Biocontrol pythium oligandrum and application

A technology of Pythium and biocontrol agents, applied in the application, biocide, microorganism and other directions, can solve limited problems, and achieve the effects of reducing environmental pollution, good application prospects, and simple cultivation conditions

Active Publication Date: 2019-10-11
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Since the research and application of Pythium oligandrum in my country started later than developed countries suc...

Method used

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  • Biocontrol pythium oligandrum and application
  • Biocontrol pythium oligandrum and application
  • Biocontrol pythium oligandrum and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The separation of embodiment 1 biocontrol Pythium oligandrum

[0035] The specific process is as follows:

[0036] 1. Collection of soil samples

[0037] The soil at a depth of 10-20 cm was collected from the ginger disease field in Lijiazhai Village, Pingli Town, Laicheng District, Laiwu City, Shandong Province, and the soil was packed and marked and brought back to the laboratory for separation.

[0038]2. Isolation of Pythium oligandrum

[0039] (1) Take the collected soil samples and use a coarse sieve to remove stones and uncorrupted plant residues.

[0040] (2) Weigh 50 g of sieved soil, add 500 ml of water, stir for 5 minutes with a glass rod, immediately filter the supernatant through sieves with apertures of 150, 61, and 38 microns, and collect the filtrate.

[0041] (3) Rinse the 38-micron sieve with a small amount of water, and collect the residue filtered on the net in a conical flask.

[0042] (4) Take 5ml of the residue suspension and add it on the sel...

Embodiment 2

[0044] Molecular identification of embodiment 2 Pythium oligandrum GAQ1

[0045] After using the CTAB method to extract the genomic DNA of the five isolated strains, use Vazyme's MaxSuper-Fidelity DNA Polymerase high-fidelity enzyme and primer pair ITS1(5'-TCCGTAGGTGAACCTGCGG-3') / ITS4(5'-TCCTCCGCTTATTGATATGC-3') to amplify the ITS region of eukaryotes, sent to Shenggong Biological Company (Shanghai ) Co., Ltd. sequenced to obtain nucleotide sequence information, and submitted the obtained sequence to the GenBank database for BLAST analysis and comparison. It was found that a strain GAQ1 isolated may be Pythium oligandrum.

[0046] In order to further verify whether the bacterial strain is Pythium oligandrum, use Vazyme's The primer pair of MaxSuper-Fidelity DNA Polymeras high-fidelity enzyme and two housekeeping genes of Pythium TUBUF2(5'-CGGTAACAACTGGGCCAAGG-3') / TUBUR1(5'-CCTGGTACTGCTGGTACTC-3') and FM66(5'-TAGGATTTCAAGATCCTGC-3' ) / FM58(5'-CCACAAATTTCACTACATTGA-3') to amp...

Embodiment 3

[0057] Embodiment 3 Plate Antagonism Detection of Pythium oligandrum GAQ1

[0058] 1. Tablet confrontation test

[0059] (1) Pythium oligandrum GAQ1 and Pythium conglomerates grown on 10% V8 solid plates for 1 day were respectively punched along the edge of the colony with a puncher with a diameter of 6 mm. The preparation method of the 10% V8 solid medium is to centrifuge the V8 juice at 5000rpm for 10min, filter through four layers of gauze, take 1ml of the filtrate, add 99ml of water, 1.5g of agar, and sterilize the culture medium at 121°C for 20min .

[0060] (2) Inoculate a plate of Pythium conglomerates at the edge of a 10% V8 solid plate with a distance of 9 cm, and inoculate a plate of Pythium oligandrum GAQ1 at the other end of the plate, and culture it at 25° C.

[0061] (3) Cover the confrontation plate with 5ml 0.1% trypan blue before contacting the two strains, partially cover, and completely cover three different stages, rotate gently to ensure that the dye is ...

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Abstract

The invention discloses a biocontrol pythium oligandrum strain for preventing and controlling pythium root rot of seedlings of cucumis sativus L., gray blight of cucumis sativus L. and meloidogyneoryzae diseases and a separation and application method thereof, and belongs to the technical field of crop disease prevention and control. The biocontrol strain is pythium oligandrum GAQ1, and the preservation number is CGMCC No.17470. The invention further discloses the separation method of the biocontrol pythium oligandrum and the application of the biocontrol pythium oligandrum in preventing and treating oomycete diseases of cucumis sativus L. and meloidogyneoryzae diseases. The pythium oligandrum CGMCC 17470 can significantly inhibit the infection of crops by pythium oxysporum, phytophthora capsici and meloidogyneoryzae. The strain GAQ1 is isolated from the soil and harmoniously integrated with the ecological environment, has no toxicity or pathogenicity, and can be prepared into a biological preparation for biological prevention and control over plant diseases.

Description

technical field [0001] The invention relates to biocontrol bacteria, in particular to Pythium oligandrum and its application. Background technique [0002] The fruit of cucumber (Cucumis sativus L.) has the characteristics of high water content and low calorie, and also has potential effects of anti-diabetes, hypolipidemic, anti-oxidation, soothing skin irritation and reducing swelling, so cucumber is planted in large areas around the world , has become an important economic crop in the world, but the yield of cucumber is threatened by a variety of pathogens every year, including oomycete pathogens from Pythium and Phytophthora. [0003] Cucumber seedlings are infected with Pythium myriotylum at the seedling stage to cause Pythium myriotylum root rot. Pythium conglomerate belongs to class Oomycetes, Pythium order, Pythium family, Pythium genus, is a class of serious soil-borne pathogens. The pathogen is mainly transmitted through irrigation water or rainwater. When the zoo...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N1/02A01N63/04A01P5/00A01P3/00C12R1/645
CPCC12N1/02C12N1/14C12R2001/645C12N1/145A01N63/30
Inventor 魏利辉张琪梦周冬梅
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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