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Ralstonia strain, and application, preparation and control effect evaluation method thereof

A technology of Ralellia bacteria and an acquisition method, which is applied in the directions of botanical equipment and methods, biochemical equipment and methods, and applications to achieve the effect of good safety

Active Publication Date: 2021-10-15
YUNNAN ACAD OF TOBACCO AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently no application of Ralstella bacteria in the prevention and treatment of tobacco bacterial wilt. The use of screening non-pathogenic R. solanacearum to develop plant vaccine engineering bacteria provides a microbial plant vaccine for the prevention and treatment of tobacco bacterial wilt. No report

Method used

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  • Ralstonia strain, and application, preparation and control effect evaluation method thereof
  • Ralstonia strain, and application, preparation and control effect evaluation method thereof
  • Ralstonia strain, and application, preparation and control effect evaluation method thereof

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Experimental program
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Effect test

Embodiment 1

[0055] Strain acquisition

[0056] Trapping, isolation and cultivation

[0057] Step 1): After removing impurities and larger lumps from the soil sample, put it into a glass petri dish with a diameter of 120 mm, the thickness of the soil layer is about 1.5 cm, and moisten the soil with distilled water using a titration bottle;

[0058] Step 2): Prepare the microorganism trapping device: first, apply glue on the edge of a microporous filter membrane with a diameter of 50 mm and a pore size of 0.45 μm, and place a stainless steel flat-bottomed gasket on the microporous filter membrane; then add 3 mL of solid to the inner cavity of the gasket Culture medium (1.2% gellan gum and 1.0% vitamin); then coat the upper surface of the metal gasket with glue, and cover another microporous filter membrane with a pore size of 0.45 μm;

[0059] Step 3): Place the microbial trapping device on the moist soil in step 1, compact it gently to ensure that the microporous filter membrane is in ful...

Embodiment 2

[0065] Evaluation of the control effect of biocontrol bacteria on tobacco bacterial wilt:

[0066] The control effect evaluation is divided into initial screening and re-screening of indoor biological testing. The specific operations are as follows:

[0067] Step S1, preliminary biometric screening

[0068] Tobacco seedling treatment: Cultivate the tobacco seedlings to the 4-5 leaf stage by the floating seedling cultivation method, take out the spare tobacco seedlings from the seedling pond 1 day before the seedlings, dry them on a floating plate, and use a sterile blade to cut the root of the tobacco seedlings 1.5cm away from the center of the tobacco plant the next day Injuries were made on both sides of the plant, and the tobacco seedlings were divided into a test group and a control group, with 2 tobacco seedlings in each group;

[0069] Biocontrol bacteria pretreatment: the test group was inoculated with 1mL of the test bacteria, and the tobacco seedlings inoculated with...

Embodiment 3

[0082] Strain Identification and Preservation

[0083] Strain identification

[0084] Conventional bacterial identification refers to the literature "Common Bacterial System Identification Manual" (edited by Dong Xiuzhu et al. Science Press. 2001).

[0085] Biolog GEN III plate was used to analyze the compound metabolism characteristics of strain 56D2.

[0086] The molecular identification method is as follows: Bacteria Genomic DNA Extraction Kit, see the kit instruction manual for the method. The 16S rDNA sequence was amplified by PCR with the general primer F27 / R1492, and amplified under conventional conditions. After the amplified product was recovered by gel, it was connected to the vector pEAZY-T5 Zero vector, and the E. coli competent cell DH5α was heat-shocked. M13R was used as primer for colony PCR identification. Positive clones were sent to Shanghai Yingjun Biotechnology Co., Ltd. for sequence determination. Through the above 16S rDNA sequence analysis, the seque...

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Abstract

The invention relates to a Ralstonia strain as well as an application, a preparation method and a control effect evaluation method thereof. The Ralstonia strain is Ralstonia sp.56D2, and the strain is preserved in the China Center for Type Culture Collection on November 12, 2019; the preservation address is Wuhan University, Bayi road No.299, Wuchang District, Wuhan City, Hubei Province; the accession number of the strain is CCTCC No: M 2019921. The invention provides Ralstonia sp. 56D2 capable of preventing and treating tobacco bacterial wilt; a preparation method of the Ralstonia sp. 56D2; and an application of the Ralstonia sp. 56D2 in tobacco bacterial wilt disease.

Description

technical field [0001] The invention belongs to the technical field of microbe application, and in particular relates to the technical field of a Rattlesia strain and its application, acquisition and control effect evaluation method. Background technique [0002] Tobacco bacterial wilt (TBW) is a vascular disease and one of the common plant bacterial diseases in tropical, subtropical and some temperate regions. When the plant is diseased, the appearance of the stem of the diseased plant can be seen as longitudinal black stripes. Sometimes the leaves on one side of the diseased plant become sick and withered first, and the roots on the corresponding side turn black and rot. In severe cases, large areas of water-soaked necrotic spots appear on the leaves. In the middle and late stages of the disease, all leaves wither. Cutting the stem of the diseased plant shows that the vascular bundles turn brown, and the part with higher water content is squeezed by hand, and it can be ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/20A01P1/00A01G22/45A01G7/06C12Q1/04C12R1/01
CPCC12N1/20A01N63/20A01G22/45A01G7/06C12Q1/04
Inventor 卢灿华马俊红盖晓彤姜宁殷红慧张朝富高朝阳雷丽萍晋艳夏振远
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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