms are significantly less than normal planting soil. Therefore, the vegetation survival rate and agricultural production efficiency in saline-alkali areas are low, seriously affecting agricultural production , forest production

Method used

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  • Preparation and application of a salt-tolerant antagonist m4-1 and its bacterial agent
  • Preparation and application of a salt-tolerant antagonist m4-1 and its bacterial agent
  • Preparation and application of a salt-tolerant antagonist m4-1 and its bacterial agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Isolation of the M4-1 strain.

[0050] Take the wheat rhizosphere soil sample from the saline-alkali land in the Yellow River Delta, take 10g of the soil sample under sterile conditions, dissolve it in a 250mL Erlenmeyer flask filled with 90mL of sterile water, 30°C, 200r / min, vibrate for 30min, and draw 1mL and 9mL of sterile water. Bacteria and water were mixed to obtain 10 -1 diluent, then take 1mL 10 -1 The diluted solution was mixed with 9mL sterile water to obtain 10 -2 Dilutions, indistinctly, to prepare 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 Wait for a series of dilutions. The chosen dilution is 10 -5 、10 -6 、10 -7 Use a pipette to draw 100mL of the soil dilution solution in the center of the LB medium plate, spread the dilution solution evenly with a spreader, and incubate at a constant temperature of 32°C for 1-3 days. Pick a single colony, streak it into a pure culture, and save it.

[0051] Several strains of pure cultures obtained above were subj...

Embodiment 2

[0054] Identification of the M4-1 strain

[0055] (1) Cell morphology and colony characteristics

[0056] The colony of the M4-1 strain on the LB solid plate was white, with a moist surface and irregular edges. With the prolongation of culture time, the surface of the colony formed tiny wrinkles, and the center of the colony was slightly raised.

[0057] (2) Physiological and biochemical characteristics

[0058] The physiological and biochemical characteristics of the M4-1 strain are shown in Table 1. The aerobic nature of the strain is facultative anaerobic; catalase, nitrate reduction, gelatin liquefaction, starch hydrolysis, citrate utilization, V-P determination, methyl red test, H 2 S test, glucose gas test, MR test and other reactions are positive; indole test and phenylalanine deaminase test are negative; it can grow in liquid LB medium with NaCl concentration of 5% and 10%.

[0059] Liquid LB medium components: peptone 10.0g, yeast powder 5.0g, NaCl 10.0g, water 1.0...

Embodiment 3

[0073] M4-1 strain rep-PCR gene fingerprint analysis.

[0074] 1. Genomic DNA extraction of M4-1 strain

[0075] Using TIANGEN Bacterial Genomic DNA Extraction Kit DP302

[0076] (1) Take 1-5 mL of bacterial culture solution, centrifuge at 10,000 rpm for 1 min, and try to suck up the supernatant;

[0077] (2) Add 110mL TE buffer solution and 70mL lysozyme (final concentration: 20μg / mL) to the cell pellet, and treat at 37°C for more than 30min;

[0078] (3) Add 20mL Proteinase K solution to the tube and mix well;

[0079] (4) Add 200mL buffer solution GB and shake for 15sec, place at 70°C for 10min, the solution should become clear, and briefly centrifuge for 10-15s to remove water droplets on the inner wall of the tube cap;

[0080] (5) Add 220mL of absolute ethanol, shake and mix well for 15sec. At this time, flocculent sediment may appear, briefly centrifuge for 10-15s to remove the water droplets on the inner wall of the tube cap;

[0081] (6) Add the solution obtained ...

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Abstract

The invention relates to the preparation and application of a salt-tolerant antagonistic bacteria M4‑1 and its agent; it is preserved in the General Microbiology Center of the China Microbiological Culture Collection Management Committee, and the preservation number is CGMCC No.15774; the bacterial strain has high salt tolerance It can colonize stably in saline-alkali soil; it can survive in liquid water-soluble fertilizer and solid water-soluble fertilizer and maintain a high survival rate; The root rot diseases of crops such as corn, vegetables and fruit trees and other soil-borne diseases have good control effects; the strain has excellent fermentation properties, and the preparation process of the bacterial agent is simple and low in cost, which is beneficial to industrial production and transportation.

Description

technical field [0001] The invention provides the preparation and application of a salt-tolerant antagonistic bacteria M4-1 and its bacterial agent, belonging to the technical field of agricultural microorganisms. Background technique [0002] The most important feature of saline-alkali soil is the high content of soluble salt and substitutable sodium. When the soil salinity exceeds 0.1% or the soil pH value is above 8.0, and the degree of sodium alkalization (ESP) is greater than or equal to 5%, It belongs to the range of saline-alkali soil. Land salinization is a kind of land desertification, which is the biggest problem in land degradation. It can lead to the decline of soil fertility, which is manifested as the reduction of soil aeration, water permeability, water retention and fertilizer retention, which makes it difficult for crop roots to absorb water, resulting in reduced production or even extinction. Produce. [0003] The area of ​​saline-alkali land in my countr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/22A01P3/00C05F11/08C12R1/07
CPCA01N63/10C12N1/20C12N1/205C12R2001/07
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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