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Bacillus velezensis LN9-2 and application thereof

A technology of Bacillus Velez and Bacillus, applied in the field of microorganisms, to achieve strong antibacterial ability and strong antibacterial effect

Active Publication Date: 2021-03-26
SHANXI AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The patent number is: CN 109266586 A, and the patent name is: Patent Bacillus Veles BMF 03 and its use and fermentation method. In this patent, a variety of pathogenic bacteria were tested, including apple rot and apple anthracnose, but did not test the effect on apple The effect of mold core and black spot fungus, the pathogen causing apple mold heart and black spot disease is polythecene pink, which is a different genus of fungus from apple rot fungi and anthracnose fungus

Method used

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  • Bacillus velezensis LN9-2 and application thereof
  • Bacillus velezensis LN9-2 and application thereof
  • Bacillus velezensis LN9-2 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Isolation, screening and purification of Bacillus Velez LN9-2

[0036] 1. Isolation and screening of Bacillus Velez LN9-2

[0037] (1) Isolation of Bacillus Velez LN9-2

[0038] Weigh the soil samples from Shenwangpo Village, Linxian County, Luliang City, Shanxi Province, put them into sterile water, shake at room temperature, perform gradient dilution, and take 0.1ml to spread on NA medium.

[0039] (2) Screening of Bacillus Velez LN9-2

[0040] Place the bacterium cake of polythecene pink in the middle of the PDA medium, and place 4 bacteria grown on the NA medium around it.

[0041] (3) Purification and preservation of strains

[0042] Plate streaking method: Streak the bacteria that have inhibitory effect on polythecene pink on a new NA plate, culture it upside down in a 28°C incubator for 48 hours, pick a single colony and add sterile glycerol to preserve it. Store at -80°C for long-term storage, coded as LN9-2.

Embodiment 2

[0043] Embodiment 2: 16S rRNA of bacterial strain LN9-2, gyrB , recN , recA genetic identification

[0044] Using bacterial 16S rRNA primers L10 5'-AGTTTGATCCTGGCTC-3' and R1451 5'-AAGGACGTGATCCAGCC-3', UP-1 5'-GAAGTCATCATGACCGTTCTGCAYGCNGG NGGNAARTTY-3' and UP-2r 5'-AGCAGGGTACGGATGTGCGAGCCRTCNARCTCNGC RTCNGAf-3',rec '-TGAGTGATCGTCAGGCAGCCTTAG-3' and recAr 5'-TTCTTCA TAAGAATACCACGAACCGC-3', recNf5'-CTTTTGCGATCAGAAGGTGCGTATCCG-3' and recNr 5'-GCCATTATAGAGGAACTGACGATTTC-3', and use the genomic DNA of strain LN9-2 as a template to prepare a 25 μL PCR reaction System (see Table 1):

[0045] Table 1 Taq polymerase chain reaction system

[0046] Element Final concentration 2×Taq mix 1×Taq mix forward primer 200nM reverse primer 200nM DNA template 100ng water Make up to 25 μl

[0047] The PCR reaction conditions were: 95°C pre-denaturation for 2 min; 95°C denaturation for 15 s, 55°C annealing for 25 s, 72°C extension for 90 s...

Embodiment 3

[0050] Example 3: Morphology and Physiological and Biochemical Identification of Bacterial Strain LN9-2

[0051] According to the morphological observation of the bacterial strain LN9-2 of the present invention, it is found that the bacterial colony of the bacterial strain LN9-2 is milky white, irregular, with rough edges and rough surface; the result presented by Gram staining is positive. The physiological and biochemical test results of the bacterial strain LN9-2 of the present invention show that the bacterial strain can utilize glucose, sucrose and mannitol. Rhamnose, citrate, lactose, arabinose, amygdalin, melibiose, sorbitol, and inositol cannot be used. V-P test, urease test, and starch hydrolysis test were positive (see Table 2).

[0052]

[0053] In conjunction with the results of the phylogenetic tree of embodiment 2 and the morphological characteristics of embodiment 3, physiology and biochemistry, bacterial strain LN9-2 is finally identified as Bacillus velei ...

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Abstract

The invention discloses bacillus velezensis LN9-2 and application thereof. The bacillus velezensis LN9-2 is classified and named as bacillus velezensis LN9-2, and is preserved in China General Microbiological Culture Collection Center on November 06, 2020, and the preservation number is CGMCC NO.21110. The bacillus velezensis LN9-2 provided by the invention is used for preventing and treating apple moldy heart disease or apple black spot disease caused by trichothecium roseum; and the result of the invention shows that the strain has relatively strong antibacterial ability to trichothecium roseum, and provides good biological control resources for controlling apple moldy heart disease and black spot disease.

Description

technical field [0001] The invention belongs to the technical field of microbes, and relates to a strain of Bacillus Velez LN9-2 and an application thereof. Background technique [0002] Apple is a worldwide fruit. It is resistant to storage and has a long supply cycle. It is rich in various vitamins and trace elements and is rich in nutrition. It is deeply loved by people. Apple moldy core disease is one of the important diseases of apples, which causes moldy core and core rot in ripe fruit, and fruit rot during storage. Investigation on the distribution status of apples[J].Plant Protection, 2016, 42(1): 175-179), which seriously affects the quality and yield of apples. The disease is caused by a variety of pathogenic fungi, and the pathogenic fungi that cause apple mold heart disease in my country are mainly Alternaria ( Alternaria spp.), Cladosporium ( Cladosporium spp.), polythecene pink ( Trichothecium roseum ) (Gao Lingling. 2010. Study on the Diversity of Patho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/22A01P3/00C12R1/07
CPCA01N63/22Y02A50/30
Inventor 房雅丽张治家高越周瑶瑶
Owner SHANXI AGRI UNIV