Bacillus pumilus and application thereof

A kind of Bacillus pumilus, germ technology, applied in the field of microbial technology application

Active Publication Date: 2021-04-09
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, there are certain limitations in...

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  • Bacillus pumilus and application thereof
  • Bacillus pumilus and application thereof
  • Bacillus pumilus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1, the acquisition of Bacillus pumilus 154-2

[0043] 1. Soil source

[0044] Soil of East Wheat Field in Wanglaoba Village, Yuanzhou District, Guyuan City, Ningxia

[0045] 2. Screening of bacterial strains

[0046] (1) Soil sample collection

[0047] Zig-shaped 5-point sampling method for soil sample collection: collect 200g soil sample at each point, mix the soil sample evenly, take 200g by quartering method and put it into a sterilization bag as a soil sample. Three soil samples were collected from each plot as replicates. Record the time, place and type of sampling. The collected soil samples were stored in a refrigerator at 4°C for use in bacterial isolation.

[0048] (2) Isolation of bacteria

[0049] Plate dilution method: Weigh 10g of soil sample into a Erlenmeyer flask, add 90mL of sterile water, then shake at 200rpm, 28°C shaker, take it out after 20min and let it stand at room temperature for 10min to make the stock solution of soil bacteria...

Embodiment 2

[0058] The 16S rRNA gene identification of embodiment 2 bacillus pumilus 154-2

[0059] Genomic DNA of strain 154-2 was extracted,

[0060] Using primers: 27F 5'-AGAGTTTGATCCTGGCTCAG-3' and 1492R5'-TACGGCTACCTTGTTACGACTT-3', the extracted DNA was used as a template for PCR amplification to obtain the target fragment. The PCR reaction system is:

[0061] Table 1 Taq polymerase chain reaction system

[0062]

[0063]

[0064] The basic conditions of the PCR reaction were: pre-denaturation at 94°C for 3 min, denaturation at 94°C for 30 s, annealing at 56°C for 30 s, extension at 72°C for 90 s (1 kb / min), pre-extension at 72°C for 8 min, and storage at 4°C for a total of 30 cycles. After the reaction, the PCR products were checked by 1% agarose gel electrophoresis, and the gel imager was used to detect and record the results. The gel electrophoresis results of 16S rRNA gene were as follows: image 3 as shown, image 3 Among them, 1 represents Marker; 2 represents the produ...

Embodiment 3

[0067] Physiological and biochemical identification of embodiment 3 Bacillus pumilus 154-2

[0068] The physiological and biochemical characteristics of Bacillus pumilus 154-2 of the present invention are: able to secrete β-galactosidase, 3-hydroxybutanone to produce acetylmethylmethanol and gelatinase; unable to hydrolyze arginine, lysine, ornithine acid, sodium citrate and sodium thiosulfate, etc.; cannot oxidize glucose, mannitol, rhamnose, and sucrose; can utilize 20 carbon sources such as L-arabinose, ribose, D-xylose, fructose, and mannose Acid production, see Table 2 and Table 3.

[0069] Table 2 Physiological and biochemical characteristics of strain 154-2 - enzyme activity, carbon source oxidation

[0070]

[0071]

[0072] +: Positive reaction; -: Negative reaction;

[0073] Table 3 Physiological and biochemical characteristics of strain 154-2 – acid production using carbon source

[0074]

[0075]

[0076] +: Positive reaction; -: Negative reaction; W...

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Abstract

The invention relates to bacillus pumilus and application thereof. The bacillus pumilus strain is preserved in China Center for Type Culture Collection on August 24th, 2020, the preservation name of the bacillus pumilus strain is bacillus pumilus 154-2, and the preservation number is CCTCC M 2020446. The bacillus pumilus disclosed by the invention has a remarkable inhibition effect on rice bacterial leaf blight pathogenic bacteria, also has broad-spectrum antibacterial activity on rice bacterial leaf streak pathogenic bacteria, banana bacterial wilt pathogenic bacteria, cowpea wilt pathogenic bacteria, pepper spot pathogenic bacteria, walnut bacterial black spot pathogenic bacteria, wheat scab pathogenic bacteria and the like (wherein the pathogenic bacteria have specific names as shown in the specification), and has an important biocontrol application prospect.

Description

technical field [0001] The invention belongs to the field of microbial technology application, and in particular relates to a strain of bacillus pumilus and its application. Background technique [0002] Rice is one of the main food crops in my country. The planting area accounts for about 1 / 4 of the country's cultivated land area, and the annual output accounts for about 1 / 2 of the country's total grain output (Abdullah AB., Ito S., Adhana K. Estimate of rice consumption in Asian countries and the world towards 2050. Proceedings for Workshop and Conference on Rice in the World at Stake[R]. 2006.), rice is also an important strategic resource for my country's food security and sustainable agricultural development. Two pathogenic strains of Xanthomonas oryzae, X.oryaze pv.oryaze (Xoo) and X.oryaze pv.oryzicola (Xoc), can infect rice and cause rice bacterial leaf blight (bacterial leafblight, BLB) and rice Bacterial leaf streak (BLS). The existence and development of these tw...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/22A01P3/00A01P1/00C12R1/07
CPCA01N63/22
Inventor 邹丽芳黄梦桑李逸朗杨瑞环闫依超方园陈功友
Owner SHANGHAI JIAO TONG UNIV
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