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Brevundimonas olei dahliae olei and application thereof

A technology of Brevundimonas and Verticillium dahliae, applied in the fields of application, bacteria, biocide, etc., can solve problems that restrict the healthy and sustainable development of the cotton industry, affect the yield of continuous cropping cotton fields and cotton quality, etc.

Active Publication Date: 2021-01-05
TARIM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0004] Cotton verticillium wilt is mainly a soil-borne fungal vascular disease caused by Verticillium dahliae, which seriously affects the yield and quality of cotton in continuous cropping cotton fields, restricting the healthy and sustainable development of the cotton industry. Cotton verticillium wilt caused by Verticillium dahliae Whether there is such a symbiotic relationship has not been reported

Method used

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  • Brevundimonas olei dahliae olei and application thereof
  • Brevundimonas olei dahliae olei and application thereof
  • Brevundimonas olei dahliae olei and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: Verticillium dahliae pathogen molecular biology identification

[0029] Extraction of DNA of Verticillium dahliae pathogenic bacteria: take Verticillium dahliae hyphae, quick-freeze in liquid nitrogen to break the wall, grind into powder, transfer to a 1.5mL centrifuge tube; add 700 μL of CTAB buffer preheated to 65°C, and bathe in 65°C water for 50 minutes, during which Turn the centrifuge tube upside down every 10 minutes to fully mix the mycelia powder and buffer; pre-cool the centrifuge to 4°C in advance, and centrifuge at 12000rpm for 15 minutes; take out the centrifuge tube and transfer the supernatant to a new centrifuge tube, add DNA Extract 600 μL, mix well and centrifuge at 12000 rpm for 15 min; transfer the supernatant to a new centrifuge tube, repeat the above steps; transfer the supernatant to a new centrifuge tube, add 600 μL DNA extract II, mix well , centrifuge at 12000rpm for 15min; take out the centrifuge tube, pipette the supernatant int...

Embodiment 2

[0037] Example 2: Detection of Verticillium dahliae diversity in 4 strains of cotton

[0038] Microbiome total DNA extraction: DNeasy PowerWater Kit was used for extraction, and primers 338F (5'-ACTCCTACGGGAGGCAGCA-3' and 806R (5'-GGACTACHVGGGTWTCTAAT-3') were used to perform PCR amplification on the bacterial 16S rDNA V3-V4 region.

[0039] PCR amplicons were purified with Agencourt AMPure Beads (Beckman Coulter, Indianapolis, IN) and quantified using the PicoGreen dsDNA Assay Kit. After a separate quantification step, amplicons were pooled in equal amounts and subjected to paired-end 2 × 300 bp sequencing using the Illumina MiSeq platform and MiSeq Reagent Kit v3.

[0040] Bioinformatics and Statistical Analysis: Sequencing data were processed using quantitative workflows in microbial ecology. Low-quality sequences were filtered by the following criteria: sequences 8 bp. Paired-end reads were assembled using FLASH. After chimera detection, UCLUST (Edgar 2010) clustered th...

Embodiment 3

[0042] Example 3: Isolation and identification of 4 strains of cotton Verticillium dahliae endosymbiotic bacteria

[0043] Isolation and morphological identification of endosymbiotic bacteria: Cultivate Verticillium dahliae in cotton for 7-10 days, scrape the Verticillium dahliae the size of a matchhead with the inoculation needle, use the general mechanical wall breaking method, put it into a frozen mortar, and grind it with liquid nitrogen quick freezing Broken. Gradient dilution, take 30 μL of different concentrations of gradient bacterial solution and spread it in LA medium, culture at 37°C for 16-24 hours, conduct preliminary identification of the isolated symbiotic bacteria strains through morphological characteristics, grow on LB medium, symbiotic bacteria The colony was milky yellow, with a raised center, shiny surface, and viscous (shown in Figure 3A).

[0044] Use an inoculation needle to pick up endosymbiont colonies, place them on a glass slide, add 2 μL of steril...

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Abstract

The invention discloses Brevundimonas olei dahliae olei and application thereof. The Brevundimonas olei dahliae olei is named as Brevundimonas olei dahliae olei, and the preservation number of the Brevundimonas olei dahliae olei is CCTCC NO: M 2020392. The strain is determined to be Brevundimonas olei dahliae olei through morphological identification and diversity sequencing analysis, and the influence of B.olei fermentation liquor on growth of the Brevundimonas oleei and microsclerosis is explored through a co-culture experiment. Through scanning electron microscope observation, as the culture time and the concentration of the fermentation liquor increase, formation of conidia of the Brevundimonas oleei is reduced, the hypha morphology is expanded, broken and deformed, the microsclerosisis gradually reduced and disappears, and it is proved through the research that the symbiotic B.olei has good bacteriostatic activity on the Brevundimonas olei.

Description

technical field [0001] The invention relates to the technical field of development and utilization of microbial germplasm resources. Specifically, it is a brevundimonas endosymbiotic bacterium of Verticillium dahliae and its application. Background technique [0002] There is a complex symbiotic relationship between microorganisms and other individuals, and this symbiotic phenomenon widely exists between microorganisms, microorganisms and plants, microorganisms and animals. Symbiotic bacteria are a common phenomenon in nature, and there are various types of hosts for symbiotic bacteria, which can co-exist with different animals, plants, corals, macrofungi, aphids, algae and other hosts, and participate in or determine certain characteristics of the host. Symbiotic bacteria can fix nitrogen for plants, provide nutrients for plants, participate in the synthesis of host secondary metabolism, and assist the host to adapt to extreme environments. Symbiotic bacteria secrete a var...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/20A01P3/00C12R1/01
CPCC12N1/20A01N63/20C12R2001/01C12N1/205
Inventor 曾红高畅华羚淇刘桂敏
Owner TARIM UNIV
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