A novel method for controlling soybean root disease by using rhizobia

A rhizobia, soybean root technology, applied in the field of root diseases, can solve the problem of slow release control efficacy

Inactive Publication Date: 2007-04-25
SHENYANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, because two kinds of bacteria live together to inoculate the soybean root system, they can colonize and reproduce in the soil and in the root system, release the control effect slowly for a long time, and overcome the problem of the short-term effect of chemical agents

Method used

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  • A novel method for controlling soybean root disease by using rhizobia
  • A novel method for controlling soybean root disease by using rhizobia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Separation and purification of rhizobia

[0024] The preserved nodules are soaked in 95% alcohol for 15 minutes, and then placed in 0.1% HgCl 2 Medium treatment for 3 to 5 minutes. The specific time varies depending on the size of the nodule. Wash 3 times with sterile water. Take 0.2ml of the last sterile water rinse and apply it to the separation medium. Observe that there are no aseptic colonies. This method verifies whether the disinfection method can kill all microorganisms on the surface of the test sample. Use a sterile filter paper to absorb the surface water, use a sterile scalpel to cut the sterile filter paper and place it on the YMA medium. After 2-3 days, take a small amount of bacteria from around the nodules and inoculate it on the medium, and cultivate at 28℃ For 48h, take a single colony and streak culture on YMA medium until a pure culture is obtained. Pick a single colony number and store it in a refrigerator at 4°C with 15% glycerol-beef peptone culture s...

Embodiment 2

[0026] Isolation of Soybean Rhizobium Endophytic Bacillus

[0027] Take 5g of soybean nodules, wrap them in sterile gauze, and soak them in 0.1% Hgcl 2 Disinfect the middle surface for 10 minutes, and then rinse with sterile water 3-5 times. Verify that the disinfection method is the same as above. The test materials were transferred into a sterile mortar and ground into a homogenate, and 5ml of sterile water was added and stirred evenly. After placing 1 mL of tissue homogenate in a water bath at 80° C. for 20 minutes, take 0.2 mL and mix it with NA separation medium to separate endophytic bacteria. Place the petri dish upside down in a constant temperature culture at 28°C for 1 to 2 days. After the plate has grown, pick a single colony with a different colony shape and purify it for 2 to 3 times. Pick a single colony number and use 15% glycerin. -The beef peptone culture solution is stored in a refrigerator at 4°C.

Embodiment 3

[0028] Example 3: Cultivation of bacterial strains Snb2 (Bacillus subtilis) and L396 (Sinorhizobium sp.)

[0029] Two strains of bacteria were inoculated on test tube agar medium by streaking method, the medium formula was NA medium, cultured at 28°C for 3 days to obtain test tube species.

[0030] Then inoculate the test tube seeds into 250mL Erlenmeyer flasks (50mL per bottle) liquid culture medium, at 25℃~28℃, the shaker speed is 150r·min -1 ~200r·min -1 , Fermentation for 168h-200h. The fermentation broth was compounded according to different proportions, and the nematode-killing and antibacterial efficacy tests were determined.

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Abstract

The invention relates to a method for using soybean root nodule and endotrophic mycorrhiza to prevent soybean root disease. Wherein, the inventive bacteria seed has been stored in national microbe storage manager center, whose storage number is sinorhizobium sp; the seed L396 storage number is CGMCC No. 1851; the bacillus subtilis seed Snb2 storage number is CGMCC No.1850. And the invention uses general liquid to ferment and cultivate, uses itself or metabolite as effective active component; mixing the ferment solutions at different ratios, with effective restrain function on soybean cyst nematization and root pathogenic fungi; the density ratio between Snb2 and L392 is 8:3, and the dead ratio of young bacteria can reach 98.92%, and the relative restrain rate on cyst cultivation can reach 99.66%; the invention prepares the metabolite into water agent, to reach 78.6% prevent rate on the soybean cyst nematization. The inventive nodule bacteria can fix nitrogen and accelerate growth.

Description

Technical field: [0001] The invention belongs to the technical field of microbial pesticides, and specifically relates to a method and application for controlling root diseases caused by plant parasitic nematodes and pathogenic fungi by synergistic action of two endophytic bacteria. Background technique: [0002] Soybean cyst nematode (SCN), also known as soybean verticillium wilt and fire dragon seedlings, has long been an important disease restricting soybean production. This disease generally reduces soybean yield by 10% to 30%, and serious plots can reach 70% to 90%, and even cause crop failure, which is more harmful than any single disease. This disease has severely occurred in the main soybean producing areas of the three northeastern provinces of my country, Inner Mongolia and Huanghuaihai. It is a soil-borne disease that is extremely difficult to control. From a global perspective, the damage and spread of SCN has a growing trend. [0003] Soybean root rot is a worldwide ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N63/02A01P5/00A01P1/00A01C1/06C09K17/00C12N1/20C12R1/125
Inventor 段玉玺王媛媛陈立杰
Owner SHENYANG AGRI UNIV
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