Serratia marcescens and separation method and application thereof

A technology of Serratia marcescens and separation method, applied in the field of microorganisms, can solve the problems of ineffective control of underground pests, etc., and achieve the effects of reducing pesticide residues, preventing damage, and achieving good results

Active Publication Date: 2016-08-17
NANYANG NORMAL UNIV
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But, above-mentioned patent Serratia marcescens is not obvi

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  • Serratia marcescens and separation method and application thereof
  • Serratia marcescens and separation method and application thereof
  • Serratia marcescens and separation method and application thereof

Examples

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Embodiment 1

[0025] Example 1 Isolation of Serratia marcescens and back-inoculation infection experiment

[0026] (1) The grubs collected from the field that died naturally were immersed in 70% ethanol for 10 seconds, and then immersed in 0.1% mercury chloride for 4 minutes to disinfect the surface of the insect body. Homogenize it under sterile conditions, add 5ml of sterile water, take 200μL, and apply Spread on NA medium plate, culture for 24 hours, pick a single colony and purify to obtain bacterial strain TC-1;

[0027] The composition of the NA medium is: 3.0 g of beef extract, 5.0 g of peptone, 3.0 g of yeast powder, 10.0 g of sucrose, 15 g of agar powder, 1000 mL of distilled water, pH 7.0, and autoclaved at 121° C. for 25 minutes;

[0028] (2) Inoculate strain TC-1 into NB culture medium, and culture it on a shaker at 28°C and 180 rpm for 2 days to obtain a bacterial suspension of strain TC-1, and adjust the concentration of the bacterial suspension to 10 9 cfu / ml, added to the s...

Embodiment 2

[0031] The identification of embodiment 2 bacterial strains

[0032] 1. Morphological analysis of strains: strain TC-1 was inoculated on NA medium and cultured at a constant temperature of 28°C for 48 hours, and the single colony shape, size and color were observed: strain TC-1 was short rod-shaped (see figure 1 ), no spores, fluorescent, Gram-negative, peri-flagellate, motile. After 48h on the NA medium, the colonies were round, smooth and moist, with neat edges and raised surfaces (see figure 2 ),rose Red;

[0033] 2. Physiological and biochemical assays of bacterial strains: Lactose, xylose, arabinose, raffinose, melibiose, sorbitol, Simon's citrate, bile aescin, malonate, Determination of 28 physiological and biochemical indicators such as phenylalanine desine, calendula alcohol, ornithine, lysine decarboxylase, lysine, arginine decarboxylase, methyl red, hydrogen sulfide, and urea.

[0034] Determination method: According to the instructions of Enterobacteriaceae bioc...

Embodiment 3

[0042] Embodiment 3 is measured to grub toxicity

[0043] Pick a ring of strain TC-1 from the NA slant, inoculate it into 50mL NB liquid medium, culture it on a shaker (180rpm) at 28°C for 12h to activate the strain, take 1ml from the activated cell suspension and inoculate it into 250ml In NB liquid medium, after 48 hours of culture in a shaker (180rpm) at 28°C, dilute to 1×10 9 cfu / ml concentration for later use.

[0044] Adopt soil-mixing inoculation method: according to the amount of soil required for the number of test worms and the estimated amount of worms and capacity, the soil is weighed, and the strain TC-1 thallus suspension (1 × 10 9 cfu / ml), diluted to a certain concentration and added to the soil, mixed evenly, so that the soil bacteria content reached 1×10 9 cfu / g, 1 / 2×10 9 cfu / g, 1 / 4×10 9 cfu / g, 1 / 8×10 9 cfu / g, and 1 / 16×10 9 cfu / g, the soil moisture content is about 17%. Put 1 grub into the rearing box, add potato pieces as feed, and fill with fungus soi...

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Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to serratia marcescens and a separation method and application thereof. The serratia marcescens is preserved in the China center for type culture collection, the preservation number is CCTCC M 2015634, and the preservation date is October 22th, 2015. Naturally dead grubs in fields are subjected to worm surface disinfection and homogenized under aseptic conditions, aquae sterilisata is added, the mixture is smeared to an NA medium plate for culture, a single colony is picked to be purified, and a serratia marcescens strain TC-1 is obtained. The serratia marcescens is used as a biological insecticide, a seed coating material and a fertilizer additive, and the problems of pesticide residues, target organism pesticide resistance, non-point source pollution and the like in agricultural production are reduced.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a Serratia marcescens and its isolation method and application. Background technique [0002] Microbial insecticides refer to preparations that use living microorganisms and their metabolites to kill insects, rodents, sterilize, weed, etc., mainly including fungal, bacterial, viral, nematode and antibiotic pesticides . Microbial-derived insecticides have the advantages of low toxicity, light pollution, and high selectivity of some drugs, and are in line with the development trend of pesticides from traditional organic chemicals to "environmentally harmonious pesticides" or "biologically rational pesticides". and users' favor. [0003] Serratia marcescens (Serratia marcescens) is an important insect pathogen, and as a pathogenic microorganism of pests, it is highly concerned by biological control workers. Serratia marcescens with insecticidal activity has been isol...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P7/04A01C1/06C12R1/425
CPCA01C1/06A01N63/00C12N1/20C12N1/205C12R2001/425
Inventor 陶爱丽王坦黄思良
Owner NANYANG NORMAL UNIV
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