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Citrobacter sp. and application thereof to prevention and treatment of scarabaeidae

A technology of Citrobacter and Scarab, applied in the field of preventing and controlling scarabs, can solve the problem that the beneficial functions of Citrobacter need to be developed and the like, and achieve the effects of being beneficial to the treatment of resistance and broadening the use.

Active Publication Date: 2021-09-03
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] More beneficial functions of Citrobacter remain to be developed

Method used

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  • Citrobacter sp. and application thereof to prevention and treatment of scarabaeidae
  • Citrobacter sp. and application thereof to prevention and treatment of scarabaeidae
  • Citrobacter sp. and application thereof to prevention and treatment of scarabaeidae

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Isolation and taxonomic identification of strains

[0025] 1.1 Field peanut planting and grub inoculation

[0026] In the experimental field of Langfang Experimental Base, Hebei Province, 1 mu of land was selected to plant peanuts, and five points were selected to bury 304 stainless steel cages in the field before sowing to limit the spread of subsequent inoculated grubs to the surrounding area. When sowing, sow 3 peanut seeds in each hole and water every 3 days. Inoculate the third-instar diablo larvae that have been starved for 48 hours at the peanut pod stage, 15 per cage, and take samples 7 days after the inoculation of diablo larvae, and collect peanut rhizosphere soil repeatedly at 3 points in each cage .

[0027] 1.2 Field rhizosphere soil collection and processing

[0028] The rhizosphere soil of peanuts was collected and processed as follows:

[0029] Take out the peanut root system completely, shake off the non-rhizosphere soil on the peanut root, leaving ...

Embodiment 2

[0034] Bioactivity Analysis

[0035] (1) Determination of biological activity of the larvae of the black beetle

[0036] Inoculate the single clone numbered IPPBiotC41 in a test tube containing 5 mL of liquid LB medium (tryptone 10.0 g / L, yeast extract 5.0 g / L, NaCl 10.0 g / L, sterilized at 121°C for 20 min), and inoculate at 30 Cultivate at ℃ for 12 hours for activation, then transfer the activated bacterial solution to solid LB medium (LB liquid medium plus agar 15g / L, culture dish diameter 15cm), cultivate at 30℃ for 12 hours, and use a sterile scraper Collect the cells with a shovel, suspend the cells evenly with PBS buffer, and count the suspended cells on plates. The counted cells were sonicated (Ampl 70%, pulsed for 3 s, paused for 5 s), so that the total protein was released from the cells, and the crushed cell liquid was obtained.

[0037] Prepare carrot shreds, rinse the carrot shreds with clear water, and let the carrot shreds dry until there is no moisture on the ...

Embodiment 3

[0064] Identification of the IPPBiotC41 strain

[0065] IPPBiotC41 genomic DNA was extracted with reference to the method described for extracting Gram-negative bacteria in molecular cloning. The 16S rDNA sequence of IPPBiotC41 was amplified with bacterial 16S rDNA universal primers: 27F (as shown in SEQ ID No. 1) and 1492R (as shown in SEQ ID No. 2), and a multi-fragment of about 1.4 kb was obtained. Purified by the kit for TA cloning, and the clone obtained by transforming Escherichia coli was sent to Sangon Bioengineering (Shanghai) Co., Ltd. for sequencing after being verified by bacterial liquid PCR. The obtained sequence was 1404bp (as shown in SEQ ID No.3) . The measured sequences were submitted to the EzBioCloud website for homology comparison, and a phylogenetic tree was constructed using MEGA6.0 ( figure 1 ). according to figure 1 It can be seen from the phylogenetic tree that it is closely related to the strain of Citrobacter brackeni of the genus Citrobacter. T...

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Abstract

The invention relates to citrobacter sp. and an application thereof to prevention and treatment of scarabaeidae. The citrobacter sp. is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.22672.

Description

technical field [0001] The invention relates to a citric acid bacterium, in particular it can be used for controlling scarabs. Background technique [0002] Citrobacter, also known as Citrobacter, was first proposed in 1932 and belongs to the family Enterobacteriaceae. Citrobacter is about 1.0 μm in diameter and 2.0-6.0 μm in length. It is a Gram-negative, rod-shaped, flagella-free facultative species that uses citrate as its carbon source to ferment glucose and carbohydrates and produce acid and gas. anaerobic bacteria. With the development of DNA hybridization technology, the genus Citrobacter has grown from the original 3 species to the current 11 species. It is known that Citrobacter grows best at 37°C and is commonly found in water, soil, esophagus and intestines of animals and humans. Although some strains can cause urinary tract infections, sepsis and infantile meningitis, they are not considered important pathogens of human diseases, but are considered opportunisti...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/20A01P7/04C12R1/01
CPCC12N1/20A01N63/20
Inventor 耿丽丽束长龙张杰徐文玥米良
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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