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Biocontrol Strain dx3 for Controlling Tomato Yellow Leaf Curl Virus and Its Application

A technology of tomato yellowing curved leaves and bio-control strains, applied in the field of agricultural microorganisms, can solve the problems of pesticide residues, complexity, difficulty in cultivating breeding lines with stable resistance, etc., and achieve the effect of saving costs and reducing residues

Inactive Publication Date: 2014-10-22
南京农大生物源农药创制有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although breeding resistant varieties is the best way to control tomato yellow leaf curl virus disease, the limited availability of commercially valuable resistant materials has greatly limited the progress of breeding efforts
Moreover, due to the complexity of the genetic mechanism and background, it is difficult to breed breeding lines with stable resistance
With the long-term use of chemical pesticides, environmental pollution, pesticide residues and other problems are intensified, which is not conducive to the sustainable development of agriculture

Method used

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  • Biocontrol Strain dx3 for Controlling Tomato Yellow Leaf Curl Virus and Its Application
  • Biocontrol Strain dx3 for Controlling Tomato Yellow Leaf Curl Virus and Its Application
  • Biocontrol Strain dx3 for Controlling Tomato Yellow Leaf Curl Virus and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Screening method: Bacillus DX3 was isolated from tomato rhizosphere soil in Ganyu County, Lianyungang City, Jiangsu Province. Gently brush the soil attached to the tomato root surface with a soft brush, weigh 3g and add it to 27ml sterilized 0.85% NaCl, vibrate on a shaker at 180rmp for 0.5h, and let it stand for 5min to obtain 10 -1 Diluent, absorb 0.1mL of supernatant and dilute in turn, take 10 -4 、10 -5 、10 -6 Apply 0.1 mL each of the three gradient dilutions on the LB plate, incubate at 28°C for 48 hours, pick a single colony and inoculate it into fresh LB solid medium, and obtain purified strains through repeated transfer. The purified strains were stored in 40% glycerol in a -70°C ultra-low temperature freezer.

[0022] Identification method: The strain was identified through morphological characteristics, physiological and biochemical experiments and 16S rDNA sequence analysis.

[0023] Morphological characteristics: non-capsulated, periosteum flagella, able...

Embodiment 2D

[0031] The preparation of embodiment 2DX3 biocontrol agent

[0032] Inoculate the Bacillus DX3 (CGMCC NO.7239) strain into the LB culture medium for 16 hours at 28°C and shake at 200 rpm for 16 hours, then take a sample in the ultra-clean bench every 2 hours to measure its OD value at 600 nm, and end when the OD value is 0.8 Cultivate and use this bacterium liquid as the seed bacterium liquid. Inoculate the seed bacteria solution into LB fermentation culture medium at a volume ratio of 1:500 for fermentation and culture, culture at 28°C and 200rpm for 48h, then centrifuge at 6000rpm for 10min, take the precipitate and dilute it with sterilized water to obtain the biocontrol agent DX3 . The total concentration of viable bacteria in the resulting biocontrol agent was 1×10 9 -1×10 10 CFU / mL.

Embodiment 3

[0033] Example 3 Greenhouse Test Determination of the Control Effect of DX3 on Tomato Yellow Leaf Curl Virus Disease

[0034] The tomato variety used in this test is "He He 903", which was transplanted at the 4-leaf stage. After 4 days of transplanting, the biocontrol agent prepared in Example 2 was diluted with water and sprayed. The spray concentration is 1×10 7 CFU / mL, the spray standard is the distribution of mist droplets on each leaf, subject to not falling. The tomato yellow leaf curl virus was inoculated 7 days after the biocontrol agent treatment. The second same treatment was carried out 15 days after the first treatment. The treatment of only inoculating virus without biocontrol bacteria was used as the control treatment. There were 24 tomato plants in each treatment group, with three repetitions. The tomatoes were cultivated in a greenhouse with a temperature of 28°C and a light of 12h / 12h. The virus began to develop 15 days after inoculation, and the disease i...

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Abstract

The invention belongs to the field of agricultural microbial technology, and relates to a bio-control strain DX3 used for preventing and controlling tomatos yellows leaf curl virus and applications thereof. It is identified that DX3 belongs to Bacillus sp.; DX3 is preserved in China General Microbiological Culture Collection Center from January, 31th, 2013; and the strain preservation number is CGMCC NO.7239. The results of a greenhouse experiment show that: the biocontrol strain DX3 is capable of reducing the frequency of toamtos yellows leaf curl virus disease significantly. A biocontrol agent is prepared by using DX3, and is diluted to 100-fold for spray processing; and bio-control effect of toamtos yellows leaf curl virus disease at room temperature can reach 57.38% by spray processing. In addition the bio-control effect under field conditions can reach 51.70 to 53.35%. Therefore, the strain and the bio-control agent prepared by using the strain can be used for preventing and controlling toamtos yellows leaf curl virus and yield increase.

Description

technical field [0001] The invention belongs to the technical field of agricultural microorganisms, and relates to a biocontrol bacterial strain DX3 for preventing and treating tomato yellow leaf curl virus disease and an application thereof. Background technique [0002] In recent years, tomato yellow leaf curl virus has been outbreaked continuously in a large area in my country. At present, the occurrence area is more than 200,000 hm2, and the economic loss is billions of yuan, which has constituted a serious threat to the sustainable development of my country's tomato industry. The disease is caused by tomato yellow leaf curl virus (TYLCV) of the genus Begomovirus in the family Geminiviridae and is transmitted by whiteflies. The viral disease has the characteristics of strong epidemic, rapid onset, rapid spread, serious damage, and difficult to control. The plots with severe disease may even cause crop failure, seriously affecting the yield and quality of tomatoes. [0...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/00A01P1/00C12R1/07
Inventor 王勇明许泉郭坚华丁雪玲
Owner 南京农大生物源农药创制有限公司
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