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Bacillus altitudinis and application thereof

A highland bacillus and pathogen technology, applied in the field of microorganisms, can solve the problems of ecological environment damage, lack of disease-resistant varieties, etc., and achieve the effect of broad-spectrum bacteriostatic ability

Active Publication Date: 2021-04-06
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the process of agricultural production, the prevention and control of these two diseases still has great limitations. In addition to the lack of good and stable disease-resistant varieties, the long-term use of chemical reagents has caused certain damage to the ecological environment. Among them, microorganisms that can antagonize rice bacterial blight and leaf spot are screened out, and they are developed into microbial agents. It is imminent to find a method that can balance the ecological environment and disease control

Method used

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  • Bacillus altitudinis and application thereof
  • Bacillus altitudinis and application thereof
  • Bacillus altitudinis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1, Isolation, Screening and Purification of Bacillus highlander 181-7

[0047] 1. Source of the strain

[0048] The pathogenic strains used are all strains preserved by our laboratory, and all of them are published strains.

[0049] 2. Acquisition of antagonistic strains

[0050] (1) Discovery of antagonistic strains

[0051] In the laboratory, a strain capable of inhibiting the growth of Xanthomonas oryzae was screened from soil samples of capsicum field in Haidian Gangwan Garden, Haikou City, Hainan Province, China.

[0052] (2) Purification and preservation of strains

[0053] Using plate streaking method: Pick out the colony that can inhibit the growth of bacterial blight, streak on a new NA plate, place it in a biochemical incubator at 28°C, and culture it upside down for more than 12 hours, pick a single colony, and number it for 181-7. Inoculate the strain 181-7 in NA liquid medium, culture it in a shaker at 28°C and 180rpm for 12h, draw 1mL of the f...

Embodiment 2

[0056] Embodiment 2, 16S rRNA gene identification of bacterial strain 181-7

[0057] Use bacterial universal primer 27F 5'-AGAGTT TGA TCCTGGCTCAG-3' and 1492R5'-TACGGCTACCTTGTTACGACTT-3', and using the genomic DNA of strain 181-7 as a template; prepare a 30 μL PCR reaction system according to the ratio in Table 1, and perform the PCR reaction:

[0058] Table 1 Taq polymerase chain reaction system

[0059]

[0060] The basic conditions of the PCR reaction were set as follows: pre-denaturation at 95°C for 5 min; denaturation at 95°C for 30 s, annealing at 56°C for 30 s, extension at 72°C for 90 s, a total of 30 cycles; pre-extension at 72°C for 10 min. After the PCR reaction, the PCR product amplified by strain 181-7 was checked by 1% agarose gel electrophoresis, and the image was detected by a gel imager, and then the original solution of the PCR product was sent to Platinum Biotechnology (Shanghai) Co., Ltd. sequencing.

[0061] The sequence of the 16S rRNA gene of Baci...

Embodiment 3

[0063] Embodiment 3, Morphological and physiological and biochemical identification of bacterial strain 181-7

[0064] According to the morphological observation of the bacterial strain 181-7 of the present invention, it was found that the colony of the bacterial strain 181-7 was milky white, with rough and irregular edges, dry and rough surface, and opaque; the result of Gram staining was positive. Physiological and biochemical test results of the bacterial strain 181-7 of the present invention show that the Bacillus highlanders can treat D-fructose-6-phosphate, D-cellobiose, D-galacturonic acid, glucuronamide, β-methyl- D-glucoside, D-salicin, N-acetyl-D-glucosamine, N-acetyl-β-D-mannosamine, a-D-glucose, D-mannose, D-fructose, D-galactose , D-melibiose, D-fucose, L-fucose, inosine, D-mannitol, glycerin, gentiobiose, sucrose, D-aspartic acid, D-trehalose, L- Alanine, L-arginine, L-aspartic acid, L-glutamic acid, L-pyroglutamic acid, L-serine, pectin, stachyose, D-gluconic a...

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Abstract

The invention relates to bacillus altitudinis and application thereof. The bacillus altitudinis is preserved in China General Microbiological Culture Collection Center on June 23, 2020, the sample name is bacillus altitudinis 181-7, the preservation number is CGMCC No.20132, and the strain is separated from a soil sample of a capsicum land in a meadow harbor garden in Haikou City in Hainan province in China. The bacillus altitudinis 181-7 disclosed by the invention shows a bacteriostatic effect on Xanthomonas oryzae pv. Oryzae and Xanthomonas oryzae pv. Oryzae, and also has a bacteriostatic effect on banana bacterial wilt pathogen, bean wilt pathogen, soybean scab pathogen and the like and some pathogenic fungi such as botrytis cinerea, phytophthora pathogen and fusarium graminearum and the like. Results show that the strain has broad-spectrum antibacterial ability, provides good biocontrol resources for preventing and treating rice bacterial leaf blight and bacterial streak, and lays a foundation for excavating novel microbial agents.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a strain of bacillus highlander and its application. Background technique [0002] There are more than ten thousand rice varieties in the world (Yu Y, Streubel J, Balzeruge S, et al. Colonization of rice leaf blades by an African strain of Xanthomonasoryzae pv. oryzae depends on a new tal effector that induces the rice nodulin-3Os11N3 gene .Molecular Plant Microbe Interaction[J].2011), nearly half of the world's population now uses rice as a staple food, and it is also the main food crop in my country. The planting area in China accounts for about 30% of the total area of ​​food crops. Rice bacterial leaf blight (BLB) and rice streak (BLS) are caused by two pathogenic strains of Xanthomonas oryzae. Two of the most serious bacterial diseases occur widely in southern rice areas such as Guangdong, Fujian, Hunan, and Jiangxi, which seriously restrict the production safety...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02A01N63/22A01P1/00A01P3/00C12R1/07
CPCC12N1/02A01N63/22
Inventor 邹丽芳黄梦桑李逸朗杨瑞环闫依超方园周琦陈功友
Owner SHANGHAI JIAO TONG UNIV
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