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A kind of mycovirus slmv1, attenuated bacterial strain and application

A technology of fungal viruses and strains, applied in the direction of viruses/bacteriophages, viruses, applications, etc., can solve the problems of continuous emergence of resistance and reduced efficacy of pharmaceuticals

Active Publication Date: 2022-05-13
HUNAN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the prevention and control of such diseases in production still mainly rely on chemical agents, but the resistance of pathogenic bacteria to commonly used fungicides continues to emerge, resulting in a significant decrease in the field control effect of most agents, and the problem of bacterial resistance is becoming more and more serious

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  • A kind of mycovirus slmv1, attenuated bacterial strain and application
  • A kind of mycovirus slmv1, attenuated bacterial strain and application
  • A kind of mycovirus slmv1, attenuated bacterial strain and application

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preparation example Construction

[0052]

[0053] The mycovirus S1MV1 of the present invention is present in a predetermined Puccinia strain, and therefore, the mycovirus can be obtained by isolating and recovering the virus from a Puccinia strain infected with the mycovirus.

[0054] Known methods can be used as methods for isolating and recovering viruses. For example, the bacterial cells can be frozen and crushed with liquid nitrogen, suspended in a predetermined buffer, and the virus can be isolated by ultracentrifugation, etc., and the virus can be recovered.

[0055] However, the mycovirus of the present invention may also exist outside the cells of the host bacteria. Therefore, for example, a phytopathogenic fungus infected with a mycovirus can be cultured in a liquid medium or the like, and the bacteria can be isolated by centrifugation, and then the virus can be isolated and recovered from the culture supernatant, whereby the virus can be recovered easily and in a relatively large amount.

[0056] ...

Embodiment 1

[0070] The preparation of embodiment 1 bacterial strain S1HN-03 of the present invention

[0071] Pick lettuce leaves with leaf spot disease from Changsha Vegetable Base in Hunan Province, use conventional tissue separation method, take the tissue at the junction of diseased and healthy leaves for separation, soak in mercuric chloride for 1 min, 70% alcohol for 5-8 seconds, rinse with sterile water three times, and then dry , transferred to a PDA plate, and cultured upside down at 28°C; after the growth of mycelium, the strain was inoculated on V8 medium (V8 fruit juice 200g, CaCO33.0g, agar 20g, supplemented with deionized water to 1L), and placed in the light Cultivate under alternating dark conditions, and after 10 days, pick the slow-growing hyphae of the colonies and transfer them to a new PDA plate. Continue to cultivate under 28 ℃ of alternating light and dark conditions, observe under a microscope for 10 days, pick a single spore on a new PDA plate, and the newly grown b...

Embodiment 2

[0072] The detection of internal double-stranded RNA of embodiment 2 bacterial strains

[0073] Intrinsic double-stranded RNA was detected from 12 strains of P. tomato from S1HN-01 to S1HN-12 of strain S1HN-03 of the present invention. Firstly, the 12 strains of Puccinia spp. that were isolated and cultured were ground, and the nucleic acid was extracted by the phenol SDS method, and DNA and single-stranded nucleic acid were selectively decomposed by DNase1 and S1 nuclease to obtain double-stranded RNA in the bacteria. Then, electrophoresis was performed on a 1% agarose gel at 120 V for 0.5 hour, and staining was performed with ethidium bromide.

[0074] see results figure 2 , the results showed that among the 12 bacterial strains, except that the bacterial strain SlHN-03 contained 4 double-stranded RNA bands of 1.4-3.6kb, the remaining 11 bacterial strains were all normal or conventional Puccinia tomato (without double-stranded RNA), However, mycoviruses have never been re...

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Abstract

A low-virulence mycovirus SlMV1 exists in SlHN-03 of Stemphylium lycopersici, and has 4 double-stranded RNAs of 1.4-3.6kb, which can reduce the pathogenicity of phytopathogenic fungi by inhibiting the synthesis of toxins related to fungal pathogenicity. pathogenicity. The fungal virus can exist outside the cell and maintain the activity of infection, so it has the advantage of directly infecting the host bacteria from the outside of the cell without relying on hyphae fusion. Through in vitro infection, SlMV1 can spread across species to fungal pathogens such as Alternaria spp., reducing their pathogenicity. Therefore, the virus can be used for the prevention and treatment of plant diseases caused by fungi such as Alternaria and Puccinia.

Description

technical field [0001] The invention belongs to the field of biological control, and relates to an attenuated bacterial strain, in particular to the mycovirus S1MV1 contained in the attenuated bacterial strain, and to the plant diseases caused by the mycovirus or the attenuated bacterial strain to fungi such as Alternaria and Puccinia preventive effect. Background technique [0002] Among plant diseases, fungi are one of the most important pathogens. About 80% of plant diseases are caused by fungi, which cause huge economic losses to agricultural production every year. At present, chemical methods are mainly used for the prevention and control of fungal diseases. Since the long-term use of chemical agents will bring about a series of related problems such as ecological balance damage, environmental pollution, and pathogenic bacteria resistance, there is an urgent need to explore green and environmentally friendly biological control methods. [0003] Mycovirus (mycovirus or ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00C12N1/14A01N63/40A01P3/00C12R1/92C12R1/645
CPCC12N7/00A01N63/40C12N2720/00021C12R2001/645C12N1/145
Inventor 周倩刘洪
Owner HUNAN AGRICULTURAL UNIV
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