CN111411050B

CN111411050B - F01 actinomycete and antiviral application thereof

Info

Publication number: CN111411050B
Application number: CN201910029773.5A
Authority: CN
Inventors: 赵磊; 吴云锋
Original assignee: Northwest A&F University
Current assignee: Northwest A&F University
Priority date: 2019-01-04
Filing date: 2019-01-04
Publication date: 2023-03-17
Anticipated expiration: 2039-01-04
Also published as: CN111411050A

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

F01 actinomycete and antiviral application thereof

Technical Field

The invention relates to the technical field of microorganisms, in particular to the field of plant virus resistance of microorganisms, and further relates to the application of actinomycetes in resisting viruses.

Background

Plant viral diseases are an important group of plant diseases, and the economic loss caused by the damage of the plant viral diseases is up to $ 600 billion annually worldwide, wherein the loss of only one food crop is up to $ 200 billion. Plant viruses do not have complete cell structures, are absolutely parasitic to plants, and lack of complete immune systems of the plants, so that the plants are infected by the viruses for a lifetime and are continuously harmed, which brings great difficulty to the control of plant virus diseases. At present, no specific anti-plant virus medicament exists in agricultural production. In the face of serious harm of plant virus diseases and shortage of plant virus resistant medicaments, the development of novel plant virus resistant active ingredients and the research on antiviral mechanisms of the novel plant virus resistant active ingredients are of great significance. In recent years, research on active ingredients such as biologically-derived polysaccharides, glycopeptides, and proteins having high activity, low toxicity, good environmental compatibility, and few side effects has been favored by many researchers. A great deal of research shows that various biological polysaccharide, glycopeptide, oligosaccharide and the like have good immune regulation and control effects on plant defense against invasion of pathogenic bacteria. These findings provide a new concept for the development of anti-plant virus agents.

Screening of biological source plant virus resistant active materials, separation and identification of active ingredients and antiviral mechanism research of the active ingredients are developed in the early stage of the laboratory, and F01 streptomyces actinomycete fermentation liquor obtained from microbial source materials has high inhibitory activity on tobacco mosaic virus, barley yellow dwarf virus and the like. The F01 streptomyces actinomycete fermentation liquid can be obtained in large quantity by fermenting the F01 streptomyces actinomycete, and has low fermentation obtaining cost, simple extraction process and no limitation of external environment conditions. More importantly, the F01 streptomyces actinomycetes is a natural product of microbial source, has the characteristics of safety to human and livestock, no side effect on plants, no residue and difficult drug resistance generation, is an antiviral substance with low toxicity and high activity which is urgently needed in agricultural production, and has great potential for industrial application.

Disclosure of Invention

The invention discloses application of Streptomyces actinomycetes F01 in resisting plant viruses.

In addition, the streptomyces actinomycete F01 has a good inhibition effect on various plant viruses, and the invention also provides an application of the streptomyces actinomycete F01 in preparing a medicine with an antiviral effect.

The streptomyces actinomycete F01 is used for preventing and treating Tobacco Mosaic Virus (TMV), wheat yellow dwarf virus (BYDV) or rice stripe disease virus (RSV), and has better effect than the existing antiviral agents, such as those taking ningnanmycin and the like as main components, and the prevention and treatment effect of the original fermentation liquor on the tobacco mosaic virus is at least 69%.

The streptomyces actinomycete F01 has the characteristics of safety to human and livestock, no side effect on plants, no residue and difficult drug resistance generation, is an antiviral substance with low toxicity and high activity urgently needed in agricultural production, and has the potential of great industrial application.

Drawings

FIG. 1: the therapeutic effect of the fermentation liquid of Streptomyces actinomycete F01 on TMV is measured by a hemiphyllic method, after TMV virus is inoculated to tobacco leaves, the left leaves are treated by the fermentation liquid of Streptomyces actinomycete F01, and the right leaves are treated by water to be used as a control.

FIG. 2: and (3) measuring the content of the TMV after the fermentation liquor of the Streptomyces actinomycete F01 is treated by real-time fluorescent quantitative PCR.

Detailed Description

The present invention is further illustrated below by way of examples for the understanding of the present invention, but the following examples do not limit the present invention.

Example 1: indoor treatment effect of Streptomyces actinomycete F01 fermentation liquor on TMV virus.

(1) The experimental conditions are as follows: selecting indoor potted tobacco as a crop, wherein the variety is heart-leaf tobacco; the target is TMV.

(2) Experimental design and arrangement: the test agent is Streptomyces actinomycete F01 fermentation liquor, and 8% ningnanmycin aqueous solution contrast agent and clear water contrast are set. 3 treatments, repeated 3 times.

(3) The application method comprises the following steps: the leaf tobacco grows to 5-6 leaf stage and is used for the test. Adopting a half-leaf withered spot method, firstly inoculating TMV virus on the whole leaf, and after inoculating for 2 hours, smearing Streptomyces actinomycete F01 fermentation liquor or a contrast medicament on the left half leaf of the leaf, and smearing clear water on the right half leaf as a contrast.

(4) The investigation method comprises the following steps: 5 days after the application of the medicament, counting the number of the scorched spots generated by the left and right half leaves, and calculating the control effect according to the following formula:

control effect (%) = (right half She Kuban number-left half She Kuban number)/left half leaf dry spot number × 100

(5) The experimental results are as follows: the therapeutic effect of the fermentation broth of Streptomyces actinomycete F01 on TMV is shown in Table 1, and the indoor therapeutic effect of the original fermentation broth of Streptomyces actinomycete F01 on TMV is 82.5 percent as shown in Table 1.

TABLE 1 indoor control effect of Streptomyces actinomycetes F01 fermentation broth on tobacco mosaic virus disease

Example 2: the field control effect of the Streptomyces actinomycete F01 fermentation liquor on tobacco mosaic virus disease.

(1) The experimental conditions are as follows: the crop is selected from field tobacco, and the variety is K326; the target is TMV.

(2) Experimental design and arrangement: the test agent is Streptomyces actinomycete F01 fermentation liquor, and 8% ningnanmycin aqueous solution contrast agent and clear water contrast are set. 3 treatments, repeated 3 times for 9 cells. The test cells are arranged in random blocks.

(3) The application method comprises the following steps: spraying the pesticide 20 days after the tobacco is transplanted, and spraying the pesticide for the second time 7 days later.

(4) The investigation method comprises the following steps: after 14 days after the second application, a pharmacodynamic survey was conducted, and the disease incidence was recorded according to the following criteria:

level 0: the whole plant is disease-free; level 1: new leaves represent mild floral leaves; stage 2: one third to one half of the leaf represents a mosaic; and 3, level: one-half to two-thirds of the leaves represent flower leaves; 4, level: whole leaf of the whole plant.

Disease index = ∑ (number of diseased plants at each stage × relative stage value)/(total number of investigated plants × highest grade value) × 100%

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

(5) The experimental results are as follows: the control effect of the fermentation broth of Streptomyces actinomycete F01 on tobacco mosaic virus is shown in Table 2, and the control effect of the original fermentation broth of Streptomyces actinomycete F01 on tobacco mosaic virus is 69.8 percent as shown in Table 2.

TABLE 2 field control effect of Streptomyces actinomycetes F01 fermentation broth on tobacco mosaic virus

Example 3: the Streptomyces actinomycete F01 fermentation liquor has field control effect on wheat yellow dwarf virus.

(1) The experimental conditions are as follows: selecting field wheat as a crop, wherein the variety is Elytrigia tritici No. 6; the target is BYDV.

(3) The application method comprises the following steps: spraying the pesticide 20 days after the wheat is turned green, and spraying the pesticide for the second time 7 days later.

(4) The investigation method comprises the following steps: after 14 days from the second application, a pharmacodynamic survey was conducted, 10 wheat plants connected to each other were surveyed per spot, 250 wheat plants were surveyed per cell, and the disease onset was recorded according to the following criteria:

level 0: the whole plant is disease-free; level 1: partial leaf tip yellowing; and 2, stage: yellowing of 1 leaf below the flag leaf; and 3, level: yellowing of 2 leaves below the flag leaf; 4, level: yellowing of 1/4 of flag leaf, and yellowing of 1 leaf below flag leaf; and 5, stage: yellowing of 1/4 of flag leaf, and yellowing of 2 leaves below flag leaf; and 6, level: yellowing of flag leaf; and 7, stage: yellowing of flag leaf, yellowing of 1 leaf below flag leaf; and 8, stage: yellowing of flag leaf, yellowing of 2 leaves below flag leaf; and 9, stage: the plant is dwarfed but can sprout; 10 level: the plant is obviously dwarfed and the ear can not be spilt.

(5) The experimental results are as follows: the control effect of the fermentation liquid of the Streptomyces actinomycete F01 on the wheat yellow dwarf virus disease is shown in Table 3, and the control effect of the fermentation liquid of the Streptomyces actinomycete F01 on the wheat yellow dwarf virus disease is 72.6 percent as shown in Table 3.

TABLE 3 field prevention and cure effect of Streptomyces actinomycetes F01 fermentation liquid on wheat yellow dwarf virus disease

Example 4: the Streptomyces actinomycete F01 fermentation liquid has field control effect on rice stripe virus.

(1) The experimental conditions are as follows: selecting field rice as the crop, wherein the variety is Eryou peiju; the target is RSV.

(3) The application method comprises the following steps: spraying the pesticide 20 days after the rice is transplanted, and spraying the pesticide for the second time 7 days later.

(4) The investigation method comprises the following steps: after 14 days from the second application, a pharmacodynamic survey was conducted, 10 rice plants connected to each other were surveyed at each site, 250 rice plants were surveyed at each site, and the disease onset was recorded according to the following criteria:

level 0: the whole plant is disease-free; level 1: the base of the heart lobe shows a small amount of chlorosis macula along the veins, and the heart lobe does not curl; and 3, level: the new leaves have yellow-green alternate stripes parallel to the veins and are slightly curled; and 5, stage: the new leaves have a large number of yellow-green alternate stripes parallel to the veins, and the leaves are curled and thin; and 7, stage: the plant is dwarfed, yellow and white stripes appear on leaves and roll up, and the newborn She Niuqu droops and cannot be normally opened; and 9, stage: the plants were severely stunted, green lost or dead.

(5) The experimental results are as follows: the control effect of the fermentation broth of Streptomyces actinomycetes F01 on rice stripe virus is shown in Table 4, and the control effect of the fermentation broth of Streptomyces actinomycetes F01 on rice stripe virus is 62.3% in Table 4.

TABLE 4 field control effect of Streptomyces actinomycetes F01 fermentation broth on rice stripe virus

Claims

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.