CN114982755A - Application of acetoin in prevention and treatment of tomato neck rot and root rot - Google Patents
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- A01N35/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical
- A01N35/02—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical containing aliphatically bound aldehyde or keto groups, or thio analogues thereof; Derivatives thereof, e.g. acetals
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Abstract
The invention discloses application of Acetoin (Acetoin) in preventing and treating tomato neck rot and root rot. The invention discovers for the first time that: acetoin (Acetoin) has obvious inhibition effect on tomato neck rot and root rot pathogenic bacteria, and can be used as an effective component for preventing and treating tomato neck rot and root rot. The invention provides a new medicinal active component for preventing and treating the tomato neck rot and root rot, and has important significance for preventing and treating the tomato neck rot and root rot.
Description
The application is a divisional application of Chinese patent application 'application of volatile substances in preventing and treating neck rot and root rot of tomatoes' (application date: 2022.01.10, application number: 202210019112.6).
Technical Field
The invention relates to the technical field of agricultural disease control, in particular to application of acetoin in controlling tomato neck rot and root rot.
Background
Tomatoes are a major vegetable crop in the world. The tomato is rich in antioxidant lycopene, vitamins and minerals. So the tomatoes have important economic value. With the acceleration of global world economy, the world tomato product trade scale is continuously enlarged in recent years. Tomatoes are affected by different pathogens, such as viruses, viroids, fungi, oomycetes, bacteria and nematodes, during their growth, which reduces tomato yield. Among them, fusarium oxysporum and root rot cause severe losses in tomato production. China is the world with the largest tomato planting area and the largest tomato yield.
Tomato neck rot root rot is a soil-borne disease caused by Fusarium oxysporum forth (Fusarium oxysporum f.sp.radialis-lycopersici). In recent years, the rapid spread of the tomato neck rot and root rot seriously affects the production of tomatoes. From seedling to adult, fusarium oxysporum and root rot can be infected. After the tomato is infected with the neck rot and root rot, the phenotypes such as vascular bundle necrosis, growth arrest, rapid wilting, withering and even death appear. When the disease is serious, the tomato neck rot and root rot can reduce the tomato yield by more than 65%. Therefore, tomato neck rot and root rot have become one of the main diseases restricting tomato production.
At present, the prevention and control of the tomato neck rot and root rot are mainly biological prevention and chemical prevention and control. Biological control is the control of pathogenic organisms by using some pathogenic antagonistic microorganisms in nature. Volatile Organic Compounds (VOCs) are one of the secondary metabolites produced by microorganisms, and are also a form of long-range action by microorganisms. They can penetrate the soil pores and regulate the growth and interaction of rhizosphere plants and microorganisms. Current research indicates that beneficial microorganisms can inhibit the growth of plant pathogens by producing Volatile Organic Compounds (VOCs). Therefore, the excavation of Volatile Organic Compounds (VOCs) has strong research significance for preventing and treating the tomato neck rot and root rot.
Disclosure of Invention
Aiming at the prior art, the invention aims to provide the application of acetoin in preventing and treating tomato neck rot and root rot. The invention discovers for the first time: acetoin (Acetoin) and 2-Heptanol (2-Heptanol) have obvious inhibition effect on tomato neck rot and root rot pathogenic bacteria, and can be used as effective components for preventing and treating tomato neck rot and root rot.
In order to realize the purpose, the invention adopts the following technical scheme:
in a first aspect of the present invention, there is provided the use of a volatile material in (1) or (2) as follows:
(1) inhibiting the growth of pathogenic bacteria of the neck rot and root rot of the tomato;
(2) preventing and controlling the neck rot and root rot of the tomato;
the volatile substance is any one of the following (a) to (c):
(a) acetoin (Acetoin);
(b) 2-Heptanol (2-Heptanol);
(c) a composition consisting of acetoin and 2-heptanol in a weight ratio of 6: 8.
In a second aspect of the present invention, there is provided a use of a volatile substance as described in any one of (a) to (c) below for the preparation of a pharmaceutical formulation for the control of tomato neck rot and root rot:
(a) acetoin (Acetoin);
(b) 2-Heptanol (2-Heptanol);
(c) a composition consisting of acetoin and 2-heptanol in a weight ratio of 6: 8.
In a third aspect of the invention, the pharmaceutical preparation for preventing and treating the tomato neck rot and root rot is provided, and the pharmaceutical preparation takes Acetoin (Acetoin), 2-Heptanol (2-Heptanol) or a composition of the Acetoin and the 2-Heptanol in a weight ratio of 6:8 as an active ingredient.
Preferably, the pharmaceutical preparation contains 6mg/mL acetoin and 0.6mg/mL 2-heptanol.
Furthermore, the pharmaceutical preparation can also comprise auxiliary materials or carriers; such adjuvants or carriers include, but are not limited to: emulsifier, dispersant, assistant, thickener, antifreeze, stabilizer, binder, etc.
Preferably, the dosage form of the pharmaceutical preparation is fumigant, powder, wettable powder, suspending agent or aqueous agent; more preferably, the pharmaceutical formulation is in the form of a fumigant.
In a fourth aspect of the invention, a method for preventing and treating tomato neck rot and root rot is provided, which comprises the following steps:
after the tomato plants are transplanted and planted or before the flowering period of the tomatoes, 6mg/mL acetoin and 0.6mg/mL 2-heptanol are applied in a root irrigation mode.
The invention has the beneficial effects that:
according to the invention, the volatile organic components of microorganisms separated from healthy tomato rhizosphere soil are analyzed, and tests show that Acetoin (Acetoin) and 2-Heptanol (2-Heptanol) in the volatile organic components have an inhibiting effect on tomato neck rot pathogenic bacteria for the first time, so that a new pharmaceutical active component is provided for preventing and treating tomato neck rot root rot.
Drawings
FIG. 1: the inhibiting effect of Acetoin with different concentrations on tomato neck rot root rot pathogenic bacteria.
FIG. 2: influence of volatile organic compounds Acetoin with different concentrations on hyphal morphology.
FIG. 3: the inhibiting effect of the 2-Heptanol with different concentrations on the tomato neck rot root rot pathogenic bacteria.
FIG. 4: influence of volatile organic compounds 2-Heptanol of different concentrations on hyphal morphology.
FIG. 5 is a schematic view of: the control effect of the plot experiment on the tomato neck rot and root rot; in the figure: "combination" means that acetoin and 2-heptanol are mixed and used, and the concentration of the acetoin in the mixed solution is 6mg/ml, and the concentration of the 2-heptanol is 0.6 mg/ml.
Detailed Description
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
As described in the background section, tomato neck rot is a soil-borne disease caused by Fusarium oxysporum forth (f.sp.radialis-lycopersici). The tomato neck rot and root rot can reduce the tomato yield by more than 65 percent, and become one of the main diseases restricting the tomato production. Therefore, the excavation of Volatile Organic Compounds (VOCs) has strong research significance for preventing and treating the tomato neck rot and root rot.
Based on the fact, the invention deeply researches the volatile organic compounds for preventing and treating the neck rot and the root rot of the tomatoes. The invention adopts a solid phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS) method to analyze volatile components of pathogenic bacteria capable of inhibiting the tomato neck rot root rot disease, and compares the obtained mass spectrum information with NIST17 and NIST17s (National institute of Standards and Technology) standard mass spectrum libraries and volatile substance data generated by the published strains to determine the substance structures corresponding to the various spectral peaks. Then, the bacteriostatic activity of the main components in the volatile substances on pathogenic bacteria of the tomato neck rot root rot disease is investigated, and Acetoin and 2-Heptanol are found to have an inhibiting effect on the pathogenic bacteria of the tomato neck rot root rot disease, so that the invention is provided.
In order to make the technical solutions of the present application more clearly understood by those skilled in the art, the technical solutions of the present application will be described in detail below with reference to specific embodiments.
The test materials used in the examples of the present invention are all conventional in the art and commercially available. The experimental procedures without specifying the detailed conditions were carried out according to the conventional experimental procedures or according to the instructions recommended by the suppliers. The tomato neck rot root rot pathogen used in the examples of the present invention is Fusarium oxysporum f.sp.radius-lycopersici, described in Journal literature "cascaded infected lipid-modified nanocapsules: Delivery efficiencyanation in soil enhanced controlled efficiency on tomato crown and root", Chemical Engineering Journal; this pathogen is known to be available from the applicant for use in repeating the present test.
Example 1: analysis of volatile organic components by solid phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS)
The volatile organic components of the strains isolated from healthy tomato rhizosphere soil were analyzed by solid phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS) as follows:
the strains were cultured in PDA medium in a headspace flask at 25 ℃ for 7 days. Volatile gases of the strain are subjected to Solid Phase Microextraction (SPME) and slightly modified. After collection, the extraction head is inserted into the sample inlet. The volatile gases were then trapped at 40 ℃ for 45 minutes and then inserted into the headspace for detection. The heating program was set to 40 ℃ for 2 minutes, then the temperature was continuously raised to 100 ℃ (10 ℃/min), 180 ℃ (5 ℃/min), 240 ℃ (10 ℃/min)/min), and the oven was maintained at 240 ℃ for 15 minutes. Volatiles collected from PDA medium without strain were used as controls. The strain was analyzed for the type and relative content of VOCs produced by the strain by SPME-GC-MS technique. The mass spectral information obtained was compared with the standard mass spectral libraries of NIST17 and NIST17s (National institute of Standards and Technology) and the volatile material data generated by the strains listed, and at least 25 volatile materials were measured for the material structure corresponding to each spectral peak (see Table 1).
Table 1: volatile gas composition analysis
The results show that the 25 volatile substances are: formamide (RT ═ 1.083), methyl isopropyl ketone (RT ═ 1.285), N-dimethylcarbamate (RT ═ 1.408), N-methyl N-octylamine (RT ═ 1.54), dimethyldiacetoxysilane (RT ═ 2.14), acetoin (RT ═ 2.346), cyclohexanol (RT ═ 2.793), toluene (RT ═ 3.021), ethylene glycol dimethyl ether (RT ═ 3.306), amyl alcohol (RT ═ 3.613), isovaleric acid (RT ═ 3.775), hexamethylcyclotrisiloxane (RT ═ 3.88), diethylnitrosamine (RT ═ 4.481), 2-heptanone (RT ═ 5.575), 2-heptanol (RT ═ 5.856), p-acetaminophenol (RT ═ 5.987), 6-methyl-2-heptanone (RT ═ 7.49), isovanillin (RT ═ 7.55), tetrakisaceton (RT ═ 56 ═ 738), acetyl cycloacetonate (RT ═ 23.738), 6-methyl-2-heptanone (RT ═ 738) P-hydroxybenzoic acid (RT ═ 13.059), decamethylcyclopentasiloxane (RT ═ 14.412), dodecamethylcyclohexasiloxane (RT ═ 20.372), tetradecylcycloheptasiloxane (RT ═ 25.257).
According to the content of the volatile substance components and the difficulty degree of obtaining, 2-Heptanol (2-Heptanol) and Acetoin (Acetoin) are selected as research objects, and the effect on the tomato neck rot root rot pathogenic bacteria is investigated.
Example 2: effect of volatile substance Acetoin (Acetoin) on the growth of tomato neck rot root rot pathogen hypha
PDA medium mixed with Acetoin at 0.2mg/ml, 0.4mg/ml, 0.6mg/ml, 1mg/ml, 1.4mg/ml, 2mg/ml, 4mg/ml, 6mg/ml, 8mg/ml, 10mg/ml and 14mg/ml was poured into the petri dishes, respectively, and PDA medium without Acetoin was used as a Control (CK). Inoculating the tomato neck rot root rot pathogen with the diameter of 8mm to the center of the culture medium. Repeat 3 times for each treatment of 3 dishes. Sealing the culture dish with a sealing film, blocking the circulation of air inside and outside the culture dish, and placing the culture dish in an incubator at 25 ℃ for culture. The growth inhibition rate is calculated by dynamically measuring the hypha diameter at 24h, 48h, 72h and 96 h.
Growth inhibition (%) × (control colony diameter-treated colony diameter)/(control colony diameter-cake diameter) × 100.
The result is shown in figure 1, and the test result shows that the inhibition rate is 1.4% when the concentration of Acetoin is 0.4 mg/ml; when the concentration is 0.6mg/ml, the inhibition rate is 5 percent; when the concentration is 1mg/ml, the inhibition rate is 12.9 percent; when the concentration is 1.4mg/ml, the inhibition rate is 18.6 percent; when the concentration is 2mg/ml, the inhibition rate is 20.4 percent; when the concentration is 4mg/ml, the inhibition rate is 38.2 percent; when the concentration is 6mg/ml, the inhibition rate is 46.1 percent; when the concentration is 8mg/ml, the inhibition rate is 60.7 percent; when the concentration is 10mg/ml, the inhibition rate is 73.6 percent; when the concentration is 14mg/ml, the inhibition rate is 85.4%. With the increase of the concentration of Acetoin, the inhibition effect on the growth of the hypha of the tomato neck rot root rot pathogenic bacteria is stronger and stronger, and the inhibition rate can reach 85.4%.
Example 3: influence of volatile organic Acetoin (Acetoin) on hyphal morphology
PDA medium mixed with Acetoin at 0.2mg/ml, 0.4mg/ml, 0.6mg/ml, 1mg/ml, 1.4mg/ml, 2mg/ml, 4mg/ml, 6mg/ml, 8mg/ml, 10mg/ml and 14mg/ml was poured into the petri dishes, respectively, and PDA medium without Acetoin was used as a Control (CK). Inoculating the tomato neck rot root rot pathogen with the diameter of 8mm to the center of the culture medium. Repeat 3 times for each treatment of 3 dishes. Sealing the culture dish with a sealing film, blocking the circulation of air inside and outside the culture dish, and placing the culture dish in an incubator at 25 ℃ for culture. Changes in the morphology and growth of fungal hyphae were observed at 96h under 40 Xlight microscopy (FIG. 2).
As a result, it was found that: with the increase of the concentration of the volatile organic compound Acetoin (Acetoin), the hypha is abnormally branched and obviously twisted, which shows that the volatile organic compound Acetoin (Acetoin) has a certain inhibition effect on the tomato neck rot root rot pathogenic bacteria by inhibiting the hypha growth.
Example 4: effect of volatile substance 2-Heptanol (2-Heptanol) on hyphal growth of plant pathogenic bacteria
PDA medium mixed with 0.2mg/ml, 0.4mg/ml, 0.6mg/ml, 0.8mg/ml, 1mg/ml, 1.2mg/ml, 1.6mg/ml 2-Heptanol was poured into each of the petri dishes, and PDA medium without 2-Heptanol was used as a Control (CK). Inoculating the tomato neck rot root rot pathogen with the diameter of 8mm to the center of the culture medium. Repeat 3 times for each treatment of 3 dishes. Sealing the culture dish with a sealing film, blocking the circulation of air inside and outside the culture dish, and placing the culture dish in an incubator at 25 ℃ for culture. The growth inhibition rate is calculated by dynamically measuring the hypha diameter at 24h, 48h, 72h and 96 h.
The growth inhibition ratio (%) (control colony diameter-treatment colony diameter)/(control colony diameter-cake diameter) × 100.
The results are shown in FIG. 3, which shows that: when the concentration of the 2-Heptanol is 0.2mg/ml, the inhibition rate is 16.6 percent; when the concentration is 0.4mg/ml, the inhibition rate is 24.6 percent; when the concentration is 0.6mg/ml, the inhibition rate is 36.4%; when the concentration is 0.8mg/ml, the inhibition rate is 50.4 percent; when the concentration is 1mg/ml, the inhibition rate is 68 percent; when the concentration is 1.2mg/ml, the inhibition rate is 87.9 percent; the inhibition rate was 100% at a concentration of 1.6 mg/ml. With the increase of the concentration of the 2-Heptano, the inhibition effect on the growth of the hyphae of the tomato neck rot root rot pathogenic bacteria is stronger and stronger, and the inhibition rate can reach 100%.
Example 5: influence of volatile organic compound 2-Heptanol (2-Heptanol) on hyphal morphology
PDA medium mixed with 0.2mg/ml, 0.4mg/ml, 0.6mg/ml, 0.8mg/ml, 1mg/ml, 1.2mg/ml, 1.6mg/ml 2-Heptanol was poured into each of the petri dishes, and PDA medium without 2-Heptanol was used as a Control (CK). Inoculating the tomato neck rot root rot pathogen with the diameter of 8mm to the center of the culture medium. Repeat 3 times for each treatment of 3 dishes. Sealing the culture dish with a sealing film, blocking the circulation of air inside and outside the culture dish, and placing the culture dish in an incubator at 25 ℃ for culture. Changes in the morphology and growth of fungal hyphae were observed at 96h under 40 Xlight microscopy (FIG. 4).
As a result, it was found that: with the increase of the concentration of the volatile organic matter 2-Heptanol, hypha is abnormally branched and obviously distorted, which shows that the volatile organic matter 2-Heptanol has certain inhibition effect on tomato neck rot root rot pathogenic bacteria by inhibiting the growth of the hypha.
Example 6: cell test
In order to further investigate the control effect of acetoin, 2-heptanol and the mixed use of acetoin and 2-heptanol on the tomato neck rot and root rot, a plot test is carried out, and the specific method comprises the following steps:
inoculating the pathogenic bacteria of tomato neck rot and root rot preserved by glycerol onto PDA culture medium by plate marking method, culturing at 25 deg.C until colony grows out, selecting a small amount of mycelia, inoculating onto PDA liquid culture medium, and culturing on shaking table at 160r/min -1 The oscillation shakes 5 d. Filtering with 8 layers of gauze, and adjusting spore suspension concentration to 1 × 10 with a hemacytometer under microscope 7 CFU/ml. When the tomato seedlings grow to have two leaves and one heart, the tomato seedlings are soaked in the root with the inoculation concentration of 1 multiplied by 10 7 CFU/ml -1 The tomato neck rot and root rot pathogenic bacteria lasts for 60 min; the tomatoes are then transplanted into the plot. One week after the tomatoes are transplanted, respectively using acetoin with the concentration of 6mg/ml, 2-heptanol with the concentration of 0.6mg/ml and acetoin and 2-heptanol in a combined way (specifically, the acetoin and the 2-heptanol are mixed, and the concentration of the acetoin in the mixed solution is 6mgPer ml, the concentration of 2-heptanol is 0.6mg/ml) is adopted to irrigate roots of the tomatoes and cover the film, and the dosage of the irrigated roots is 50ml per plant. 3 replicates per treatment, each treatment area 18m 2 (2 m.times.9 m). And (90 d) counting the prevention and treatment effects.
Control effect (%) [ (control disease index-treatment disease index)/control disease index ] × 100
Disease index (%) [ Σ (disease grade × number of plants of the same grade)/(maximum disease grade × total number of plants) ] × 100.
Reference documents: simplified and representational bacterial community of mail roots.DOI:10.1073/pnas.1616148114.
The occurrence degree of diseases: grade 0, healthy and disease-free; grade 1, the root is browned, soft rot and constriction are avoided, the leaves are healthy, and no obvious scab exists on the root; 2, the root is browned, and has obvious shrinkage, the leaf tip or the leaf is yellowish, and the root is browned; grade 3, the root is browned and rotten, the leaves are yellow, and the root is browned or even blacked; grade 4, rotten roots and roots, and necrotic seedlings.
The results are shown in FIG. 5, which shows that: the control rates of controlling the tomato neck rot and root rot by using a 2-heptanol plot test with the concentration of 6mg/ml acetoin and the concentration of 0.6mg/ml are respectively 34% and 13%; and the combined use of acetoin and 2-heptanol has the control effect of 51 percent.
The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims (7)
1. Use of Acetoin (Acetoin) in the following (1) or (2):
(1) inhibiting the growth of pathogenic bacteria of the neck rot and root rot of the tomato;
(2) preventing and treating the neck rot and root rot of the tomato.
2. Application of Acetoin (Acetoin) in preparation of pharmaceutical preparation for preventing and treating tomato neck rot and root rot.
3. A pharmaceutical preparation for preventing and treating tomato neck rot and root rot takes acetoin as an active ingredient.
4. The pharmaceutical formulation of claim 3, further comprising an adjuvant or carrier.
5. The pharmaceutical formulation according to claim 3 or 4, wherein the formulation of the pharmaceutical formulation is a fumigant, a powder, a wettable powder, a suspension or an aqueous agent.
6. The pharmaceutical formulation of claim 5, wherein the pharmaceutical formulation is in the form of a fumigant.
7. A method for preventing and treating tomato neck rot and root rot is characterized by comprising the following steps: after the tomato plants are transplanted and planted or before the flowering period of the tomatoes, 6mg/mL acetoin is applied in a root irrigation mode.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210677619.0A CN114982755A (en) | 2022-01-10 | 2022-01-10 | Application of acetoin in prevention and treatment of tomato neck rot and root rot |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210677619.0A CN114982755A (en) | 2022-01-10 | 2022-01-10 | Application of acetoin in prevention and treatment of tomato neck rot and root rot |
| CN202210019112.6A CN114287424B (en) | 2022-01-10 | 2022-01-10 | Application of volatile substances in prevention and treatment of tomato neck rot and root rot |
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| CN202210019112.6A Active CN114287424B (en) | 2022-01-10 | 2022-01-10 | Application of volatile substances in prevention and treatment of tomato neck rot and root rot |
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| JP2835598B2 (en) * | 1996-05-20 | 1998-12-14 | 多木化学株式会社 | Seedling cultivation soil, method for producing the same, and method for growing disease-resistant seedlings |
| WO2016044563A1 (en) * | 2014-09-17 | 2016-03-24 | Bayer Cropscience Lp | Compositions comprising recombinant bacillus cells and a fungicide |
| US20170251664A1 (en) * | 2016-03-07 | 2017-09-07 | The United States Of America, As Represented By The Secretary Of Agriculture | Methods of Attracting Drosophila Suzukii Using Acetoin Blend |
| PE20211401A1 (en) * | 2018-09-28 | 2021-08-02 | Fmc Corp | COMPOSITIONS CONTAINING BACILLUS AMYLOLIQUEFACIENS FCC1256 AND METHODS FOR CONTROLLING PLANT PATHOGENS |
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