Saprolegnia antagonist and application thereof

An antagonistic antibacterial and water mold technology, which is applied in the application field as a biosafety drug, can solve problems such as unfavorable purification, poor protein thermal stability, protease, etc., and achieves the effect of low cost of promotion and use.

Inactive Publication Date: 2011-03-16
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the previously reported antagonistic substances of various Bacillus species with antifungal activity have been confirmed to have protein properties (see Li Yonggang, Hao Zhongna, Yang Mingxiu, et al. Identification of rice disease biocontrol strain L1 in cold regions and research on its antibacterial mechanism[J] .Journal of Northeast Agricultural University, 2008, 39(7): 9-12), due to the poor thermal stability of protein and its susceptibility to protease degradation in the environment, it is not conducive to the large-scale purification of its active ingredients for pharmaceutical companies for production practice

Method used

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  • Saprolegnia antagonist and application thereof
  • Saprolegnia antagonist and application thereof
  • Saprolegnia antagonist and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Screening of Bacillus Sh1

[0029] 1. Plate confrontation culture method to screen Saprolegnia antagonistic strains

[0030] The bacteria to be selected and Saprolegnia were inoculated on the PDA plate respectively, and cultured at a constant temperature of 26°C for 3 days. When the mycelium of Saprolegnia was handed over around the antagonistic bacteria and an obvious inhibition zone appeared, the distance from the edge of Saprolegnia mycelium to the edge of the bacterial colony was used as Criteria for judging the magnitude of bacterial antagonism. Record the number of antagonistic strains.

[0031] 2. Discovery of Bacillus Sh1 that antagonizes Saprolegnia

[0032] The Saprolegnia used in this experiment grows well on the PDA plate at 26°C. Generally, after the agar block containing Saprolegnia mycelium is inoculated in the middle of the plate, the entire plate can be covered in three to four days. When using the flat plate confrontation culture method t...

Embodiment 2

[0035] Embodiment two: the identification of bacillus Sh1

[0036] 1. Identification of Bacillus Sh1

[0037] In this example, the molecular identification of Sh1 is carried out according to the sequence of bacterial 16S rDNA. The strain genomic DNA was extracted using the Genome Extraction Kit of Shanghai Tiangen. Using bacterial genomic DNA as a template, bacterial 16S rDNA universal primers: 27f: 5'-GAGAGTTTGATCCTGGCTCAG-3' and 1541r: 5'-AAGGAGGTGATCCAGCCGCA-3'; were selected to amplify the 16S rDNA sequence of Sh1. The PCR reaction conditions were set according to the product instructions of Shanghai Sangon's PCR kit. The amplified band was recovered from the DNA electrophoresis gel using the DNA recovery kit from Shanghai Tiangen. The recovered samples were sent to Shanghai Sangong for sequence determination. The type of Sh1 was determined by Blast similarity comparison with known bacterial 16S rDNA in Genbank.

[0038] 2. 16S rDNA sequencing identified Sh1 as Bacill...

Embodiment 3

[0042] Embodiment three: the bacteriostasis of bacillus Sh1

[0043] 1. Pathological observation of hyphae inhibition

[0044] Saprolegnia and Bacillus Sh1 were cultured on PDA medium at 26°C. When Saprolegnia formed an inhibition zone around Sh1, pick the hyphae at the edge of the inhibition zone and observe the changes of Saprolegniasis hyphae under a microscope.

[0045] 2. Inhibitory effect of Sh1 on saprolegniasis hyphae

[0046] In the antibacterial plate, the hyphae growing normally and the hyphae at the edge of the inhibition zone were compared and observed under the microscope. It can be found that the hyphae of the saprolegniasis at the edge of the inhibition zone are distorted, the hyphae are broken in many places or the middle part is constricted, and the growth of the mycelium is obviously inhibited. compare the two image 3 , A is the hyphae around the inhibition zone, B is the normal hyphae.

[0047] This example shows that Bacillus Sh1 has a strong inhibito...

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Abstract

The invention discloses a saprolegnia antagonist and application thereof. The saprolegnia antagonist can be used as a biological medicinal source of a mountain-green-substituted medicament, can reduce environmental pollutions and can meet the requirements of biological prevention and control of fish water mildew. The saprolegnia antagonist Sh1 provided by the invention belongs to Bacillus amyloliquefaciens, and has the collection number of CCTCC NO: M2010129. The active ingredients of the novel antagonist provided by the invention are high in stability, high in antagonist capability, safe, effective and stable; moreover, the antagonist can be produced by a conventional production process and a conventional production flow, and is low in popularization and use cost.

Description

technical field [0001] The invention relates to a bacillus, in particular to a bacillus which has an antagonistic effect on fish saprolegniasis, and its application as a biosafety medicine. Background technique [0002] Saprolegniasis is a general term for a class of parasitic diseases caused by Saprolegnia or Achlya in aquaculture. Saprolegniasis is the most common pathogen causing saprolegniasis in aquatic animals at present, and Saprolegniasis is traditionally regarded as a kind of fish fungal disease. Because Saprolegnia is lower than fungi, in taxonomy, this class of organisms has been divided from the original fungi kingdom into the kingdom of velvet, so now it is also considered that Saprolegnia belongs to Oomycetes. Freshwater fish roe, fry, and adult fish, including grass carp and other four major domestic fish, may suffer from saprolegniasis, and once it occurs, it is difficult to treat. In late winter and early spring, the disease is most serious when the water ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K35/74A61P31/10C12R1/07A61K35/742
Inventor 刘丽玲曹海鹏吕利群
Owner SHANGHAI OCEAN UNIV
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