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Serratia marcescens SfSm-1 with broad-spectrum toxicity and application thereof

A technology of Serratia marcescens and sfsm-1, which is applied in the field of agricultural microorganisms, can solve problems such as difficult prevention and control, achieve good biological control effects, change microbial populations, and be environmentally friendly.

Active Publication Date: 2022-03-15
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, not only need to reduce the use of chemical pesticides, but also need to take different control measures for prevention and control. In biological control, screening microbial strains that are pathogenic or have control effects on agricultural pests is a current research hotspot, as disclosed in Chinese patent CN103571778A A strain of Serratia marcescens PS-1 from the flea beetle has strong pathogenicity to the larvae and adults of the flea beetle and the larvae and adults of the beet armyworm, and can effectively Control the development of the experimental population of beet armyworm; and it is disclosed in the prior art that Serratia marcescens OPT01 can be used to control pest potato tuber moth, but this bacterial strain is only used for the control of potato tuber moth, and some existing microbial strains It only has a good control effect on one or two types of pests, but there are many kinds of agricultural pests. At present, there are still few strains with broad-spectrum insecticidal activity, especially for pests that already have resistance, it is difficult to control
Therefore, it is still necessary to adopt environmentally friendly green control methods to replace chemical pesticides to prevent environmental pollution. At present, there is still a lack of more microbial strains with broad-spectrum insecticidal activity or better control effects to strengthen the control of pests.

Method used

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  • Serratia marcescens SfSm-1 with broad-spectrum toxicity and application thereof
  • Serratia marcescens SfSm-1 with broad-spectrum toxicity and application thereof
  • Serratia marcescens SfSm-1 with broad-spectrum toxicity and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The isolation and cultivation of embodiment 1 bacterial strain

[0036] Under sterile conditions, put Spodoptera frugiperda infected with bacteria into a 1.5mL centrifuge tube, add 100μL PBS buffer solution and grind for 1min at intervals of 10s, after grinding, add 500μL LB liquid medium for cultivation, in a shaker at 30°C, 200rpm Cultivate for 2h. Take 100 μL of the culture solution after the above cultivation in LB solid medium, spread on the plate, and culture at 30° C. for 24 hours.

[0037] After the cultivation, pick the red single colony in the LB solid medium to streak culture in the new LB solid medium, and continue to pick the red single colony for passage until the strain is purified. The slant medium can be stored at 4°C for one month, and the prepared 20% glycerol bacteria can be stored for a long time at -80°C.

Embodiment 2

[0038] Identification and Phylogenetic Analysis of Example 2 Bacterial Strains

[0039] (1) Traditional biological identification:

[0040] The above-mentioned isolated strains were identified as Gram-negative bacteria. The bacteria were rod-shaped, the colonies were round, and the surface was shiny. They were milky white colonies at the beginning of the culture at 30°C, and the prodigiosin was produced by the bacteria in the later stage. However, it is red, and no or less prodigiosin is produced under higher temperature conditions, and the colony morphology is as follows figure 1 shown.

[0041] (2) Determination of physiological and biochemical characteristics:

[0042] The determination of API 20E physiological and biochemical indicators of the strains is shown in Table 1.

[0043] Determination of physiological and biochemical indicators of strains in table 1

[0044]

[0045] Note: "+" is positive reaction, "-" is negative reaction.

[0046] (3) Molecular biologic...

Embodiment 3

[0052] Example 3 Toxicity determination of Serratia marcescens SfSm-1 to different pests

[0053] Preparation of bacterial solution: Using the strain SfSm-1 isolated in the above example, the activated strain was transferred to 30°C at a ratio of 1:100, cultured at 140rpm for 20h and activated for 10h. Centrifuge at 8000g for 5min, discard the supernatant, wash the bacteria twice with PBS, and dilute to a certain multiple (1×10 8 cfu / ml) for use.

[0054] Select 2nd instar beet armyworm larvae, fall armyworm larvae, diamondback moth larvae, cabbage caterpillar larvae, cotton bollworm larvae, Asian corn borer larvae, and Spodoptera litura larvae that are relatively consistent in growth. Each type of worm was divided into 6 groups, each group had three replicates, and each replicate had 24 worms, and starvation was performed for 2 hours.

[0055] Mix the prepared SfSm-1 bacterial solution and feed at a ratio of 1:4 (1ml bacterial solution is mixed with 4g feed, and 80% water i...

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Abstract

The invention discloses a serratia marcescens SfSm-1 strain with broad-spectrum toxicity, the strain is preserved in Guangdong Microbial Culture Collection Center on September 9, 2021, and the preservation number is GDMCC No: 61923. Studies show that the SfSm-1 strain has a broad-spectrum insecticidal effect on agricultural pests and has a good control effect on beet armyworms, spodoptera frugiperda, plutella xylostella, cabbage caterpillars, cotton bollworms, ostrinia furnacalis, phyllotreta striolata, brown planthopper and prodenia litura, the insecticidal effect on the beet armyworms is best, and the death rate reaches up to 94% or above; after the SfSm-1 strain infects beet armyworms, intestinal tracts of pests can be obviously destroyed, so that the beet armyworms show food refusal, and the activity is reduced until the beet armyworms die. The strain can be used as an environment-friendly green medicament to replace a chemical pesticide, provides a new microbial strain and a prevention and control method strategy for prevention and control of agricultural pests, and has a very good biological prevention and control effect.

Description

technical field [0001] The invention belongs to the technical field of agricultural microorganisms. More specifically, it relates to a strain of Serratia marcescens SfSm-1 with broad-spectrum toxicity and its application. Background technique [0002] Serratia marcescens (Serratia marcescens), widely present in nature, also known as prodigal bacilli, Gram-negative bacteria, belongs to the genus Serratia in the family Enterobacteriaceae, is sensitive to temperature, and can secrete bacteria at 20-28°C Prodigiosin, a small red molecular substance, the ability to produce pigment gradually disappears when the temperature continues to rise, and almost no pigment is produced at 37°C, and pigment can be produced again when the temperature is restored. Serratia marcescens can be divided into pigment-producing strains and non-pigmenting strains, which can be transformed under certain conditions. At present, the pathogenicity of Serratia marcescens to insects is still unclear, but t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/20A01P7/04C12R1/43
CPCC12N1/20A01N63/20Y02A50/30
Inventor 许小霞金丰良吴洪鑫林博达
Owner SOUTH CHINA AGRI UNIV
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