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Method for screening multifunctional bacteria for degrading organophosphorus pesticides

A technology of organophosphorus pesticides and screening methods, which is applied in the methods of using microorganisms, methods based on microorganisms, bacteria, etc., can solve problems such as no strain screening methods involved, and achieve the effects of easy survival and reproduction, simple operation, and cost reduction.

Inactive Publication Date: 2013-06-12
SHANDONG INST OF POMOLOGY
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Problems solved by technology

However, so far, the growth-promoting bacteria in the rhizosphere of plants (Yang Rong et al., 2012; Guo Changhong et al., 2012) and the microorganisms used in the bioremediation of organophosphorus pesticide-contaminated soils (Peng Xiawei et al., 2011; Jiang Jian et al., 2012) have been screened separately. Even if a certain strain is found to have the functions of nutrition, disease suppression and degradation of organophosphorus pesticides (Wu Haoqiong et al., 2012), there is no specific screening method for strains with these functions at the same time

Method used

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  • Method for screening multifunctional bacteria for degrading organophosphorus pesticides
  • Method for screening multifunctional bacteria for degrading organophosphorus pesticides
  • Method for screening multifunctional bacteria for degrading organophosphorus pesticides

Examples

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preparation example Construction

[0069] Preparation of medium:

[0070] 1) Preparation of LB liquid medium (pH7.0): mix 10g of tryptone, 5g of yeast extract, 10g of sodium chloride and double distilled water, and adjust the volume to 1000mL with double distilled water, adjust the pH with sodium hydroxide When the value reaches 7.0, sterilize at 121°C for 20 minutes after aliquoting.

[0071] 2) Preparation of LB solid medium (pH7.0): mix 10g tryptone, 5g yeast extract, 10g sodium chloride, 17-20g agar powder and double distilled water, and dilute to 1000mL with double distilled water, Adjust the pH value to 7.0 with sodium hydroxide, and sterilize at 121°C for 20 minutes after aliquoting.

[0072] 3) Preparation of improved MM medium: 40mmol / L 3-(N-morpholino) sodium propanesulfonate (adjust pH 7.4 with KOH), 2mmo / L K 3 PO 4 (pH7.0), glucose 2% (W / V), (NH 4 ) 2 SO 4 2.0g / L, MgSO 4 ·7H 2 O0.2g / L, sodium citrate 2H 2 O1.0g / L, potassium glutamate 1.0g / L, tryptophan 8.0mg / L, 3.0nmol / L ammonium molybdate, 4...

Embodiment 1

[0082] Example 1: Degradation of glyphosate by siderophilic bacteria Bacillus subtilis Bs-15

[0083] 1. Disease prevention and growth promotion of siderophilic bacteria B. subtilis Bs-15

[0084] B. subtilis Bs-15 (originally numbered as CAS15) is screened from the rhizosphere soil of rubber trees, and has strong siderophilic ability. Plant pathogens such as base rot, mango anthracnose, and Pythium melon fruit have a strong inhibitory effect [Yu Xianmei, Zheng Fucong, Lin Chao, et al. Isolation and identification of soil siderophilic antagonistic bacterium CAS15. Journal of Plant Protection, 2009, 36(2): 129-135].

[0085] The pot experiment was used to study the biocontrol effect of siderophilic bacteria B. subtilis Bs-15 on sweet pepper wilt and the promotion effect on sweet pepper growth. This part has been published [Yu Xianmei, Zhou Guangfang, Xin Li. The conditions for Bacillus subtilis Bs-15 to produce siderophore and its effect on disease prevention and growth pro...

Embodiment 2

[0179] Example 2: Isolation and identification of siderophilic bacteria CAS17 and degradation of chlorpyrifos

[0180] 1. Isolation and screening of strain CAS17

[0181] The rhizosphere soil of fruit trees such as apples, pears, and walnuts was collected, brought back to the laboratory, and sieved through a 20-mesh sieve after natural air-drying.

[0182] Take 1g of soil sample, add 50mL of sterilized distilled water, shake and culture at room temperature 150r / min for 2h, stand still to get supernatant, and separate and screen by CAS detection plate method and plate dilution method. According to the presence and size of the orange halo, the strains with strong siderophilic ability were screened out. CAS17 has a strong siderophilic ability and produces a larger orange halo ( Figure 13 ). Store at 4°C after purification.

[0183] 2. The domestication of strain CAS17 to chlorpyrifos

[0184] Inoculate strain CAS17 in LB liquid medium containing chlorpyrifos (concentration:...

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Abstract

The invention discloses a method for screening multifunctional bacterial for degrading organophosphorus pesticides. The method comprises the following steps of: (1) activating or screening plant growth-promoting rhizobacteria; (2) domesticating strains; (3) degrading the activity of the organophosphorus pesticides by using the strains; (4) optimizing the influence of environmental factors on the capacity of degrading the organophosphorus pesticides of the strains, and optimizing the condition; and (5) researching the degrading effect on the organophosphorus pesticides in soil of the strains through basin and earthen bowl tests to analyze the biological remediation potential of the strains to the soil contaminated by the organophosphorus pesticides. The bacterial obtained through the method can keep the stabilities of disease-preventing and growth-promoting functions and capacity of degrading the organophosphorus pesticides for a long time; due to the diversity of the functions, the bacterial can survive and propagate easily after being applied into soil, so that the functions of the bacterial can be given into full play; and besides, the method is simple to operate, low in cost and time-saving and labor-saving. Therefore, the method can have an extensive application prospect in the fields of screening the multifunctional bacteria, performing microbial remediation on the soil contaminated by the organophosphorus pesticides, and the like.

Description

Technical field: [0001] The invention relates to a screening method for multifunctional organophosphorus pesticide-degrading bacteria, which is mainly used for the domestication of obtained plant rhizosphere growth-promoting bacteria, and the multifunctional organophosphorus pesticide-degrading bacteria with the ability of plant disease prevention and growth promotion are obtained through domestication. Background technique: [0002] Pesticides have played a huge role in agricultural production. According to statistics from the Food and Agriculture Organization of the United Nations, if pesticides are not used, the annual losses caused by diseases, insect pests and weeds in the world account for about 36% of the total crop output, and the annual loss is as high as 125 billion U.S. dollars (Cooper and Dobson, 2007; Eddleston and Bateman, 2007). The use of pesticides ensures the stable and abundant agricultural production and human food supply. Organophosphorus pesticides (Or...

Claims

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Application Information

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IPC IPC(8): C12N1/36C12N1/20C12N1/14C12R1/125
Inventor 余贤美艾呈祥于婷董庆龙安淼王海荣刘嘉芬
Owner SHANDONG INST OF POMOLOGY
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