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Biocontrol strain HW2 for preventing and treating cucumber green mottle mosaic virus and application of biocontrol strain HW2

A technology for green mottled flowers and biocontrol strains is applied in the field of agricultural microorganisms, which can solve the problems of difficult prevention and control, complex genetic mechanism and background, and difficulty in cultivating varieties with stable resistance, so as to achieve the effect of saving expenses and reducing residues.

Inactive Publication Date: 2016-01-06
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although cultivating disease-resistant varieties is the fastest and most effective method, due to the complexity of its genetic mechanism and background, it is difficult to breed varieties with stable resistance. At the same time, due to the high price of disease-resistant varieties, ordinary farmers do not have the ability to purchase them. The promotion and application of disease-resistant varieties
On the other hand, cucumber green mottle mosaic virus disease is mainly transmitted by seed transmission and contact infection. The virus can enter the tissue through the host's tiny wounds, and it can also infect healthy plants with the virus through grafting operations. The virus can survive the winter in the soil , and the activity can be maintained for more than 1 year, becoming the source of infection in the next year, making it difficult to prevent and control

Method used

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  • Biocontrol strain HW2 for preventing and treating cucumber green mottle mosaic virus and application of biocontrol strain HW2
  • Biocontrol strain HW2 for preventing and treating cucumber green mottle mosaic virus and application of biocontrol strain HW2
  • Biocontrol strain HW2 for preventing and treating cucumber green mottle mosaic virus and application of biocontrol strain HW2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Isolation method: Stenotrophomonas maltophilia HW2 (CGMCC NO.11024) was isolated from healthy watermelon leaves in a watermelon greenhouse in Dingji Town, Huaian City, Jiangsu Province. Cut the watermelon leaf tissue into small pieces of 1 cm, soak in 1% sodium hypochlorite for 5 min, soak in 70% alcohol for 1-2 min, and wash with sterile water for 3 times (take the last washing solution 100 μl and apply it on R 2 A solid culture medium, if sterile growth after 48h, it is considered that the surface is clean and sterile). Take 3g of the sterilized sample and place it in a sterile mortar, add 27ml of sterile 0.85% NaCl, macerate the tissue, grind it, and dilute with sterile 0.85% NaCl gradient. take 10 -1 、10 -2 、10 -3 100 μl each of the three gradient dilutions were applied to R 2 On A, culture at 28°C for 48h. Pick the largest single colony to inoculate into fresh R 2 A solid medium, through repeated transfer to obtain purified strains. The purified strains were...

Embodiment 2

[0046] The preparation of embodiment 2HW2 biocontrol agent

[0047] Inoculate Stenotrophomonas maltophilia HW2 (CGMCC NO.11024) strain into LB culture medium for 16 hours at 28°C with shaking at 200rpm, then take samples every 2 hours in a clean bench to measure its OD value at 600nm, when OD When the value is 0.8, the culture is terminated, and this bacterial solution is used as the seed bacterial solution. The seed bacteria solution was inoculated into LB fermentation broth at a volume ratio of 1:500 for fermentation and culture, cultured at 28°C and 200 rpm for 48 hours, then centrifuged at 6000 rpm for 10 minutes, and the precipitate was diluted with sterilized water to obtain the biocontrol agent HW2. The total concentration of viable bacteria in the resulting biocontrol agent was 1×10 9 ~1×10 10 CFU / mL.

Embodiment 3

[0048] Example 3 Greenhouse Test Determination of the Control Effect of HW2 on Cucumber Green Mottled Mosaic Virus

[0049] The cucumbers are transplanted at the 2-3 leaf stage, and the roots are irrigated and sprayed at the same time. Example 2 preparation of HW2 (CGMCCNO.11024) bacterial agent root irrigation concentration is 1 × 10 8 CFU / mL, the dosage for each seedling is 30ml. The spray concentration is 1×10 7 CFU / mL, the spray standard is that there is a mist-like droplet distribution on each leaf, subject to not falling, spray evenly. After 7 days of biocontrol agent treatment, inoculate cucumber green mottle mosaic virus (the pathogenic bacteria used is the virus CGMMV, donated by Zhejiang Academy of Agricultural Sciences, it is an infectious clone Agrobacterium, the strain name is called EHA105 in the experiment, and the transformed plasmid is pCB-CGMMV Prepare EHA105 in LB culture solution containing rifampicin and kanamycin (the final concentration is 50mg / mL) to...

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Abstract

The invention belongs to the field of agricultural microbiology, and relates to a biocontrol strain HW2 for preventing and treating cucumber green mottle mosaic virus and an application of the biocontrol strain HW2. Through authentication, the biocontrol strain HW2 belongs to stenotrophomonas maltophilia, and is preserved in the China General Microbiological Culture Collection Center (CGMCC) on June 29, 2015, wherein the preservation number of the biocontrol strain HW2 is CGMCC NO.11024. Meanwhile, the greenhouse experimental result shows that the biocontrol strain HW2 can obviously reduce the disease severity of the cucumber green mottle mosaic virus, and has an obvious effect in preventing and treating cucumber green mottle mosaic virus, the control effect is within 53.85%-57.02%, the microbial inoculum has a very good growth promoting effect for cucumber, the biomass increase can achieve 42.76%, and the content of chlorophyll of the cucumber leaf is obviously increased. Therefore, the strain and the microbial inoculum prepared by the strain can be applied to the prevention and treatment of the cucumber green mottle mosaic virus and growth promotion.

Description

technical field [0001] The invention belongs to the field of agricultural microorganisms, and relates to a biocontrol bacterial strain HW2 for preventing and treating cucumber green mottle mosaic virus disease and an application thereof. Background technique [0002] Cucumber green mottlemosaic virus (CGMMV) is a member of the Tobamovirus genus in the family Tymoviridae, and is an important quarantine virus on Cucurbitaceae plants in many countries and regions in the world. It seriously threatens the production of watermelons, melons, cucumbers and other crops. In 1935, Ainsworth of the United Kingdom reported the harm of CGMMV on cucumber for the first time, and then it was spread to more than 20 countries in Europe, Asia and South America. The disease was first discovered in Gaizhou, Liaoning, my country in 2005. Because of its serious damage, the Ministry of Agriculture issued Announcement No. 788 on December 21, 2006, identifying CGMMV as a national agricultural plant q...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P1/00A01P21/00C12R1/01
Inventor 刘红霞李宏伟
Owner NANJING AGRICULTURAL UNIVERSITY
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