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A kind of Pseudomonas putida and its bacterial agent and application

A technology of Pseudomonas putida and microbial inoculum, which is applied in the field of plant protection, can solve problems such as affecting the soil ecological environment, and achieve the effects of alleviating obstacles to continuous cropping and good prospects for development and application.

Active Publication Date: 2020-03-17
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, previous studies on the microbial degradation of self-toxic substances mainly focused on herbaceous crops such as strawberries and cucumbers, while apples were more studied on woody fruit trees, while the related research on peaches was rarely reported.
At the same time, the types of autotoxic substances in different plants and the content in the rhizosphere soil are different, so the degrading bacteria screened by other crops may not be suitable for the autotoxic substances of peach trees
On the other hand, since most of the bacteria in microbial fertilizers belong to exogenous microorganisms, they may affect the original ecological environment of the soil.

Method used

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  • A kind of Pseudomonas putida and its bacterial agent and application
  • A kind of Pseudomonas putida and its bacterial agent and application
  • A kind of Pseudomonas putida and its bacterial agent and application

Examples

Experimental program
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Effect test

Embodiment 1

[0032] The screening and identification of embodiment 1 benzoic acid degrading bacteria

[0033] 1. Isolation and purification of benzoic acid degrading bacteria

[0034] Weigh 10 g of soil and add it to 100 mL of sterile water, place it on a magnetic stirrer and stir for 30 min, then let it stand, and the supernatant liquid is the soil leachate. Take 5mL of soil extract and put it into 45mL of enrichment medium, culture in a 150mL Erlenmeyer flask at 30°C and 150r / min on a shaker for 3 days. Subsequently, in a 150mL Erlenmeyer flask, add 45mL of new enrichment medium (containing benzoic acid 100mg / L), and then add 5mL of the previous enrichment medium, place at 30°C, 150r / min shaker culture, 3d is an acclimatization cycle. After each cycle, 10% of the inoculum was added to the fresh enrichment medium, and the concentration of benzoic acid was gradually increased from 100mg / L, 200mg / L to 500mg / L. Then replace the enrichment medium with MSM medium containing 100 mg / L benzoic ...

Embodiment 2

[0044] Degradation characteristics of embodiment 2 Pseudomonas putida WH-B3 (Pseudomonas putida WH-B3)

[0045] a. Degradation test in liquid culture: inoculate bacteria into 50mL MSM medium containing 500mg / L benzoic acid, culture on a shaker at 30°C and 150r / min, and take the treatment without adding bacteria as a control. Measure the OD value of the bacterial liquid with a spectrophotometer at 600nm wavelength every 4h, and draw the growth curve, the results are as follows: figure 2 As shown in A. Simultaneously, every 4h, adopt high performance liquid chromatography to measure the concentration of benzoic acid, draw the degradation curve of benzoic acid, the result is as follows figure 2 Shown in B. Each time, 5 mL of the bacterial liquid was centrifuged at 2000 rpm for 10 min, and the supernatant was extracted twice with 10 mL of ether, then evaporated to dryness with a vacuum concentrator, and then 5 mL of methanol was added to dissolve the solid matter for the deter...

Embodiment 3

[0047] Example 3 Toxicity analysis of Pseudomonas putida WH-B3 to benzoic acid degradation products

[0048] Lettuce, an internationally accepted allelopathic model plant, was used as the bioassay receptor for the experiment. Soak the three filter papers with 0.2% oxalic acid in advance, then rinse the air-dried filter papers with distilled water and put them into a 9cm-diameter, high-temperature sterilized petri dish for use. Cultivate the degrading bacteria with MSM containing 500 mg / L benzoic acid for 12 hours (the degradation of benzoic acid is basically completed), take 10 mL of the bacterial liquid and centrifuge at 2000 rpm for 10 min, extract the supernatant with 20 mL of ether twice, and then concentrate it with a vacuum concentrator To dryness, add 5 mL of methanol to completely dissolve the solid residue, add it to a petri dish with filter paper, and add 10 mL of sterilized distilled water after the methanol is completely volatilized. The lettuce seeds were disinfect...

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Abstract

The invention relates to a Pseudomonas putida WH‑B3 (Pseudomonas putida WH‑B3), which is preserved in the China Center for Type Culture Collection (CCTCC), and the preservation number is CCTCC M 2019048. The invention provides a preparation method of the microbial bacterium containing the above-mentioned Pseudomonas putida and a microbial bacterium agent, and provides an application method of the above-mentioned microbial bacterium agent in degrading benzoic acid and alleviating peach continuous cropping obstacles. Pseudomonas putida WH-B3 of the present invention has extremely strong degradability to benzoic acid, to 500mg kg in soil in 24 hours ‑1 The degradation efficiency of benzoic acid can reach more than 99%, and the degradation product is non-toxic, and has good development and application prospects.

Description

technical field [0001] The invention relates to the field of plant protection, in particular to a pseudomonas putida and its bacterial agent and its application in degrading benzoic acid and alleviating peach continuous cropping obstacles. Background technique [0002] Peach is one of the four major fruits in my country. It has a wide range of cultivation areas in various provinces and autonomous regions of my country, and is also planted all over the world. Peach fruit is beautiful in appearance, delicious in taste and rich in nutritional value, and is very popular among consumers. In recent years, my country's peach production and consumption have ranked first in the world. However, with the increase in demand for peaches, some orchards have common obstacles to continuous cropping due to land shortages, lack of management systems, and improper soil restoration techniques, which have become constraints. An important factor for the healthy and sustainable development of the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/27C09K17/14A01P21/00C12R1/40
CPCA01N63/00C09K17/14C12N1/20C12N1/205C12R2001/40
Inventor 刘军伟李国怀何昊叶俊丽朱炜
Owner HUAZHONG AGRI UNIV
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