CN111411050B - F01 actinomycete and antiviral application thereof - Google Patents

Abstract

The invention discloses a streptomyces actinomycete F01, wherein the Streptomyces actinomycete F01 is preserved in China general microbiological culture preservation management center in 2018, 12 and 24 months, and the preservation number is as follows: CGMCC No.17030. The invention also discloses application of the F01 streptomyces actinomycetes in resisting crop viruses. The F01 Streptomyces actinomycetes has a control effect on various plant virus diseases, such as tobacco mosaic virus disease, wheat yellow dwarf virus disease or rice stripe disease virus disease.

Description

A F01 actinomycete and its antiviral application

technical field

The invention relates to the technical field of microbes, in particular to the field of anti-plant viruses of microorganisms, and further relates to the use of actinomycetes for anti-viruses.

Background technique

Plant virus diseases are an important class of plant diseases. The global economic losses caused by plant virus diseases are as high as 60 billion U.S. dollars every year, of which the loss of food crops alone is as high as 20 billion U.S. dollars. Plant viruses do not have a complete cell structure, and they are absolutely parasitic to plants. In addition, plants themselves lack a complete immune system. Once a plant is infected by a virus, it will carry the virus for life and suffer continuous damage. Prevention and control posed great difficulties. At present, there is no specific anti-plant virus agent in agricultural production. Facing the serious damage of plant virus diseases and the shortage of anti-plant virus agents, it is of great significance to develop new anti-plant virus active ingredients and study their anti-virus mechanism. In recent years, the research on bio-derived polysaccharides, glycopeptides, proteins and other active ingredients with high activity, low toxicity, good environmental compatibility and few side effects has been favored by many scientific researchers. A large number of studies have shown that many biological polysaccharides, glycopeptides, and oligosaccharides have good immune regulation effects on plant defense against the invasion of pathogenic bacteria. These findings provide new ideas for the development of anti-plant virus agents.

In the early stage, our laboratory carried out the screening of bio-sourced anti-plant virus active materials, the isolation and identification of active ingredients and the research on its anti-virus mechanism. Yellow dwarf virus, etc. have high inhibitory activity. F01 Streptomyces actinomycetes fermented liquid can be obtained in large quantities through fermentation of F01 Streptomyces actinomycetes, the fermentation acquisition cost is low, the extraction process is simple and is not restricted by external environmental conditions. More importantly, F01 Streptomyces actinomycetes are natural products of microbial origin, which are safe for humans and animals, have no side effects on plants, have no residues, and are not easy to produce drug resistance. The antiviral substance has great potential for industrial application.

Contents of the invention

The invention discloses the use of a streptomyces actinomycete F01 for resisting plant viruses.

In addition, because the Streptomyces Actinomyces F01 of the present invention has a good inhibitory effect on various plant viruses, the present invention also provides a use of the Streptomyces Actinomyces F01 for preparing medicines with antiviral effects.

The above Streptomyces actinomycetes F01 are used for preventing and treating tobacco mosaic virus (tobacco mosaic virus, TMV), wheat yellow dwarf virus (barley yellow dwarf virus, BYDV) or rice stripe leaf blight virus (ricestipe virus, RSV) , compared with existing antiviral agents, such as antiviral agents with Ningnanmycin etc. as main components, it has a better effect, and the control effect of its original fermentation liquid on tobacco mosaic virus is at least 69%.

The above-mentioned Streptomyces actinomycetes F01 has the characteristics of being safe for humans and animals, having no side effects on plants, no residue, and not easy to produce drug resistance. It is an antiviral substance with low toxicity and high activity that is urgently needed in agricultural production and has great industrial applications. potential.

Description of drawings

Figure 1: Determination of the therapeutic effect of Streptomyces Actinomyces F01 fermentation broth on TMV by the half-leaf method. After tobacco leaves were inoculated with TMV virus, the left leaves were treated with Streptomyces Actinomyces F01 fermentation broth, and the right leaves were treated with water as control.

Figure 2: Real-time fluorescent quantitative PCR assay for the content of TMV virus after treatment of Streptomyces actinomyces F01 fermentation broth.

Detailed ways

In order to understand the present invention, the following examples further illustrate the present invention, but the following examples do not limit the present invention.

Embodiment 1: Indoor treatment effect of Streptomyces Actinomyces F01 fermented liquid on TMV virus.

(1) Experimental conditions: indoor potted tobacco was selected as the crop, and the variety was Nicotiana cordata; the target was TMV.

(2) Experimental design and arrangement: the experimental agent was Streptomyces Actinomyces F01 fermentation broth, and 8% Ningnanmycin aqueous solution control agent and clear water control were set up. 3 treatments, repeated 3 times.

(3) Application method: Tobacco heart leaf grows to the 5-6 leaf stage and is used for the test. Using the half-leaf dead spot method, the whole leaf was first inoculated with TMV virus. After 2 hours of inoculation, the left half of the leaf was smeared with Streptomyces Actinomyces F01 fermentation broth or a control agent, and the right half of the leaf was smeared with water as a control.

(4) Investigation method: 5 days after applying the agent, count the number of dead spots on the left and right half leaves, and calculate the control effect according to the following formula:

Control effect (%)=(number of dead spots on the right half leaf-number of dead spots on the left half leaf)/number of dead spots on the left half leaf×100

(5) Experimental results: the therapeutic effect of Streptomyces Actinomyces F01 fermented liquid on TMV is shown in Table 1, as seen from Table 1, the indoor therapeutic effect of Streptomyces Actinomyces F01 original fermented liquid on TMV is 82.5%.

Table 1 Indoor control effect of streptomyces actinomycetes F01 fermentation liquid on tobacco mosaic virus disease

Example 2: Field Control Effect of Streptomyces Actinomyces F01 Fermented Liquid on Tobacco Mosaic Virus Disease.

(1) Experimental conditions: field tobacco was selected as the crop, and the variety was K326; the target was TMV.

(2) Experimental design and arrangement: the experimental agent was Streptomyces Actinomyces F01 fermentation broth, and 8% Ningnanmycin aqueous solution control agent and clear water control were set up. 3 treatments, repeated 3 times, a total of 9 plots. The experimental plots were arranged in random blocks.

(3) Application method: Spray the pesticide 20 days after transplanting the tobacco, and spray the pesticide for the second time after every 7 days.

(4) Investigation method: Conduct drug efficacy investigation 14 days after the second application of the drug, and record the incidence according to the following standards:

Grade 0: The whole plant is disease-free; Grade 1: The new leaves show slight mosaic; Grade 2: One-third to one-half of the leaves show mosaic; Grade 3: One-half to two-thirds of the leaves show flowers Leaves; Grade 4: leaves and mosaic leaves of the whole plant.

Disease index = ∑ (number of diseased plants at all levels × relative level value) / (total number of investigated plants × highest grading value) × 100%

Control effect (%)=(control disease index-treatment disease index)/control disease index×100

(5) Experimental result: the control effect of Streptomyces Actinomyces F01 fermented liquid to tobacco mosaic virus disease is shown in Table 2, and by Table 2, the control of Streptomyces actinomyces F01 original fermented liquid to tobacco mosaic virus disease The effect is 69.8%.

Table 2 Field control effect of Streptomyces actinomyces F01 fermentation liquid on tobacco mosaic virus disease

Example 3: Field Control Effect of Streptomyces Actinomyces F01 Fermented Liquid on Wheat Yellow Dwarf Virus Disease.

(1) Experimental conditions: the crop is field wheat, the variety is Xiaoyan 6; the target is BYDV.

(2) Experimental design and arrangement: the experimental agent was Streptomyces Actinomyces F01 fermentation broth, and 8% Ningnanmycin aqueous solution control agent and clear water control were set up. 3 treatments, repeated 3 times, a total of 9 plots. The experimental plots were arranged in random blocks.

(3) Application method: Spray the pesticide 20 days after the wheat turns green, and spray the pesticide for the second time after every 7 days.

(4) Investigation method: Conduct drug efficacy investigation 14 days after the second spraying, investigate 10 connected wheat plants at each point, investigate 250 plants in each plot, and record the incidence according to the following standards:

Grade 0: No disease in the whole plant; Grade 1: Yellowing of some leaf tips; Grade 2: Yellowing of 1 leaf under the flag leaf; Grade 3: Yellowing of 2 leaves under the flag leaf; Grade 4: Yellowing of 1/2 flag leaf 4. Yellowing of 1 lower leaf of the flag leaf; Grade 5: yellowing of 1/4 of the flag leaf, yellowing of 2 lower leaves of the flag leaf; Grade 6: yellowing of the flag leaf; Grade 7: yellowing of the flag leaf, yellowing of the lower flag leaf 1 leaf yellowing; Grade 8: flag leaf yellowing, and 2 leaves below the flag leaf yellowing; Grade 9: plant dwarfing, but earing is possible; Grade 10: plant dwarfing significantly, earing not possible.

Disease index = ∑ (number of diseased plants at all levels × relative level value) / (total number of investigated plants × highest grading value) × 100%

Control effect (%)=(control disease index-treatment disease index)/control disease index×100

(5) Experimental result: the control effect of Streptomyces Actinomyces F01 fermented liquid to wheat yellow dwarf virus disease is shown in Table 3, as seen in Table 3, the control effect of Streptomyces actinomyces F01 fermented liquid to wheat yellow dwarf virus disease was 72.6%.

Table 3 Field control effect of Streptomyces actinomycetes F01 fermentation broth on wheat yellow dwarf virus disease

Example 4: Field Control Effect of Streptomyces Actinomyces F01 Fermented Liquid on Rice Stripe Blight Virus Disease.

(1) Experimental conditions: field rice was selected as the crop, and the variety was Liangyoupeijiu; the target was RSV.

(2) Experimental design and arrangement: the experimental agent was Streptomyces Actinomyces F01 fermentation broth, and 8% Ningnanmycin aqueous solution control agent and clear water control were set up. 3 treatments, repeated 3 times, a total of 9 plots. The experimental plots were arranged in random blocks.

(3) Application method: Spray application 20 days after rice transplanting, and spray for the second time after 7 days.

(4) Investigation method: Conduct drug efficacy investigation 14 days after the second spraying, investigate 10 connected rice plants at each point, investigate 250 plants in each plot, and record the disease according to the following standards:

Grade 0: The whole plant is disease-free; Grade 1: A small amount of chlorotic yellow spots appear at the base of the heart leaf along the veins without curling; Grade 3: The new leaves have yellow and green stripes parallel to the veins and are slightly curled; Grade 5: There are a lot of new leaves Yellow and green stripes parallel to the veins, the leaves are curled and thin; grade 7: the plant is dwarfed, the leaves appear yellow and white stripes rolled up, the new leaves are twisted and drooping, and cannot open normally; grade 9: the plant is severely dwarfed, chlorotic or dead .

Disease index = ∑ (number of diseased plants at all levels × relative level value) / (total number of investigated plants × highest grading value) × 100%

Control effect (%)=(control disease index-treatment disease index)/control disease index×100

(5) Experimental result: the control effect of Streptomyces Actinomyces F01 fermented liquid to rice stripe leaf blight virus disease is shown in Table 4, as seen from Table 4 the effect of Streptomyces actinomyces F01 fermented liquid on rice stripe leaf blight virus disease The control effect is 62.3%.

Table 4 Field control effect of Streptomyces actinomycetes F01 fermentation liquid on rice stripe leaf blight virus disease

Claims (2)

1. The streptomyces is strain F01, and is preserved in China general microbiological culture collection management center 24.12.2018 with the preservation number: CGMCC No.17030.

2. The application of the streptomyces F01 strain as claimed in claim 1, wherein the streptomyces F01 strain is used for preventing and treating crop virus diseases, and the crop virus diseases are tobacco mosaic disease, wheat yellow dwarf disease or rice stripe disease.

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