CN113498781A - Application of nano manganese dioxide in preventing and treating bacterial leaf blight of rice - Google Patents
Application of nano manganese dioxide in preventing and treating bacterial leaf blight of rice Download PDFInfo
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- CN113498781A CN113498781A CN202110937128.0A CN202110937128A CN113498781A CN 113498781 A CN113498781 A CN 113498781A CN 202110937128 A CN202110937128 A CN 202110937128A CN 113498781 A CN113498781 A CN 113498781A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/02—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing liquids as carriers, diluents or solvents
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N59/00—Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
- A01N59/16—Heavy metals; Compounds thereof
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Abstract
The invention provides an application of nano manganese dioxide in preventing and treating bacterial blight of rice, which comprises the following steps: (1) fully mixing nano manganese dioxide particles with water to form a nano pesticide preparation in a suspension form, wherein the concentration of the nano manganese dioxide particles in the nano pesticide preparation is 50-500 mg/L; (2) and uniformly spraying the nano pesticide preparation on rice leaves. The nano pesticide preparation prepared by the invention has obvious effect on inhibiting bacterial leaf blight infection, and can play a role of fertilizer to promote the growth of plants.
Description
Technical Field
The invention belongs to the field of plant pathogen prevention and control, and particularly relates to a novel nano pesticide preparation which has a good effect of preventing and controlling bacterial blight of rice and can be used as a fertilizer for promoting plant growth. In particular to a nano pesticide preparation for preventing and treating bacterial leaf blight of rice, which comprises the main components, a preparation method and an application method thereof.
Background
The rice bacterial leaf blight is commonly called fire wind, local fire burn, bacterial leaf blight and the like, and is caused by pathogenic bacteria of the bacterial leaf blight, and the harmful parts mainly comprise leaves and leaf sheaths. The disease is frequently found in low-lying and waterlogging-prone areas such as lake-coast and the like, and is mainly concentrated in main rice production areas such as east China and south China. The infected rice plant is mainly characterized by 'dehydration, withering, curling and wilting', so that the leaves of the crop are necrotic, thereby being incapable of photosynthesis to synthesize nutrient substances and having great harm to rice seedlings. Therefore, the rice has low seed setting rate, more green grains and crisp rice quality, seriously harms the food yield and the nutritional quality of the main rice production area in China, causes huge economic loss to the grain industry, and is a major challenge in the current grain safety problem in China.
The bacterial blight of rice (scientific name: Xanthomonas oryzae) belongs to the order Pseudomonas, the family Pseudomonas, the genus Xanthomonas. The bacterial cells are single, short rod-shaped, round at both ends, and about 0.6 μm × 1.5 μm in size. 1 linear flagellum is grown at one end of the thallus, and has a length of about 6-9 μm and a width of about 30 nm.
The nano material has huge application potential because the huge specific surface area, flexible surface functionalization and inherent physicochemical properties of the nano material are listed as one of 8 cross fields in the aspect of nano agriculture by the American nano technology action. Research shows that the foliage application of the low-concentration nano material does not influence the normal growth and development of plants, can promote the growth of the plants, improves the photosynthesis efficiency under abiotic stress, and improves the biomass and the fruit quality. The antibacterial activity of the traditional copper-containing antibacterial agent widely applied to agricultural production at present is greatly influenced by the characteristics of the valence state, the particle size, the crystal phase and the like of copper element. The nanometer material has higher antibacterial activity than the traditional commercial copper-based antibacterial agent due to the huge specific surface area, and the antibacterial mechanism of the nanometer material is mainly that the nanometer material destroys cell membranes to generate a large amount of active oxygen in bacteria.
The manganese dioxide nano material used in the invention has a nano structure, can effectively destroy cell membranes of bacteria under the action of oxidative stress generated by the nano material, has a large amount of +2 and +3 manganese ions on the surface of the nano manganese dioxide, can cause metal toxicity to fungi, and can effectively sterilize the fungi.
Plant pathogen inhibitors are currently widely used worldwide, and have great potential in inhibiting a variety of plant diseases. And because manganese is an important element required by the growth of plants, the proper addition of the element can not only effectively promote the growth of the plants, but also improve the stress resistance of the plants and effectively resist the invasion of various agricultural pathogenic bacteria.
Disclosure of Invention
The invention aims to provide application of nano manganese dioxide in preventing and treating bacterial blight of rice, and the application comprises main components, a preparation method and an application method of the nano manganese dioxide.
According to a first aspect of the present invention, there is provided a use of nano manganese dioxide in controlling bacterial blight of rice, comprising:
(1) fully mixing nano manganese dioxide particles with water to form a nano pesticide preparation in a suspension form, wherein the concentration of the nano manganese dioxide particles in the nano pesticide preparation is 50-500 mg/L;
(2) and uniformly spraying the nano pesticide preparation on rice leaves.
Preferably, the particle size of the nano manganese dioxide particles is 20-30nm, and the purity is more than 99.9%.
Preferably, the single application rate in step (2) is 3-5ml per strain.
According to the second aspect of the invention, the nano pesticide preparation with double effects of preventing and treating the bacterial blight of rice and the fertilizer is prepared by uniformly mixing nano manganese dioxide particles and water, and the concentration of the nano manganese dioxide particles in the finally formed nano pesticide preparation is 50-500 mg/L.
The important components of the novel bacterial blight disease inhibitor comprise two components of nano manganese dioxide and deionized water.
Manganese dioxide is an oxide with the molecular formula of MnO2The chlorine gas is black amorphous powder or black orthorhombic crystal, is insoluble in water, weak acid, weak base, nitric acid and cold sulfuric acid, and is dissolved in concentrated hydrochloric acid under heating condition to generate chlorine gas. It is used for preparing manganese salt, and also used as oxidant and rust remover. Because of their unique structural features, low toxicity, active oxygen scavenging properties, and the like, they are widely used in the biomedical field. Research shows that the nano manganese dioxide with a proper concentration has no obvious toxicological effect on plants, and can promote the growth of plant seedlings. Meanwhile, under the abiotic stress conditions such as heavy metal stress, heat stress and the like, the nano manganese dioxide can effectively help plants to resist the stress from the outside by regulating and increasing the synthesis of chlorophyll and proline. In addition, in the invention, the manganese dioxide nanoparticles are found to inhibit the growth and reproduction of xanthomonas oryzae and limit the diffusion of xanthomonas oryzae on leaves of rice.
The two main components, namely the manganese dioxide nano particles and the deionized water, jointly act from two angles of inhibiting the growth and the propagation of the bacterial blight and limiting the diffusion of the bacterial blight in the leaves of the plants so as to inhibit the infection of the bacterial blight to the rice and comprehensively improve the growth condition of the infected plants.
Based on the severe current situation that rice bacterial leaf blight is abused in China and the knowledge that the important manganese dioxide nano pesticide preparation inhibits bacterial blight infection and promotes plant growth, the novel bacterial leaf blight nano pesticide preparation is invented by adjusting the proportion to relieve the attack of bacterial blight on rice, can play a role of a fertilizer to promote plant growth, effectively promotes the stress resistance of rice infected by bacterial leaf blight, and has great feasibility and great application potential on inhibiting the bacterial leaf blight of rice.
Drawings
Fig. 1 is a transmission electron microscope image of nano manganese dioxide used in the present invention.
Fig. 2 shows the growth of rice after applying various concentrations of the nano-pesticide formulation of example 1.
FIG. 3 shows the plant height, root length, net leaf weight and root weight of rice after applying the nano pesticide formulations of example 1 at different concentrations.
Fig. 4 shows various root morphology parameters of rice, including total root length, total surface area, mean root diameter, and total root volume, after application of different concentrations of the nano-pesticide formulation of example 1.
Fig. 5 shows the relative chlorophyll content of rice leaves after application of different concentrations of the nano-pesticide formulation of example 1.
Fig. 6 shows the MDA content and activities of various antioxidant system enzymes (including SOD enzyme, POD enzyme, and CAT enzyme) in rice leaves after applying nano pesticide formulations of different concentrations in example 1.
Fig. 7 shows the manganese content in rice leaves and roots after application of various concentrations of the nano-pesticide formulation of example 1.
FIG. 8 shows the change of absorbance of bacterial liquid of Xanthomonas oryzae-containing bacteria after applying the nano-pesticide formulations of different concentrations in example 1.
FIG. 9 shows the infection on leaves of rice after applying different concentrations of the nano-pesticide formulation artificially inoculated with P.albuginea in example 1.
Detailed Description
The preparation of a nano pesticide preparation having the effect of inhibiting bacterial blight infection and the method of using the same according to the present invention will be described in detail below by way of specific example 1, which is intended to describe the invention in more detail and should not be construed as limiting the invention in any way.
The reagents used in the following examples are as follows:
the nano manganese dioxide is purchased from Shanghai Pantian powder company (see figure 1, the particle size is 20-30nm, the purity is more than 99.9%), and other chemical reagents in the experiment are purchased from Beijing chemical plants.
Example 1
The present example illustrates that the nano pesticide preparation of the present invention has no obvious toxicological effect on rice at a proper concentration and can effectively inhibit the infection of the bacterial blight.
The rice (Y Liangyou) needed by the experiment is purchased from Chinese academy of agricultural sciences, is disinfected with 3% hydrogen peroxide for 20min before germination, is washed with deionized water for 3 times, after seeds with similar sizes are selected and put into a culture dish, a layer of filter paper is covered on the surface of the seeds, a proper amount of deionized water is sprinkled, and the seeds germinate for one week at the temperature of 25 ℃. Then selecting rice seedlings with similar sizes, transferring the rice seedlings into a 250ml centrifuge tube containing 1/4 concentration Mucun nutrient solution, and culturing at 150 mu mol m-2s-1The Photosynthetic Active Radiation (PAR) of (1) is prepared for experiments after culturing for one week in an environment with the relative humidity of 60-65% and the temperature of 25-28 ℃.
In order to determine the toxicological effect of the nano pesticide preparation, 50, 250 and 500mg/L of the nano pesticide preparation (the invention) with different concentrations and the nano pesticide preparation B are added into a centrifuge tube containing the full-concentration Mucun nutrient solution (the control group is not added with any nano pesticide preparation, and the ingredient of the nano pesticide preparation B is nano Mn3O4) And transplanting the seedlings which germinate and are cultured for two weeks, continuously culturing for two weeks, observing the toxicological effect, and replacing the nutrient solution twice in the culture period in order to prevent the shortage of oxygen and nutrients in the nutrient solution in the culture period. After the culture, the indexes are measured according to the following methods:
phenotypic parameters of rice: plant height, root length, biomass (fresh and dry weight) were measured using a ruler and an electronic balance. Plant root morphological parameters: scanning was performed with a root scanner (Epson expression1680, Japan), and after the rice roots were washed with distilled water, the differently treated rice roots were placed on a tray filled with distilled water, and scanned after being sufficiently stretched. The scanned pictures were used to calculate root morphology related parameters using a root image analysis system (version Pro 2007d, Quebec, Canada) of image analysis software Win RHIZO. Determination of leaf relative chlorophyll: the relative chlorophyll content of rice leaves was determined using a hand-held relative chlorophyll meter. Leaf hormone, enzyme assay: the concentrations of Salicylic Acid (SA), abscisic acid (ABA) and Catalase (CAT), Peroxidase (POD), Malondialdehyde (MDA) and superoxide dismutase (SOD) in virus-infected leaves were measured using a biological kit. And (3) measuring the manganese content of each part of the rice: after drying the plants, grinding and weighing 0.2g, and measuring the manganese content of each part of the rice by using ICP-OES.
After the experiments confirm that the nano manganese dioxide with a proper concentration has no obvious toxicological effect on plants, antibacterial experiments are carried out in order to continuously explore the control effect of the nano pesticide preparation on rice bacterial leaf blight.
The following two experiments were performed on NA solid medium by removing Paddy rice bacterial blight PXO99A from a freezer at-80 ℃. First, streaking was performed, and after 3 to 5 days of inverted culture at 28 ℃, a small amount of PXO99A was taken from the NA medium by a pipette gun in a super clean bench to 5mL of NA liquid medium, and 5 μ l of cephalexin antibiotic (antibiotic: medium: 1: 1000) was added. Placing the liquid culture medium on a shaker, culturing at 28 deg.C and 200rpm for 24 hr, taking out 10 μ L of the shaken bacterial solution in a clean bench, adding 90 μ L of sterile water, and mixing with pipette. In a spectrophotometer, the OD value after the bacterial liquid is diluted by 10 times is measured under the condition of the wavelength of 600 nm. According to the OD value measured in the previous step, the prepared PXO99A bacterial liquid is diluted with sterile water to obtain the OD600 of 0.1. In a sterile tube with OD600 ═ 1, 50, 250 and 500mg/L of nano pesticide preparation B (the control group is not added with any nano pesticide preparation, only added with deionized water with the same amount, and the nano pesticide preparation B is added with nano Mn with the same amount) and 50, 250 and 500mg/L of nano pesticide preparation B after ultrasonic oscillation are added3O4). After 24h of incubation, the inhibition of bacterial growth was determined by measuring the optical density at OD600 nm with an ultraviolet spectrophotometer. Each treatment was set to 3 replicates.
After leaves of similar size were cut from 4 weeks old plants, they were soaked in 70% alcohol for 10 seconds in an ultraclean bench, rinsed 3 times with sterile water, and then soaked in 2% sodium hypochlorite for 10 minutes for sterilization. The treated leaves were laid flat on agar medium and both ends of the leaves were pressed with sterilized wet cotton to provide moisture to keep the leaves fresh and to fix the leaves. Then, 3 wounds on the leaf vein were evenly scratched with a sterile needle, each wound was inoculated with 10 μ L of fungus (OD600 ═ 0.1), and the next day after inoculation with xanthomonas, 1mL of three different concentrations of 50, 250, 500mg/L of the nano-pesticide preparation was sprayed. The time interval between inoculation and treatment was 24 hours. Thereafter, the leaves were placed in agar medium at 26 ℃ and the change of the wound of the leaves was observed and recorded.
Data processing: all experiments were performed in 4 replicates and the data for each treatment represented mean ± Standard Deviation (SD). Analysis of variance (ANOVA) and T-test were performed using Statistix 8.1, an analytical software, to calculate statistical differences between experimental data (P.ltoreq.0.05).
The results show that the 50mg/L nano pesticide preparation has no inhibiting effect on the plant height, the root length and the biomass of the rice, but slightly promotes the growth of the rice, particularly remarkably promotes the root length of the rice (figure 2). The nano pesticide formulation of the present invention is significantly less toxic to crops than another nano pesticide formulation B, wherein the emphasis is on the root weight of rice (fig. 3); the root is an important organ for plants to absorb nutrients, and the roots of the plants are more divergent and the nutrients are more well absorbed and obtained. By scanning the plant root system, we prove that the nano pesticide preparation in the invention has no obvious influence on the total root length, the total root surface area, the average root diameter and the total root volume of the root of rice, and on the contrary, the nano pesticide preparation under partial concentration also promotes the increase of the root system index (figure 4). Furthermore, we found that the nano pesticide formulation had no significant effect on the relative chlorophyll content of rice leaves at low concentrations (fig. 5). We also analyzed the change of antioxidant system in rice leaves (fig. 6), MDA (malondialdehyde) is an index reflecting the peroxidation of plant cell membrane lipid, we found that the low concentration nano pesticide preparation has no significant influence on the content of MDA and the activity of various antioxidant system enzymes, and the above results are enough to prove that the nano pesticide preparation of the present invention has little toxicity to plants. Finally, we also analyzed the Mn content in the roots and leaves of rice, and the results showed that the nano pesticide formulation of the present invention significantly reduced the Mn accumulation in various parts of the plant compared to formulation B (fig. 7).
In the antibacterial experiment, after culturing for 24h, we found that the bacterial liquid OD treated by the nano pesticide preparation is significantly lower than that of the control group without the bacteriostatic agent (fig. 8). In addition, in the rice in-situ culture experiment, the phenomenon that the rice leaves inoculated with the bacterial blight bacteria obviously turn yellow and become necrotic can be found through pictures, and meanwhile, the bacteriostatic effect is extremely obvious under the action of 50mg/L of the nano pesticide preparation (figure 9). The reason is that the nanometer pesticide preparation inhibits the activity and the swimming capacity of the biological membrane and reduces the propagation speed of pathogenic bacteria, and meanwhile, the nanometer pesticide preparation can influence a regulation loop formed by the movement participation of the pathogenic bacteria and the biological membrane, so that the propagation of the bacterial blight on the one hand is inhibited, the number of the bacterial blight on the other hand is increased slowly, and the capability of infecting a host on the other hand is damaged, thereby playing the role of inhibiting the pathogenic bacteria.
Claims (4)
1. The application of nano manganese dioxide in preventing and treating bacterial blight of rice is characterized by comprising the following steps:
(1) fully mixing nano manganese dioxide particles with water to form a nano pesticide preparation in a suspension form, wherein the concentration of the nano manganese dioxide particles in the nano pesticide preparation is 50-500 mg/L;
(2) and uniformly spraying the nano pesticide preparation on rice leaves.
2. Use according to claim 1, wherein the nano manganese dioxide particles have a particle size of 20-30nm and a purity of > 99.9%.
3. The use according to claim 1, wherein the single application rate in step (2) is 3-5ml per strain.
4. A nano-pesticide preparation is characterized in that the nano-pesticide preparation is prepared by uniformly mixing nano-manganese dioxide particles and water, and the concentration of the nano-manganese dioxide particles in the finally formed nano-pesticide preparation is 50-500 mg/L.
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Cited By (1)
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| CN114794148A (en) * | 2022-05-11 | 2022-07-29 | 福建省烟草公司三明市公司 | Protective agent for enhancing cold resistance of plants, preparation method and application thereof |
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Application publication date: 20211015 |