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Salt-tolerant biocontrol bacterium B268 for bacterial wilt of horsetail beefwood and application of salt-tolerant biocontrol bacterium B268

A biocontrol technology of bacteria and casuarina, which is applied in the direction of application, bacteria, biocide, etc., to achieve the effects of strong biofilm formation ability, strong growth and colonization ability, and wide tolerance range

Active Publication Date: 2017-05-10
ZHEJIANG FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The new bacterial strain B268 involved in this patent has the closest kinship relationship with Siamese bacillus (B.siamensis). Although existing literature has predicted the biocontrol potential of this kind of bacillus from the perspective of genome, it has not been used in the prevention and treatment of any diseases at present. have been reported in the study

Method used

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  • Salt-tolerant biocontrol bacterium B268 for bacterial wilt of horsetail beefwood and application of salt-tolerant biocontrol bacterium B268
  • Salt-tolerant biocontrol bacterium B268 for bacterial wilt of horsetail beefwood and application of salt-tolerant biocontrol bacterium B268
  • Salt-tolerant biocontrol bacterium B268 for bacterial wilt of horsetail beefwood and application of salt-tolerant biocontrol bacterium B268

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Isolation and activity screening of bacterial strain Bacillus siamensis B268

[0031] The samples were collected from coastal casuarina plantations in Yueqing City, Zhejiang Province, where a verification experiment on the control effect of casuarina bacterial wilt by watering with seawater had been carried out. Select disease-free and diseased plants, and collect soil below 10 cm in root circumference.

[0032] Before the strains were isolated, the salinity of the samples was verified, that is, the metal element composition and content of the soil samples were analyzed by Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES). The results showed that the The concentrations of sodium and magnesium ions are 0.5-0.7% and 0.3-0.5%, respectively, which are obviously higher than the common soil sample as a control.

[0033] Take 10g of soil samples and suspend them in 20ml of sterile saline, and extract them for 2 days at 28°C and 140rpm. The suspens...

Embodiment 2

[0039] Example 2: Phylogenetic identification of strain Bacillus siamensis B268

[0040] Pick a single colony on the plate of strain B268, transfer it to nutrient-rich artificial seawater medium, and collect the cells after culturing at 28°C and 140rpm for 24 hours. At the same time, use the bacterial genome rapid extraction kit (Dongsheng Bio) to extract the genome template. . Bacterial universal primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-ACGGTTACCTTGTTACGACTT-3') were used to amplify the 16S rRNA gene. Vector connection (TaKaRa), and transform Trans1-T1 competent cells (TransGen Biotech), pick positive recombinants and send them to the sequencing company for sequencing.

[0041] Under the microscope, strain B268 becomes rod-shaped, and the formation of spores can be observed ( figure 1 ).

[0042] The full-length 16S rRNA gene of strain B268 is 1513nt (see the sequence listing for details), and the sequence has been submitted to the GenBank database with the ...

Embodiment 3

[0045] Example 3: Verification of antagonistic activity of bacterial strain Bacillus siamensis B268

[0046] Use nutrient-enriched artificial seawater medium and Modified YPG salt-free medium to prepare confrontation plates, and use Ralstonia solanacearum YQ isolated from casuarina diseased strains in our laboratory and Ralstonia solanacearum LMG 2299 isolated from tomato diseased strains purchased from BCCM T Validation of antagonistic activity for the target. Strain B268 was activated by streaking on a plate and cultured at 28°C for 24 hours. The two pathogenic strains were activated by liquid shaking tubes. When using nutrient-rich artificial seawater medium, culture at 28°C and 140rpm for 48 hours. When using Modified YPG salt-free medium , cultivated at 28° C., 140 rpm for 24 hours. The preparation method of the confrontation plate was the same as in Example 1, and the formation of the inhibition zone was observed after culturing at 28° C. for 3 days.

[0047] Such as ...

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Abstract

The invention provides a salt-tolerant biocontrol bacterium B268 for the bacterial wilt of horsetail beefwood. The salt-tolerant biocontrol bacterium B268 for the bacterial wilt of horsetail beefwood is classified and named as Bacillus siamensis, and is preserved in China General Microbiological Collection Center, the preservation number is CGMCC No.13225, and the preservation date is November, 1, 2016. The strain is separated from seawater immersed root soil of horsetail beefwood, has an antagonistic activity for pathogenic bacterial strains of the bacterial wilt of horsetail beefwood and tomatoes, and also has the an activity of dissolving phosphorus, an activity of synthesizing a siderophore and a plant growth hormone and other activities of promoting plant growth. In addition, the tolerance scope of sea salt of the strain is wide, a stable biological film can be formed at high salt content, and the capabilities of growth and colonization are high, and therefore, the strain B268 can be applied to the biological prevention and control of the bacterial wilt of horsetail beefwood, is particularly applied to the preparation of a biological control agent or a pesticide for the bacterial wilt of horsetail beefwood, and is also applied to the preparation of a bacterial manure for promoting the growth and improving the disease resistance of horsetail beefwood.

Description

technical field [0001] The invention belongs to microbial strains and applications thereof, and relates to a strain of salt-tolerant casuarina bacterial wilt biocontrol bacteria B268, which is a new strain of halo-tolerant bacillus Bacillus siamensis, and the strain preservation number is CGMCC No.13225. The invention also relates to the biocontrol activity of the bacterial strain, including antibacterial activity and plant growth-promoting activity, and its application in the prevention and treatment of casuarina bacterial wilt. Background technique [0002] Bacterial wilt, also known as bacterial wilt, is a worldwide soil-borne disease caused by Ralstonia solanacearum, which occurs widely in tropical, subtropical and temperate regions. Due to the influence of the greenhouse effect, there has been a tendency to spread to high latitudes. R. solanacearum contains 6 biochemical types (biovar I-VI) and 5 physiological races (race 1-5). The complex intraspecific diversity enabl...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P1/00A01P21/00C05F11/08C12R1/07
CPCA01N63/00C05F11/08C12N1/205C12R2001/07
Inventor 张心齐张昕林海萍陈旭华
Owner ZHEJIANG FORESTRY UNIVERSITY
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