Method for inhibiting tomato brown wrinkled fruit virus by using beta-glucan and application of beta-glucan

Abstract

The invention discloses a method for inhibiting tomato brown wrinkled fruit virus by using beta-glucan and application. The method comprises the following steps: spraying a prepared beta-glucan solution with the concentration of 25mg / L to 100mg / L to Bensi tobacco leaves; after 48 hours after the beta-glucan solution is sprayed, agrobacterium tumefaciens obtained through ToBRFV-GFP infectious cloning is used for inoculation to the lower leaves of the nicotiana benthamiana; 11 days after virus inoculation, the virus amount of ToBRFV is reduced, and the fluorescence intensity of GFP is weakened.

Description

Technical Field

[0001] The present invention relates to the technical field of plant anti-virus, and particularly relates to a method for inhibiting Tomato brown rugose fruit virus by β-glucan and its application. Background Art

[0002] The most important factor for the epidemic occurrence of traditional plant virus diseases and threatening global crops is that once a plant is infected, it is almost impossible to get rid of the virus, so it will continue to exist as an infection source. Tomatobrown rugose fruit virus (ToBRFV), as an emerging and rapidly spreading plant RNA virus, threatens global tomato production. The infectivity of its strains is highly stable, and it can be transmitted by seeds or through various mechanical contact methods. High survival rate, latency and easy transmissibility have made the crop damage caused by ToBRFV increasingly serious, and global warming has also become a major driving force for the epidemic spread of ToBRFV. Therefore, effectively preventing and controlling its harm has become the research direction of researchers. At present, although chemical pesticides still play an irreplaceable role in the control of plant virus diseases, they are always accompanied by the pressure of environmental and ecological safety.

[0003] β-glucan is a large class of natural polysaccharides composed of D-glucose monomers connected by β-glycosidic bonds. They naturally exist in the cell walls of various sources, such as yeast, bacteria, seaweed, grains, legumes and many medicinal mushrooms. We found in the experiment that β-glucan can effectively inhibit the infection of ToBRFV at different concentrations and can be used as an immune inducer to induce plant disease resistance. Summary of the Invention

[0004] To solve the above technical problems, the purpose of the present invention is to provide a method for inhibiting Tomato brown rugose fruit virus by β-glucan and its application.

[0005] The purpose of the present invention is achieved by the following technical solutions:

[0006] A kind of β-glucan, the configured concentration of the β-glucan is 25mg / L to 100mg / L.

[0007] Further, the configured concentration of the β-glucan is 50mg / L.

[0008] A method for inhibiting Tomato brown rugose fruit virus by β-glucan, comprising:

[0009] Spraying the β-glucan solution with a configured concentration of 25mg / L to 100mg / L onto Nicotiana benthamiana leaves;

[0010] 48 h after spraying the β-glucan solution, the lower leaves of Nicotiana benthamiana were inoculated with the Agrobacterium tumefaciens of the ToBRFV-GFP infectious clone;

[0011] 11 days after virus inoculation, the virus amount of ToBRFV decreased and the fluorescence intensity of GFP weakened.

[0012] Application of β-glucan in inhibiting Tomato brown rugose fruit virus ToBRFV.

[0013] Compared with the prior art, one or more embodiments of the present invention may have the following advantages:

[0014] β-glucan delays or reduces the infection of ToBRFV on tobacco by reducing the replication of ToBRFV in plants. Description of the Drawings

[0015] Figure 1 is the preventive effect diagram of different concentrations of β-glucan on the infection of ToBRFV in plants;

[0016] Figure 2 is the relative expression amount diagram of the ToBRFV cp gene at different concentrations;

[0017] Figure 3 is the symptom diagram of Nicotiana benthamiana infected with ToBRFV under different treatments 11 days after inoculating β-glucan. Detailed Embodiments

[0018] To make the objectives, technical solutions and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with embodiments and the accompanying drawings.

[0019] Example 1

[0020] A kind of β-glucan can effectively inhibit the infection of ToBRFV by pretreating Nicotiana benthamiana with different concentrations of β-glucan. The concentration of the β-glucan provided in this example is 25 mg / L to 100 mg / L. Respectively take β-glucan with the prepared concentrations of 25 mg / L, 50 mg / L and 100 mg / L.

[0021] Example 2

[0022] The β-glucan solutions with the prepared concentrations of 25 mg / L, 50 mg / L and 100 mg / L were sprayed on Nicotiana benthamiana with 3-4 leaves for treatment, and the β-glucan solution was used for foliar spraying until the liquid drops were evenly distributed on the leaves; 48 h after spraying the β-glucan solution, the lower leaves of Nicotiana benthamiana were inoculated with the Agrobacterium tumefaciens of the ToBRFV-GFP infectious clone, and dark treatment was carried out overnight. 11 days after virus inoculation, it was found that the fluorescence intensity of GFP weakened with the increase of β-glucan ( Figure 1)。The relative accumulation of ToBRFV in Nicotiana benthamiana plants pretreated with these three concentrations of β-glucan solutions at 25 mg / L, 50 mg / L, and 100 mg / L was lower than that in Nicotiana benthamiana directly inoculated with ToBRFV ( Figure 2 )。

[0023] The results showed that β-glucan delayed or reduced the infection of ToBRFV on tobacco by reducing the replication of ToBRFV in plants.

[0024] This example also found that the activity of plants induced to resist ToBRFV after pretreatment with β-glucan was higher than that of oligosaccharins. Pretreatment was carried out using β-glucan and oligosaccharins at the same concentration, specifically, solutions with a concentration of 25%-100% were diluted 300 times. After 48 h, the lower leaves of Nicotiana benthamiana were inoculated with the Agrobacterium tumefaciens of the ToBRFV-GFP infectious clone and treated in the dark overnight. The experiment found that the severity of the disease in Nicotiana benthamiana plants pretreated with β-glucan solution after inoculation with ToBRFV was lower than that in Nicotiana benthamiana plants pretreated with oligosaccharins and then inoculated. Figure 3 Figure 10 shows the symptoms of Nicotiana benthamiana infected with ToBRFV after different treatments 11 days after inoculation.

[0025] As found in the above examples, pretreatment of Nicotiana benthamiana with β-glucan at different concentrations such as 25%, 50%, and 100% could effectively inhibit the infection of ToBRFV, and it was found that its antiviral activity was better than that of the commercial pesticide oligosaccharins.

[0026] Field application method: Spray an appropriate concentration of β-glucan solution at 25 mg / L - 100 mg / L on each plant in the field.

[0027] Although the disclosed embodiments of the present invention are as above, the described content is only an embodiment adopted for easy understanding of the present invention and is not intended to limit the present invention. Any person skilled in the art within the technical field to which the present invention pertains can make any modifications and changes in the form of implementation and details without departing from the spirit and scope disclosed by the present invention. However, the scope of patent protection of the present invention shall still be subject to the scope defined by the appended claims.

Claims

1. A β-glucan, characterized in that, The configured concentration of the β-glucan is 25 mg / L to 100 mg / L.

2. The β-glucan according to claim 1, characterized in that, The configured concentration of the β-glucan is 50 mg / L.

3. A method for inhibiting Tomato brown rugose fruit virus by β-glucan, characterized in that spraying the β-glucan solution with a configured concentration of 25 mg / L to 100 mg / L onto Nicotiana benthamiana leaves; 48 hours after spraying the β-glucan solution, inoculating the lower leaves of Nicotiana benthamiana with the Agrobacterium of the ToBRFV-GFP infectious clone; 11 days after inoculating the virus, the virus amount of ToBRFV decreases and the fluorescence intensity of GFP weakens.

4. The method for inhibiting tomato brown rugose fruit virus by β-glucan according to claim 3, wherein The configured concentration of the β-glucan solution is 50 mg / L.

5. Use of the β-glucan according to claim 1 or claim 2 in inhibiting Tomato brown rugose fruit virus ToBRFV.

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