Stenotrophomonas (sp.) and application thereof

A technique of oligotrophomonas and culture medium, applied in the field of oligotrophs, can solve the problems of limited degradation effect of plant residues of transgenic Bt plants, inability to degrade transgenic Bt plants, etc., and achieves the effect of good application prospect.

Active Publication Date: 2011-07-06
中农科合肥食品营养与健康创新研究院有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] 1. Insect consumption: the Bt insecticidal protein in the soil is degraded after entering the body of insects (including target insects and non-target insects). Bt protein;
[0005] 2. The effect of light: Bt insecticidal protein is degraded and ineffective under the irradiation of ultraviolet rays in the sun, but the effect on the degradation of plant residues of Bt-transformed plants is limited;

Method used

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  • Stenotrophomonas (sp.) and application thereof
  • Stenotrophomonas (sp.) and application thereof
  • Stenotrophomonas (sp.) and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1, the isolation and identification of bacteria

[0036] 1. Isolation of bacteria

[0037] In March 2010, in the ultra-clean workbench, the soil obtained from the rice of Fujian Academy of Agricultural Sciences was shaken in sterile distilled water for 15 minutes to prepare the bacterial suspension, and the shaking table speed was 180rpm; the bacterial suspension was washed with sterile water Spread it on the NA medium plate after doubling dilution, and cultivate it at 37°C. After about 24 hours, the colonies cover the entire plate. Use an inoculation loop to pick several different colonies on the plate and place them in the NA liquid medium. Shake culture for 20h. Then, the bacterial cells obtained by centrifuging the above-mentioned fermentation broth were applied to the experiment of degrading the crude Bt protein, thereby screening out the bacterial strain with the strongest degrading ability to the Bt protein, and performing Gram staining. The specific...

Embodiment 2

[0052] Embodiment 2, the cultivation of bacteria

[0053] Method I

[0054] Stenotrophomonas sp. FJSB3CGMCC №4344 was inoculated in liquid medium I, shaken and cultivated for 20 hours at a temperature of 37°C and a rotation speed of 200 rpm (radius of rotation: 20 mm) to obtain a bacterial liquid, counted by hemocytometer Plate count bacterial solution concentration. Under this culture condition, the concentration of bacteria in the bacteria liquid is 2.0×10 9 CFU / mL; all the substances in the culture container are recorded as bacterial liquid.

[0055] 1 liter of liquid medium I consists of peptone, beef extract, NaCl and water; the concentration of peptone in the medium is 1%, the concentration of beef extract in the medium is 0.3%, and the concentration of NaCl in the medium The concentration in the medium is 0.5%, and the percentages are all mass percentages; the pH value of the culture medium is 7.4.

[0056] Method II

[0057] Stenotrophomonas sp. FJSB3CGMCC №4344 w...

Embodiment 3

[0062] Embodiment 3, bacteria are to the degradation of crude anti-insect (Bt) protein

[0063] Bacillus thuringiensis subsp. mellonella was purchased from China Agricultural Microorganism Culture Collection Center (ACCC for short), and its number is ACCC 10029.

[0064] The preparation method of the crude anti-insect protein supernatant: Bacillus thuringiensis was inoculated into NB liquid medium, and cultured in liquid state at 37° C. and 200 rpm on a shaker for 48 hours. Centrifuge 500mL of Bacillus thuringiensis liquid at 5000rpm for 5min, take the precipitate, add 250mL of lysate to it, and dissolve the spore crystal mixture in a 4°C refrigerator for 4h; then centrifuge at 5000rpm for 5min, and adjust the pH of the supernatant to anti- The isoelectric point of the insecticidal protein is 4.5, and the anti-insect protein is deposited, and the crystal protein is deposited in a refrigerator at 4°C overnight. After settling, centrifuge at 4°C and 11,000 rpm for 10 minutes to...

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Abstract

The invention discloses stenotrophomonas (sp.) and application thereof. The stenotrophomonas (sp.) provided by the invention is stenotrophomonas (sp.) FJSB3 with the collection number of CGMCC No.4344. As an experiment proves, the stenotrophomonas (sp.) FJSB3 with the collection number of CGMCC No.4344 can degrade pure Bt proteins and the Bt proteins in trans-Bt rice straw. The stenotrophomonas (sp.) strain has good application prospect in the fields of degradation of Bt proteins in soil and improvement on the soil environment.

Description

technical field [0001] The invention relates to a strain of Stenotrophomonas and its application. Background technique [0002] Bt (Bacillus thuringiensis) is a Gram-positive aerobic bacillus that can produce more than one insecticidal crystal protein during its spore formation. In 2009, transgenic rice obtained the safety certificate issued by the Ministry of Agriculture, which shows that my country has taken a key step towards the commercial cultivation of genetically modified rice. However, when the toxic proteins released by transgenic Bt crops enter the soil, they may enrich Bt proteins in the soil ecosystem, affecting soil specific biological populations, functional groups, soil biodiversity and soil ecological processes. Therefore, there is an urgent need for an efficient and safe degradation method to eliminate the potential harm of such toxic proteins to the ecological environment and humans. [0003] At present, there are mainly the following methods for Bt prote...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20B09C1/10
Inventor 刘阳赵辉欣邢福国
Owner 中农科合肥食品营养与健康创新研究院有限公司
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