CN110612988A - Application of Arthrobacter ureafaciens in prevention and treatment of corn diseases - Google Patents

Application of Arthrobacter ureafaciens in prevention and treatment of corn diseases Download PDF

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CN110612988A
CN110612988A CN201911044205.9A CN201911044205A CN110612988A CN 110612988 A CN110612988 A CN 110612988A CN 201911044205 A CN201911044205 A CN 201911044205A CN 110612988 A CN110612988 A CN 110612988A
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corn
arthrobacter ureafaciens
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arthrobacter
ureafaciens
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吴晓青
张新建
周方园
周红姿
赵晓燕
张广志
谢雪迎
范素素
王加宁
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Ecology Institute Of Shandong Academy Of Sciences (the Sino-Japanese Friendship Biotechnology Research Center Shandong Academy Of Sciences)
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Ecology Institute Of Shandong Academy Of Sciences (the Sino-Japanese Friendship Biotechnology Research Center Shandong Academy Of Sciences)
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
    • A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
    • A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
    • A01G24/12—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material containing soil minerals
    • A01G24/15—Calcined rock, e.g. perlite, vermiculite or clay aggregates
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
    • A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
    • A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
    • A01G24/22—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing plant material
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
    • A01G24/30—Growth substrates; Culture media; Apparatus or methods therefor based on or containing synthetic organic compounds
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K17/00—Soil-conditioning materials or soil-stabilising materials
    • C09K17/14—Soil-conditioning materials or soil-stabilising materials containing organic compounds only
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K2101/00—Agricultural use

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  • Environmental Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Soil Sciences (AREA)
  • Inorganic Chemistry (AREA)
  • General Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Materials Engineering (AREA)
  • Organic Chemistry (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Pretreatment Of Seeds And Plants (AREA)

Abstract

The invention discloses application of Arthrobacter ureafaciens in preventing and treating corn diseases. The Arthrobacter ureafaciens (Arthrobacter ureafaciens) Fp 64. The bacteriostasis rate of the Arthrobacter ureafaciens on pathogenic bacteria of basal rot of corn stalk and Pythium irregulare and Fusarium oxysporum is 93.82% and 54.76%, the bacteriostasis rate on rhizoctonia solani (AG1-IA fusion group) as a pathogenic bacterium of corn sheath blight is 88.02%, and the bacteriostasis rate on Curvularia lunata as a pathogenic bacterium of corn leaf spot is 78.85%. The strain is prepared into a spraying microbial inoculum, a seed coating agent or an active seedling raising matrix, can be used for preventing and treating the corn stalk basal rot, the corn sheath blight and the corn leaf spot independently or simultaneously, reduces the compatibility cost of various strains, and can realize the application reduction and the efficiency improvement.

Description

Application of Arthrobacter ureafaciens in prevention and treatment of corn diseases
Technical Field
The invention relates to the technical field of biological control, in particular to application of Arthrobacter ureafaciens in control of corn diseases.
Background
Corn is an important food crop in China, and in the corn planting process, corn stalk rot, corn sheath blight and corn leaf spot are typical diseases which harm the quality and the yield of corn. The corn stalk rot, also called corn stalk rot and corn bacterial wilt, is a typical soil-borne disease which can occur in the whole growth period of plants and mainly damages the roots and the stalk bases of the plants, so that the stalk bases are browned and softened, the fruit clusters droop, and finally the whole plants die due to wilting. The pathogenic bacteria causing the basal rot of the corn stalk are Pythium spp and Fusarium spp, and the Pythium spp and the Fusarium spp are used for causing diseases independently or cooperatively. The corn sheath blight mainly occurs at the later growth stage of the corn, and damages leaf sheaths and stems of a plurality of sections of the corn near the ground, so that stem base is rotten, and large loss is easily caused. The pathogenic bacterium of corn sheath blight is Rhizoctonia sp. Corn leaf spot mainly harms leaves, leaf sheaths and bracts, and influences photosynthesis to reduce corn yield. The pathogenic bacteria of corn leaf spot are Curvularia spp. Aiming at the basal rot, sheath blight and leaf spot of corn, a common medicament control means is to spray a large amount of chemical pesticides such as carbendazim, chlorothalonil and the like or prepare a chemical seed coating agent, so that the potential safety risks of grains and the hidden dangers of water and soil environmental pollution are brought. In order to relieve the continuous innovation and breakthrough of the biological control technology of chemical pesticides on corn diseases, currently developed biocontrol bacteria comprise bacillus, pseudomonas, trichoderma, actinomycetes and the like, and the prior art for controlling the corn diseases by utilizing arthrobacter is not available.
Disclosure of Invention
The invention aims to solve the technical problem of providing the application of Arthrobacter ureafaciens in preventing and treating corn diseases aiming at typical corn diseases, in particular to the application of Arthrobacter ureafaciens in preventing and treating corn basal rot, corn sheath blight and corn leaf spot.
The application technical scheme of the Arthrobacter ureafaciens in preventing and treating corn diseases is as follows,
the Arthrobacter ureafaciens Fp64 is preserved in China general microbiological culture Collection center (CGMCC) in 26.04.2018, has the address of No. 3 of Xilu No.1 of Beijing Korean district, and has the preservation number of CGMCC No. 15644.
The strain is disclosed in patent application with publication number CN 108795799A, publication number 2018.11.13 and invention name of multifunctional arthrobacter Fp64 for controlling soil phosphorus loss and application thereof.
The Arthrobacter ureafaciens can tolerate the NaCl salt concentration of 100g/L, so that a biocontrol product prepared by the Arthrobacter ureafaciens can be used in saline-alkali soil.
The bacteriostasis rate of the Arthrobacter ureafaciens on the maize stalk basal rot pathogenic bacterium Pythium irregulare (Pythium irregulare) and Fusarium oxysporum (Fusarium oxysporum) is 93.82 percent and 54.76 percent, the bacteriostasis rate on the maize sheath blight pathogenic bacterium Rhizoctonia solani (AG1-IA fusion group) is 88.02 percent, and the bacteriostasis rate on the maize leaf spot pathogenic bacterium Curvularia lunata (Curvularia lunata) is 78.85 percent. The invention also provides a spraying microbial inoculum, a seed coating agent or an active seedling substrate for preventing and treating the corn stalk-based rot, the corn sheath blight and the corn leaf spot, wherein the spraying microbial inoculum, the seed coating agent or the active seedling substrate for preventing and treating the corn stalk-based rot and the corn sheath blight are prepared by adopting Arthrobacter ureafaciens Fp 64.
The spraying microbial inoculum is a water aqua and comprises the following components in parts by weight: 50-99 parts of active ingredients, 0.2-15 parts of dispersing agents, 0.2-15 parts of penetration enhancers, 0.2-20 parts of surfactants, 0.1-0.5 part of preservatives and 6.5-8.0 parts of pHs;
wherein the active ingredient is Arthrobacter ureafaciens Fp64 fermentation product; the dispersant is dispersant NNO; the penetration enhancer is at least one of azone and thiaketone; the surfactant is at least one of sodium dodecyl sulfate, sodium lignosulfonate and Chinese honeylocust powder; the preservative is at least one of potassium sorbate and calcium propionate;
the preparation method of the spraying microbial inoculum comprises the following steps:
(1) activating Arthrobacter ureafaciens (Arthrobacter ureafaciens) Fp64 to obtain Arthrobacter ureafaciens Fp64 seed liquid;
(2) carrying out amplification culture on Arthrobacter ureafaciens Fp64 seed liquid to obtain Fp64 fermentation product;
(3) the Arthrobacter ureafaciens Fp64 fermentation product is used as an active ingredient to be combined with other additives to form an Fp64 spraying microbial inoculum.
The method for using the spraying microbial inoculum for preventing and treating the corn stalk base rot, the corn sheath blight and the corn leaf spot comprises the steps of diluting the spraying microbial inoculum by 100 times, and spraying stalk roots or irrigating roots in a large-horn mouth period, a grouting period and a milk stage or when sporadic plants are found in a field; spraying once every 7-15 days, and continuously spraying for 2-3 times.
The seed coating agent comprises the following components in parts by weight: 10-40 parts of active ingredients, 40-60 parts of fillers, 0.1-0.2 part of dispersing agents, 0.1-0.5 part of preservatives and 4-20 parts of film forming agents;
wherein the active ingredient is mother powder of Arthrobacter ureafaciens Fp 64; the filler is diatomite; the dispersant is dispersant NNO; the preservative is at least one of potassium sorbate and calcium propionate; the film forming agent is a commercially available film forming agent;
the preparation method of the seed coating agent comprises the following steps:
(1) activating Arthrobacter ureafaciens (Arthrobacter ureafaciens) Fp64 to obtain Arthrobacter ureafaciens Fp64 seed liquid;
(2) carrying out amplification culture on Arthrobacter ureafaciens Fp64 seed liquid to obtain Arthrobacter ureafaciens Fp64 fermentation liquid;
(3) drying the Arthrobacter ureafaciens Fp64 fermentation liquor to obtain dry powder serving as Arthrobacter ureafaciens Fp64 mother powder;
(4) the Arthrobacter ureafaciens Fp64 mother powder is used as an active ingredient to be combined with other additives into the Arthrobacter ureafaciens Fp64 seed coating agent.
The application method of the seed coating agent for preventing and treating the corn stalk base rot, the corn sheath blight and the corn leaf spot comprises 20-25% of coating by mass. The coated corn seeds are sown in a conventional manner.
The active seedling raising substrate comprises the following components in parts by weight of 20-40 parts of active components; 10-15 parts of decomposed wheat straw, 20-30 parts of vermiculite, 5-10 parts of perlite, 5-10 parts of humic acid, 3-5 parts of plant ash and 0.1-0.2 part of inorganic salt;
wherein the active ingredient is Arthrobacter ureafaciens Fp64 fermentation product; the inorganic salt comprises ammonium dihydrogen phosphate, calcium chloride, magnesium chloride and trace elements;
the preparation steps of the active seedling substrate comprise:
(1) activating Arthrobacter ureafaciens Fp64 to obtain Arthrobacter ureafaciens Fp64 seed liquid;
(2) carrying out amplification culture on Arthrobacter ureafaciens Fp64 seed liquid to obtain an Arthrobacter ureafaciens Fp64 fermentation product;
(3) the Arthrobacter ureafaciens Fp64 fermentation product is used as an active ingredient to be combined with other matrix ingredients to form the Arthrobacter ureafaciens Fp64 active seedling culture matrix.
An active seedling substrate for preventing and treating the basal rot, sheath blight and leaf spot of corn is prepared through putting the active seedling substrate in conventional seedling tray, forcing the corn seeds by conventional method, sowing one seedling in each hole. Transplanting the seedlings according to a conventional mode after seedling cultivation.
The invention has the beneficial effects that: the arthrobacter strain has antagonistic action on corn pythium, fusarium, rhizoctonia and curvularia, can be used for independently or simultaneously preventing and treating corn basal rot, corn sheath blight and corn leaf spot, reduces the compatibility cost of various strains, and can realize application reduction and efficiency improvement;
the arthrobacter strains utilized by the invention are obtained by separating and screening healthy farmland soil, and are environment-friendly; the strain also has the characteristic of regulating soil phosphorus, and can effectively relieve the loss of phosphorus to adjacent water bodies and reduce the application amount of chemical fertilizers after being applied to corn farmlands for a long time; the strain also has the characteristic of high salt tolerance, and has strong adaptability to corn farmlands in saline-alkali and salinized fields. Therefore, the spraying microbial inoculum, the coating agent and the seedling raising substrate prepared by the strain have the advantages of environmental friendliness, reduction of soil phosphorus loss and strong adaptability to saline lands;
the invention provides a spraying microbial inoculum and a seed coating agent (seed coating agent) for sowed corn varieties, provides a disease-preventing active seedling raising matrix for corn varieties (such as fruit corn) which are not suitable for sowing, and has wide application range and strong practicability.
Description of the drawings:
FIG. 1 is a photograph showing the antagonistic effect of Fp64 on Pythium irregulare treatment and a control;
FIG. 2 shows photographs of antagonism treatment and control of Fp64 against Fusarium oxysporum;
FIG. 3 is a photograph showing the antagonistic action of Fp64 against Rhizoctonia solani treatment and a control;
FIG. 4 is a photograph showing the antagonistic action of Fp64 against Curvularia lunata.
The specific implementation mode is as follows:
for better understanding of the present invention, the technical solution of the present invention will be described in detail with specific examples, but the present invention is not limited thereto.
Example 1
Antagonism of Arthrobacter ureafaciens Fp64 on pathogenic bacteria of basal rot, sheath blight and leaf spot of corn
(1) Source of pathogenic bacteria
The pathogenic bacteria are collected from the diseased part of the corn plant, and the collected part is the corn field of Manzhuanzhu, Taian, Shandong province. Wherein the pathogenic bacteria of the basal rot of corn stalk are identified as Pythium irregulare and fusarium oxysporum (fusarium oxysporum), the pathogenic bacteria of the sheath blight of corn are identified as AG1-IA fusion group of Rhizoctonia solani (Rhizoctonia solani), and the pathogenic bacteria of the leaf spot of corn are identified as Curvularia lunata (Curvularia lunata).
(2) Confrontation test
Activating Arthrobacter ureafaciens Fp64 and the four pathogenic bacteria on a culture medium commonly used by bacteria and fungi, respectively, and mixing the activated Fp64 with sterile distilled water to obtain a mixture with a concentration of 1 × 109The bacterial suspension of (4). Preparing PDA culture medium, cooling to below 50 deg.C, mixing with 2mL bacterial suspension per 100mL PDA, shaking, pouring into 8.5cm flat plate, standing, and solidifying to obtain the final product. And (3) beating bacterial cakes on the activated four pathogenic bacteria colonies by using a 5mm puncher, respectively inoculating each pathogenic bacteria cake to the central position of a counter plate, carrying out dark culture at 25 ℃ for 5-9 days, and measuring the growth diameter of the strain. The test is carried out by taking a sterile PDA plate to be inoculated into each pathogenic bacteria cake as a control and setting three experiments for repetition.
1L of PDA culture medium formula: 200g of potato, 20g of glucose, 10g of agar and 1L of water. The medium was autoclaved at 115 ℃ for 30 min.
The results showed that the rate of inhibition of Arthrobacter ureafaciens Fp64 against Pythium irregulare was 93.82% (FIG. 1), that against Fusarium oxysporum was 54.76% (FIG. 2), that against Rhizoctonia solani was 88.02% (FIG. 3), and that against Curvularia lunata was 78.85% (FIG. 4). The result shows that Fp64 has obvious antagonistic action on pathogenic bacteria of corn stalk basal rot, corn sheath blight and corn leaf spot.
Example 2
Preparation of spraying microbial inoculum of Arthrobacter ureafaciens Fp64
(1) Obtaining seed liquid of Arthrobacter ureafaciens Fp 64;
arthrobacter ureafaciens Fp64 was activated on LB medium, transferred to 100mL of LB medium, and cultured at 30 ℃ and 180rpm for 16hr with shaking to obtain a seed solution.
1L LB medium formula: 10g of peptone, 5g of yeast extract, 10g of NaCl, and distilled water to make up to 1L, 15g of agar powder (no liquid medium added), and pH 7.0. The medium was autoclaved at 120 ℃ under 0.1MPa for 20 min.
(2) Obtaining of Arthrobacter ureafaciens Fp64 fermentation product
Transferring the Arthrobacter ureafaciens Fp64 seed solution into an amplification culture medium according to the ratio of 1:1000, and performing amplification culture under the conditions of 30 ℃, stirring speed of 200rpm, ventilation of 1:2 and fermentation period of 24hr to obtain Fp64 fermentation liquid.
1L of the amplification medium formula: 10g of soybean meal, 22g of corn steep liquor, 10g of yeast extract, 2g of ammonium sulfate, 4g of sodium chloride and distilled water, wherein the total amount is 1L, and the pH value is 7.0. The medium was autoclaved at 120 ℃ under 0.1MPa for 20 min.
(3) Acquisition of spraying microbial inoculum of Arthrobacter ureafaciens Fp64
The Arthrobacter ureafaciens Fp64 fermentation product and other additives are combined into Fp64 spraying microbial inoculum. The microbial inoculum comprises the following components in percentage by weight: 80 parts of Arthrobacter ureafaciens Fp64 fermentation product, 5 parts of dispersant NNO, 5 parts of azone, 10 parts of sodium dodecyl sulfate and 0.4 part of potassium sorbate. The viable count of the obtained spraying microbial inoculum is (5-10) multiplied by 108cfu/mL。
Example 3
Field control effect determination of Arthrobacter ureafaciens Fp64 spraying microbial inoculum
The test variety is the summer maize variety Zhengdan 958.
Test and control agents: spraying the inoculum of Arthrobacter ureafaciens Fp64 was provided in example 1. The contrast agent is 50% carbendazim, is purchased from local agricultural markets, is a common chemical agent for preventing and treating the basal rot of corn stems, sheath blight of corn and leaf spot of corn in production, and is used according to the recommended dosage in commercial fields.
The experiment is repeated for 3 times after 3 treatments, and 100 times of diluent for spraying the bactericide of Arthrobacter ureafaciens Fp64, 500 times of diluent for 50 percent of carbendazim and clear water are compared. Respectively spraying stem root parts in the large-horn mouth period, the grouting period and the milk stage of the corn to uniformly cover the stems, and spraying for 2-3 times at intervals of 7-15 days. Setting a cell, surveying a method, setting the morbidity and grading the disease condition according to the technical specification for identifying the disease and insect resistance of corn, NY/T1248.6-2016, 1248.9-2016 and 1248.10-2016 of the Ministry of agriculture.
The corn stalk base rot disease incidence plant rate calculation formula:wherein X represents the diseased plant percentage (%), Y represents the number of diseased plants (plant), and N represents the total number of examined plants (plant).
The disease index calculation formula of the corn sheath blight disease and the corn leaf spot disease is as follows:wherein X represents disease index, SiIndicates disease level, j indicates corresponding to SiThe number of the plants in the level is the number of the plants; n represents the total number of investigated plants, and the unit is plant.
The prevention effect calculation formula is as follows:
the test results are shown in Table 1.
The control effects of the spraying microbial inoculum of the Arthrobacter ureafaciens Fp64 on the basal rot, the sheath blight and the leaf spot of corn in the field are 75.07%, 80.54% and 68.45% respectively, and the corresponding control effects of the contrast agent are 61.25%, 62.58% and 64.36% respectively, as shown in Table 1. The result shows that the Fp64 spraying microbial inoculum has obvious control effect on the basal rot, the sheath blight and the leaf spot of the corn, so that the spraying microbial inoculum can be used for controlling three diseases independently or simultaneously, and has good field application value.
TABLE 1 Fp64 determination of field control effect of spraying fungicide on corn stalk base rot and corn sheath blight
Example 4
Preparation of Arthrobacter ureafaciens Fp64 seed coating agent
(1) Obtaining seed liquid of Arthrobacter ureafaciens Fp 64;
same as example 1
(2) Obtaining of Arthrobacter ureafaciens Fp64 fermentation product
Same as example 1
(3) Obtaining of Arthrobacter ureafaciens Fp64 powder
Taking the Arthrobacter ureafaciens Fp64 fermentation product, and adding gelatin as a thermal protective agent in a weight percentage of 3%. And (3) drying by adopting a microbial agent spray dryer to obtain dry powder, namely the arthrobacter Fp64 mother powder. The viable count of the obtained bacterial powder is (100-500) multiplied by 108cfu/g。
(4) Preparation of seed coating agent of Arthrobacter ureafaciens Fp64
The Arthrobacter ureafaciens Fp64 powder and other additives are combined into the Arthrobacter ureafaciens Fp64 seed coating agent. The seed coating agent comprises the following components in percentage by weight: 20 parts of Arthrobacter ureafaciens Fp64 mother powder, 50 parts of diatomite, 0.2 part of dispersant NNO, 0.2 part of potassium sorbate and 0.4 part of film forming agent (German Bayer).
Example 5
Determination of field control effect of Arthrobacter ureafaciens Fp64 seed coating agent
The test variety is the summer maize variety Zhengdan 958.
Test and control agents: fp64 seed coating is provided by example 3. The contrast agent was prepared by mixing 5 parts by weight of 50% carbendazim, 50 parts by weight of diatomaceous earth, 0.2 parts by weight of NNO, 0.2 parts by weight of potassium sorbate and 0.4 parts by weight of film-forming agent (german bayer) in parts by weight to obtain a carbendazim seed coating agent.
Coating the corn seeds by a seed coating machine (Nanjing agriculture and animal husbandry machinery factory) according to the coating mass percentage of 23 percent.
The experiment was run for 3 replicates of 3 treatments, Fp64 seed coating, carbendazim seed coating and clear water controls. The seeding mode is a conventional mode. Setting a cell, surveying a method, setting the morbidity and grading the disease condition according to the technical specification for identifying the disease and insect resistance of corn, NY/T1248.6-2016, 1248.9-2016 and 1248.10-2016 of the Ministry of agriculture.
The corn stalk base rot disease incidence plant rate calculation formula:wherein X represents the incidence of disease(%), Y represents the number of diseased plants (plants), and N represents the total number of examined plants (plants).
The disease index calculation formula of the corn sheath blight disease and the corn leaf spot disease is as follows:wherein X represents disease index, SiIndicates disease level, j indicates corresponding to SiThe number of the plants in the level is the number of the plants; n represents the total number of investigated plants, and the unit is plant.
The prevention effect calculation formula is as follows:
the test results are shown in Table 2.
The seed coating agent of the Arthrobacter ureafaciens Fp64 provided by the invention has control effects on basal rot and sheath blight of corn in fields of 68.11%, 71.55% and 58.63% respectively, and the control agents have corresponding control effects of 56.15%, 59.90% and 52.00% respectively. The result shows that the Fp64 seed coating agent has obvious control effect on the corn stalk basal rot, the sheath blight and the leaf spot, can be used for controlling three diseases independently or simultaneously, and has good field application value.
TABLE 2 Fp64 determination of field control effect of seed coating agent on corn stalk rot and corn sheath blight
Example 6
Preparation of Arthrobacter ureafaciens Fp64 active seedling culture medium
(1) Obtaining seed liquid of Arthrobacter ureafaciens Fp 64;
same as example 1
(2) Obtaining of Arthrobacter ureafaciens Fp64 fermentation product
Same as example 1
(3) Obtaining of Arthrobacter ureafaciens Fp64 active matrix
The Arthrobacter ureafaciens Fp64 fermentation product and other additives are combined into an Fp64 active matrix.
The active seedling raising matrix comprises, by weight, 40 parts of a urea-producing arthrobacter Fp64 fermented product, 12 parts of decomposed wheat straw, 25 parts of vermiculite, 8 parts of perlite, 8 parts of humic acid, 4 parts of plant ash and 0.2 part of inorganic salt (ammonium dihydrogen phosphate, calcium chloride, magnesium chloride and trace element fertilizer).
Example 7
Determination of field control effect of urea-producing node rod Fp64 active seedling culture medium
The test variety is fruit corn.
Test and control matrices: urea-producing node rod Fp64 active matrix is provided by example 5. Mixing 3 parts by weight of 50% carbendazim, 12 parts by weight of decomposed wheat straw, 25 parts by weight of vermiculite, 8 parts by weight of perlite, 8 parts by weight of humic acid, 4 parts by weight of plant ash and 0.2 part by weight of inorganic salt (ammonium dihydrogen phosphate, calcium chloride, magnesium chloride and trace element fertilizer) to form a carbendazim matrix serving as a control matrix.
The experiment was repeated 3 times with 3 treatments in total, producing urea internode Fp64 active matrix, carbendazim matrix and clear water control. The corn seedling raising mode is a conventional mode. Setting a cell, surveying a method, setting the morbidity and grading the disease condition according to the technical specification for identifying the disease and insect resistance of corn, NY/T1248.6-2016, 1248.9-2016 and 1248.10-2016 of the Ministry of agriculture.
The corn stalk base rot disease incidence plant rate calculation formula:wherein X represents the diseased plant percentage (%), Y represents the number of diseased plants (plant), and N represents the total number of examined plants (plant).
The disease index calculation formula of the corn sheath blight disease and the corn leaf spot disease is as follows:wherein X represents disease index, SiIndicates disease level, j indicates corresponding to SiThe number of the plants in the level is the number of the plants; n representsThe total number of plants was investigated in units of plants.
The prevention effect calculation formula is as follows:
the test results are shown in Table 3.
The control effects of the urea-producing nodal rod Fp64 active matrix on corn stalk basal rot, corn sheath blight and corn leaf spot in the field are 64.36%, 69.94% and 63.28 respectively, and the corresponding control effects of the contrast agent are 52.81%, 51.82% and 65.25% respectively. The result shows that the Fp64 active matrix has obvious control effect on the basal rot, the sheath blight and the leaf spot of the corn, and the active matrix can be used for controlling three diseases independently or simultaneously, thereby having good field application value.
TABLE 3 Fp64 determination of field control effect of active matrix on corn stalk rot and corn sheath blight
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (9)

1. The application of the Arthrobacter ureafaciens in preventing and treating corn diseases is characterized in that the Arthrobacter ureafaciens Fp64 is preserved in China general microbiological culture Collection center (CGMCC) in 26.04.2018, the address is No. 3 of Xilu No.1 of Beijing Korean area, and the preservation number is CGMCC No. 15644.
2. The use according to claim 1, wherein the Arthrobacter ureafaciens is tolerant to NaCl salt concentrations of 100 g/L.
3. The use according to claim 1, wherein the Arthrobacter ureafaciens has a bacterial inhibition rate of 93.82% and 54.76% against Pythium irregulare (Pythium irregularium) and Fusarium oxysporum (Fusarium oxysporum) which are pathogens of basal rot of corn, 88.02% against Rhizoctonia solani (AG1-IA fusion group) which is a pathogen of sheath blight of corn, and 78.85% against Curvularia lunata (Curvularia lunata).
4. The application of the Arthrobacter ureafaciens Fp64 as a spraying microbial agent, a seed coating agent or an active seedling substrate for preventing and treating basal rot, sheath blight and leaf spot of corn.
5. The spraying microbial inoculum for preventing and treating the basal rot of corn stalk, the sheath blight of corn and the leaf spot of corn is characterized in that the spraying microbial inoculum is an aqueous agent, and the contents of the components in parts by weight are as follows: 50-99 parts of active ingredients, 0.2-15 parts of dispersing agents, 0.2-15 parts of penetration enhancers, 0.2-20 parts of surfactants, 0.1-0.5 part of preservatives and 6.5-8.0 parts of pHs;
wherein the active ingredient is Arthrobacter ureafaciens Fp64 fermentation product; the dispersant is dispersant NNO; the penetration enhancer is at least one of azone and thiaketone; the surfactant is at least one of sodium dodecyl sulfate, sodium lignosulfonate and Chinese honeylocust powder; the preservative is at least one of potassium sorbate and calcium propionate;
the preparation method of the spraying microbial inoculum comprises the following steps:
(1) activating Arthrobacter ureafaciens (Arthrobacter ureafaciens) Fp64 to obtain Arthrobacter ureafaciens Fp64 seed liquid;
(2) carrying out amplification culture on Arthrobacter ureafaciens Fp64 seed liquid to obtain Fp64 fermentation product;
(3) the Arthrobacter ureafaciens Fp64 fermentation product is used as an active ingredient to be combined with other additives to form an Fp64 spraying microbial inoculum.
6. The spraying microbial inoculum for preventing and treating the stem rot, the sheath blight and the leaf spot of the corn as claimed in claim 5, wherein the spraying microbial inoculum is diluted by 100 times, and the stem roots are sprayed or irrigated when a sporadic plant is found in a large horn mouth period, a grouting period and a milk stage or in a field; spraying once every 7-15 days, and continuously spraying for 2-3 times.
7. The seed coating agent for preventing and treating the corn stalk base rot, the corn sheath blight and the corn leaf spot is characterized by comprising the following components in parts by weight: 10-40 parts of active ingredients, 40-60 parts of fillers, 0.1-0.2 part of dispersing agents, 0.1-0.5 part of preservatives and 4-20 parts of film forming agents;
wherein the active ingredient is mother powder of Arthrobacter ureafaciens Fp 64; the filler is diatomite; the dispersant is dispersant NNO; the preservative is at least one of potassium sorbate and calcium propionate; the film forming agent is a commercially available film forming agent;
the preparation method of the seed coating agent comprises the following steps:
(1) activating Arthrobacter ureafaciens (Arthrobacter ureafaciens) Fp64 to obtain Arthrobacter ureafaciens Fp64 seed liquid;
(2) carrying out amplification culture on Arthrobacter ureafaciens Fp64 seed liquid to obtain Arthrobacter ureafaciens Fp64 fermentation liquid;
(3) drying the Arthrobacter ureafaciens Fp64 fermentation liquor to obtain dry powder serving as Arthrobacter ureafaciens Fp64 mother powder;
(4) the Arthrobacter ureafaciens Fp64 mother powder is used as an active ingredient to be combined with other additives into the Arthrobacter ureafaciens Fp64 seed coating agent.
8. The seed coating agent for preventing and treating the basal rot of corn stalk, the sheath blight of corn and the leaf spot of corn as claimed in claim 7, wherein the coating mass percentage is 20% -25%. The coated corn seeds are sown in a conventional manner.
9. An active seedling raising matrix for preventing and treating corn stalk rot, corn sheath blight and corn leaf spot is characterized in that the active seedling raising matrix comprises 20-40 parts by weight of active ingredients; 10-15 parts of decomposed wheat straw, 20-30 parts of vermiculite, 5-10 parts of perlite, 5-10 parts of humic acid, 3-5 parts of plant ash and 0.1-0.2 part of inorganic salt;
wherein the active ingredient is Arthrobacter ureafaciens Fp64 fermentation product; the inorganic salt comprises ammonium dihydrogen phosphate, calcium chloride, magnesium chloride and trace elements;
the preparation steps of the active seedling substrate comprise:
(1) activating Arthrobacter ureafaciens Fp64 to obtain Arthrobacter ureafaciens Fp64 seed liquid;
(2) carrying out amplification culture on Arthrobacter ureafaciens Fp64 seed liquid to obtain an Arthrobacter ureafaciens Fp64 fermentation product;
(3) the Arthrobacter ureafaciens Fp64 fermentation product is used as an active ingredient to be combined with other matrix ingredients to form the Arthrobacter ureafaciens Fp64 active seedling culture matrix.
CN201911044205.9A 2019-10-30 2019-10-30 Application of Arthrobacter ureafaciens in prevention and treatment of corn diseases Pending CN110612988A (en)

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