CN115537369A - Application of 2-dodecenedioic acid in promoting growth and colonization of pseudomonas monteilii - Google Patents

Application of 2-dodecenedioic acid in promoting growth and colonization of pseudomonas monteilii Download PDF

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CN115537369A
CN115537369A CN202211397712.2A CN202211397712A CN115537369A CN 115537369 A CN115537369 A CN 115537369A CN 202211397712 A CN202211397712 A CN 202211397712A CN 115537369 A CN115537369 A CN 115537369A
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pseudomonas
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pseudomonas monteilii
dodecenedioic acid
monteilii
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CN115537369B (en
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杨敏
朱书生
邓琳梅
罗丽芬
李玥
叶辰
黄惠川
刘屹湘
梅馨月
杜飞
邓维萍
何霞红
朱有勇
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Yunnan Agricultural University
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Abstract

The invention provides application of 2-dodecenedioic acid in promoting growth and colonization of pseudomonas monteilii. According to the invention, 2-dodecenedioic acid is used as an exogenous additive of Pseudomonas montmorii PM-41, the effect of the 2-dodecenedioic acid in the growth and colonization process of Pseudomonas montmorii PM-41 is evaluated, and the Pseudomonas montmorii PM-41 has an obvious chemotactic effect on the 2-dodecenedioic acid; and 2-dodecenedioic acid can obviously promote the growth of the Pseudomonas montmorii PM-41 and the formation of a biological membrane, and promote the colonization of the Pseudomonas montmorii PM-41 in the rhizosphere of the pseudo-ginseng.

Description

Application of 2-dodecenedioic acid in promoting growth and colonization of pseudomonas monteilii
Technical Field
The invention relates to the field of agriculture, in particular to application of panax notoginseng root secretion 2-dodecenedioic acid in promoting growth and colonization of panax notoginseng rhizosphere beneficial pseudomonas monteilii.
Background
Yunnan is the main producing area of the traditional Chinese medicine, the pseudo-ginseng likes a warm and humid growing environment, continuous cropping obstacles are key factors limiting the healthy development of the pseudo-ginseng industry, and the yield and the quality of the pseudo-ginseng are seriously influenced. The current main means for relieving the continuous cropping obstacle of the panax notoginseng comprise soil fumigation, crop rotation, biological control and the like. Soil fumigation usually uses some high-toxicity chemical fumigants, and the harm to the environment limits the application of the fumigants in actual production; the crop rotation period is long, and the crop suitable for relieving the continuous cropping obstacle of the panax notoginseng is not applied to production at present. The beneficial microorganisms are utilized to relieve the continuous cropping obstacle of the panax notoginseng, so that the method has the advantages of low toxicity, no pollution, environmental friendliness and the like, is an effective way for green ecological prevention and control of the panax notoginseng, but the effective colonization of the microorganisms at the rhizosphere of the panax notoginseng limits the application of the microorganisms in production.
Pseudomonas monteilii PM-41 is isolated from rhizosphere soil of panax notoginseng in the earlier stage by the applicant, can degrade autotoxin saponin secreted into the rhizosphere soil in the growth process of panax notoginseng, can obviously antagonize beneficial microorganisms of rhizopus bacteria, and can obviously relieve continuous cropping obstacles of panax notoginseng (CN 201910935485.6). However, how to enhance the colonization of the PM-41 of the Pseudomonas menhadenii (Pseudomonas monteilii) on the rhizosphere of the panax notoginseng is still one of the problems of whether the strain can be applied to the control of the panax notoginseng root rot.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides application of panax notoginseng root secretion 2-dodecenedioic acid in promoting growth and colonization of panax notoginseng rhizosphere beneficial pseudomonas monteilii.
2-dodecenedioic acid is a monounsaturated dicarboxylic acid isolated from kidney beans. White to beige crystalline powder, stable at normal temperature and pressure, soluble in water, ethanol, methanol, petroleum ether, and chloroform. Can cause the reduction of membrane phospholipid peroxidation and is a potential drug agent for treating a plurality of skin diseases related to collagen biosynthesis disorder and oxidative stress. 2-dodecenedioic acid can also be found from root exudates of Panax notoginseng.
The invention finds the application potential of 2-dodecenoic acid in root secretion of panax notoginseng in promoting colonization and growth of beneficial microorganisms of Pseudomonas monteilii (Pseudomonas monteilii) PM-41, and has the effects of promoting growth of Pseudomonas monteilii (Pseudomonas monteilii) PM-41, promoting formation of a biofilm of Pseudomonas monteilii (Pseudomonas monteilii) PM-41, and promoting colonization of Pseudomonas monteilii (Pseudomonas monteilii) PM-41 in rhizosphere of panax notoginseng.
In a first aspect, the present invention provides the use of 2-dodecenedioic acid to promote the growth and colonization of Pseudomonas monteilii (Pseudomonas monteilii).
In a preferred embodiment, the use comprises the treatment of Pseudomonas monteilii (Pseudomonas monteilii) with 2-dodecenedioic acid.
In a preferred embodiment, the 2-dodecenedioic acid treating Pseudomonas monteilii (Pseudomonas monteilii) is administered in the form of a solution at a concentration of 1 to 2000. Mu.g/mL.
Preferably, the concentration is 1. Mu.g/mL, 5. Mu.g/mL, 10. Mu.g/mL, 20. Mu.g/mL, 30. Mu.g/mL, 40. Mu.g/mL, 50. Mu.g/mL, 100. Mu.g/mL, 200. Mu.g/mL, 300. Mu.g/mL, 500. Mu.g/mL, 1000. Mu.g/mL, 2000. Mu.g/mL. The concentration is preferably 1 to 1000. Mu.g/mL, more preferably 1 to 100. Mu.g/mL, and still more preferably 5 to 50. Mu.g/mL.
In a preferred embodiment, the treatment comprises contacting Pseudomonas monteilii (Pseudomonas monteilii) with 2-dodecenedioic acid for a period of time. Preferably, the contacting comprises incubating Pseudomonas montelulis (Pseudomonas montelulii) with 2-dodecenedioic acid, or adding 2-dodecenedioic acid together with Pseudomonas montelulii to the environment to be colonized, such as soil, particularly Panax notoginseng rhizosphere soil. Preferably, the time is greater than 12h, more preferably greater than 24h.
In a preferred embodiment, the growth includes strain propagation and biofilm formation.
In a preferred embodiment, said colonization comprises colonization in soil, in particular in the rhizosphere soil of notoginseng.
Therefore, preferably, the application comprises adding Pseudomonas mendocina (Pseudomonas monteilii) and 2-dodecenedioic acid to the panax notoginseng rhizosphere soil together, thereby promoting colonization and growth of Pseudomonas mendocina (Pseudomonas monteilii) in the panax notoginseng rhizosphere soil.
In a second aspect, the present invention provides a method of promoting growth and colonization by Pseudomonas mendii (Pseudomonas monteilii), the method comprising treating Pseudomonas mendii (Pseudomonas monteilii) with 2-dodecenoic acid.
In a preferred embodiment, the 2-dodecenedioic acid treating Pseudomonas monteilii (Pseudomonas monteilii) is administered in the form of a solution at a concentration of 1 to 2000. Mu.g/mL.
Preferably, the concentration is 1. Mu.g/mL, 10. Mu.g/mL, 20. Mu.g/mL, 30. Mu.g/mL, 40. Mu.g/mL, 50. Mu.g/mL, 100. Mu.g/mL, 200. Mu.g/mL, 300. Mu.g/mL, 500. Mu.g/mL, 1000. Mu.g/mL, 2000. Mu.g/mL. The concentration is preferably 1 to 1000. Mu.g/mL, more preferably 1 to 10. Mu.g/mL.
In a preferred embodiment, the growth includes strain propagation and biofilm formation.
In a preferred embodiment, said colonization comprises colonization in soil, in particular in the rhizosphere soil of panax notoginseng.
Therefore, preferably, the method comprises adding Pseudomonas mendocina (Pseudomonas monteilii) and 2-dodecenedioic acid together to the notoginseng rhizosphere soil, thereby promoting the growth and colonization of Pseudomonas mendocina (Pseudomonas monteilii) in the notoginseng rhizosphere soil.
In the invention, 2-dodecenedioic acid can be prepared into solution formulations such as aqueous solution, aqueous solvent, drops, missible oil, microemulsion, aqueous emulsion, microcapsule, aerosol, spray and the like with auxiliary materials when being applied. Wherein the concentration of 2-dodecenedioic acid is 1 to 2000. Mu.g/mL, specifically 1. Mu.g/mL, 10. Mu.g/mL, 20. Mu.g/mL, 30. Mu.g/mL, 40. Mu.g/mL, 50. Mu.g/mL, 100. Mu.g/mL, 200. Mu.g/mL, 300. Mu.g/mL, 500. Mu.g/mL, 1000. Mu.g/mL, 2000. Mu.g/mL. The concentration is preferably 1 to 1000. Mu.g/mL, more preferably 1 to 10. Mu.g/mL.
In a third aspect, the present invention provides a solution formulation comprising 2-dodecenedioic acid, wherein the concentration of 2-dodecenedioic acid is 1 to 2000. Mu.g/mL, specifically 1. Mu.g/mL, 10. Mu.g/mL, 20. Mu.g/mL, 30. Mu.g/mL, 40. Mu.g/mL, 50. Mu.g/mL, 100. Mu.g/mL, 200. Mu.g/mL, 300. Mu.g/mL, 500. Mu.g/mL, 1000. Mu.g/mL, 2000. Mu.g/mL. The concentration is preferably 1 to 1000. Mu.g/mL, more preferably 1 to 10. Mu.g/mL.
The preparation formulation comprises aqueous solution, aqueous solvent, drop, missible oil, microemulsion, aqueous emulsion, microcapsule, aerosol, spray and the like.
In a fourth aspect, the present invention provides the use of 2-dodecenedioic acid in the preparation of a formulation for promoting the growth and colonization of Pseudomonas montelukast (Pseudomonas monteilii).
In a preferred embodiment, the growth includes strain propagation and biofilm formation.
In a preferred embodiment, said colonization comprises colonization in soil, in particular in the rhizosphere soil of notoginseng.
In a preferred embodiment, the formulation is a solution formulation wherein the concentration of 2-dodecenedioic acid is 1 to 2000. Mu.g/mL, specifically 1. Mu.g/mL, 10. Mu.g/mL, 20. Mu.g/mL, 30. Mu.g/mL, 40. Mu.g/mL, 50. Mu.g/mL, 100. Mu.g/mL, 200. Mu.g/mL, 300. Mu.g/mL, 500. Mu.g/mL, 1000. Mu.g/mL, 2000. Mu.g/mL. The concentration is preferably 1 to 1000. Mu.g/mL, more preferably 1 to 10. Mu.g/mL.
The preparation formulation comprises aqueous solution, aqueous solvent, drop, missible oil, microemulsion, aqueous emulsion, microcapsule, aerosol, spray and the like.
In the invention, the Pseudomonas monteilii (Pseudomonas monteilii) comprises Pseudomonas monteilii (Pseudomonas monteilii) PM-41, and the preservation information is as follows: the preservation number is CGMCC No.18491; the classification is named as: pseudomonas monteilii; the preservation unit is China general microbiological culture Collection center; the preservation address is No. 3 of Xilu No.1 of Beijing, chaoyang, the area of the morning, the zip code: 100101; the preservation date is 2019, 9 and 16.
Has the advantages that:
the invention provides application of 2-dodecenedioic acid serving as a root secretion of panax notoginseng in promoting growth and colonization of beneficial Pseudomonas monteilii (Pseudomonas monteilii) of panax notoginseng rhizosphere. The invention evaluates the function of 2-dodecenedioic acid in the growth and colonization process of the PM-41 of the Pseudomonas monteilii (Pseudomonas monteilii) by using the 2-dodecenedioic acid as the exogenous additive of the PM-41 of the Pseudomonas monteilii, and is favorable for analyzing the function of the 2-dodecenedioic acid in the root secretion of the panax notoginseng in promoting the growth and colonization of the PM-41 of the useful microorganism Pseudomonas monteilii (Pseudomonas monteilii). In chemotaxis test, the PM-41 of the Pseudomonas mendii (Pseudomonas monteilii) has obvious chemotactic effect on 2-dodecenedioic acid with the concentration of 1-1000 mu g/mL, in growth promotion test, the PM-41 of the Pseudomonas mendii (Pseudomonas monteilii) can be obviously promoted at the concentration of 10-1000 mu g/mL, the formation of a biomembrane of the PM-41 of the Pseudomonas mendii (Pseudomonas monteilii) can be obviously promoted at the concentration of 1-1000 mu g/mL, and in colonization test, the PM-41 of the Pseudomonas mendii (Pseudomonas monteilii) can be obviously promoted at the rhizosphere of panax notoginseng at the concentration of 10 mu g/mL. Therefore, in the use of the beneficial microorganism, namely the Pseudomonas mentii PM-41, the 2-dodecenedioic acid with the concentration of 10 mu g/mL can promote the growth and the biofilm formation of the Pseudomonas mentii PM-41 and the colonization of the rhizosphere of the pseudo-ginseng.
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FIG. 1: determination of chemotactic Effect of P.mundtii (Pseudomonas monteilii) PM-41 on 2-dodecenedioic acid (different lower case letters indicate significant differences at P <0.05 level as determined by Duncan's New double-offset method);
FIG. 2: measuring the effect of 2-dodecenedioic acid on the growth of Pseudomonas monteilii PM-41;
FIG. 3: determination of the Effect of 2-dodecenedioic acid on the formation of biofilms by Pseudomonas monteilii PM-41 (different lower case letters indicate significant differences at P <0.05 levels as determined by Duncan's New Bipolar Difference method);
FIG. 4: the effect of 2-dodecenedioic acid on the colonization ability of pseudomonas monteilii PM-41 in soil was determined (indicating a significant difference at a P <0.05 level as compared to the control by t-test).
Detailed Description
The present invention is described in more detail below to facilitate an understanding of the present invention.
It should be understood that the terms or words used in the specification and claims should not be construed as having a meaning defined in dictionary, but should be construed as having a meaning consistent with their meaning in the context of the present invention on the basis of the following principles: the concept of terms may be defined appropriately by the inventors for the best explanation of the invention.
Example 1: chemotactic Effect of P.mundtii PM-41 on 2-dodecenedioic acid
Culturing the strain in 900 μ l LB liquid medium for 2 days to obtain bacterial concentration OD 590 Is up toTo 0.6, prepare a sterile yellow tip, 1ml syringe, LB plate medium. Preparing 2-dodecenedioic acid into solutions with different concentrations (1 mug/mL, 10 mug/mL, 50 mug/mL, 100 mug/mL and 1000 mug/mL) by using sterile water, transferring 100 mug of bacteria liquid into a yellow gun head by using a liquid transferring gun under the sterile condition of an ultra-clean workbench, sucking 100 mug of 2-dodecenedioic acid solution into the syringe, inserting a syringe needle into the yellow gun head, flatly placing the syringe needle on the workbench, standing for 40min, transferring the liquid in the syringe into a sterile clean 2mL tube, diluting by 10 times, 100 times and 1000 times by using the sterile water, then taking 100 mug for plating, culturing at 28 ℃ for 1 day, and observing and counting the colony number.
The test results in figure 1 show that the strain PM-41 has remarkable chemotactic effect on 2-dodecenedioic acid with the concentration of 1 mug/mL, 10 mug/mL, 50 mug/mL, 100 mug/mL and 1000 mug/mL. The strain PM-41 has obvious chemotactic effect on 2-dodecenedioic acid.
Example 2: 2-dodecenedioic acid promoting pseudomonas monteilii PM-41 growth and biofilm formation
100 mul of the bacterial liquid is taken to be put into 900 mul of LB liquid culture medium and cultured overnight at 28 ℃ to ensure that the bacterial concentration OD 590 The value reaches 0.4, 20 mul of bacterial liquid is taken to be added into 180 mul of 2-dodecenedioic acid 1/10LB solution with different concentrations (1 mu g/mL,10 mu g/mL,50 mu g/mL,100 mu g/mL,1000 mu g/mL) of a 96-well plate, standing and culturing are carried out for 52h at 28 ℃, and the OD of the strain is measured during the culture period 590 Value, drawing a growth curve; and after 52h of culture, the formation of the biofilm of the strain under different concentrations of metabolites is measured by a crystal violet method.
The test result of FIG. 2 shows that after 48 hours of culture, 2-dodecenedioic acid of 10-1000 mug/mL can obviously promote the growth of the strain. The test result in FIG. 3 shows that after 52h of culture, 1, 50, 100 and 1000. Mu.g/mL of 2-dodecenedioic acid can promote the formation of the strain biofilm, wherein the concentration of 1000. Mu.g/mL has a remarkable promoting effect.
Example 3: 2-dodecenedioic acid promotes colonization of pseudomonas monteilii PM-41 in rhizosphere soil
Pseudomonas monteilii PM-41 for screening chloramphenicol and rifampicin resistance markers, and culturing for 2 days to make its bacteria concentration OD 590 The value reached 0.6. Each one of which isThe tissue culture bottle is filled with pseudo-ginseng rhizosphere soil, 17mL of solution (5 mL of bacterial solution, 7mL of 2-dodecenedioic acid solution with the concentration of 10 mu g/mL and 5mL of water/antibiotics) is added, and the mixture is stirred uniformly and then covered with a cover for culture for 10 days. On the 5 th day during the culture period, 5g of soil was taken, 50mL of sterile water was added, and the mixture was shaken on a shaker for 30min, diluted 10-fold, 100-fold, and 1000-fold, and 100. Mu.L of the mixture was aspirated by a pipette and plated on a TSA medium to which antibiotics were added, and after overnight culture at 28 ℃, the colony formation number was recorded, and the colonization effect of P.montmordii PM-41 in soil was evaluated.
The test result of FIG. 4 shows that, compared with the blank control, the number of colonized colonies of the Pseudomonas mendii PM-41 in the soil after 3 days, 5 days and 10 days of inoculation is obviously increased under the condition of the existence of 2-dodecenedioic acid, which indicates that the synergistic addition of 10 mug/mL of 2-dodecenedioic acid is beneficial to promoting the colonization of the Pseudomonas mendii at the rhizosphere of pseudo-ginseng.
The above description is of the preferred embodiment of the present invention, but it is not intended to limit the present invention. Those skilled in the art may make modifications and variations to the embodiments disclosed herein without departing from the scope and spirit of the invention.

Claims (9)

1. Application of 2-dodecenedioic acid in promoting growth and colonization of Pseudomonas monteilii (Pseudomonas monteilii) is provided.
2. Use according to claim 1, characterized in that it comprises the treatment of Pseudomonas menhadiensis (Pseudomonas monteilii) with 2-dodecenoic acid.
3. A method of promoting growth and colonization of Pseudomonas mendii (Pseudomonas monteilii), the method comprising treating Pseudomonas mendii (Pseudomonas monteilii) with 2-dodecenoic acid.
4. Use, method according to claim 2 or 3, characterized in that the 2-dodecenedioic acid of Pseudomonas monteilii (Pseudomonas monteilii) is administered in the form of a solution at a concentration of 1 to 2000 μ g/mL at the time of treatment.
5. The use or method according to any one of claims 2 to 4, wherein Pseudomonas monteilii (Pseudomonas monteilii) and 2-dodecenedioic acid are added together to the rhizosphere soil of Panax notoginseng.
Use of 2-dodecenedioic acid for the preparation of a formulation for promoting the growth and colonization of Pseudomonas monteilii (Pseudomonas monteilii).
7. The use, method according to any one of claims 1-6, wherein said growth comprises strain propagation and biofilm formation; the colonization includes colonization in soil, particularly in the rhizosphere soil of panax notoginseng.
8. A solution formulation comprising 2-dodecenedioic acid, wherein the concentration of the 2-dodecenedioic acid is 1 to 2000 μ g/mL.
9. The solution formulation of claim 8, wherein the formulation is in the form of aqueous solution, aqueous solvent, drops, emulsifiable concentrate, microemulsion, aqueous emulsion, microcapsule, aerosol, spray.
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CN116064364A (en) * 2022-11-09 2023-05-05 云南农业大学 12-oxo-plant dienoic acid promotes pseudomonas mongolica growth and colonization

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