CN112143679A - Marseillella foldata, composite coating agent, and preparation method and application thereof - Google Patents
Marseillella foldata, composite coating agent, and preparation method and application thereof Download PDFInfo
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Abstract
The invention provides a folding mosaic bacterium (Massilia plicata), a composite coating agent, a preparation method and application thereof. The strain preservation number of the folding mosaic bacterium is CGMCC No. 20439. The composite coating agent comprises an agent A, an agent B and an agent C; the agent A comprises one or more of ground phosphate rock, potassium mineral powder, boron mineral powder, iron silicate mineral powder, diatomite and straw powder; the agent B comprises folded mosaic bacteria; the agent C comprises carboxymethyl cellulose and molasses. The folded mosaic bacteria can promote the growth of stress-resistant tuber planting blocks of potatoes; the prepared compound coating agent is coated on potato tubers for planting and cutting, can remarkably promote the development and the establishment of the root system of potato seedlings, enables the root system to be more developed, thickens the stems, thickens the leaves, increases the chlorophyll, enhances the resistance to soil-borne diseases, enhances the resistance to drought and salt stress, and remarkably improves the yield and the quality of the potato tubers.
Description
Technical Field
The invention belongs to the technical field of microbial resource utilization and ecological agriculture, and relates to a folding mosaic bacterium (Massilia plicata), a composite coating agent, a preparation method and application thereof; in particular to folded mosaic bacteria for promoting the growth resistance of potato tuber planting blocks, a compound coating agent, a preparation method and application thereof, which are suitable for planting potatoes.
Background
Potatoes are global grain and vegetable dual-purpose crops, and are the fourth main grain crop except rice, wheat and corn. China is the world where the most potatoes are planted and produced, and is located in provinces such as inner Mongolia, Gansu and Qinghai in northwest of China and is the most main production area of potatoes in China. The drought and rain-less climate makes the potatoes produced in the northwest region have better taste and are rather popular with people. However, drought and soil impoverishment result in low potato yields, limiting the associated production. In recent years, with the large-scale and basic potato planting, diseases and insect pests of potatoes frequently occur, a large amount of pesticides have to be used in the planting process, the quality of the potatoes is seriously influenced, the potato planting method has great pressure on the ecological environment, and the potato planting method does not accord with the sustainable agricultural development trend.
The plant nutrient supply is improved, particularly the levels of potassium and silicate are increased, and the drought resistance and salt stress resistance of the plants are facilitated. The microorganisms can promote the growth, yield and quality of plants under stress through a series of functions and actions: various plant hormones such as auxin and the like are generated to promote the development and the establishment of the root system of the plant and increase the nutrition and the water absorption; can also stimulate plants to synthesize soluble sugar and proline and improve the activity of antioxidant enzymes such as SOD to reduce stress such as drought; the photosynthetic performance is improved, and the plant productivity is enhanced; ACC deaminase is produced to inhibit premature senescence or premature apoptosis of plants; inhibiting the occurrence and development of diseases; and so on. Nitrogen, phosphorus and potassium are large amount of nutrient elements required for plant growth, and the microorganism can increase supply by phosphate and potassium dissolving and nitrogen fixing functions and effects, especially for potato which is a high-potassium-demand crop. Stress such as drought has the greatest influence on plant emergence and seedling growth, and is an important means for promoting plant emergence and seedling growth for plant seeds and coating of propagation roots or stems.
At present, most of the related domestic patent applications for seed coating relate to the preparation of related coating agents for controlling plant diseases and insect pests by using chemical agents, and a small amount of related coating agents are related to chemical substances and microorganisms. For example, a patent application entitled "coating composition for pathogen control in vegetables" (application No. 201280030540.3 ") describes the combination of wax and some microorganisms to form a particulate carrier to control pests; a patent application entitled "a potato seed coating for controlling pests" (application No. 201510840969.4) describes the combination of starch, weathered coal, and pectin to form a coating powder to control the occurrence of potato/potato pests; the patent application named as 'disease-resistant, insect-repellent and biological seed coating agent and the preparation method thereof' (application number '201710206032.0') describes that pseudomonas fluorescens, bacillus subtilis, chitin and the like are mixed to form a liquid coating agent for preventing and treating plant diseases and insect pests; the patent application entitled "a preparation method of microbial seed coating and its application" (application number "201910299546.4") emphasizes the coating technology using chlamydospore of trichoderma, bonding with diatomite, cyclodextrin and the like, and mainly solves the problem of short preservation time of trichoderma spores; the patent application entitled "a microbial seed pelleting coating composition" (application number "201910124557.9") describes the formation of a microbial coating composition by combining various bacillus species and mold species with calcium peroxide to achieve disease control and enhanced seed germination in marginal lands; and so on.
Disclosure of Invention
Based on the defects in the prior art, the invention aims to provide a folded mosaic bacterium (Massilia plicata) for promoting the growth resistance of potato tuber planted blocks; the second purpose of the invention is to provide the application of the folding mosaic bacteria in promoting the anti-reverse growth of potato tuber planting and cutting blocks; the third purpose of the invention is to provide a compound coating agent containing the folding mosaic bacteria, which can promote the stress-resistant growth of potatoes after being coated on tubers of the potatoes; the fourth purpose of the invention is to provide a preparation method of the compound coating agent; the fifth purpose of the invention is to provide the application of the compound coating agent in the coating of potato tuber planting blocks; the sixth purpose of the invention is to provide a method for coating and planting potato tuber planting cuts by using the compound coating agent.
The purpose of the invention is realized by the following technical scheme:
on one hand, the invention provides a folding mosaic bacterium (Massilia plicata) for promoting the growth resistance of potato tuber planting blocks, which comprises the following components: the strain preservation number of the folding mosaic bacterium B260(Massilia plicata B260) is CGMCC No. 20439; the preservation date is as follows: year 2020, month 07, 23; the preservation unit is as follows: china general microbiological culture Collection center (CGMCC); the address of the preservation unit is as follows: the microbial research institute of Chinese academy of sciences, Beijing, China, the postal code: 100101; the classification is named as: marseillea plicata (Massilia plicata).
The inventor separates the functions of plant growth promoting traits such as nitrogen fixation, phosphorus dissolution, potassium dissolution, plant hormone production, siderophore production, ACC deaminase production and the like of the rhizosphere by collecting the roots of field potato seedlings and adopting a coating and scribing technology; a strain capable of promoting the anti-reverse growth of potato tuber planting cut pieces is obtained through separation and purification, and is identified as folding mosaic bacteria (Massilia plicata) through species identification, and the strain preservation is carried out.
The folding mosaic bacterium B260(Massilia plicata B260) has the functions of producing phytohormones, auxin, gibberellin, calcium phosphate-phosphorus (phosphate rock powder-phosphorus), potassium phosphate (potassium ore powder-potassium), self-generated nitrogen fixation, ACC (para 2, 4-diacetyl gamboge phenol) deaminase, siderophore, HCN (hydrocyanic acid) or DAPG (para 2, 4-diacetyl gamboge phenol) and the like; the method can obviously promote the development and the establishment of the root system of the potato seedling, the root system is developed, the stem is thickened, the leaves are thickened, the chlorophyll is increased, the resistance to soil-borne diseases is enhanced, the resistance to drought, plant diseases and insect pests and salt stress is enhanced, the growth is promoted, and the yield and the quality of the potato tuber are obviously improved.
On the other hand, the invention also provides application of the folding mosaic bacteria in promoting the anti-reverse growth of potato tuber planting and cutting blocks.
In still another aspect, the present invention also provides a composite coating agent, wherein the composite coating agent comprises an agent a, an agent B and an agent C;
the agent A comprises one or more of phosphate rock powder, potassium mineral powder, boron mineral powder, iron silicate mineral powder, diatomite and straw powder;
the agent B comprises the folded mosaic bacteria;
the agent C comprises carboxymethyl cellulose and molasses.
The inventor researches and discovers that after the compound coating agent disclosed by the invention is adopted to coat potato tuber planting blocks, the establishment of potato root systems can be remarkably promoted, the resistance to drought and diseases is enhanced, the growth is promoted, and the yield and the quality of potato tuber are improved.
In the composite coating agent, the weight ratio of the agent A, the agent B and the agent C is preferably 1 kg: (10-50) g: (120-360) g; the effective viable count concentration of the folded mosaic bacteria in the agent B is 109～1011CFU/g or 109～1011CFU/ml。
In the composite coating agent, preferably, the agent A consists of ground phosphate rock, potassium mineral powder, boron mineral powder, iron silicate mineral powder, diatomite and straw powder;
wherein the weight ratio of the phosphate rock powder, the potassium mineral powder, the boron mineral powder, the iron silicate mineral powder, the diatomite to the straw powder is (40-60): (40-50): (1-5): (0.5-2): (5-10): (0.01-0.05).
In the above composite coating agent, preferably, the weight ratio of the potassium mineral powder, the boron mineral powder, the iron silicate mineral powder, the diatomite and the straw powder is 50: 40: 3: 1: 7: 0.03.
In the compound coating agent A, powdered rock phosphate, potassium mineral powder, boron mineral powder, iron silicate mineral powder, diatomite and the like are all obtained from markets and are mixed according to a specific proportion, so that the coating of tubers can be completed; on the other hand, the nutrition of the tubers can be increased, the nutrition supply of the tubers in the sprouting period is improved, and meanwhile, the surface of the tubers is protected, so that strains are prevented from falling off and losing, and a natural protective film is formed; the straw powder is obtained by crushing waste plants of crops such as potatoes, has the granularity of 0.05-0.2 mm, can form a film with good air permeability together with polysaccharide and other materials produced by folding mosaic bacteria, and is sterilized by high-temperature steaming or ammonia water fumigation before compounding.
In the composite coating agent, the agent B preferably comprises a liquid microbial inoculum or a solid powdery microbial inoculum.
In the composite coating agent, preferably, the liquid microbial inoculum comprises a protective agent and folded mosaic bacteria; the effective viable count concentration of the folded mosaic bacteria in the liquid microbial inoculum is 109～1011CFU/mL。
In the composite coating agent, preferably, the solid powdered microbial inoculum comprises folded mosaic bacteria and an additive (which is a conventional additive in the field) for maintaining the survival of strains, and the effective viable count concentration of the folded mosaic bacteria in the solid powdered microbial inoculum is 109～1011CFU/g。
In the above composite coating agent, preferably, the protecting agent comprises K having pH of 7.22HPO4-KH2PO4Buffer and/or glycerol.
In the composite coating agent, the carboxymethyl cellulose is preferably mixed with the molasses in the agent C in equal mass.
In the compound coating agent C, carboxymethyl cellulose and molasses can be used as adhesives, and simultaneously, the molasses contains a large amount of carbon sources, vitamins and other nutrient substances.
In still another aspect, the present invention also provides a method for preparing a composite coating agent, comprising the steps of:
performing strain activation, shake flask culture and fermentation tank amplification culture on the folded mosaic bacteria by using a modified R2A culture medium, centrifuging to collect cells, freeze-drying, adding additives to prepare a solid powdery microbial inoculum, or directly adding a liquid protective agent (such as glycerol, buffer solution and the like) to prepare a liquid microbial inoculum to obtain the agent B;
respectively preparing the agent A and the agent C according to the formula of the agent A and the agent C;
and independently packaging the agent A, the agent B and the agent C to obtain the compound coating agent.
In the compound coating agent, the agent A, the agent B and the agent C can be respectively and independently packaged, and water is added according to the proportion for mixing and blending when the compound coating agent is used, so that the independent packaging is more favorable for ensuring the survival of microorganisms, prolonging the storage time and ensuring the coating effect. In addition, although the material components and the proportion of the composite coating agent have outstanding effects particularly on potato tuber cutting, the composite coating agent can also be expanded to the application of other crops.
In the preparation method, in the process of preparing the agent A, the straw powder needs to be steamed at high temperature or fumigated by ammonia water for sterilization, and then is mixed with other mineral powder in proportion.
In the above preparation method, preferably, the composite coating agent further comprises the following preparation steps:
adding the agent B into the agent C, adding sterile water (sterile water without viable bacteria after filtration sterilization or high-temperature sterilization), and stirring uniformly to obtain a mixed solution;
then adding the agent A into the mixed solution and mixing uniformly to prepare the composite coating agent.
In another aspect, the invention also provides the application of the compound coating agent in the coating of potato tuber planting blocks.
In another aspect, the invention also provides a potato tuber planting, cutting and coating planting method, which comprises the following steps:
step one, adding the agent B into the agent C, adding sterile water, and uniformly stirring to obtain a first mixed solution; then adding the agent A into the first mixed solution to obtain a composite coating agent liquid solution for standby;
wherein, in the liquid solution of the composite coating agent, the dosage ratio of the agent A to the sterile water is 1 kg: (40-60) L; the effective viable count concentration of the folded mosaic bacteria in the liquid solution of the composite coating agent is 106～108CFU/mL；
Uniformly spraying the liquid solution of the compound coating agent on the surfaces of the potato tuber planting blocks which are aired for 12-24 hours, turning and stirring uniformly, airing for 5-10 min every time of spraying for ensuring full adsorption, and finally airing for 30-60 min for sowing, or sealing in a clean packaging bag, storing in a cool place for no more than 2 days and sowing;
wherein the dosage ratio of the potato tuber planting cut blocks to the liquid solution of the composite coating agent is (50-100) kg: (1-5) L.
In the above planting method, preferably, the dosage ratio of the agent a to the sterile water is 1 kg: 50L.
In the planting method, the ratio of the potato tuber planting cut block to the liquid solution of the compound coating agent is preferably 70 kg: 4L.
In the planting method, preferably, the seeds are aired for 8min every time of spraying 1 time to ensure full adsorption, and finally, the seeds are sowed immediately after airing for 50 min.
The invention has the beneficial effects that:
the folded mosaic bacteria can promote the growth of stress-resistant tuber planting blocks of potatoes; the prepared compound coating agent is coated on potato tubers for planting and cutting, can remarkably promote the development and the establishment of the root system of potato seedlings, enables the root system to be more developed, thickens the stems, thickens the leaves, increases the chlorophyll, enhances the resistance to soil-borne diseases, enhances the resistance to drought and salt stress, and remarkably improves the yield and the quality of the potato tubers.
Drawings
FIG. 1 is a plot of a field planted with or without potato tubers coated with the compound coating agent of example 5 of the present invention (A in FIG. 1 is a field panorama; B in FIG. 1 is tuber yield per unit area (coated); C in FIG. 1 is tuber yield per unit area (uncoated)).
FIG. 2 shows the compound coating agent of the present invention (A in FIG. 2 is the dry powder mixture of agent A; B in FIG. 2 is the liquid B containing the folded mosaic bacteria of the present invention; C in FIG. 2 is the liquid mixture of agent C; D in FIG. 2 is the finished product of potato coated tubers).
Culture deposit for patent procedure:
the folded Marseillea B260(Massilia plicata B260) of the present invention;
the preservation date is as follows: year 2020, month 07, 23;
the preservation unit: china general microbiological culture Collection center (CGMCC);
the address of the depository: west road No. 1, north chen, chaoyang district, beijing, zip code: 100101;
the preservation number is: CGMCC No. 20439;
and (3) classification and naming: marseillea plicata (Massilia plicata).
Detailed Description
The technical solutions of the present invention will be described in detail below in order to clearly understand the technical features, objects, and advantages of the present invention, but the present invention is not limited to the practical scope of the present invention.
The formula of the invention in the following embodiments is:
the media formulations used in the following examples:
1. modified R2A medium: glucose 1.3g, bacteriological peptone 1.3g, acid hydrolyzed casein 1.3g, yeast extract 1.3g, K2HPO3·3H2O0.8 g, sodium pyruvate 0.8g, MgSO4·7H20.15g of O and 1000mL of distilled water; agar solid medium was added with 15g of agar.
2. Nitrogen fixation culture medium: KH (Perkin Elmer)2PO4 0.2g，CaCO3 5g，MgSO4·7H20.2g of O, 10g of glucose, 1.2g of NaCl, CaSO4·2H20.1g of O, 20g of agar and 1000mL of distilled water, and the pH value is natural.
3. Inorganic phosphorus culture medium: glucose 10g, (NH)4)2SO4 0.5g，NaCl 0.3g，KCl 0.3g，MgSO4·7H2O 0.3g，Ca3(PO4)2 25g，FeSO4·7H2O 0.03g，MnSO4·4H20.03g of O, 20g of agar and 1000mL of distilled water, and the pH value is natural.
4. Organic phosphorus culture medium: glucose 10g, (NH)4)2SO4 0.5g，NaCl 0.3g，KCl 0.3g，MgSO4·7H20.3g of O, 25g of calcium phytate and FeSO4·7H2O 0.03g，MnSO4·4H20.03g of O, 20g of agar and 1000mL of distilled water, and the pH value is natural.
5. Potassium-dissolving culture medium: sucrose 5.0g, NH4NO3 1.7g，Na2HPO4 2.0g，CaCO3 0.1g，MgSO40.5g，FeCl30.005g of potassium feldspar, 1.0g of potassium feldspar; 1000mL of distilled water.
6. LB culture medium: 10g of tryptone, 5g of yeast extract, 10g of NaCl, 15g of agar, 1000mL of deionized water and pH 7.4.
7. DF culture medium: KH (Perkin Elmer)2PO4 4g，Na2HPO4 6g，MgSO4·7H20.2g of O, 2g of glucose, 1.62mL of gluconic acid, 2g of citric acid, (NH)4)2SO42g, 1000mL of deionized water, pH 7.2.
8. ADF culture medium: adding (NH) in DF medium4)2SO4And removing to obtain the ADF culture medium.
Example 1: screening, separating and identifying strain
Collecting field potato seedling roots, separating the functions of strains of plant growth promoting characters such as nitrogen fixation, phosphorus dissolution, potassium dissolution, plant hormone production, siderophore production, ACC deaminase production and the like in the rhizosphere by a coating and scribing technology, performing qualitative or quantitative determination on the plant growth promoting capacity by using culture media such as nitrogen fixation, phosphorus dissolution, potassium dissolution and the like, and screening to obtain 5 strains shown in table 1.
Table 1:
according to the experimental results of table 1, the strain with the strain number B260 has various growth promoting abilities, and is more suitable for the climatic or geological conditions (large temperature difference, drought, saline alkali, etc.) in northern arid regions, so that it is used as an excellent strain obtained by screening.
The genomic DNA of the strain obtained by separation and purification was used as a template, and 27F and 1492R were used as primers to PCR amplify the 16S rRNA gene of the strain, and the amplified product was sequenced, and the obtained sequences were classified by on-line EzTaxon (the 16S rRNA gene sequence was initially determined to be homologous with a degree of similarity of more than 97%), and the results are shown in table 2.
Table 2:
| strain numbering | The most closely related species | Similarity (%) |
| B260 | Massilia plicata | 99 |
The results in Table 2 show that the similarity between the 16S rRNA gene sequence of the excellent strain B260 obtained by screening and the mosaic bacterium of the model species reaches 99%.
Example 2: activation, culture and collection of bacteria
Absorbing 100 mu L of glycerol-conserving liquid (the folded mosaic bacteria B260 of the example 1), coating the glycerol-conserving liquid on a modified R2A culture medium, culturing at 25 ℃, picking a single colony position modified R2A corresponding liquid culture medium which has typical colony characteristics and grows rapidly, and culturing at 25 ℃; culturing to late logarithmic phase, centrifuging at 8000g centrifugal force, and centrifuging with K2HPO4-KH2PO4The buffer (pH7.2) repeated 3 times after washing, with the same buffer heavy suspension to cell concentration to 109～1011CFU/mL (as in B in FIG. 2), i.e., to obtain the B agent.
Example 3: optimization of non-biological coating components
Preparing a composite coating agent liquid solution:
step one, 5g of the agent B in the example 2 is added into the agent C (the agent C comprises carboxymethyl cellulose, molasses and the like in mass, and the mass concentration of the agent C in the liquid solution of the composite coating agent is shown in the following table 3), 5L of sterile water is added, and the mixture is uniformly stirred to obtain a first mixed solution; then adding 100g of the agent A into the first mixed solution to obtain a composite coating agent liquid solution for standby;
wherein the agent A consists of ground phosphate rock, potassium mineral powder, boron mineral powder, iron silicate mineral powder, diatomite and straw powder; the effective viable count concentration of the folded mosaic bacteria in the liquid solution of the composite coating agent is 106～108CFU/mL。
Uniformly spraying the prepared liquid solution of the composite coating agent on the surfaces of the potato tuber planting cut blocks which are aired for 12 hours, turning and stirring uniformly, airing for 10min every 1 time to ensure full adsorption, and finally airing for 30min and then sowing;
wherein the dosage ratio of the potato tuber planting cut blocks to the liquid solution of the composite coating agent is 70 kg: 4L.
The PEG6000 simulated drought stress potted potato is used for comprehensively selecting and optimizing the distribution ratio of the abiotic coating groups on the basis of single-factor research by the contents of the components of the agent A (shown in the following table 3), the concentration of the agent C, the seedling emergence speed of the potato, the size of the potato in 60 days and the adhesion degree (the falling rate after drying and shaking for 10min at the rotating speed of 100r.p.m. in a vertically reversed manner, namely the falling weight of the coating materials in blocks/the total weight of the coating materials before falling), and the result is shown in the table 3.
Table 3:
note: the non-biological component proportion:
the mass ratio of the agent A to the raw materials is as follows: powdered rock phosphate: potassium mineral powder: boron mineral powder: iron silicate mineral powder: diatomite: straw powder.
The mass concentration of the agent C is as follows: for example 0.40% (about 20g), the expression C means that the mass concentrations of carboxymethyl cellulose and molasses are both 0.20%.
As can be seen from table 3: the combination 1 is excellent in the overall effect of coating and growth promotion.
Example 4: optimized determination of effective bacteria number proportion
Simulating drought stress potted potatoes by PEG6000, and fixing the dosage of the agent A: the weight ratio of the phosphate rock powder, the potassium mineral powder, the boron mineral powder, the iron silicate mineral powder, the diatomite and the straw powder is 50: 40: 3: 1: 7: 0.03, the proportion of the effective bacteria number is comprehensively selected and optimized according to the total viable bacteria number, the potato emergence speed and the 60-day time size of the folded mosaic bacteria B260, and the result is shown in Table 4.
Table 4:
note:
an agent A: the weight ratio of the phosphate rock powder, the potassium mineral powder, the boron mineral powder, the iron silicate mineral powder, the diatomite and the straw powder is 50: 40: 3: 1: 7: 0.03;
and (2) agent B: the effective viable count concentration of the folded mosaic bacteria in the B agent is 109～1011CFU/mL；
C, agent C: the mass concentration of the cellulose and the molasses in the liquid solution of the composite coating agent is as follows: 0.4 percent;
effective viable count concentration range (CFU/mL) in the composite coating agent liquid solution: the amount of bacteria is maintained at 10 when adding agent B9～1011The effective viable count concentration in the liquid solution of the composite coating agent prepared by adding the agent B in the CFU/mL range is shown in Table 4.
As can be seen from table 4: the combination 1 is excellent in the overall effect of coating and growth promotion.
Example 5: comparative experiment of coating on growth of potatoes under drought stress
Preparing a composite coating agent liquid solution:
step one, 50g of the agent B in the embodiment 2 is added into 200g of the agent C (the agent C is composed of carboxymethyl cellulose, molasses and the like), and 50L of sterile water is added and stirred uniformly to obtain a first mixed solution; then, 1000g of the agent A is added into the first mixed solution to obtain a composite coating agent liquid solution for standby;
wherein the agent A consists of ground phosphate rock, potassium mineral powder, boron mineral powder, iron silicate mineral powder, diatomite and straw powder; the effective viable count concentration of the folded mosaic bacteria in the liquid solution of the composite coating agent is 106～108CFU/mL。
Uniformly spraying the prepared liquid solution of the composite coating agent on the surfaces of the potato tuber planting cut blocks which are aired for 12 hours, turning and stirring uniformly, airing for 10min every 1 time to ensure full adsorption, and finally airing for 30min and then sowing;
wherein the dosage ratio of the potato tuber planting cut blocks to the liquid solution of the composite coating agent is 70 kg: 4L.
And C, processing.
The PEG6000 was used to simulate drought stress in potted potatoes and the potato growth index was determined and the results are shown in table 5 and fig. 1.
Table 5:
note: b260: the compound coating agent liquid solution of the embodiment;
only replacing the strain B260 with Massilia plicata (CCTCC AB 207820) strain, and keeping the other operations consistent with the group B260;
CK-control without coating treatment.
As can be seen from the experimental data in table 5: the composite coating agent prepared by the excellent strain B260 preserved by the invention has better promotion capability on the growth and development of the potatoes than the folded mosaic bacteria preserved by the same-genus preservation center.
As can be seen from fig. 1: the potatoes coated by the composite coating agent of the embodiment have higher yield, larger size and smooth shape compared with the potatoes which are not coated.
Example 6: microbial inoculum for antagonizing potato pathogenic microorganisms
The folded mosaic bacterium B260 disclosed by the invention has a certain inhibition effect on pathogenic microorganisms which harm potatoes and are relatively heavy in production, such as late blight, scab, black nevus and the like, and as a result, as shown in Table 6, the effect of inhibiting late blight, scab, black nevus and blight pathogenic bacteria by inoculating the folded mosaic bacterium B260 disclosed by the invention is improved compared with that of blank treatment of non-inoculated bacteria.
Table 6:
the results of the plate confrontation experiment in Table 6 show that: the inhibition rate of the folded mosaic bacteria B260 can reach 38-90%, and certain potential of preventing and controlling potato soil-borne diseases is shown.
Claims (10)
1. A folded mosaic bacterium (Massilia plicata) for promoting growth of potato tuber and cut piece and resisting growth has a strain preservation number of CGMCC No. 20439.
2. Use of the folded mosaic bacteria of claim 1 for promoting the anti-adversity growth of potato tuber-planted cuts.
3. A compound coating agent, wherein the compound coating agent comprises an agent A, an agent B and an agent C;
the agent A comprises one or more of phosphate rock powder, potassium mineral powder, boron mineral powder, iron silicate mineral powder, diatomite and straw powder;
the agent B comprises the folded mosaic bacterium of claim 1;
the agent C comprises carboxymethyl cellulose and molasses.
4. The composite coating agent according to claim 3, wherein the weight ratio of the agent A to the agent B to the agent C is 1 kg: (10-50) g: (120-360) g; the effective viable count concentration of the folded mosaic bacteria in the agent B is 109～1011CFU/g or 109～1011CFU/mL。
5. The composite coating agent of claim 3 or 4, wherein the agent A consists of ground phosphate rock, potassium ore, boron ore, iron silicate ore, diatomite and straw powder;
wherein the weight ratio of the phosphate rock powder to the potassium mineral powder to the boron mineral powder to the iron silicate mineral powder to the diatomite to the straw powder is (40-60) to (40-50) to (1-5) to (0.5-2) to (5-10) to (0.01-0.05);
preferably, the weight ratio of the potassium mineral powder, the boron mineral powder, the iron silicate mineral powder, the diatomite to the straw powder is 50: 40: 3: 1: 7: 0.03.
6. The compound coating agent of claim 3 or 4, wherein the agent B comprises a liquid microbial agent or a solid powdery microbial agent;
preferably, the liquid microbial inoculum comprises a protective agent and folding mosaic bacteria; the effective viable count concentration of the folded mosaic bacteria in the liquid microbial inoculum is 109～1011CFU/mL；
Preferably, the solid powdery microbial inoculum comprises folded mosaic bacteria and an additive for maintaining survival of strains, and the effective viable count concentration of the folded mosaic bacteria in the solid powdery microbial inoculum is 109～1011CFU/g；
Preferably, the protective agent comprises K at pH7.22HPO4-KH2PO4Buffer and/or glycerol.
7. The compound coating agent according to claim 3 or 4, wherein in the agent C, the carboxymethyl cellulose is mixed with the molasses in equal mass.
8. A preparation method of a composite coating agent comprises the following steps:
carrying out strain activation, shake flask culture and fermentation tank amplification culture on the folded mosaic bacteria of claim 1 by using a modified R2A culture medium, centrifugally collecting cells, freeze-drying, adding an additive to prepare a solid powdery bacteria agent, or directly adding a liquid protective agent to prepare a liquid bacteria agent, thus preparing the agent B of any one of claims 3-7;
preparing the agent A and the agent C respectively according to the formula of the agent A and the agent C in any one of claims 3 to 7;
independently packaging the agent A, the agent B and the agent C to obtain a compound coating agent;
preferably, the compound coating agent further comprises the following preparation steps when in use:
adding the agent B into the agent C, adding sterile water, and uniformly stirring to obtain a mixed solution;
then adding the agent A into the mixed solution and mixing uniformly to prepare the composite coating agent.
9. Use of the compound coating agent of any one of claims 3 to 7 in coating of potato tuber planting cut pieces.
10. A potato tuber planting, cutting, coating and planting method comprises the following steps:
step one, adding the agent B in any one of claims 3 to 7 into the agent C, adding sterile water, and uniformly stirring to obtain a first mixed solution; then adding the agent A into the first mixed solution to obtain a composite coating agent liquid solution for standby;
wherein, in the liquid solution of the composite coating agent, the dosage ratio of the agent A to the sterile water is 1 kg: (40-60) L; the effective viable count concentration of the folded mosaic bacteria in the liquid solution of the composite coating agent is 106～108CFU/mL；
Uniformly spraying the liquid solution of the compound coating agent on the surfaces of the potato tuber planting blocks which are aired for 12-24 hours, turning and stirring uniformly, airing for 5-10 min every time of spraying for ensuring full adsorption, and finally airing for 30-60 min for sowing, or sealing in a clean packaging bag, storing in a cool place for no more than 2 days and sowing;
wherein the dosage ratio of the potato tuber planting cut blocks to the liquid solution of the composite coating agent is (50-100) kg: (1-5) L.
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