CN115226580B - Biological source method for preventing and treating banded sclerotial blight of wheat and application thereof - Google Patents
Biological source method for preventing and treating banded sclerotial blight of wheat and application thereof Download PDFInfo
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Abstract
Provided is a biogenic method for controlling banded sclerotial blight of wheat, comprising: sequentially vernalizing and sterilizing the wheat seeds; treating the wheat seeds with the pyriform spore liquid and germinating the wheat seeds; selecting wheat seeds with good germination to transplant in soil; after the wheat seeds are broken, spraying sodium nitroprusside solution to the wheat leaves. The method has the advantages that through the double action of root endophytic fungi colonization and leaf signal molecule induction on wheat, the two cooperatively activate the immune reaction in plants, and the pathogen resistance of the wheat is enhanced; the sheath blight resistance effect is better, the pollution of the medicament to the environment or the increase of the plant drug resistance problem are avoided, the environment is protected, the operation is simple, and the cost is low.
Description
Technical Field
The application belongs to the technical field of microbial fertilizers, and particularly relates to a biological source method for preventing and treating wheat sheath blight and application thereof.
Background
Wheat is one of the most widely planted cereal crops worldwide and plays an important role in agricultural economy. Global wheat yields of 2015-2016 were about 7.368 billion tons, with a value of about 1450 billion dollars. As the world population increases, the demand for crop products and the demands for food safety and nutritional balance continue to increase, and by 2050, the demand for wheat is expected to increase by 60%.
The sheath blight of wheat is a soil-borne disease caused by rhizoctonia solani infection, and can be infected in the whole growth and development period, so that the symptoms of rotted buds, withered seedlings, rotted stems of flowers, white ears of dried plants and the like of the wheat are caused. Some wheat with serious illness often cannot heading to form dry booting, and some wheat can heading, but is firm, the seeds are shrunken, dry white ears are formed, and high quality and high yield of wheat are seriously threatened.
Aiming at the problem of pathogenic bacteria infection of wheat, two measures of disease-resistant variety planting and chemical agent prevention and treatment are generally relied on. However, the limited variety of wheat with natural resistance to the Rhizoctonia cerealis pathogen severely hampers the progress of resistance breeding using conventional methods. Therefore, at present, the mode of controlling plant diseases mainly depends on chemical agents, but chemical control has serious environmental and ecological safety hazards, such as untargeted effects and increased pathogen resistance.
In addition, the use of chemical fertilizers in large quantities is a major way of achieving yield increase in agriculture.
Therefore, new agents and new methods which are green, efficient, economical and environment-friendly are required to be searched for the yield increase and disease prevention and control of wheat.
Disclosure of Invention
The primary aim of the application is to overcome the defects and shortcomings of the prior art and provide a novel biological source disease-resistant growth-promoting method for preventing and treating banded sclerotial blight of wheat and improving wheat yield.
The application also aims to provide application of the biological source disease-resistant growth-promoting method for preventing and treating the sheath blight of wheat.
The aim of the application is achieved by the following technical scheme:
a biological source method for controlling banded sclerotial blight of wheat, comprising:
(1) Sequentially vernalizing and sterilizing the wheat seeds;
(2) Treating the wheat seeds with the pyriform spore liquid and germinating the wheat seeds;
(3) Selecting wheat seeds with good germination to transplant in soil;
(4) After the wheat seeds are broken, spraying sodium nitroprusside solution to the wheat leaves.
Wherein, in the step (1), the vernalization processing includes: the wheat seeds are vernalized overnight in a refrigerator at 3-5℃ (preferably 4℃).
Wherein, in the step (1), the sterilization treatment comprises: sequentially soaking wheat seeds in an ethanol solution and a sodium hypochlorite solution respectively to sterilize the wheat seeds, and then uniformly placing the sterilized wheat seeds in a culture dish.
Wherein, in the step (1), the sterilization treatment comprises: soaking wheat seeds in 65-75vol% (preferably 70vol%) ethanol solution for 2-5min (preferably 3 min); rinsing with water for 4-6 times (preferably 5 times); soaking wheat seeds in sodium hypochlorite solution with effective chlorine concentration of 15-25% (preferably 20 vol%) for 8-12min (preferably 10 min), and washing with water for 5-9 times (preferably 7 times); the sterilized wheat seeds were uniformly placed in a sterilized petri dish with two layers of filter paper laid on it.
In the step (1), the sterilization treatment is preferably performed while stirring, preferably magnetic stirring, the wheat seeds are soaked in an ethanol solution or a sodium hypochlorite solution.
In the step (1), the sterilization treatment is preferably a washing step with distilled water.
Wherein, in the step (2), the method comprises the following steps: scraping Pityrosporum indicum spore liquid with Tween water at concentration of 0.015-0.025% (preferably 0.02 vol%) to obtain density of about 10 4 -10 6 personal/mL (preferably 10 5 individual/mL) Pityrosporum indicum spore liquid; pouring the Pityrosporum indicum spore liquid into the culture dish containing wheat seeds in the step (1), and culturing at room temperature (20-25deg.C, preferably 25deg.C) under dark condition with relative humidity of 65% until it germinates.
In the step (2), the Pityrosporum indicum spore liquid can not submerge wheat seeds, so that the seeds are fully soaked.
Wherein, in the step (3), the method comprises the following steps: seeds with good germination are selected to be transplanted into soil, and a proper amount of water is sprayed daily to ensure survival of plants.
Wherein, in the step (4), the method comprises the following steps: after the wheat seeds are broken into the soil for 2-4 days (preferably 3 days), 0.20-0.30mmol/L (preferably 0.25 mmol/L) of sodium nitroprusside solution is sprayed to each leaf of the wheat, and 0.20-0.30mmol/L (preferably 0.25 mmol/L) of sodium nitroprusside solution is sprayed to each leaf of the wheat again at intervals of 1-2 days (preferably 1 day), wherein the spraying amount of the sodium nitroprusside solution sprayed each time is 2-5 ml/strain (preferably 3 ml/strain).
Wherein in the step (2), the tween is tween 20.
Wherein the water of the preparation reagent used in each step is distilled water.
Wherein, in the step (4), the sodium nitroprusside provides exogenous NO; and spraying the sodium nitroprusside leaves after the piriform Piraculosa is colonized.
Wherein the culture dish, filter paper and Tween water are sterilized by sterilizing with high pressure steam sterilizing pot at 120-125deg.C (preferably 121deg.C) for 15-25min (preferably 20 min).
The application also provides application of the biological source method for preventing and treating the sheath blight of the wheat in preventing and treating the sheath blight of the wheat.
The beneficial effects of the application are as follows:
(1) The method for preventing and controlling the sheath blight of the wheat can effectively cause the Pityrosporum indicum serving as a beneficial fungus to colonize the root of the wheat by adopting a seed soaking method.
Pityrosporum indicum and plant root symbiosis can mediate plant disease resistance enhancement through three aspects:
(a) Pityrosporum indicum achieves resistance to pathogenic bacteria by maintaining the integrity of the biofilm system, maintaining low levels of membrane lipid peroxidation and stabilizing intracellular osmotic pressure, and regulating enzyme activity;
(b) Pityrosporum indicum enhances the resistance of plants to pathogenic bacteria by inducing resistance gene expression;
(c) Pityrosporum indicum affects hormone metabolism by rearranging gene expression profiles to increase plant resistance to pathogenic bacteria.
(2) According to the method for preventing and treating the wheat sheath blight, after the Pityrosporum indicum successfully colonizes the root system of the wheat, the exogenous signal molecule nitric oxide, namely sodium nitroprusside, is adopted to spray the upper part, namely the leaves. NO as a signal molecule for plants can participate in the defense reaction during plant-pathogen interactions by activating defense genes and inducing allergic cell death as well as producing phytoalexins.
(3) In general, the application uses the Pityrosporum indicum root to colonize, then uses the NO leaf to spray, which can realize the complete immune induction of the plant system with the root and the leaf, and fully ensures that the sodium nitroprusside spray does not influence the prior colonization of Pityrosporum indicum.
(4) The application uses root endophytic fungi colonization and leaf signal molecule induction to act on wheat in double, and the two cooperate to activate immune reaction in plants, so as to enhance the capability of the wheat for resisting pathogens. Research shows that compared with wheat treated by the biological source method for preventing and treating the sheath blight of the wheat, compared with wheat infected by sheath blight alone, piriform sporulation fertilizer alone or NO signal substance alone, the wheat treated by the biological source method for preventing and treating the sheath blight of the wheat has the advantages that the plant height of the wheat is respectively increased by 15%, 13% and 11%, the root length is respectively increased by 16%, 7.4% and 6.7%, and the fresh weight is respectively increased by 32.3%,18.9% and 22% after 14 days; compared with the method for treating infected wheat by independently applying Pityrosporum indicum biological bacterial fertilizer or NO signal substance, the method has the advantages that the superoxide dismutase activity is respectively improved by 15% and 53%, the catalase activity is respectively improved by 54% and 16%, the peroxidase activity is respectively improved by 16% and 17%, the relative conductivity (membrane stability characterization index) and the malondialdehyde content are obviously improved, and the photosynthesis index chlorophyll content and the relative water content are also obviously improved, so that the method has better sheath blight resistance effect, avoids the pollution of the medicament to the environment or the problem of increasing the plant drug resistance, is environment-friendly, and is simple to operate and low in cost.
(5) The biological source disease-resistant growth-promoting combination method is green and environment-friendly and can effectively resist the demand of wheat sheath blight in the market, effectively prevent and treat the damage of pathogens to wheat plants, overcome the defects of environmental pollution, strong drug resistance and the like in the prior art, and has the advantages of stable effect, low cost, simple operation and market development prospect.
Detailed Description
The present application will be described in further detail with reference to specific examples. It is to be understood that these examples are for illustration only and are not intended to limit the scope of the application. Further, it is understood that various changes and modifications may be made by those skilled in the art after reading the teachings of the present application, and such equivalents are intended to fall within the scope of the application as defined in the claims.
Example 1
The endophytic fungi are selected from Pityrosporum pekinensis and stored in China Committee for culture Collection of microorganisms (CGMCC: 10325).
The plant is selected from wheat, the wheat variety is break wheat, seeds are purchased in the market, and the seeds are stored in a refrigerator at 4 ℃.
The soil is mixed soil with the mass ratio of vermiculite to perlite being 2:1.
To compare the effect of the method of the application, seven treatment groups were set, respectively:
(1) Blank control (Mock);
(2) Rhizoctonia solani (Rh);
(3) Sodium mono Shi Xiaopu (NO);
(4) Single inoculated piriform Piri (Piri);
(5) Sodium nitroprusside + sheath blight germ (NO + Rh);
(6) Piriopsis cumingii (Piri+Rh);
(7) Piriosa+sodium nitroprusside+Rhizoctonia solani (Piri+NO+Rh).
The specific application method comprises the following steps:
(1) Placing the wheat seeds in a refrigerator at 4 ℃ for vernalization overnight; soaking the seeds in 70% ethanol for 3min, washing with distilled water for 5 times, soaking the seeds in 20% sodium hypochlorite for 10min, and washing with distilled water for 7 times; uniformly placing the sterilized seeds in a culture dish which is sterilized and paved with two layers of filter paper;
(2) Using 0.02% Tween waterScraping Pityrosporum indicum spores to obtain spore liquid, and regulating spore liquid density to about 10 5 individual/mL; spraying proper amount of spore liquid into Piri, piri+Rh, piri+NO+Rh treatment group culture dishes, placing the seeds which cannot be immersed in the spore liquid, culturing in dark condition with 25 ℃ and relative humidity of 65%, spraying once again after 24 hours, and sprouting;
(3) Transplanting the seeds with good germination into soil, culturing the flowerpot at 25 ℃, simulating natural condition illumination, and making the flowerpot dark for 12 hours;
(4) After the seeds germinate for 3 days, uniformly spraying 0.25mmol/L sodium nitroprusside on each leaf of the wheat in the NO, NO+Rh and Piri+NO+Rh treatment groups, and carrying out secondary enhanced spraying on the 5 th day of growth every other day;
(5) And (3) when the wheat germinates, treating the Rhizoctonia cerealis of the wheat Rh, NO+Rh, piri+Rh and Piri+NO+Rh, scraping germs by using a toothpick, inserting the germs into the basal part of the wheat stem, and spraying a proper amount of water every day to ensure the normal growth of the wheat.
Tables 1-9 provide a comparison of the performance data for the different treatment groups.
Tables 17 d, 14d comparison of growth parameters for different treatment groups
Table 2.7 d, comparison of superoxide dismutase Activity of 14d different treatment groups
Tables 3.7 d, 14d comparison of catalase Activity of different treatment groups
Tables 4.7 d, 14d comparison of peroxidase Activity in different treatment groups
Tables 5.7 d, 14d comparative relative conductivities of different treatment groups
Tables 6.7 d, 14d malondialdehyde content comparison of different treatment groups
Tables 7.7 d, 14d comparison of chlorophyll content of different treatment groups
Tables 8.7 d, 14d comparison of relative moisture content for different treatment groups
Tables 9.7 d, 14d comparison of soluble sugar content for different treatment groups
Disease resistance tests were performed using the methods described in the above examples, and the activities of superoxide dismutase, peroxidase, and catalase in 7 treatment groups were measured, and resistance comparisons were made; determining membrane stability indexes including relative conductivity and malondialdehyde content, and judging membrane oxidation stability conditions of different treatment groups; and determining the growth indexes such as chlorophyll content, relative water content photosynthesis index, plant height, root length and fresh weight growth parameter index, and comparing the growth conditions of different treatments. The experiment is carried out for 7 days and 14 days, the effect of the implementation method is stable and effective, the method 1-3 is repeated three times, and the data are basically stable each time, so that the experiment is proved to have no error.
From the data, the activities of superoxide dismutase, peroxidase and catalase in the Piri+NO+Rh treated groups were lower than those in the Rh treated groups at 7 days and 14 days; but higher than the no+rh, piri+rh treated group, suggesting that Piri and NO act synergistically to further activate the oxidative defense system in plants, but it is different from the extent of injury to the host by pathogenic bacteria. According to the malondialdehyde content of the oxidation degree of the membrane and the relative conductivity of the damage degree of the membrane, the dual effects of Piri and NO effectively relieve the damage of bacteria to cell membranes and further enhance the chlorophyll and soluble sugar content in plants.
According to the experimental results, the biological yield of wheat plants under the infection of banded sclerotial blight can be obviously improved, the cell membrane stability is enhanced, and the chlorophyll and soluble sugar contents in plants are improved by adopting the method disclosed by the application; effectively relieves disease symptoms. The method is simple to operate, low in cost, green and environment-friendly, and can achieve the effect of effectively preventing and treating the sheath blight of wheat.
It is apparent that the above examples are given by way of illustration only and are not limiting of the embodiments. Other variations or modifications of the above teachings will be apparent to those of ordinary skill in the art. It is not necessary here nor is it exhaustive of all embodiments. While still being apparent from variations or modifications that may be made by those skilled in the art are within the scope of the application.
Claims (7)
1. A biological source method for controlling banded sclerotial blight of wheat, comprising:
(1) Sequentially vernalizing and sterilizing the wheat seeds;
(2) Treating the wheat seeds with the pyriform spore liquid and germinating the wheat seeds;
(3) Selecting wheat seeds with good germination to transplant in soil;
(4) Spraying sodium nitroprusside solution to the wheat leaves after the wheat seeds break the soil;
wherein, in the step (2), the method comprises the following steps: scraping Pityrosporum indicum spore liquid with Tween water with concentration of 0.015-0.025% (v/v), and concocting to obtain density of 10 4 -10 6 individual/mL of Pityrosporum indicum spore liquid;
in the step (4), the method comprises the following steps: spraying 0.20-0.30mmol/L sodium nitroprusside solution to each leaf of the wheat after the wheat seeds are broken into earth for 2-4 days, and spraying 0.20-0.30mmol/L sodium nitroprusside solution to each leaf of the wheat again after 1-2 days;
wherein, the spraying amount of the sodium nitroprusside solution sprayed each time is 2-5 ml/plant.
2. The biogenic method for controlling wheat sheath blight according to claim 1, wherein in step (1), the sterilization treatment comprises: the wheat seeds are soaked in ethanol solution and sodium hypochlorite solution respectively to sterilize the wheat seeds, and then the sterilized wheat seeds are placed in a culture dish.
3. The biogenic method for controlling wheat sheath blight according to claim 1, wherein the piriform spore liquid is poured into the culture dish containing the wheat seeds in step (1), and the culture dish is placed under dark condition of room temperature and relative humidity of 65% for germination.
4. The biogenic method for controlling wheat sheath blight according to claim 1, wherein in step (2), the pyriform spore liquid does not submerge the wheat seeds so that the seeds are sufficiently infiltrated.
5. The biogenic method for controlling wheat sheath blight according to claim 1, wherein in step (4), the nitroprusside solution is sprayed onto the wheat leaves after the piriform in the Indian is colonized.
6. The biogenic method for controlling banded sclerotial blight of wheat as claimed in claim 2, wherein the petri dish, filter paper, tween water are sterilized.
7. Use of a biogenic method for controlling banded sclerotial blight in wheat as claimed in any one of claims 1 to 6 for controlling banded sclerotial blight in wheat.
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