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A kind of peanut endogenous Burkholderia cepacia and application thereof

A technology of Burkholderia and onion, which is applied in the field of microorganisms, can solve the problems such as failure to prevent and control peanut fruit rot and high-efficiency biocontrol bacteria, and achieve the effects of remarkable control effect, high safety in use and environmental friendliness.

Active Publication Date: 2022-03-22
SHANDONG PEANUT RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no high-efficiency biocontrol bacteria for preventing and treating peanut fruit rot in the prior art

Method used

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  • A kind of peanut endogenous Burkholderia cepacia and application thereof
  • A kind of peanut endogenous Burkholderia cepacia and application thereof
  • A kind of peanut endogenous Burkholderia cepacia and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Isolation and purification of strains

[0034] Healthy peanut plants were collected from the experimental field of Shandong Peanut Research Institute in Laixi City, Shandong Province, and the root, stem and leaf tissues were cut out, rinsed with tap water, and then washed 3 times with sterile water. Then soak it in 75% ethanol for 30 seconds, then soak it in 2% sodium hypochlorite solution for 2.5 minutes, take it out, soak it in sterile water and rinse it five times. Put it in a sterilized mortar, add 1mL sterile normal saline and 1g sterilized quartz sand in turn, grind fully at room temperature, take homogenate, and carry out 10-10 3 Take 100 μl of 10 for serial dilution -4 and 10 -5 The dilutions of the above-mentioned nutrient agar medium were spread on the nutrient agar medium, and each dilution was repeated 3 times. After being cultured in a constant temperature incubator at 28°C for 48 hours, a single colony with different cultural characteristics was picked a...

Embodiment 2

[0036] Screening of strains with antibacterial activity

[0037] In order to obtain the endophytic strain with the highest antagonistic activity, all candidate isolates were tested for their antagonism on Pythium conglomerate mycelium growth by disc diffusion method.

[0038] Through experiments, 5 strains of active bacteria were screened out, namely bacterial strain PRI08, bacterial strain PRI20, bacterial strain PRI16, bacterial strain PRI19, bacterial strain PRI17, and the above-mentioned 5 strains of active bacteria all showed different mycelia growth inhibition effects (see figure 1 ), where each experiment was repeated 3 times, and CK was the blank control group.

[0039] All the 5 strains of active bacteria showed different mycelial growth inhibition effects. In the mycelial growth inhibition test, the PRI08 strain had the highest inhibition rate (PIRG) of radial growth 2 days after inoculation (54% ± 2%) (P<0.05); Since the PRI08 strain had higher antagonistic activit...

Embodiment 3

[0041] Identification of the PRI08 strain

[0042] Morphological identification: PRI08 strains were cultured on nutrient agar medium at 28°C, and the colonies were round, yellow, smooth, raised, and entire. Bacteria were Gram-negative, strictly aerobic, and both catalase and oxidase were positive. At the same time, peanut seedlings inoculated with the cell suspension of the strain did not show obvious disease symptoms.

[0043] Molecular identification: Pick a single colony on the LB plate, dissolve it in 10 μL sterile water, use the bacterial suspension as a template, and amplify the 16S sequence with bacterial 16S universal primers. Upstream sequence, as shown in SEQ ID No.2, downstream sequence: as shown in SEQ ID No.3, a fragment of the bacterium was amplified.

[0044] The PCR system is a 50 μL system, 2 μL of bacterial suspension, 1.5 μL of upstream primers, 1.5 μL of downstream primers (primer concentration is 10 μmol / L), 5 μL of 10×PCR buffer, 1 μL of dNTPs, 2 μL of ...

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Abstract

The invention discloses Burkholderia cepacia endogenous to peanuts and application thereof, and relates to the technical field of microbes. The Burkholderia cepacia (Burkholderia cepacia) PRI08 is preserved in the General Microbiology Center (CGMCC) of the China Microbiological Culture Collection Management Committee, the preservation date is September 29, 2020, and the preservation number is CGMCC No.20842 . The present invention successfully isolates and screens a strain of Burkholderia cepacia (Burkholderia cepacia) PRI08 from healthy peanut plants, which is preserved in the General Microorganism Center (CGMCC) of the China Microbiological Culture Collection Management Committee (CGMCC), and the preservation number is CGMCC No.20842; Burkholderia cepacia PRI08 and the microbial agent prepared from the suspension of Burkholderia cepacia PRI08 can be used for the prevention and treatment of peanut fruit rot, the control effect is remarkable, green and environmentally friendly, Friendly to the environment.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to Burkholderia cepacia endogenous to peanuts and an application thereof. Background technique [0002] Peanut is an important oil crop and economic crop in the world, and it is also an important crop for my country's export and foreign exchange earnings, occupying an important position in the national economy. Peanut fruit rot, also known as fruit rot, is a worldwide disease that mainly harms peanut pods and causes pods to rot. [0003] At present, regarding the prevention and treatment of peanut fruit rot, it is mainly to use the control measures such as chemical agent sterilization and crop rotation cultivation. Due to the wide range of pathogenic hosts of this disease, it is difficult to control to a certain extent. It is highly valued, and it is urgent to find a new way to safely prevent and control peanut fruit rot. [0004] Biological control plays an increasingly i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/20A01P3/00C12R1/01
CPCC12N1/20A01N63/20
Inventor 于静迟玉成许曼琳吴菊香张霞郭志青李莹宋新颖何康
Owner SHANDONG PEANUT RES INST
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