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Method for identifying brevibacillus laterosporus in microbial fertilizer

A technology of Brevibacillus and Laterosporus, which is applied in the detection of Brevibacillus Laterosporus and primers in microbial fertilizers, and can solve the problems of cumbersome operation steps, long detection time and poor specificity

Inactive Publication Date: 2011-01-26
INST OF AGRI RESOURCES & REGIONAL PLANNING CHINESE ACADEMY OF AGRI SCI
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

The key to the commonly used plate count detection method is to accurately identify effective bacteria. The traditional strain identification currently used is mainly based on morphological characteristics and various physiological and biochemical methods. The operation steps are cumbersome, the detection time is long, and the specificity is poor.

Method used

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  • Method for identifying brevibacillus laterosporus in microbial fertilizer
  • Method for identifying brevibacillus laterosporus in microbial fertilizer
  • Method for identifying brevibacillus laterosporus in microbial fertilizer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0049] Detection of standard strains by the PCR method using primers specific for Brevibacillus spp.

[0050] (1) Material

[0051] Test bacteria: the strains listed in Table 1 below are the standard strains to be tested

[0052] Table 1

[0053]

[0054] (2) Design and synthesis of primers

[0055] According to the 16S rDNA nucleic acid sequence of Brevibacillus spp., the specific primers of Bacillus spp. were designed. The primers are shown in Table 2. The primers were synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd.

[0056] Table 2

[0057] type

[0058](3) DNA extraction: Take 1.5 mL of bacterial liquid in an Eppendorf tube, centrifuge at 9000 r / min for 4 min to collect bacterial cells, and store at -20°C. Wash the collected cells twice with 1×TE buffer solution, then add 500 μL GUTC buffer solution, mix well, let stand for 20 minutes, add 20 μL ~ 30 μL diatomaceous earth adsorption solution to each tube, oscillate evenly and p...

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Abstract

The invention provides technology for simply and rapidly detecting and identifying brevibacillus laterosporus in a microbial fertilizer. In the technology, a specific primer for the brevibacillus laterosporus is designed, a PCR identification method for the brevibacillus laterosporus is constructed, and the brevibacillus laterosporus in the microbial fertilizer can be detected and identified according to a PCR amplification result of sample genomic DNA.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for detecting Brevibacillus laterosporus in microbial fertilizers through PCR reaction using appropriate primers, and the primers used in the method. technical background [0002] Brevibacillus lateralsporosa is a bacterium that is widely distributed in nature and exists in soil, ocean, animals, and insects. With the in-depth research on Bacillus brevis, it has been found that it can produce a variety of metabolites with potential applications, such as insecticidal proteins, antibacterial substances, and anticancer immunomodulatory substances. In addition, the functions of Brevibacillus lateralosporus in dissolving inorganic phosphorus, degrading organic phosphorus, controlling pests, and improving crop defense capabilities have also been gradually discovered, researched, developed and utilized. [0003] In view of the versatility of Brevibacillus lateralosporus, many compa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
Inventor 曹凤明李俊沈德龙姜昕关大伟李力杨小红陈慧君葛一凡
Owner INST OF AGRI RESOURCES & REGIONAL PLANNING CHINESE ACADEMY OF AGRI SCI
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