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Endophytic Bacillus velesi and its application

A technology of bacillus and safflower gentian, applied in the application, bacteria, biocide and other directions, can solve the problems of increasing control difficulty, environmental pollution, threats to human health, etc., and achieves high safety, convenient use, and easy mass production. production effect

Active Publication Date: 2022-04-01
LIUPANSHUI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In order to prevent the occurrence of post-harvest anthracnose in mangoes, chemical fungicides are mainly added to preservatives or fumigants in production, but long-term use will not only lead to the development of drug resistance of many anthracnose bacteria, making the prevention and control more difficult, but also easily causing environmental pollution. threat to human health

Method used

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  • Endophytic Bacillus velesi and its application
  • Endophytic Bacillus velesi and its application
  • Endophytic Bacillus velesi and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1, Isolation and identification of Gentiana safflower endophytic Bacillus Velez SB023 strain

[0031] Bacillus Velez SB023 was obtained by separating from healthy stalks of G. rhodantha by tissue block separation and purified by streaking on a plate; the morphological characteristics of Bacillus Velez SB023 are as follows: Bacillus Velez Bacillus SB023 can grow on NB medium and PDA medium, and the colony morphology of cultured at 32°C for 48 hours is as follows: figure 1 shown. The colony on NB medium is small, 0.5-2mm, transparent to white, with moist surface, wrinkled, and irregular edges; it grows rapidly on PDA plate, 3-12mm, fried egg-like, surface wrinkled and raised, and edges are irregular tidy. Observed by staining, the strain is long rod-shaped, with a size of 0.46-0.63×1.4-3.3μm ( n=30), Gram-positive, endo-spore, peri-flagellate.

Embodiment 2

[0032] Embodiment 2, the physiological and biochemical characteristics of Bacillus Velez SB023

[0033] According to the method provided by "Bergey's Bacteria Identification Manual", different culture media were prepared for physiological and biochemical identification of the SB023 strain. The identification results are shown in Table 1. According to conventional physiological and biochemical indicators, it is a close species of Bacillus subtilis. The lecithin hydrolysis of the strain was positive, and "Common Bacterial System Identification Manual" (Dong Xiuzhu, Cai Miaoying. Common Bacterial System Identification Manual [M]. Beijing: Science Press, 2001) reported the reconciliation of Bacillus subtilis (B.subtilis) B. amyloliquefaciens cannot hydrolyze lecithin, so the strain SB023 was initially identified as B. velezensis.

[0034] Table 1. Physiological and biochemical characteristics of B. velezensis SB023 strain

[0035]

[0036]

[0037] Note: + means positive; ...

Embodiment 3

[0038] Embodiment 3, Bacillus Velez SB023 molecular biology identification

[0039] Using the DNA of Bacillus Velez SB023 as a template, PCR amplification was carried out using bacterial 16S universal primers, and the amplification primers were:

[0040] 27F: 5'-caggcctaacacatgcaagtc-3 (SEQ ID NO.1);

[0041] 1492R: 5'-gggcggwgtgtacaaggc-3 (SEQ ID NO. 2);

[0042] The PCR reaction system is 20 μL, 2×PCR Mix 10 μL, upstream and downstream primers 0.5 μL, template DNA 1 μL, ddH 2 O8μL; PCR reaction conditions: pre-denaturation at 95°C for 5min; denaturation at 95°C for 30s, annealing at 55°C for 30s, extension at 72°C for 90s, 30 cycles; extension at 72°C for 7min. The PCR products were detected by agarose gel electrophoresis and sent to Beijing Qingke Biotechnology Co., Ltd. for purification and sequencing. The resulting DNA sequence has been submitted to GenBank, accession number: MT509797.

[0043]The sequence was input into BLAST in GenBank for homology comparison. After...

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Abstract

The invention discloses an endophytic Bacillus velezensis strain SB023 (Bacillus velezensisSB023) isolated from the stalk of Gentianarhodantha and its application; The strain has a significant inhibitory effect on the mycelial growth and spore germination of the pathogenic fungus of mango anthracnose, and can efficiently inhibit the infection of the fruit by the pathogenic fungus, achieving a good control effect. The biocontrol bacterial strain provided by the invention is safe, green, environment-friendly and efficient, and can provide a new approach for the biological control of mango anthracnose during storage.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an endophytic Bacillus velesi SB023 of Gentiana safflower and its application. Background technique [0002] Anthracnose is one of the most serious diseases of mangoes during storage after harvesting. At the beginning of the disease, dark brown round lesions are formed on the fruit, and after expanding, they become round or irregular, black, with a dry and sunken center, which seriously affects the appearance of the fruit. The value is reduced, often resulting in greater economic losses. Mango is a tropical fruit and should not be refrigerated for a long time. If the fruit surface, container or storage environment are not thoroughly disinfected during storage at room temperature, anthrax outbreaks are very easy. Mango anthracnose is mainly caused by several pathogenic bacteria such as Colletotrichum.asianum, C.gloeosporioides and C.acutatum (Yang Bo. Investigation and Pathogens of P...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23B7/16A23B7/155A23B7/154A01N63/22A01P3/00C12R1/07
CPCA23B7/16A23B7/155A23B7/154A01N63/22A23V2002/00C12R2001/07C12N1/205A23V2200/10A23V2200/22A23V2250/30A23V2250/5026
Inventor 张晓勇杨友联罗凯李树江严凯周雪林
Owner LIUPANSHUI NORMAL UNIV
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